HK40005287A

HK40005287A - Human antigen binding proteins that bind to a complex comprising beta-klotho and an fgf receptor

Info

Publication number: HK40005287A
Application number: HK19128675.6A
Authority: HK
Inventors: Yang Li; Jennitte STEVENS; Chadwick Terence King; Ian Foltz; Gunasekaran Kannan; Junming Yie; Shaw-Fen Sylvia Hu
Original assignee: Amgen Inc.
Priority date: 2011-06-06
Filing date: 2019-08-26
Publication date: 2020-05-08

Description

The present application is a divisional application of European patent application EP 12 727 726.7 (Filing Date: June 05, 2012 ) the content of which is hereby incorporated in its entirety. The application claims priority to U.S. Application No. 13/487,061 (filed June 01, 2012 ), which claims benefit to U.S. Provisional applications 61/493,933 (filed June 6, 2011 ), 61/501,133 (filed June 24, 2011 ), and 61/537,998 (filed September 22, 2011 ), the contents of each of which are hereby incorporated in their entirety.

FIELD OF THE INVENTION

The present disclosure relates to nucleic acid molecules encoding antigen binding proteins that bind to a complex comprising β-Klotho and at least one of (i) FGFR1c, (ii) FGFR2c and (iii) FGFR3c, including antigen binding proteins that induce FGF21-like signaling, as well as pharmaceutical compositions comprising antigen binding proteins that bind to a complex comprising β-Klotho and at least one of (i) FGFR1c, (ii) FGFR2c and (iii) FGFR3c, including antigen binding proteins that induce FGF21-like signaling, and methods for treating metabolic disorders using such nucleic acids, polypeptides, or pharmaceutical compositions. Diagnostic methods using the antigen binding proteins are also provided.

BACKGROUND

Fibroblast Growth Factor 21 (FGF21) is a secreted polypeptide that belongs to a subfamily of Fibroblast Growth Factors (FGFs) that includes FGF19, FGF21, and FGF23 (Itoh et al., (2004) Trend Genet. 20:563-69). FGF21 is an atypical FGF in that it is heparin independent and functions as a hormone in the regulation of glucose, lipid, and energy metabolism.

It is highly expressed in liver and pancreas and is the only member of the FGF family to be primarily expressed in liver. Transgenic mice overexpressing FGF21 exhibit metabolic phenotypes of slow growth rate, low plasma glucose and triglyceride levels, and an absence of age-associated type 2 diabetes, islet hyperplasia, and obesity. Pharmacological administration of recombinant FGF21 protein in rodent and primate models results in normalized levels of plasma glucose, reduced triglyceride and cholesterol levels, and improved glucose tolerance and insulin sensitivity. In addition, FGF21 reduces body weight and body fat by increasing energy expenditure, physical activity, and metabolic rate. Experimental research provides support for the pharmacological administration of FGF21 for the treatment of type 2 diabetes, obesity, dyslipidemia, and other metabolic conditions or disorders in humans.

FGF21 is a liver derived endocrine hormone that stimulates glucose uptake in adipocytes and lipid homeostasis through the activation of its receptor. Interestingly, in addition to the canonical FGF receptor, the FGF21 receptor also comprises the membrane associated β-Klotho as an essential cofactor. Activation of the FGF21 receptor leads to multiple effects on a variety of metabolic parameters.

In mammals, FGFs mediate their action via a set of four FGF receptors, FGFR1-4, that in turn are expressed in multiple spliced variants, e.g., FGFR1c, FGFR2c, FGFR3c and FGFR4. Each FGF receptor contains an intracellular tyrosine kinase domain that is activated upon ligand binding, leading to downstream signaling pathways involving MAPKs (Erk1/2), RAF1, AKT1 and STATs. (Kharitonenkov et al., (2008) BioDrugs 22:37-44). Several reports suggested that the "c"-reporter splice variants of FGFR1-3 exhibit specific affinity to β-Klotho and could act as endogenous receptor for FGF21 (Kurosu et al., (2007) J. Biol. Chem. 282:26687-95); Ogawa et al., (2007) Proc. Natl. Acad. Sci. USA 104:7432-37); Kharitonenkov et al., (2008) J. Cell Physiol. 215:1-7). In the liver, which abundantly expresses both β-Klotho and FGFR4, FGF21 does not induce phosphorylation of MAPK albeit the strong binding of FGF21 to the β--Klotho-FGFR4 complex. In 3T3-L1 cells and white adipose tissue, FGFR1 is by far the most abundant receptor, and it is therefore most likely that FGF21's main functional receptors in this tissue are the β--Klotho/FGFR1c complexes.

The present disclosure provides a human (or humanized) antigen binding protein, such as a monoclonal antibody, that induces FGF21-like signaling, e.g., an agonistic antibody that mimics the function of FGF21. Such an antibody is a molecule with FGF21-like activity and selectivity but with added therapeutically desirable characteristics typical for an antibody such as protein stability, lack of immunogenicity, ease of production and long half-life in vivo.

SUMMARY

The instant disclosure provides antigen binding proteins that bind a complex comprising β-Klotho and at least one of (i) FGFR1c, (ii) FGFR2c and (iii) FGFR3c, including antigen binding proteins that induce FGF21-like signaling, as well as pharmaceutical compositions comprising antigen binding proteins that bind to a complex comprising β-Klotho and at least one of (i) FGFR1c, (ii) FGFR2c and (iii) FGFR3c, including antigen binding proteins that induce FGF21-like signaling. In another aspect, also provided are expression vectors and host cells transformed or transfected with the expression vectors that comprise the aforementioned isolated nucleic acid molecules that encode the antigen binding proteins disclosed herein. Representative heavy and light chains are provided in Tables 1A and 1B; representative variable region heavy chain and light chain sequences are provided in Tables 2A and 2B; coding sequences for the variable region of the heavy and light chains are provided in Tables 2C and 2D; Tables 3A and 3B provide CDR regions of the disclosed variable heavy and light chains, and Tables 3C and 3D provide coding sequences for the disclosed CDRs.

In another aspect, also provided are methods of preparing antigen binding proteins that specifically or selectively bind a complex comprising β-Klotho and at least one of (i) FGFR1c, (ii) FGFR2c and (iii) FGFR3c and comprise the step of preparing the antigen binding protein from a host cell that secretes the antigen binding protein.

Other embodiments provide a method of preventing or treating a condition in a subject in need of such treatment comprising administering a therapeutically effective amount of a pharmaceutical composition provided herein to a subject, wherein the condition is treatable by lowering blood glucose, insulin or serum lipid levels. In embodiments, the condition is type 2 diabetes, obesity, dyslipidemia, NASH, cardiovascular disease or metabolic syndrome.

These and other aspects are described in greater detail herein. Each of the aspects provided can encompass various embodiments provided herein. It is therefore anticipated that each of the embodiments involving one element or combinations of elements can be included in each aspect described, and all such combinations of the above aspects and embodiments are expressly considered. Other features, objects, and advantages of the disclosed antigen binding proteins and associated methods and compositions are apparent in the detailed description that follows.

BRIEF DESCRIPTION OF THE FIGURES

Figure 1a-1b is an alignment showing the sequence homology between human FGFR1c (GenBank Accession No P11362; SEQ ID NO: 4) and murine FGFR1c (GenBank Accession No NP_034336; SEQ ID NO: 1832); various features are highlighted, including the signal peptide, transmembrane sequence, heparin binding region, and a consensus sequence (SEQ ID NO: 1833) is provided.
Figure 2a-2c is an alignment showing the sequence homology between human β-Klotho (GenBank Accession No NP_783864; SEQ ID NO: 7) and murine β-Klotho (GenBank Accession No NP_112457; SEQ ID NO: 10); various features are highlighted, including the transmembrane sequence and two glycosyl hydrolase domains, and a consensus sequence (SEQ ID NO: 1834) is provided.
Figure 3 is a plot showing the representative data from Luciferase reporter activity screens of the antibodies disclosed herein with FGF21 and a reference antibody 16H7.1 as positive controls (insert); these hybridomas were generated by immunization with cell-bound receptor of 293T transfectants expressing full length human β-Klotho and an N-terminal truncated form of human FGFR1c encompassing amino acid residue #141 to #822 polypeptide of SEQ ID NO:4. X- and Y- axis in the plot are % FGF21 activity from two independent assays (n=1 and n=2) of the same set of hybridoma samples (gray circles) showing the consistency of the assays; several hybridoma samples were also included as negative controls (black circles);
Figure 4 shows a schematic representation of the chimeras constructed in relation to present invention.
Figure 5 shows the ability of the antigen binding proteins, as well as human FGF21, to activate chimeras in L6 cells.
Figures 6a-e show the amino acid alignment of heavy and light chains of the antibodies compared to the corresponding germline V-gene sequence.

DETAILED DESCRIPTION

The section headings used herein are for organizational purposes only and are not to be construed as limiting the subject matter described.

Unless otherwise defined herein, scientific and technical terms used in connection with the present application shall have the meanings that are commonly understood by those of ordinary skill in the art. Further, unless otherwise required by context, singular terms shall include pluralities and plural terms shall include the singular.

Generally, nomenclatures used in connection with, and techniques of, cell and tissue culture, molecular biology, immunology, microbiology, genetics and protein and nucleic acid chemistry and hybridization described herein are those well known and commonly used in the art. The methods and techniques of the present application are generally performed according to conventional methods well known in the art and as described in various general and more specific references that are cited and discussed throughout the present specification unless otherwise indicated. See, e.g., Sambrook et al., Molecular Cloning: A Laboratory Manual, 3rd ed., Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y. (2001) and subsequent editions, Ausubel et al., Current Protocols in Molecular Biology, Greene Publishing Associates (1992), and Harlow & Lane, Antibodies: A Laboratory Manual, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y. (1988), which are incorporated herein by reference. Enzymatic reactions and purification techniques are performed according to manufacturer's specifications, as commonly accomplished in the art or as described herein. The terminology used in connection with, and the laboratory procedures and techniques of, analytical chemistry, synthetic organic chemistry, and medicinal and pharmaceutical chemistry described herein are those well known and commonly used in the art. Standard techniques can be used for chemical syntheses, chemical analyses, pharmaceutical preparation, formulation, and delivery, and treatment of patients.

It should be understood that the instant disclosure is not limited to the particular methodology, protocols, and reagents, etc., described herein and as such can vary. The terminology used herein is for the purpose of describing particular embodiments only, and is not intended to limit the scope of the present disclosure.

Other than in the operating examples, or where otherwise indicated, all numbers expressing quantities of ingredients or reaction conditions used herein should be understood as modified in all instances by the term "about." The term "about" when used in connection with percentages can mean ±5%, e.g., 1%, 2%, 3%, or 4%.

I. DEFINITIONS

As used herein, the terms "a" and "an" mean "one or more" unless specifically stated otherwise.

As used herein, an "antigen binding protein" is a protein comprising a portion that binds to an antigen or target and, optionally, a scaffold or framework portion that allows the antigen binding portion to adopt a conformation that promotes binding of the antigen binding protein to the antigen. Examples of antigen binding proteins include a human antibody, a humanized antibody; a chimeric antibody; a recombinant antibody; a single chain antibody; a diabody; a triabody; a tetrabody; a Fab fragment; a F(ab')₂ fragment; an IgD antibody; an IgE antibody; an IgM antibody; an IgG1 antibody; an IgG2 antibody; an IgG3 antibody; or an IgG4 antibody, and fragments thereof. The antigen binding protein can comprise, for example, an alternative protein scaffold or artificial scaffold with grafted CDRs or CDR derivatives. Such scaffolds include, but are not limited to, antibody-derived scaffolds comprising mutations introduced to, for example, stabilize the three-dimensional structure of the antigen binding protein as well as wholly synthetic scaffolds comprising, for example, a biocompatible polymer. See, e.g., Korndorfer et al., (2003) Proteins: Structure, Function, and Bioinformatics, 53(1):121-129; Roque et al., (2004) Biotechnol. Prog. 20:639-654. In addition, peptide antibody mimetics ("PAMs") can be used, as well as scaffolds based on antibody mimetics utilizing fibronectin components as a scaffold.

An antigen binding protein can have, for example, the structure of a naturally occurring immunoglobulin. An "immunoglobulin" is a tetrameric molecule. In a naturally occurring immunoglobulin, each tetramer is composed of two identical pairs of polypeptide chains, each pair having one "light" (about 25 kDa) and one "heavy" chain (about 50-70 kDa). The amino-terminal portion of each chain includes a variable region of about 100 to 110 or more amino acids primarily responsible for antigen recognition. The carboxy-terminal portion of each chain defines a constant region primarily responsible for effector function. Human light chains are classified as kappa and lambda light chains. Heavy chains are classified as mu, delta, gamma, alpha, or epsilon, and define the antibody's isotype as IgM, IgD, IgG, IgA, and IgE, respectively. Within light and heavy chains, the variable and constant regions are joined by a "J" region of about 12 or more amino acids, with the heavy chain also including a "D" region of about 10 more amino acids. See generally, Fundamental Immunology 2nd ed. Ch. 7 (Paul, W., ed., Raven Press, N.Y. (1989)), incorporated by reference in its entirety for all purposes. The variable regions of each light/heavy chain pair form the antibody binding site such that an intact immunoglobulin has two binding sites.

Naturally occurring immunoglobulin chains exhibit the same general structure of relatively conserved framework regions (FR) joined by three hypervariable regions, also called complementarity determining regions or CDRs. From N-terminus to C-terminus, both light and heavy chains comprise the domains FR1, CDR1, FR2, CDR2, FR3, CDR3 and FR4. The assignment of amino acids to each domain can be done in accordance with the definitions of Kabat et al., (1991) "Sequences of Proteins of Immunological Interest", 5th Ed., US Dept. of Health and Human Services, PHS, NIH, NIH Publication no. 91-3242. Although presented herein using the Kabat nomenclature system, as desired, the CDRs disclosed herein can also be redefined according an alternative nomenclature scheme, such as that of Chothia (see Chothia & Lesk, (1987) J. Mol. Biol. 196:901-917; Chothia et al., (1989) Nature 342:878-883 or Honegger & Pluckthun, (2001) J. Mol. Biol. 309:657-670).

In the context of the instant disclosure an antigen binding protein is said to "specifically bind" or "selectively bind" its target antigen when the dissociation constant (K_D) is ≤10^-8 M. The antibody specifically binds antigen with "high affinity" when the K_D is ≤5x 10^-9 M, and with "very high affinity" when the K_D is ≤5x 10^-10 M. In one embodiment, the antibodies will bind to a complex comprising β-Klotho and an FGFR, including a complex comprising both human FGFR1c and human β-Klotho, with a K_D of between about 10^-7 M and 10^-12 M, and in yet another embodiment the antibodies will bind with a K_D ≤5x 10^-9.

An "antibody" refers to an intact immunoglobulin or to an antigen binding portion thereof that competes with the intact antibody for specific binding, unless otherwise specified. Antigen binding portions can be produced by recombinant DNA techniques or by enzymatic or chemical cleavage of intact antibodies. Antigen binding portions include, inter alia, Fab, Fab', F(ab')₂, Fv, domain antibodies (dAbs), fragments including complementarity determining regions (CDRs), single-chain antibodies (scFv), chimeric antibodies, diabodies, triabodies, tetrabodies, and polypeptides that contain at least a portion of an immunoglobulin that is sufficient to confer specific antigen binding to the polypeptide.

A Fab fragment is a monovalent fragment having the V_L, V_H, C_L and C_H1 domains; a F(ab')₂ fragment is a bivalent fragment having two Fab fragments linked by a disulfide bridge at the hinge region; a Fd fragment has the V_H and C_H1 domains; an Fv fragment has the V_L and V_H domains of a single arm of an antibody; and a dAb fragment has a V_H domain, a V_L domain, or an antigen-binding fragment of a V_H or V_L domain ( US Patent Nos. 6,846,634 , and 6,696,245 ; and US App. Pub. Nos. 05/0202512 , 04/0202995 , 04/0038291 , 04/0009507 , 03/0039958 , Ward et al., Nature 341:544-546 (1989)).

A single-chain antibody (scFv) is an antibody in which a V_L and a V_H region are joined via a linker (e.g., a synthetic sequence of amino acid residues) to form a continuous protein chain wherein the linker is long enough to allow the protein chain to fold back on itself and form a monovalent antigen binding site (see, e.g., Bird et al., (1988) Science 242:423-26 and Huston et al., (1988) Proc. Natl. Acad. Sci. USA 85:5879-83). Diabodies are bivalent antibodies comprising two polypeptide chains, wherein each polypeptide chain comprises V_H and V_L domains joined by a linker that is too short to allow for pairing between two domains on the same chain, thus allowing each domain to pair with a complementary domain on another polypeptide chain (see, e.g., Holliger et al., (1993) Proc. Natl. Acad. Sci. USA 90:6444-48, and Poljak et al., (1994) Structure 2:1121-23). If the two polypeptide chains of a diabody are identical, then a diabody resulting from their pairing will have two identical antigen binding sites. Polypeptide chains having different sequences can be used to make a diabody with two different antigen binding sites. Similarly, tribodies and tetrabodies are antibodies comprising three and four polypeptide chains, respectively, and forming three and four antigen binding sites, respectively, which can be the same or different.

Complementarity determining regions (CDRs) and framework regions (FR) of a given antibody can be identified using the system described by Kabat et al., (1991) "Sequences of Proteins of Immunological Interest", 5th Ed., US Dept. of Health and Human Services, PHS, NIH, NIH Publication no. 91-3242. Although presented using the Kabat nomenclature system, as desired, the CDRs disclosed herein can also be redefined according an alternative nomenclature scheme, such as that of Chothia (see Chothia & Lesk, (1987) J. Mol. Biol. 196:901-917; Chothia et al., (1989) Nature 342:878-883 or Honegger & Pluckthun, (2001) J. Mol. Biol. 309:657-670). One or more CDRs can be incorporated into a molecule either covalently or noncovalently to make it an antigen binding protein. An antigen binding protein can incorporate the CDR(s) as part of a larger polypeptide chain, can covalently link the CDR(s) to another polypeptide chain, or can incorporate the CDR(s) noncovalently. The CDRs permit the antigen binding protein to specifically bind to a particular antigen of interest.

An antigen binding protein can but need not have one or more binding sites. If there is more than one binding site, the binding sites can be identical to one another or can be different. For example, a naturally occurring human immunoglobulin typically has two identical binding sites, while a "bispecific" or "bifunctional" antibody has two different binding sites. Antigen binding proteins of this bispecific form (e.g., those comprising various heavy and light chain CDRs provided herein) comprise aspects of the instant disclosure.

The term "human antibody" includes all antibodies that have one or more variable and constant regions derived from human immunoglobulin sequences. In one embodiment, all of the variable and constant domains are derived from human immunoglobulin sequences (a fully human antibody). These antibodies can be prepared in a variety of ways, examples of which are described below, including through the immunization with an antigen of interest of a mouse that is genetically modified to express antibodies derived from human heavy and/or light chain-encoding genes, such as a mouse derived from a XENOMOUSE®, ULTIMAB™, HUMAB-MOUSE®, VELOCIMOUSE®, VELOCIMMUNE®, KYMOUSE, or ALIVAMAB system, or derived from human heavy chain transgenic mouse, transgenic rat human antibody repertoire, transgenic rabbit human antibody repertoire or cow human antibody repertoire or HUTARG™ technology. Phage-based approaches can also be employed.

A humanized antibody has a sequence that differs from the sequence of an antibody derived from a non-human species by one or more amino acid substitutions, deletions, and/or additions, such that the humanized antibody is less likely to induce an immune response, and/or induces a less severe immune response, as compared to the non-human species antibody, when it is administered to a human subject. In one embodiment, certain amino acids in the framework and constant domains of the heavy and/or light chains of the non-human species antibody are mutated to produce the humanized antibody. In another embodiment, the constant domain(s) from a human antibody are fused to the variable domain(s) of a non-human species. In another embodiment, one or more amino acid residues in one or more CDR sequences of a non-human antibody are changed to reduce the likely immunogenicity of the non-human antibody when it is administered to a human subject, wherein the changed amino acid residues either are not critical for immunospecific binding of the antibody to its antigen, or the changes to the amino acid sequence that are made are conservative changes, such that the binding of the humanized antibody to the antigen is not significantly worse than the binding of the non-human antibody to the antigen. Examples of how to make humanized antibodies can be found in U.S. Patent Nos. 6,054,297 , 5,886,152 and 5,877,293 .

The term "chimeric antibody" refers to an antibody that contains one or more regions from one antibody and one or more regions from one or more other antibodies. In one embodiment, one or more of the CDRs are derived from a human antibody that binds to a complex comprising β-Klotho and at least one of (i) FGFR1c, (ii) FGFR2c and (iii) FGFR3c. In another embodiment, all of the CDRs are derived from a human antibody that binds to a complex β-Klotho and at least one of (i) FGFR1c, (ii) FGFR2c and (iii) FGFR3c. In another embodiment, the CDRs from more than one human antibody that binds to a complex comprising β-Klotho and at least one of (i) FGFR1c, (ii) FGFR2c and (iii) FGFR3c are mixed and matched in a chimeric antibody. For instance, a chimeric antibody can comprise a CDR1 from the light chain of a first human antibody that binds to a complex comprising β-Klotho and at least one of (i) FGFR1c, (ii) FGFR2c and (iii) FGFR3c, a CDR2 and a CDR3 from the light chain of a second human antibody that binds to a complex comprising β-Klotho and at least one of (i) FGFR1c, (ii) FGFR2c and (iii) FGFR3c, and the CDRs from the heavy chain from a third antibody that binds to a complex comprising β-Klotho and at least one of (i) FGFR1c, (ii) FGFR2c and (iii) FGFR3c. Further, the framework regions can be derived from one of the same antibodies that bind to a complex comprising β-Klotho and at least one of (i) FGFR1c, (ii) FGFR2c and (iii) FGFR3c, from one or more different antibodies, such as a human antibody, or from a humanized antibody. In one example of a chimeric antibody, a portion of the heavy and/or light chain is identical with, homologous to, or derived from an antibody from a particular species or belonging to a particular antibody class or subclass, while the remainder of the chain(s) is/are identical with, homologous to, or derived from an antibody or antibodies from another species or belonging to another antibody class or subclass. Also included are fragments of such antibodies that exhibit the desired biological activity (e.g., the ability to specifically bind to a complex comprising β-Klotho and at least one of (i) FGFR1c, (ii) FGFR2c and (iii) FGFR3c).

The term "light chain" includes a full-length light chain and fragments thereof having sufficient variable region sequence to confer binding specificity. A full-length light chain includes a variable region domain, V_L, and a constant region domain, C_L. The variable region domain of the light chain is at the amino-terminus of the polypeptide. Light chains include kappa ("κ") chains and lambda ("λ") chains.

The term "heavy chain" includes a full-length heavy chain and fragments thereof having sufficient variable region sequence to confer binding specificity. A full-length heavy chain includes a variable region domain, V_H, and three constant region domains, C_H1, C_H2, and C_H3. The V_H domain is at the amino-terminus of the polypeptide, and the C_H domains are at the carboxyl-terminus, with the C_H3 being closest to the carboxy-terminus of the polypeptide. Heavy chains can be of any isotype, including IgG (including IgG1, IgG2, IgG3 and IgG4 subtypes), IgA (including IgA1 and IgA2 subtypes), IgM and IgE.

The term "immunologically functional fragment" (or simply "fragment") of an antigen binding protein, e.g., an antibody or immunoglobulin chain (heavy or light chain), as used herein, is an antigen binding protein comprising a portion (regardless of how that portion is obtained or synthesized) of an antibody that lacks at least some of the amino acids present in a full-length chain but which is capable of specifically binding to an antigen. Such fragments are biologically active in that they bind specifically to the target antigen and can compete with other antigen binding proteins, including intact antibodies, for specific binding to a given epitope. In one aspect, such a fragment will retain at least one CDR present in the full-length light or heavy chain, and in some embodiments will comprise a single heavy chain and/or light chain or portion thereof. These biologically active fragments can be produced by recombinant DNA techniques, or can be produced by enzymatic or chemical cleavage of antigen binding proteins, including intact antibodies. Immunologically functional immunoglobulin fragments include, but are not limited to, Fab, Fab', F(ab')₂, Fv, domain antibodies and single-chain antibodies, and can be derived from any mammalian source, including but not limited to human, mouse, rat, camelid or rabbit. It is contemplated further that a functional portion of the antigen binding proteins disclosed herein, for example, one or more CDRs, could be covalently bound to a second protein or to a small molecule to create a therapeutic agent directed to a particular target in the body, possessing bifunctional therapeutic properties, or having a prolonged serum half-life.

An "Fc" region contains two heavy chain fragments comprising the C_H2 and C_H3 domains of an antibody. The two heavy chain fragments are held together by two or more disulfide bonds and by hydrophobic interactions of the C_H3 domains.

An "Fab' fragment" contains one light chain and a portion of one heavy chain that contains the V_H domain and the C_H1 domain and also the region between the C_H1 and C_H2 domains, such that an interchain disulfide bond can be formed between the two heavy chains of two Fab' fragments to form an F(ab')₂ molecule.

An "F(ab')₂ fragment" contains two light chains and two heavy chains containing a portion of the constant region between the C_H1 and C_H2 domains, such that an interchain disulfide bond is formed between the two heavy chains. A F(ab')₂ fragment thus is composed of two Fab' fragments that are held together by a disulfide bond between the two heavy chains.

The "Fv region" comprises the variable regions from both the heavy and light chains, but lacks the constant regions.

A "domain antibody" is an immunologically functional immunoglobulin fragment containing only the variable region of a heavy chain or the variable region of a light chain. In some instances, two or more V_H regions are covalently joined with a peptide linker to create a bivalent domain antibody. The two V_H regions of a bivalent domain antibody can target the same or different antigens.

A "hemibody" is an immunologically-functional immunoglobulin construct comprising a complete heavy chain, a complete light chain and a second heavy chain Fc region paired with the Fc region of the complete heavy chain. A linker can, but need not, be employed to join the heavy chain Fc region and the second heavy chain Fc region. In particular embodiments a hemibody is a monovalent form of an antigen binding protein disclosed herein. In other embodiments, pairs of charged residues can be employed to associate one Fc region with the second Fc region.

A "bivalent antigen binding protein" or "bivalent antibody" comprises two antigen binding sites. In some instances, the two binding sites have the same antigen specificities. Bivalent antigen binding proteins and bivalent antibodies can be bispecific, as described herein, and form aspects of the instant disclosure.

A "multispecific antigen binding protein" or "multispecific antibody" is one that targets more than one antigen or epitope, and forms another aspect of the instant disclosure.

A "bispecific," "dual-specific" or "bifunctional" antigen binding protein or antibody is a hybrid antigen binding protein or antibody, respectively, having two different antigen binding sites. Bispecific antigen binding proteins and antibodies are a species of multispecific antigen binding protein or multispecific antibody and can be produced by a variety of methods including, but not limited to, fusion of hybridomas or linking of Fab' fragments. See, e.g., Songsivilai and Lachmann, (1990) Clin. Exp. Immunol. 79:315-321; Kostelny et al., (1992) J. Immunol. 148:1547-1553. The two binding sites of a bispecific antigen binding protein or antibody will bind to two different epitopes, which can reside on the same (e.g., β-Klotho, FGFR1c, FGFR2c, or FGFR3c) or different protein targets (e.g., β-Klotho and one of (i) FGFR1c, (ii) FGFR2c, and (iii) FGFR3c).

The terms "FGF21-like signaling" and "induces FGF21-like signaling," when applied to an antigen binding protein of the present disclosure, means that the antigen binding protein mimics, or modulates, an in vivo biological effect induced by the binding to a complex comprising β-Klotho and at least one of (i) FGFR1c, (ii) FGFR2c and (iii) FGFR3c and induces a biological response that otherwise would result from FGF21 binding to a complex comprising β-Klotho and at least one of (i) FGFR1c, (ii) FGFR2c and (iii) FGFR3c in vivo. In assessing the binding and specificity of an antigen binding protein, e.g., an antibody or immunologically functional fragment thereof, an antibody or fragment is deemed to induce a biological response when the response is equal to or greater than 5%, and preferably equal to or greater than 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90% or 95%, of the activity of a wild type FGF21 standard comprising the mature form of SEQ ID NO: 2 (i.e., the mature form of the human FGF21 sequence) and has the following properties: exhibiting an efficacy level of equal to or more than 5% of an FGF21 standard, with an EC₅₀ of equal to or less than 100nM, e.g., 90 nM, 80 nM, 70nM, 60nM, 50nM, 40nM, 30nM, 20nM or 10 nM in (1) the recombinant FGF21 receptor-mediated luciferase reporter cell assay of Example 4; (2) ERK-phosphorylation in the recombinant FGF21 receptor mediated cell assay of Example 4; and (3) ERK-phosphorylation in human adipocytes as described in Example 4. The "potency" of an antigen binding protein is defined as exhibiting an EC50 of equal to or less than 100nM, e.g., 90nM, 80nM, 70nM, 60nM, 50nM, 40nM, 30nM, 20nM, 10 nM and preferably less than 10nM of the antigen binding protein in the following assays: (1) the recombinant FGF21 receptor mediated luciferase-reporter cell assay of Example 4; (2) the ERK-phosphorylation in the recombinant FGF21 receptor mediated cell assay of Example 4; and (3) ERK-phosphorylation in human adipocytes as described in Example 4.

It is noted that not all of the antigen binding proteins of the present disclosure induce FGF21-mediated signaling (e.g., that induce agonistic activity), nor is this property desirable in all circumstances. Nevertheless, antigen binding proteins that do not induce FGF21-mediated signaling form aspects of the present disclosure and may be useful as diagnostic reagents or other applications.

As used herein, the term "FGF21R" means a multimeric receptor complex that FGF21 is known or suspected to form in vivo. In various embodiments, FGF21R comprises (i) an FGFR, e.g., FGFR1c, FGFR2c, FGFR3c or FGFR4, and (ii) β-Klotho.

The term "polynucleotide" or "nucleic acid" includes both single-stranded and double-stranded nucleotide polymers. The nucleotides comprising the polynucleotide can be ribonucleotides or deoxyribonucleotides or a modified form of either type of nucleotide. Said modifications include base modifications such as bromouridine and inosine derivatives, ribose modifications such as 2', 3'-dideoxyribose, and internucleotide linkage modifications such as phosphorothioate, phosphorodithioate, phosphoroselenoate, phosphorodiselenoate, phosphoroanilothioate, phoshoraniladate and phosphoroamidate.

The term "oligonucleotide" means a polynucleotide comprising 200 or fewer nucleotides. In some embodiments, oligonucleotides are 10 to 60 bases in length. In other embodiments, oligonucleotides are 12, 13, 14, 15, 16, 17, 18, 19, or 20 to 40 nucleotides in length. Oligonucleotides can be single stranded or double stranded, e.g., for use in the construction of a mutant gene. Oligonucleotides can be sense or antisense oligonucleotides. An oligonucleotide can include a label, including a radiolabel, a fluorescent label, a hapten or an antigenic label, for detection assays. Oligonucleotides can be used, for example, as PCR primers, cloning primers or hybridization probes.

An "isolated nucleic acid molecule" means a DNA or RNA of genomic, mRNA, cDNA, or synthetic origin or some combination thereof which is not associated with all or a portion of a polynucleotide in which the isolated polynucleotide is found in nature, or is linked to a polynucleotide to which it is not linked in nature. For purposes of this disclosure, it is understood that "a nucleic acid molecule comprising" a particular nucleotide sequence does not encompass intact chromosomes. Isolated nucleic acid molecules "comprising" specified nucleic acid sequences can include, in addition to the specified sequences, coding sequences for up to ten or even up to twenty other proteins or portions thereof, or can include operably linked regulatory sequences that control expression of the coding region of the recited nucleic acid sequences, and/or can include vector sequences.

Unless specified otherwise, the left-hand end of any single-stranded polynucleotide sequence discussed herein is the 5' end; the left-hand direction of double-stranded polynucleotide sequences is referred to as the 5' direction. The direction of 5' to 3' addition of nascent RNA transcripts is referred to as the transcription direction; sequence regions on the DNA strand having the same sequence as the RNA transcript that are 5' to the 5' end of the RNA transcript are referred to as "upstream sequences;" sequence regions on the DNA strand having the same sequence as the RNA transcript that are 3' to the 3' end of the RNA transcript are referred to as "downstream sequences."

The term "control sequence" refers to a polynucleotide sequence that can affect the expression and processing of coding sequences to which it is ligated. The nature of such control sequences can depend upon the host organism. In particular embodiments, control sequences for prokaryotes can include a promoter, a ribosomal binding site, and a transcription termination sequence. For example, control sequences for eukaryotes can include promoters comprising one or a plurality of recognition sites for transcription factors, transcription enhancer sequences, and transcription termination sequence. "Control sequences" can include leader sequences and/or fusion partner sequences.

The term "vector" means any molecule or entity (e.g., nucleic acid, plasmid, bacteriophage or virus) used to transfer protein coding information into a host cell.

The term "expression vector" or "expression construct" refers to a vector that is suitable for transformation of a host cell and contains nucleic acid sequences that direct and/or control (in conjunction with the host cell) expression of one or more heterologous coding regions operatively linked thereto. An expression construct can include, but is not limited to, sequences that affect or control transcription, translation, and, if introns are present, affect RNA splicing of a coding region operably linked thereto.

As used herein, "operably linked" means that the components to which the term is applied are in a relationship that allows them to carry out their inherent functions under suitable conditions. For example, a control sequence in a vector that is "operably linked" to a protein coding sequence is ligated thereto so that expression of the protein coding sequence is achieved under conditions compatible with the transcriptional activity of the control sequences.

The term "host cell" means a cell that has been transformed, or is capable of being transformed, with a nucleic acid sequence and thereby expresses a gene of interest. The term includes the progeny of the parent cell, whether or not the progeny is identical in morphology or in genetic make-up to the original parent cell, so long as the gene of interest is present.

The term "transduction" means the transfer of genes from one bacterium to another, usually by bacteriophage. "Transduction" also refers to the acquisition and transfer of eukaryotic cellular sequences by replication-defective retroviruses.

The term "transfection" means the uptake of foreign or exogenous DNA by a cell, and a cell has been "transfected" when the exogenous DNA has been introduced inside the cell membrane. A number of transfection techniques are well known in the art and are disclosed herein. See, e.g., Graham et al., (1973) Virology 52:456; Sambrook et al., (2001), supra; Davis et al., (1986) Basic Methods in Molecular Biology, Elsevier; Chu et al., (1981) Gene 13:197. Such techniques can be used to introduce one or more exogenous DNA moieties into suitable host cells.

The term "transformation" refers to a change in a cell's genetic characteristics, and a cell has been transformed when it has been modified to contain new DNA or RNA. For example, a cell is transformed where it is genetically modified from its native state by introducing new genetic material via transfection, transduction, or other techniques. Following transfection or transduction, the transforming DNA can recombine with that of the cell by physically integrating into a chromosome of the cell, or can be maintained transiently as an episomal element without being replicated, or can replicate independently as a plasmid. A cell is considered to have been "stably transformed" when the transforming DNA is replicated with the division of the cell.

The terms "polypeptide" or "protein" are used interchangeably herein to refer to a polymer of amino acid residues. The terms also apply to amino acid polymers in which one or more amino acid residues is an analog or mimetic of a corresponding naturally occurring amino acid, as well as to naturally occurring amino acid polymers. The terms can also encompass amino acid polymers that have been modified, e.g., by the addition of carbohydrate residues to form glycoproteins, or phosphorylated. Polypeptides and proteins can be produced by a naturally-occurring and non-recombinant cell, or polypeptides and proteins can be produced by a genetically-engineered or recombinant cell. Polypeptides and proteins can comprise molecules having the amino acid sequence of a native protein, or molecules having deletions from, additions to, and/or substitutions of one or more amino acids of the native sequence. The terms "polypeptide" and "protein" encompass antigen binding proteins that specifically or selectively bind to a complex comprising β-Klotho and an FGFR (e.g., FGFR1c, FGFR2c or FGFR3c), or sequences that have deletions from, additions to, and/or substitutions of one or more amino acids of an antigen binding protein that specifically or selectively binds to a complex comprising β-Klotho and at least one of (i) FGFR1c, (ii) FGFR2c and (iii) FGFR3c. The term "polypeptide fragment" refers to a polypeptide that has an amino-terminal deletion, a carboxyl-terminal deletion, and/or an internal deletion as compared with the full-length protein. Such fragments can also contain modified amino acids as compared with the full-length protein. In certain embodiments, fragments are about five to 500 amino acids long. For example, fragments can be at least 5, 6, 8, 10, 14, 20, 50, 70, 100, 110, 150, 200, 250, 300, 350, 400, or 450 amino acids long. Useful polypeptide fragments include immunologically functional fragments of antibodies, including binding domains. In the case of an antigen binding protein that binds to a complex β-Klotho and at least one of (i) FGFR1c, (ii) FGFR2c and (iii) FGFR3c, useful fragments include but are not limited to a CDR region, a variable domain of a heavy or light chain, a portion of an antibody chain or just its variable region including two CDRs, and the like.

The term "isolated protein" referred means that a subject protein (1) is free of at least some other proteins with which it would normally be found, (2) is essentially free of other proteins from the same source, e.g., from the same species, (3) is expressed by a cell from a different species, (4) has been separated from at least about 50 percent of polynucleotides, lipids, carbohydrates, or other materials with which it is associated in nature, (5) is operably associated (by covalent or noncovalent interaction) with a polypeptide with which it is not associated in nature, or (6) does not occur in nature. Typically, an "isolated protein" constitutes at least about 5%, at least about 10%, at least about 25%, or at least about 50% of a given sample. Genomic DNA, cDNA, mRNA or other RNA, of synthetic origin, or any combination thereof can encode such an isolated protein. Preferably, the isolated protein is substantially free from proteins or polypeptides or other contaminants that are found in its natural environment that would interfere with its therapeutic, diagnostic, prophylactic, research or other use.

A "variant" of a polypeptide (e.g., an antigen binding protein, or an antibody) comprises an amino acid sequence wherein one or more amino acid residues are inserted into, deleted from and/or substituted into the amino acid sequence relative to another polypeptide sequence. Variants include fusion proteins.

A "derivative" of a polypeptide is a polypeptide (e.g., an antigen binding protein, or an antibody) that has been chemically modified in some manner distinct from insertion, deletion, or substitution variants, e.g., by conjugation to another chemical moiety.

The term "naturally occurring" as used throughout the specification in connection with biological materials such as polypeptides, nucleic acids, host cells, and the like, refers to materials which are found in nature.

"Antigen binding region" means a protein, or a portion of a protein, that specifically binds a specified antigen, e.g., a complex comprising β-Klotho and anβ-Klotho and at least one of (i) FGFR1c, (ii) FGFR2c and (iii) FGFR3c. For example, that portion of an antigen binding protein that contains the amino acid residues that interact with an antigen and confer on the antigen binding protein its specificity and affinity for the antigen is referred to as "antigen binding region." An antigen binding region typically includes one or more "complementary binding regions" ("CDRs"). Certain antigen binding regions also include one or more "framework" regions. A "CDR" is an amino acid sequence that contributes to antigen binding specificity and affinity. "Framework" regions can aid in maintaining the proper conformation of the CDRs to promote binding between the antigen binding region and an antigen.

In certain aspects, recombinant antigen binding proteins that bind to a complex comprising β-Klotho and at least one of (i) FGFR1c, (ii) FGFR2c and (iii) FGFR3c, are provided. In this context, a "recombinant protein" is a protein made using recombinant techniques, i.e., through the expression of a recombinant nucleic acid as described herein. Methods and techniques for the production of recombinant proteins are well known in the art.

The term "compete" when used in the context of antigen binding proteins (e.g., neutralizing antigen binding proteins, neutralizing antibodies, agonistic antigen binding proteins, agonistic antibodies and binding proteins that bind to a complex comprising β-Klotho and at least one of (i) FGFR1c, (ii) FGFR2c and (iii) FGFR3c) that compete for the same epitope or binding site on a target means competition between antigen binding proteins as determined by an assay in which the antigen binding protein (e.g., antibody or immunologically functional fragment thereof) under study prevents or inhibits the specific binding of a reference molecule (e.g., a reference ligand, or reference antigen binding protein, such as a reference antibody) to a common antigen (e.g., FGFR1c, FGFR2c, FGFR3c, β-Klotho or a fragment thereof, or a complex comprising β-Klotho and at least one of (i) FGFR1c, (ii) FGFR2c and (iii) FGFR3c). Numerous types of competitive binding assays can be used to determine if a test molecule competes with a reference molecule for binding. Examples of assays that can be employed include solid phase direct or indirect radioimmunoassay (RIA), solid phase direct or indirect enzyme immunoassay (EIA), sandwich competition assay (see, e.g., Stahli et al., (1983) Methods in Enzymology 9:242-253); solid phase direct biotin-avidin EIA (see, e.g., Kirkland et al., (1986) J. Immunol. 137:3614-3619) solid phase direct labeled assay, solid phase direct labeled sandwich assay (see, e.g., Harlow and Lane, (1988) supra); solid phase direct label RIA using I-125 label (see, e.g., Morel et al., (1988) Molec. Immunol. 25:7-15); solid phase direct biotin-avidin EIA (see, e.g., Cheung, et al., (1990) Virology 176:546-552); and direct labeled RIA (Moldenhauer et al., (1990) Scand. J. Immunol. 32:77-82). Typically, such an assay involves the use of a purified antigen bound to a solid surface or cells bearing either of an unlabelled test antigen binding protein or a labeled reference antigen binding protein. Competitive inhibition is measured by determining the amount of label bound to the solid surface or cells in the presence of the test antigen binding protein. Usually the test antigen binding protein is present in excess. Antigen binding proteins identified by competition assay (competing antigen binding proteins) include antigen binding proteins binding to the same epitope as the reference antigen binding proteins and antigen binding proteins binding to an adjacent epitope sufficiently proximal to the epitope bound by the reference antigen binding protein for steric hindrance to occur. Additional details regarding methods for determining competitive binding are provided in the examples herein. Usually, when a competing antigen binding protein is present in excess, it will inhibit specific binding of a reference antigen binding protein to a common antigen by at least 40%, 45%, 50%, 55%, 60%, 65%, 70% or 75%. In some instance, binding is inhibited by at least 80%, 85%, 90%, 95%, or 97% or more.

The term "antigen" refers to a molecule or a portion of a molecule capable of being bound by a selective binding agent, such as an antigen binding protein (including, e.g., an antibody or immunological functional fragment thereof), and may also be capable of being used in an animal to produce antibodies capable of binding to that antigen. An antigen can possess one or more epitopes that are capable of interacting with different antigen binding proteins, e.g., antibodies.

The term "epitope" means the amino acids of a target molecule that are contacted by an antigen binding protein (for example, an antibody) when the antigen binding protein is bound to the target molecule. The term includes any subset of the complete list of amino acids of the target molecule that are contacted when an antigen binding protein, such as an antibody, is bound to the target molecule. An epitope can be contiguous or non-contiguous (e.g., (i) in a single-chain polypeptide, amino acid residues that are not contiguous to one another in the polypeptide sequence but that within in context of the target molecule are bound by the antigen binding protein, or (ii) in a multimeric receptor comprising two or more individual components, e.g., a complex comprising β-Klotho and at least one of (i) FGFR1c, (ii) FGFR2c and (iii) FGFR3c, amino acid residues that are present on one or more of the individual components, but which are still bound by the antigen binding protein). In certain embodiments, epitopes can be mimetic in that they comprise a three dimensional structure that is similar to an antigenic epitope used to generate the antigen binding protein, yet comprise none or only some of the amino acid residues found in that epitope used to generate the antigen binding protein. Most often, epitopes reside on proteins, but in some instances can reside on other kinds of molecules, such as nucleic acids. Epitope determinants can include chemically active surface groupings of molecules such as amino acids, sugar side chains, phosphoryl or sulfonyl groups, and can have specific three dimensional structural characteristics, and/or specific charge characteristics. Generally, antigen binding proteins specific for a particular target molecule will preferentially recognize an epitope on the target molecule in a complex mixture of proteins and/or macromolecules.

The term "identity" refers to a relationship between the sequences of two or more polypeptide molecules or two or more nucleic acid molecules, as determined by aligning and comparing the sequences. "Percent identity" means the percent of identical residues between the amino acids or nucleotides in the compared molecules and is calculated based on the size of the smallest of the molecules being compared. For these calculations, gaps in alignments (if any) must be addressed by a particular mathematical model or computer program (i.e., an "algorithm"). Methods that can be used to calculate the identity of the aligned nucleic acids or polypeptides include those described in Computational Molecular Biology, (Lesk, A. M., ed.), (1988) New York: Oxford University Press; Biocomputing Informatics and Genome Projects, (Smith, D. W., ed.), 1993, New York: Academic Press; Computer Analysis of Sequence Data, Part I, (Griffin, A. M., and Griffin, H. G., eds.), 1994, New Jersey: Humana Press; von Heinje, G., (1987) Sequence Analysis in Molecular Biology, New York: Academic Press; Sequence Analysis Primer, (Gribskov, M. and Devereux, J., eds.), 1991, New York: M. Stockton Press; and Carillo et al., (1988) J. Applied Math. 48:1073.

In calculating percent identity, the sequences being compared are aligned in a way that gives the largest match between the sequences. The computer program used to determine percent identity is the GCG program package, which includes GAP (Devereux et al., (1984) Nucl. Acid Res. 12:387; Genetics Computer Group, University of Wisconsin, Madison, WI). The computer algorithm GAP is used to align the two polypeptides or polynucleotides for which the percent sequence identity is to be determined. The sequences are aligned for optimal matching of their respective amino acid or nucleotide (the "matched span", as determined by the algorithm). A gap opening penalty (which is calculated as 3x the average diagonal, wherein the "average diagonal" is the average of the diagonal of the comparison matrix being used; the "diagonal" is the score or number assigned to each perfect amino acid match by the particular comparison matrix) and a gap extension penalty (which is usually 1/10 times the gap opening penalty), as well as a comparison matrix such as PAM 250 or BLOSUM 62 are used in conjunction with the algorithm. In certain embodiments, a standard comparison matrix (see, Dayhoff et al., (1978) Atlas of Protein Sequence and Structure 5:345-352 for the PAM 250 comparison matrix; Henikoff et al., (1992) Proc. Natl. Acad. Sci. U.S.A. 89:10915-10919 for the BLOSUM 62 comparison matrix) is also used by the algorithm.

Recommended parameters for determining percent identity for polypeptides or nucleotide sequences using the GAP program are the following:

Algorithm: Needleman et al., 1970, J. Mol. Biol. 48:443-453;
Comparison matrix: BLOSUM 62 from Henikoff et al., 1992, supra;
Gap Penalty: 12 (but with no penalty for end gaps)
Gap Length Penalty: 4
Threshold of Similarity: 0

Certain alignment schemes for aligning two amino acid sequences can result in matching of only a short region of the two sequences, and this small aligned region can have very high sequence identity even though there is no significant relationship between the two full-length sequences. Accordingly, the selected alignment method (e.g., the GAP program) can be adjusted if so desired to result in an alignment that spans at least 50 contiguous amino acids of the target polypeptide.

As used herein, "substantially pure" means that the described species of molecule is the predominant species present, that is, on a molar basis it is more abundant than any other individual species in the same mixture. In certain embodiments, a substantially pure molecule is a composition wherein the object species comprises at least 50% (on a molar basis) of all macromolecular species present. In other embodiments, a substantially pure composition will comprise at least 80%, 85%, 90%, 95%, or 99% of all macromolecular species present in the composition. In other embodiments, the object species is purified to essential homogeneity wherein contaminating species cannot be detected in the composition by conventional detection methods and thus the composition consists of a single detectable macromolecular species.

The terms "treat" and "treating" refer to any indicia of success in the treatment or amelioration of an injury, pathology or condition, including any objective or subjective parameter such as abatement; remission; diminishing of symptoms or making the injury, pathology or condition more tolerable to the patient; slowing in the rate of degeneration or decline; making the final point of degeneration less debilitating; improving a patient's physical or mental well-being. The treatment or amelioration of symptoms can be based on objective or subjective parameters; including the results of a physical examination, neuropsychiatric exams, and/or a psychiatric evaluation. For example, certain methods presented herein can be employed to treat Type 2 diabetes, obesity and/or dyslipidemia, either prophylactically or as an acute treatment, to decrease plasma glucose levels, to decrease circulating triglyceride levels, to decrease circulating cholesterol levels and/or ameliorate a symptom associated with type 2 diabetes, obesity and dyslipidemia.

An "effective amount" is generally an amount sufficient to reduce the severity and/or frequency of symptoms, eliminate the symptoms and/or underlying cause, prevent the occurrence of symptoms and/or their underlying cause, and/or improve or remediate the damage that results from or is associated with diabetes, obesity and dyslipidemia. In some embodiments, the effective amount is a therapeutically effective amount or a prophylactically effective amount. A "therapeutically effective amount" is an amount sufficient to remedy a disease state (e.g., diabetes, obesity or dyslipidemia) or symptoms, particularly a state or symptoms associated with the disease state, or otherwise prevent, hinder, retard or reverse the progression of the disease state or any other undesirable symptom associated with the disease in any way whatsoever. A "prophylactically effective amount" is an amount of a pharmaceutical composition that, when administered to a subject, will have the intended prophylactic effect, e.g., preventing or delaying the onset (or reoccurrence) of diabetes, obesity or dyslipidemia, or reducing the likelihood of the onset (or reoccurrence) of diabetes, obesity or dyslipidemia or associated symptoms. The full therapeutic or prophylactic effect does not necessarily occur by administration of one dose, and may occur only after administration of a series of doses. Thus, a therapeutically or prophylactically effective amount can be administered in one or more administrations.

"Amino acid" takes its normal meaning in the art. The twenty naturally-occurring amino acids and their abbreviations follow conventional usage. See, Immunology-A Synthesis, 2nd Edition, (E. S. Golub and D. R. Green, eds.), Sinauer Associates: Sunderland, Mass. (1991), incorporated herein by reference for any purpose. Stereoisomers (e.g., D-amino acids) of the twenty conventional amino acids, unnatural or non-naturally occurring or encoded amino acids such as α-,α-disubstituted amino acids, N-alkyl amino acids, and other unconventional amino acids can also be suitable components for polypeptides and are included in the phrase "amino acid." Examples of non-natural and non-naturally encoded amino acids (which can be substituted for any naturally-occurring amino acid found in any sequence disclosed herein, as desired) include: 4-hydroxyproline, γ-carboxyglutamate, ε-N,N,N-trimethyllysine, ε-N-acetyllysine, O-phosphoserine, N-acetylserine, N-formylmethionine, 3-methylhistidine, 5-hydroxylysine, σ-N-methylarginine, and other similar amino acids and imino acids (e.g., 4-hydroxyproline). In the polypeptide notation used herein, the left-hand direction is the amino terminal direction and the right-hand direction is the carboxyl-terminal direction, in accordance with standard usage and convention. A nonlimiting lists of examples of non-naturally occurring/encoded amino acids that can be inserted into an antigen binding protein sequence or substituted for a wild-type residue in an antigen binding sequence include β-amino acids, homoamino acids, cyclic amino acids and amino acids with derivatized side chains. Examples include (in the L-form or D-form; abbreviated as in parentheses): citrulline (Cit), homocitrulline (hCit), Nα-methylcitrulline (NMeCit), Nα-methylhomocitrulline (Nα-MeHoCit), ornithine (Orn), Nα-Methylornithine (Nα-MeOrn or NMeOrn), sarcosine (Sar), homolysine (hLys or hK), homoarginine (hArg or hR), homoglutamine (hQ), Nα-methylarginine (NMeR), Nα-methylleucine (Nα-MeL or NMeL), N-methylhomolysine (NMeHoK), Nα-methylglutamine (NMeQ), norleucine (Nle), norvaline (Nva), 1,2,3,4-tetrahydroisoquinoline (Tic), Octahydroindole-2-carboxylic acid (Oic), 3-(1-naphthyl)alanine (1-Nal), 3-(2-naphthyl)alanine (2-Nal), 1,2,3,4-tetrahydroisoquinoline (Tic), 2-indanylglycine (IgI), para-iodophenylalanine (pI-Phe), para-aminophenylalanine (4AmP or 4-Amino-Phe), 4-guanidino phenylalanine (Guf), glycyllysine (abbreviated "K(Nε-glycyl)" or "K(glycyl)" or "K(gly)"), nitrophenylalanine (nitrophe), aminophenylalanine (aminophe or Amino-Phe), benzylphenylalanine (benzylphe), γ-carboxyglutamic acid (γ-carboxyglu), hydroxyproline (hydroxypro), p-carboxyl-phenylalanine (Cpa), α-aminoadipic acid (Aad), Nα-methyl valine (NMeVal), N-α-methyl leucine (NMeLeu), Nα-methylnorleucine (NMeNle), cyclopentylglycine (Cpg), cyclohexylglycine (Chg), acetylarginine (acetylarg), α, β-diaminopropionoic acid (Dpr), α, γ-diaminobutyric acid (Dab), diaminopropionic acid (Dap), cyclohexylalanine (Cha), 4-methyl-phenylalanine (MePhe), β, β-diphenyl-alanine (BiPhA), aminobutyric acid (Abu), 4-phenyl-phenylalanine (or biphenylalanine; 4Bip), α-amino-isobutyric acid (Aib), beta-alanine, beta-aminopropionic acid, piperidinic acid, aminocaprioic acid, aminoheptanoic acid, aminopimelic acid, desmosine, diaminopimelic acid, N-ethylglycine, N-ethylaspargine, hydroxylysine, allo-hydroxylysine, isodesmosine, allo-isoleucine, N-methylglycine, N-methylisoleucine, N-methylvaline, 4-hydroxyproline (Hyp), γ-carboxyglutamate, ε-N,N,N-trimethyllysine, ε-N-acetyllysine, O-phosphoserine, N-acetylserine, N-formylmethionine, 3-methylhistidine, 5-hydroxylysine, ω-methylarginine, 4-Amino-O-Phthalic Acid (4APA), and other similar amino acids, and derivatized forms of any of those specifically listed.

II. GENERAL OVERVIEW

Antigen-binding proteins that bind to a complex comprising β-Klotho and at least one of (i) FGFR1c, (ii) FGFR2c and (iii) FGFR3c are provided herein. A unique property of the antigen binding proteins disclosed herein is the agonistic nature of these proteins, specifically the ability to mimic the in vivo effect of FGF21 and to induce FGF21-like signaling. More remarkably and specifically, some of the antigen binding proteins disclosed herein induce FGF21-like signaling in several in vitro cell-based assay, including the ELK-luciferase reporter assay of Example 4 under the following conditions: (1) the binding to and activity of the FGF21 receptor is β-Klotho dependent; (2) the activity is selective to the FGFR/β--Klotho complex; (3) the binding to the FGFR1c/βKlotho complex triggers FGF21-like signaling pathways; and (4) the potency (EC50) is comparable to a wild-type FGF21 standard comprising the mature form of SEQ ID NO: 2, as measured in the following cell-based assays: (1) the recombinant FGF21 receptor mediated luciferase-reporter cell assay of Example 4; (2) the ERK-phosphorylation in the recombinant FGF21 receptor mediated cell assay of Example 4; and (3) ERK-phosphorylation in human adipocytes as described in more details in Example 6. The disclosed antigen binding proteins, therefore, are expected to exhibit activities in vivo that are consistent with the natural biological function of FGF21. This property makes the disclosed antigen binding proteins viable therapeutics for the treatment of metabolic diseases such as type 2 diabetes, obesity, dyslipidemia, NASH, cardiovascular disease, metabolic syndrome and broadly any disease or condition in which it is desirable to mimic or augment the in vivo effects of FGF21.

In some embodiments of the present disclosure the antigen binding proteins provided can comprise polypeptides into which one or more complementary determining regions (CDRs) can be embedded and/or joined. In such antigen binding proteins, the CDRs can be embedded into a "framework" region, which orients the CDR(s) such that the proper antigen binding properties of the CDR(s) is achieved. In general, such antigen binding proteins that are provided can facilitate or enhance the interaction between an FGFR (e.g., FGFR1c, FGFR2c or FGFR3c) and β-Klotho, and can substantially induce FGF21-like signaling. Accordingly, the antigen binding proteins provided herein mimic the in vivo role of FGF21 and are thus "agonistic" and offer potential therapeutic benefit for the range of conditions which benefit from FGF21 therapy, including type 2 diabetes, obesity, dyslipidemia, NASH, cardiovascular disease, metabolic syndrome and broadly any disease or condition in which it is desirable to mimic or augment the in vivo effects of FGF21.

Certain antigen binding proteins described herein are antibodies or are derived from antibodies. In certain embodiments, the polypeptide structure of the antigen binding proteins is based on antibodies, including, but not limited to, monoclonal antibodies, bispecific antibodies, minibodies, domain antibodies, synthetic antibodies (sometimes referred to herein as "antibody mimetics"), chimeric antibodies, humanized antibodies, human antibodies, antibody fusions (sometimes referred to herein as "antibody conjugates"), hemibodies and fragments thereof. The various structures are further described herein below.

The antigen binding proteins provided herein have been demonstrated to bind to a complex comprising β-Klotho and at least one of (i) FGFR1c, (ii) FGFR2c and (iii) FGFR3c, and particularly to a complex comprising human β-Klotho and a human FGFR (e.g., a human FGFR1c, a human FGFR2c or a human FGFR3c). As described and shown in the Examples presented herein, based Western blot results, known commercially-available anti-β-Klotho or anti-FGFR1c antibodies bind to denatured β-Klotho or FGFR1c whereas the antigen binding protein (which are agonistic antibodies) do not. Conversely, the provided antigen binding proteins recognize the native structure of the FGFR1c and β-Klotho on the cell surface whereas the commercial antibodies do not. The antigen binding proteins that are provided therefore mimic the natural in vivo biological activity of FGF21. As a consequence, the antigen binding proteins provided herein are capable of activating FGF21-like signaling activity. In particular, the disclosed antigen binding proteins can have one or more of the following activities in vivo: induction of FGF21-like signal transduction pathways, lowering blood glucose levels, lowering circulating lipid levels, improving metabolic parameters and other physiological effects induced in vivo by the formation of the ternary complex of an FGFR (e.g., FGFR1c, FGFR2c or FGFR3c), β-Klotho and FGF21, for example conditions such as type 2 diabetes, obesity, dyslipidemia, NASH, cardiovascular disease, and metabolic syndrome.

The antigen binding proteins that specifically bind to a complex comprising β-Klotho and at least one of (i) FGFR1c, (ii) FGFR2c and (iii) FGFR3c that are disclosed herein have a variety of utilities. Some of the antigen binding proteins, for instance, are useful in specific binding assays, in the affinity purification of a complex comprising β-Klotho and at least one of (i) FGFR1c, (ii) FGFR2c and (iii) FGFR3c, including the human forms of these disclosed proteins, and in screening assays to identify other agonists of FGF21-like signaling activity.

The antigen binding proteins that specifically bind to a complex comprising β-Klotho and at least one of (i) FGFR1c, (ii) FGFR2c and (iii) FGFR3c that are disclosed herein can be used in a variety of treatment applications, as explained herein. For example, certain antigen binding proteins are useful for treating conditions associated with FGF21-like signaling processes in a patient, such as reducing, alleviating, or treating type 2 diabetes, obesity, dyslipidemia, NASH, cardiovascular disease, and metabolic syndrome. Other uses for the antigen binding proteins include, for example, diagnosis of diseases or conditions associated with β-Klotho, FGFR1c, FGFR2c, FGFR3c, FGFR4 or FGF21, and screening assays to determine the presence or absence of these molecules. Some of the antigen binding proteins described herein can be useful in treating conditions, symptoms and/or the pathology associated with decreased FGF21-like signaling activity. Exemplary conditions include, but are not limited to, diabetes, obesity, NASH and dyslipidemia.

FGF21

The antigen binding proteins disclosed herein induce FGF21-mediated signaling, as defined herein. In vivo, the mature form of FGF21 is the active form of the molecule. The nucleotide sequence encoding full length FGF21 is provided; the nucleotides encoding the signal sequence are underlined.

The amino acid sequence of full length FGF21 is provided; the amino acids that make up the signal sequence are underlined:

FGFR1c

The antigen binding proteins disclosed herein bind to FGFR1c, in particular human FGFR1c, when associated with β-Klotho. The nucleotide sequence encoding human FGFR1c (GenBank Accession Number NM_023110) is provided:

The amino acid sequence of human FGFR1c (GenBank Accession Number NP_075598) is provided:

The antigen binding proteins described herein bind the extracellular portion of FGFR1c. An example of an extracellular region of FGFR1c is:

As described herein, FGFR1c proteins can also include fragments. As used herein, the terms are used interchangeably to mean a receptor, in particular and unless otherwise specified, a human receptor, that upon association with β-Klotho and FGF21 induces FGF21-like signaling activity.

The term FGFR1c also includes post-translational modifications of the FGFR1c amino acid sequence, for example, possible N-linked glycosylation sites. Thus, the antigen binding proteins can bind to or be generated from proteins glycosylated at one or more of the positions.

β-Klotho

The antigen binding proteins disclosed herein bind to β-Klotho, in particular human β-Klotho. The nucleotide sequence encoding human β-Klotho (GenBank Accession Number NM_175737) is provided:

The amino acid sequence of full length human β-Klotho (GenBank Accession Number NP_783864) is provided:

The antigen binding proteins described herein bind the extracellular portion of β-Klotho. An example of an extracellular region of β-Klotho is:

The murine form of • -Klotho, and fragments and subsequences thereof, can be of use in studying and/or constructing the molecules provided herein. The nucleotide sequence encoding murine • -Klotho (GenBank Accession Number NM_031180) is provided:

The amino acid sequence of full length murine • -Klotho (GenBank Accession Number NP_112457) is provided:

As described herein, β-Klotho proteins can also include fragments. As used herein, the terms are used interchangeably to mean a co-receptor, in particular and unless otherwise specified, a human co-receptor, that upon association with FGFR1c and FGF21 induces FGF21-like signaling activity.

The term β-Klotho also includes post-translational modifications of the β-Klotho amino acid sequence, for example, possible N-linked glycosylation sites. Thus, the antigen binding proteins can bind to or be generated from proteins glycosylated at one or more of the positions.

Antigen Binding Proteins that Specifically Bind to a Complex Comprising β-Klotho and an FGFR (e.g., FGFR1c, FGFR2c or FGFR3c)

A variety of selective binding agents useful for modulating FGF21-like signaling are provided. These agents include, for instance, antigen binding proteins that contain an antigen binding domain (e.g., single chain antibodies, domain antibodies, hemibodies, immunoadhesions, and polypeptides with an antigen binding region) and specifically bind to a complex comprising β-Klotho and at least one of (i) FGFR1c, (ii) FGFR2c and (iii) FGFR3c, in particular a complex comprising human β-Klotho and a human FGFR (e.g., human FGFR1c, human FGFR2c or human FGFR3c). Some of the agents, for example, are useful in mimicking the signaling effect generated in vivo by the association of an FGFR (e.g., FGFR1c, FGFR2c or FGFR3c) with β-Klotho and with FGF21, and can thus be used to enhance or modulate one or more activities associated with FGF21-like signaling.

In general, the antigen binding proteins that are provided typically comprise one or more CDRs as described herein (e.g., 1, 2, 3, 4, 5 or 6 CDRs). In some embodiments the antigen binding proteins are naturally expressed by clones, while in other embodiments, the antigen binding protein can comprise (a) a polypeptide framework structure and (b) one or more CDRs that are inserted into and/or joined to the polypeptide framework structure. In some of these embodiments a CDR forms a component of a heavy or light chains expressed by the clones described herein; in other embodiments a CDR can be inserted into a framework in which the CDR is not naturally expressed. A polypeptide framework structure can take a variety of different forms. For example, a polypeptide framework structure can be, or comprise, the framework of a naturally occurring antibody, or fragment or variant thereof, or it can be completely synthetic in nature. Examples of various antigen binding protein structures are further described below.

In some embodiments in which the antigen binding protein comprises (a) a polypeptide framework structure and (b) one or more CDRs that are inserted into and/or joined to the polypeptide framework structure, the polypeptide framework structure of an antigen binding protein is an antibody or is derived from an antibody, including, but not limited to, monoclonal antibodies, bispecific antibodies, minibodies, domain antibodies, synthetic antibodies (sometimes referred to herein as "antibody mimetics"), chimeric antibodies, humanized antibodies, antibody fusions (sometimes referred to as "antibody conjugates"), and portions or fragments of each, respectively. In some instances, the antigen binding protein is an immunological fragment of an antibody (e.g., a Fab, a Fab', a F(ab')₂, or a scFv).

Certain of the antigen binding proteins as provided herein specifically bind to a complex comprising β-Klotho and at least one of (i) FGFR1c, (ii) FGFR2c and (iii) FGFR3c, including the human forms of these proteins. In one embodiment, an antigen binding protein specifically binds to both human FGFR1c comprising the amino acid sequence of SEQ ID NO: 4, and human β-Klotho comprising the amino acid sequence of SEQ ID NO: 7, and in another embodiment an antigen binding protein specifically binds to both human FGFR1c comprising the amino acid sequence of SEQ ID NO: 4 and human β-Klotho having the amino acid sequence of SEQ ID NO: 7 and induces FGF21-like signaling. Thus, an antigen binding protein can, but need not, induce FGF21-like signaling.

Antigen Binding Protein Structure

Some of the antigen binding proteins that specifically bind to a complex comprising β-Klotho and at least one of (i) FGFR1c, (ii) FGFR2c and (iii) FGFR3c, including the human forms of these proteins, provided herein have a structure typically associated with naturally occurring antibodies. The structural units of these antibodies typically comprise one or more tetramers, each composed of two identical couplets of polypeptide chains, though some species of mammals also produce antibodies having only a single heavy chain. In a typical antibody, each pair or couplet includes one full-length "light" chain (in certain embodiments, about 25 kDa) and one full-length "heavy" chain (in certain embodiments, about 50-70 kDa). Each individual immunoglobulin chain is composed of several "immunoglobulin domains," each consisting of roughly 90 to 110 amino acids and expressing a characteristic folding pattern. These domains are the basic units of which antibody polypeptides are composed. The amino-terminal portion of each chain typically includes a variable domain that is responsible for antigen recognition. The carboxy-terminal portion is more conserved evolutionarily than the other end of the chain and is referred to as the "constant region" or "C region". Human light chains generally are classified as kappa ("κ") and lambda ("λ") light chains, and each of these contains one variable domain and one constant domain. Heavy chains are typically classified as mu, delta, gamma, alpha, or epsilon chains, and these define the antibody's isotype as IgM, IgD, IgG, IgA, and IgE, respectively. IgG has several subtypes, including, but not limited to, IgG1, IgG2, IgG3, and IgG4. IgM subtypes include IgM, and IgM2. IgA subtypes include IgA1 and IgA2. In humans, the IgA and IgD isotypes contain four heavy chains and four light chains; the IgG and IgE isotypes contain two heavy chains and two light chains; and the IgM isotype contains five heavy chains and five light chains. The heavy chain C region typically comprises one or more domains that can be responsible for effector function. The number of heavy chain constant region domains will depend on the isotype. IgG heavy chains, for example, each contain three C region domains known as C_H1, C_H2 and C_H3. The antibodies that are provided can have any of these isotypes and subtypes. In certain embodiments, an antigen binding protein that specifically binds to a complex comprising β-Klotho and at least one of (i) FGFR1c, (ii) FGFR2c and (iii) FGFR3c is an antibody of the IgG1, IgG2, or IgG4 subtype.

In full-length light and heavy chains, the variable and constant regions are joined by a "J" region of about twelve or more amino acids, with the heavy chain also including a "D" region of about ten more amino acids. See, e.g., Fundamental Immunology, 2nd ed., Ch. 7 (Paul, W., ed.) 1989, New York: Raven Press (hereby incorporated by reference in its entirety for all purposes). The variable regions of each light/heavy chain pair typically form the antigen binding site.

One example of an IgG2 heavy constant domain of an exemplary monoclonal antibody that specifically binds to a complex comprising β-Klotho and at least one of (i) FGFR1c, (ii) FGFR2c and (iii) FGFR3c has the amino acid sequence:

One example of a kappa light constant domain of an exemplary monoclonal antibody that binds to a complex comprising β-Klotho and at least one of (i) FGFR1c, (ii) FGFR2c and (iii) FGFR3c has the amino acid sequence:

One example of a lambda light constant domain of an exemplary monoclonal antibody that binds to a complex comprising β-Klotho and at least one of (i) FGFR1c, (ii) FGFR2c and (iii) FGFR3c has the amino acid sequence:

Variable regions of immunoglobulin chains generally exhibit the same overall structure, comprising relatively conserved framework regions (FR) joined by three hypervariable regions, more often called "complementarity determining regions" or CDRs. The CDRs from the two chains of each heavy chain/light chain pair mentioned above typically are aligned by the framework regions to form a structure that binds specifically with a specific epitope on the target protein (e.g., a complex comprising β-Klotho and at least one of (i) FGFR1c, (ii) FGFR2c and (iii) FGFR3c. From N-terminal to C-terminal, naturally-occurring light and heavy chain variable regions both typically conform with the following order of these elements: FR1, CDR1, FR2, CDR2, FR3, CDR3 and FR4. A numbering system has been devised for assigning numbers to amino acids that occupy positions in each of these domains. This numbering system is defined in Kabat et al., (1991) "Sequences of Proteins of Immunological Interest", 5th Ed., US Dept. of Health and Human Services, PHS, NIH, NIH Publication no. 91-3242. Although presented using the Kabat nomenclature system, as desired, the CDRs disclosed herein can also be redefined according an alternative nomenclature scheme, such as that of Chothia (see Chothia & Lesk, (1987) J. Mol. Biol. 196:901-917; Chothia et al., (1989) Nature 342:878-883 or Honegger & Pluckthun, (2001) J. Mol. Biol. 309:657-670).

The various heavy chain and light chain variable regions of antigen binding proteins provided herein are depicted in Table 2. Each of these variable regions can be attached to the disclosed heavy and light chain constant regions to form a complete antibody heavy and light chain, respectively. Further, each of the so-generated heavy and light chain sequences can be combined to form a complete antibody structure. It should be understood that the heavy chain and light chain variable regions provided herein can also be attached to other constant domains having different sequences than the exemplary sequences listed above.

Specific examples of some of the full length light and heavy chains of the antibodies that are provided and their corresponding amino acid sequences are summarized in Tables 1A and 1B. Table 1A shows exemplary light chain sequences, and Table 1B shows exemplary heavy chain sequences.



63E6		14
66F7		15
66D4		16
66B4		17
65B1		18
65B4		19

67A4		20
63A10v1		21
63A10v2		1835
63A10v3		1836
65H11v1		22
65H11v2		1837
67G10v1		23

67G10v2		24
64C8		25
64A8		26
67B4
63G8v1		1838
63G8v2		1839
63G8v3		1840
66G2		27
68D3v1		28
68D3v2
65D1		29
64H5		30
65G4
65D4		31
65E3		32
67G8		33

65B7v1		34
65B7v2		1841
63B6		35
64D4
63F5		36
65E8		37
63H11
64E6
67G7
65F11
65C1		38
66F6		39
64A6		40
65F9		41
64A7		42
65C3		43
68D5
67F5		44
64B10v1		45
64B10v2
68C8		46
67A5		47
67C10		48
64H6		49
63F9		50
67F6v1		51
67F6v2		1842
48C9		52
49A12
51E2
48F3		53

48F8		54
53B9
56B4
57E7
57F11
48H11		55
49A10		56
48D4
49C8		57
52H1
49G2		58
50C12
55G11
49G3		59
49H12		60
51A8		61
51C10.1		62
51C10.2		63
51E5		64
51G2		65
52A8		66
52B8		67
52C1		68
52F8		69
52H2		70
53F6		71

53H5.2		72
53H5.3		73
54A1		74
55G9
54H10.1		75
55D1
48H3
53C11
55D3		76
55E4		77
49B11
50H10
53C1
55E9		78
55G5		79
56A7		80
56E4
56C11		81
56E7		82
56G1		83

56G3.3		84
55B10
57B12		85
57D9		86
59A10		87
49H4
59C9		88
58A5
57A4
57F9
59G10.2		89
59G10.3		90
60D7		91
60F9		92
48B4
52D6
60G5.2		93
61G5		94
52C5		95
61H5		96
52B9
59D10v1		97
59D10v2		98
56G3.2		99
68G5		100
60G5.1		1843

48G4		101
53C3.1
50G1		102
58C2		103
50D4		104
50G5v1		105
50G5v2		106
51C1		107
53C3.2		108
54H10.3		109
55A7		110
55E6		111
61E1		112


63E6		113
66F7
66D4		114
66B4		115
65B1		116

65B4		117
67A4		118
63A10v1		119
63A10v2
63A10v3
65H11v1	120
65H11v2
67G10v1		121
67G10v2
64C8		122
63G8v1		123
63G8v2
63G8v3
68D3v1
64A8
67B4

68D3v2		1844
66G2		124
65D1		125
64H5		126
65D4		127
65E3		128
65G4		129
68G5		130
67G8		131
65B7v1		132
65B7v2
63B6 64D4		133
63F5		134
63H11		135
64E6		136
65E8
65F11
67G7
65C1		137
66F6		138
64A6		139
65F9		140
64A7		141
65C3		142
68D5
67F5		143
64B10v1		144
64B10v2		1845
68C8		145
67A5		146
67C10		147
64H6		148
63F9		149
67F6v1		150
67F6v2
48C9		151
49A12
51E2
48F3		152
48F8		153
53B9
56B4
57E7
57F11
48H11		154
49A10		155
48D4
49C8		156
52H1
49G2		157
50C12
55G11
49G3		158
49H12		159
51A8		160
51C10.2		161
51E5		162
51G2		163
52A8		164
52B8		165
52C1		166
52F8		167
52H2		168
53F6		169
53H5.2		170
53H5.3		171
54A1		172
55G9
54H10.1		173
55D1
48H3
53C11
55D3		174
55E4		175
52C5
60G5.1
49B11
50H10
53C1
55E9		176
55G5		177
50G1		178
56A7		179
56E4
56C11		180
56E7		181
56G1		182
56G3.3		183
55B10
57B12		184
57D9		185
58C2		186
59A10		187
49H4
59C9		188
58A5
57A4
57F9
59G10.2		189
59G10.3		190
60D7		191
60F9		192
48B4
52D6
60G5.2		193
61G5		194
59D10v1		195
59D10v2
51C10.1
56G3.2		196
48G4		197
53C3.1
61H5		198
52B9
50D4		199
50G5v1		200
50G5v2
51C1		201
53C3.2		202
54H10.3		203
55A7		204
55E6		205
61E1		206

Each of the exemplary heavy chains (H1, H2, H3 etc.) listed in Table 1B, infra, can be combined with any of the exemplary light chains shown in Table 1A, infra, to form an antibody. Examples of such combinations include H1 combined with any of L1 through L100; H2 combined with any of L1 through L100; H3 combined with any of L1 through L100, and so on. In some instances, the antibodies include at least one heavy chain and one light chain from those listed in Tables 1A and 1B, infra; particular examples pairings of light chains and heavy chains include L1 with H1, L2 with H1, L3 with H2 or H3, L4 with H4, L5 with H5, L6 with H6, L7 with H6, L8 with H7 or H8, L9 with H9, L10 with H9, L11 with H10, L12 with H11, L13 with H12, L13 with H14, L14 with H13, L15 with H14, L16 with H15, L17 with H16, L18 with H17, L19 with H18, L20 with H19, L21 with H20, L22 with H21, L23 with H22, L24 with H23, L25 with H24, L26 with H25, L27 with H26, L28 with H27, L29 with H28, L30 with H29, L31 with H30, L32 with H31, L33 with H32, L34 with H33, L35 with H34, L36 with H35, L37 with H36, L38 with H37, L39 with H38, L40 with H39, L41 with H40, L42 with H41, L43 with H42, L44 with H43, L45 with H44, L46 with H45, L47 with H46, L48 with H47, L49 with H48, L50 with H49, L51 with H50, L52 with H51, L53 with H52, L54 with H53, L55 with H54, and L56 with H54, L57 with H54, L58 with H55, L59 with H56, L60 with H57, L61 with H58, L62 with H59, L63 with H60, L64 with H1, L65 with H62, L66 with H63, L67 with H64, L68 with H65, L69 with H66, L70 with H67, L71 with H68, L72 with H69, L73 with H70, L74 with H70, and L75 with H70, L76 with H71, L77 with H72, L78 with H73, L79 with H74, L80 with H75, L81 with H76, L82 with H77, L83 with H78, L84 with H79, L85 with H80, L86 with H81, L87 with H82, L88 with H86, L89 with H83, L90 with H84, L91 with H85, L92 with H87, L93 with H88, L94 with H88, L95 with H89, L96 with H90, L97 with H91, L98 with H92, L99 with H93, and L100 with H94. In addition to antigen binding proteins comprising a heavy and a light chain from the same clone, a heavy chain from a first clone can be paired with a light chain from a second clone (e.g., a heavy chain from a first clone paired with a light chain from a second clone or a heavy chain from a first clone paired with a light chain from a second clone). Generally, such pairings can include VL with 90% or greater homology can be paired with the heavy chain of the naturally occurring clone.

In some instances, the antibodies comprise two different heavy chains and two different light chains listed in Tables 1A and 1B, infra. In other instances, the antibodies contain two identical light chains and two identical heavy chains. As an example, an antibody or immunologically functional fragment can include two L1 light chains with two H1 heavy chains, two L2 light chains with two H1 heavy chains, two L3 light chains with two H2 heavy chains or two H3 heavy chains, two L4 light chains with two H4 heavy chains, two L5 light chains with two H5 heavy chains, two L6 light chains with two H6 heavy chains, two L7 light chains with two H6 heavy chains, two L8 light chains with two H7 heavy chains or two H8 heavy chains, two L9 light chains with two H9 heavy chains, two L10 light chains with two H9 heavy chains, two L11 light chains with two H10 heavy chains, two L12 light chains with two H11 heavy chains, two L13 light chains with two H12 heavy chains, two L13 light chains with two H14 heavy chains, two L14 light chains with two H13 heavy chains, two L15 light chains with two H14 heavy chains, two L16 light chains with two H15 heavy chains, two L17 light chains with two H16 heavy chains, two L18 light chains with two H17 heavy chains, two L19 light chains with two H18 heavy chains, two L20 light chains with two H19 heavy chains, two L21 light chains with two H20 heavy chains, two L22 light chains with two H21 heavy chains, two L23 light chains with two H22 heavy chains, two L24 light chains with two H23 heavy chains, two L25 light chains with two H24 heavy chains, two L26 light chains with two H25 heavy chains, two L27 light chains with two H26 heavy chains, two L28 light chains with two H27 heavy chains, two L29 light chains with two H28 heavy chains, two L30 light chains with two H29 heavy chains, two L31 light chains with two H30 heavy chains, two L32 light chains with two H31 heavy chains, two L33 light chains with two H32 heavy chains, two L34 light chains with two H33 heavy chains, two L35 chains with two H34 heavy chains, two L36 chains with two H35 heavy chains, two L37 light chains with two H36 heavy chains, two L38 light chains with two H37 heavy chains, two L39 light chains with two H38 heavy chains, two L40 light chains with two H39 heavy chains, two L41 light chains with two H40 heavy chains, two L42 light chains with two H41 heavy chains, two L43 light chains with two H42 heavy chains, two L44 light chains with two H43 heavy chains, two L45 light chains with two H44 heavy chains, two L46 light chains with two H45 heavy chains, two L47 light chains with two H46 heavy chains, two L48 light chains with two H47 heavy chains, two L49 light chains with two H48 heavy chains, two L50 light chains with two H49 heavy chains, two L51 light chains with two H50 heavy chains, two L52 light chains with two H51 heavy chains, two L53 light chains with two H52 heavy chains, two L54 light chains with two H53 heavy chains, two L55 light chains with two H54 heavy chains, and two L56 light chains with two H54 heavy chains, two L57 light chains with two H54 heavy chains, two L58 light chains with two H55 heavy chains, two L59 light chains with two H56 heavy chains, two L60 light chains with two H57 heavy chains, two L61 light chains with two H58 heavy chains, two L62 light chains with two H59 heavy chains, two L63 light chains with two H60 heavy chains, two L64 light chains with two H1 heavy chains, two L65 light chains with two H62 heavy chains, two L66 light chains with two H63 heavy chains, two L67 light chains with two H64 heavy chains, two L68 light chains with two H65 heavy chains, two L69 light chains with two H66 heavy chains, two L70 light chains with two H67 heavy chains, two L71 light chains with two H68 heavy chains, two L72 light chains with two H69 heavy chains, two L73 light chains with two H70 heavy chains, two L74 light chains with two H70 heavy chains, and two L75 light chains with two H70 heavy chains, two L76 light chains with two H71 heavy chains, two L77 light chains with two H72 heavy chains, two L78 light chains with two H73 heavy chains, two L79 light chains with two H74 heavy chains, two L80 light chains with two H75 heavy chains, two L81 light chains with two H76 heavy chains, two L82 light chains with two H77 heavy chains, two L83 light chains with two H78 heavy chains, two L84 light chains with two H79 heavy chains, two L85 light chains with two H80 heavy chains, two L86 light chains with two H81 heavy chains, two L87 light chains with two H82 heavy chains, two L88 light chains with two H86 heavy chains, two L89 light chains with two H83 heavy chains, two L90 light chains with two H84 heavy chains, two L91 light chains with two H85 heavy chains, two L92 light chains with two H87 heavy chains, two L93 light chains with two H88 heavy chains, two L94 light chains with two H88 heavy chains, two L95 light chains with two H89 heavy chains, two L96 light chains with two H90 heavy chains, two L97 light chains with two H91 heavy chains, two L98 light chains with two H92 heavy chains, two L99 light chains with two H93 heavy chains, and two L100 light chains with two H94 heavy chains, as well as other similar combinations of pairs comprising the light chains and pairs of heavy chains as listed in Tables 1A and 1B, infra.

In another aspect of the instant disclosure, "hemibodies" are provided. A hemibody is a monovalent antigen binding protein comprising (i) an intact light chain, and (ii) a heavy chain fused to an Fc region (e.g., an IgG2 Fc region of SEQ ID NO: 11), optionally via a linker, The linker can be a (G₄S)_x linker (SEQ ID NO: 207) where "x" is a non-zero integer (e.g., (G₄S)₂, (G₄S)₃, (G₄S)₄, (G₄S)₅, (G₄S)₆, (G₄S)₇, (G₄S)₈, (G₄S)₉, (G₄S)_10,; SEQ ID NOs: 208-216, respectively). Hemibodies can be constructed using the provided heavy and light chain components.

Other antigen binding proteins that are provided are variants of antibodies formed by combination of the heavy and light chains shown in Tables 1A and 1B, infra and comprise light and/or heavy chains that each have at least 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% identity to the amino acid sequences of these chains. In some instances, such antibodies include at least one heavy chain and one light chain, whereas in other instances the variant forms contain two identical light chains and two identical heavy chains.

Variable Domains of Antigen Binding Proteins

Also provided are antigen binding proteins that contain an antibody heavy chain variable region selected from the group consisting of V_H1, V_H2, V_H3, V_H4, V_H5, V_H6, V_H7, V_H8, V_H9, V_H10, V_H11, V_H12, V_H13, V_H14, V_H15, V_H16, V_H17, V_H18, V_H19, V_H20, V_H21 V_H22, V_H23, V_H24, V_H25, V_H26, V_H27, V_H28, V_H29, V_H30, V_H31, V_H32, V_H33, V_H34, V_H35, V_H36, V_H37, V_H38, V_H39, V_H40, V_H41, V_H42, V_H43, V_H44, V_H45, V_H46, V_H47, V_H48, V_H49, V_H50, V_H51, V_H52, V_H53, V_H54, V_H55, V_H56, V_H57, V_H58, V_H59, V_H60, V_H61, V_H62, V_H63, V_H64, V_H65, V_H66, V_H67, V_H68, V_H69, V_H70, V_H71, V_H72, V_H73, V_H74, V_H75, V_H76, V_H77, V_H78, V_H79, V_H80, 81, V_H82, V_H83, V_H84, V_H85, V_H86, V_H87, V_H88, V_H89, V_H90, V_H91, V_H92, V_H93, and V_H94 as shown in Table 2B and/or an antibody light chain variable region selected from the group consisting of V_L1, V_L2, V_L3, V_L4, V_L5, V_L6, V_L7, V_L8, V_L9, V_L10, V_L11, V_L12, V_L13, V_L14, V_L15, V_L16, V_L17, V_L18, V_L19, V_L20, V_L21, V_L22, V_L23, V_L24, V_L25, V_L26, V_L27, V_L28, V_L29, V_L30, V_L31, V_L32, V_L33, V_L34, V_L35, V_L36, V_L37, V_L38, V_L39, V_L40, V_L41, V_L42, V_L43, V_L44, V_L45, V_L46, V_L47, V_L48, V_L49, V_L50, V_L51, V_L52, V_L53, V_L54, V_L55, V_L56, V_L57, V_L58, V_L59, V_L60, V_L61, V_L62, V_L63, V_L64, V_L65, V_L66, V_L67, V_L68, V_L69, V_L70, V_L71, V_L72, V_L73, V_L74, V_L75, V_L76, V_L77, V_L78, V_L79, V_L80, V_L81, V_L82, V_L83, V_L84, V_L85, V_L86, V_L87, V_L88, V_L89, V_L90, V_L91, V_L92, V_L93, V_L94, V_L95, V_L96, V_L97, V_L98, V_L99 and V_L100 as shown in Table 2A, and immunologically functional fragments, derivatives, muteins and variants of these light chain and heavy chain variable regions.



63E6		217
66F7		218
66D4		219
66B4		220
65B1		221
65B4		222
67A4		223
63A10v1		224
63A10v2		1846
63A10v3		1847
65H11v1		225
65H11v2		1848
67G10v1		226
67G10v2		227
64C8		228
64A8		229
67B4
63G8v1		1849
63G8v2		1850
63G8v3		1851
66G2		230
68D3v1		231
68D3v2
65D1		232
64H5		233
65G4
65D4		234
65E3		235
68G5		236
67G8		237
65B7v1		238
65B7v2		1852
63B6		239
64D4
63F5		240
65E8		241
63H11
64E6
65F11
67G7
65C1		242
66F6		243
64A6		244
65F9		245
64A7		246
65C3		247
68D5
67F5		248
64B10v1		249
64B10v2
68C8		250
67A5		251
67C10		252
64H6		253
63F9		254
67F6v1		255
67F6v2		1853
48C9		256
49A12
51E2
48F3		257
48F8		258
53B9
56B4
57E7
57F11
48H11		259
49A10		260
48D4
49C8		261
52H1
49G2		262
50C12
55G11
49G3		263
49H12		264
51A8		265
51C10.1		266
51C10.2		267
51E5		268
51G2		269
52A8		270
52B8		271
52C1		272
52F8		273
52H2		274
53F6		275
53H5.2		276
53H5.3		277
54A1		278
55G9
54H10.1		279
55D1
48H3
53C11
55D3		280
55E4		281
49B11
50H10
53C1
55E9		282
55G5		283
56A7		284
56E4
56C11		285
56E7		286
56G1		287
56G3.3		288
55B10
57B12		289
57D9		290
59A10		291
49H4
59C9		292
58A5
57A4
57F9
59G10.2		293
59G10.3		294
60D7		295
60F9		296
48B4
52D6
60G5.2		297
61G5		298
52C5		299
61H5		300
52B9
59D10v1		301
59D10v2		302
56G3.2		303
48G4		304
53C3.1
50G1		305
58C2		306
60G5.1		1854
50D4		307
50G5v1		308
50G5 v2		309
51C1		310
53C3.2		311
54H10.3		312
55A7		313
55E6		314
61E1		315



63E6		316
66F7
66D4		317
66B4		318
65B1		319
65B4		320
67A4		321
63A10v1		322
63A10v2
63A10v3
65H11v1		323
65H11v2
67G10v1		324
67G10v2
64C8		325
63G8v1		326
63G8v2
63G8v3
68D3v1
64A8
67B4
68D3v2		1855
66G2		327
65D1		328
64H5		329
65D4		330
65E3		331
65G4		332
68G5		333
67G8		334
65B7v1		335
65B7v2
63B6 64D4		336
63F5		337
63H11		338
65E8		339
64E6
65F11
67G7
65C1		340
66F6		341
64A6		342
65F9		343
64A7		344
65C3		345
68D5
67F5		346
64B10v1		347
64B10v2		1856
68C8		348
67A5		349
67C10		350
64H6		351
63F9		352
67F6v1		353
67F6v2
48C9		354
49A12
51E2
48F3		355
48F8		356
53B9
56B4
57E7
57F11
48H11		357
49A10		358
48D4
49C8		359
52H1
49G2		360
50C12
55G11
49G3		361
49H12		362
51A8		363
51C10.1		364
59D10v1
59D10v2
51C10.2		365
51E5		366
51G2		367
52A8		368
52B8		369
52C1		370
52F8		371
52H2		372
53F6		373
53H5.2		374
53H5.3		375
54A1 55G9		376
54H10.1		377
55D1
48H3
53C11
55D3		378
55E4		379
49B11
50H10
53C1
52C5
60G5.1
55E9		380
55G5		381
50G1		382
56A7		383
56E4
56C11		384
56E7		385
56G1		386
56G3.3		387
55B10
57B12		388
57D9		389
58C2		390
59A10 49H4		391
59C9		392
58A5
57A4
57F9
59G10.2		393
59G10.3		394
60D7		395
60F9		396
48B4
52D6
60G5.2		397
61G5		398
56G3.2		399
48G4		400
53C3.1
61H5		401
52B9
50D4		402
50G5v1		403
50G5v2
51C1		404
53C3.2		405
54H10.3		406
55A7		407
55E6		408
61E1		409



63E6		410
66D4		411
66B4		412
65B1		413
65B4		414
67A4		415
63A10v1		416
63A10v2		1857
63A10v3		1858
65H11v1		417
65H11v2		1859
67G10v1		418
67G10v2		419
64C8		420
64A8		421
67B4
63G8v1		1860
63G8v2		1861
63G8v3		1862
66G2		422
68D3v1		423
68D3v2
65D1		424
65G4		425
64H5
65D4		426
65E3		427
68G5		428
67G8		429
65B7v1		430
65B7v2		1863
63B6		431
64D4
63F5		432
65E8		433
63H11
64E6
65F11
67G7
65C1		434
66F6		435
64A6		436
65F9		437
64A7		438
65C3		439
68D5
67F5		440
64B10v1		441
64B10v2
68C8		442
67A5		443
67C10		444
64H6		445
63F9		446
67F6v1		447
67F6v2		1864
48C9		448
49A12
51E2


48F3		449
48F8		450
53B9
56B4
57E7
57F11
48H11		451
49A10		452
48D4
49C8		453
52H1
49G2		454
50C12
55G11
49G3		455
49H12		456
51A8		457
51C10.1		458
51C10.2		459
51E5		460
51G2		461
52A8		462
52B8		463
52C1		464
52F8		465
52H2		466
53F6		467
53H5.2		468
53H5.3		469
54A1 55G9		470
54H10.1		471
55D1
48H3
53C11
55D3		472
55E4		473
49B11
50H10
53C1
55E9		474
55G5		475
56A7		476
56E4
56C11		477
56E7		478
56G1		479
56G3.3		480
55B10
57B12		481
57D9		482
59A10		483
49H4
59C9		484
58A5
57A4
57F9
59G10.2		485
59G10.3		486
60D7		487
60F9		488
48B4
52D6
60G5.2		489
61G5		490
52C5		491
61H5		492
52B9
59D10v1		493
59D10v2		494
56G3.2		495
66F7		496
48G4		497
53C3.1
50G1		498
58C2		499
60G5.1		1865
50D4		500
50G5v1		501
50G5v2		502
51C1		503
53C3.2		504
54H10.3		505
55A7		506
55E6		507
61E1		508



63E6 66F7		509
66D4		510
66B4		511
65B1		512
65B4		513
67A4		514
63A10v1 63A10v2 63A10v3		515
65H11v1 65H11v2		516
67G10v1 67G10v2		517
64C8		518
63G8v1 63G8v2 63G8v3 68D3v1 64A8 67B4		519
68D3v2		1866
66G2		520
65D1		521
64H5		522
65D4		523
65E3		524
65G4		525
68G5		526
67G8		527
65B7v1 65B7v2		528
63B6 64D4		529
63F5		530
63H11		531
65E8 64E6 65F11 67G7		532
65C1		533
66F6		534
64A6		535
65F9		536
64A7		537
65C3 68D5		538
67F5		539
64B10v1		540
64B10v2		1867
68C8		541
67A5		542
67C10		543
64H6		544
63F9		545
67F6v1 67F6v2		546
48C9 49A12 51E2		547
48F3		548
48F8 53B9 56B4 57E7 57F11		549
48H11		550
49A10 48D4		551
49C8 52H1		552
49G2 50C12 55G11		553
49G3		554
49H12		555
51A8		556
51C10.1 59D10v1 59D10v2		557
51C10.2		558
51E5		559
51G2		560
52A8		561
52B8		562
52C1		563
52F8		564
52H2		565
53F6		566
53H5.2		567
53H5.3		568
54A1 55G9		569
54H10.1 55D1 48H3 53C11		570
55D3		571
55E4 49B11 50H10 53C1 52C5 60G5.1		572
55E9		573
55G5		574
50G1		575
56A7 56E4		576
56C11		577
56E7		578
56G1		579
56G3.3 55B10		580
57B12		581
57D9		582
58C2		583
59A10 49H4		584
59C9 58A5 57A4 57F9		585
59G10.2		586
59G10.3		587
60D7		588
60F9 48B4 52D6		589
60G5.2		590
61G5		591
56G3.2		592
48G4 53C3.1		593
61H5 52B9		594
50D4		595
50G5v1 50G5v2		596
51C1		597
53C3.2		598
54H10.3		599
55A7		600
55E6		601
61E1		602

Each of the heavy chain variable regions listed in Table 2B can be combined with any of the light chain variable regions shown in Table 2A to form an antigen binding protein. Examples of such combinations include V_H1 combined with any of V_L1, V_L2, V_L3, V_L4, V_L5, V_L6, V_L7, V_L8, V_L9, V_L10, V_L11, V_L12, V_L13, V_L14, V_L15, V_L16, V_L17, V_L18, V_L19, V_L20, V_L21, V_L22, V_L23, V_L24, V_L25, V_L26, V_L27, V_L28, V_L29, V_L30, V_L31, V_L32, V_L33, V_L34, V_L35, V_L36, V_L37, V_L38, V_L39, V_L40, V_L41, V_L42, V_L43, V_L44, V_L45, V_L46, V_L47, V_L48, V_L49, V_L50, V_L51, V_L52, V_L53, V_L54, V_L55, V_L56, V_L57, V_L58, V_L59, V_L60, V_L61, V_L62, V_L63, V_L64, V_L65, V_L66, V_L67, V_L68, V_L69, V_L70, V_L71, V_L72, V_L73, V_L74, V_L75, V_L76, V_L77, V_L78, V_L79, V_L80, V_L81, V_L82, V_L83, V_L84, V_L85, V_L86, V_L87, V_L88, V_L89, V_L90, V_L91, V_L92, V_L93, V_L94, V_L95, V_L96, V_L97, V_L98, V_L99 and V_L100; V_H2 combined with any of V_L1, V_L2, V_L3, V_L4, V_L5, V_L6, V_L7, V_L8, V_L9, V_L10, V_L11, V_L12, V_L13, V_L14, V_L15, V_L16, V_L17, V_L18, V_L19, V_L20, V_L21, V_L22, V_L23, V_L24, V_L25, V_L26, V_L27, V_L28, V_L29, V_L30, V_L31, V_L32, V_L33, V_L34, V_L35, V_L36, V_L37, V_L38, V_L39, V_L40, V_L41, V_L42, V_L43, V_L44, V_L45, V_L46, V_L47, V_L48, V_L49, V_L50, V_L51, V_L52, V_L53, V_L54, V_L55, V_L56, V_L57, V_L58, V_L59, V_L60, V_L61, V_L62, V_L63, V_L64, V_L65, V_L66, V_L67, V_L68, V_L69, V_L70, V_L71, V_L72, V_L73, V_L74, V_L75, V_L76, V_L77, V_L78, V_L79, V_L80, V_L81, V_L82, V_L83, V_L84, V_L85, V_L86, V_L87, V_L88, V_L89, V_L90, V_L91, V_L92, V_L93, V_L94, V_L95, V_L96, V_L97, V_L98, V_L99 and V_L100; V_H3 combined with any of V_L1, V_L2, V_L3, V_L4, V_L5, V_L6, V_L7, V_L8, V_L9, V_L10, V_L11, V_L12, V_L13, V_L14, V_L15, V_L16, V_L17, V_L18, V_L19, V_L20, V_L21, V_L22, V_L23, V_L24, V_L25, V_L26, V_L27, V_L28, V_L29, V_L30, V_L31, V_L32, V_L33, V_L34, V_L35, V_L36, V_L37, V_L38, V_L39, V_L40, V_L41, V_L42, V_L43, V_L44, V_L45, V_L46, V_L47, V_L48, V_L49, V_L50, V_L51, V_L52, V_L53, V_L54, V_L55, V_L56, V_L57, V_L58, V_L59, V_L60, V_L61, V_L62, V_L63, V_L64, V_L65, V_L66, V_L67, V_L68, V_L69, V_L70, V_L71, V_L72, V_L73, V_L74, V_L75, V_L76, V_L77, V_L78, V_L79, V_L80, V_L81, V_L82, V_L83, V_L84, V_L85, V_L86, V_L87, V_L88, V_L89, V_L90, V_L91, V_L92, V_L93, V_L94, V_L95, V_L96, V_L97, V_L98, V_L99 and V_L100; and so on.

In some instances, the antigen binding protein includes at least one heavy chain variable region and/or one light chain variable region from those listed in Tables 2A and 2B. In some instances, the antigen binding protein includes at least two different heavy chain variable regions and/or light chain variable regions from those listed in Table 2B. An example of such an antigen binding protein comprises (a) one V_H1, and (b) one of V_H2, V_H3, V_H4, V_H5, V_H6, V_H7, V_H8, V_H9, V_H10, V_H11, V_H12, V_H13, V_H14, V_H15, V_H16, V_H17, V_H18, V_H19, V_H20, V_H21 V_H22, V_H23, V_H24, V_H25, V_H26, V_H27, V_H28, V_H29, V_H30, V_H31, V_H32, V_H33, V_H34, V_H35, V_H36, V_H37, V_H38, V_H39, V_H40, V_H41, V_H42, V_H43, V_H44, V_H45, V_H46, V_H47, V_H48, V_H49, V_H50, V_H51, V_H52, V_H53, V_H54, V_H55, V_H56, V_H57, V_H58, V_H59, V_H60, V_H61, V_H62, V_H63, V_H64, V_H65, V_H66, V_H67, V_H68, V_H69, V_H70, V_H71, V_H72, V_H73, V_H74, V_H75, V_H76, V_H77, V_H78, V_H79, V_H80, 81, V_H82, V_H83, V_H84, V_H85, V_H 86, V_H 87, V_H88, V_H89, V_H90, V_H91, V_H92, V_H93, and V_H94. Another example comprises (a) one V_H2, and (b) one of V_H1, V_H3, V_H4, V_H5, V_H6, V_H7, V_H8, V_H9, V_H10, V_H11, V_H12, V_H13, V_H14, V_H15, V_H16, V_H17, V_H18, V_H19, V_H20, V_H21 V_H22, V_H23, V_H24, V_H25, V_H26, V_H27, V_H28, V_H29, V_H30, V_H31, V_H32, V_H33, V_H34, V_H35, V_H36, V_H37, V_H38, V_H39, V_H40, V_H41, V_H42, V_H43, V_H44, V_H45, V_H46, V_H47, V_H48, V_H49, V_H50, V_H51, V_H52, V_H53, V_H54, V_H55, V_H56, V_H57, V_H58, V_H59, V_H60, V_H61, V_H62, V_H63, V_H64, V_H65, V_H66, V_H67, V_H68, V_H69, V_H70, V_H71, V_H72, V_H73, V_H74, V_H75, V_H76, V_H77, V_H78, V_H79, V_H80, 81, V_H82, V_H83, V_H84, V_H85, V_H 86, V_H 87, V_H88, V_H89, V_H90, V_H91, V_H92, V_H93, and V_H94. Yet another example comprises (a) one V_H3, and (b) one of V_H1, V_H2, V_H4, V_H5, V_H6, V_H7, V_H8, V_H9, V_H10, V_H11, V_H12, V_H13, V_H14, V_H15, V_H16, V_H17, V_H18, V_H19, V_H20, V_H21 V_H22, V_H23, V_H24, V_H25, V_H26, V_H27, V_H28, V_H29, V_H30, V_H31, V_H32, V_H33, V_H34, V_H35, V_H36, V_H37, V_H38, V_H39, V_H40, V_H41, V_H42, V_H43, V_H44, V_H45, V_H46, V_H47, V_H48, V_H49, V_H50, V_H51, V_H52, V_H53, V_H54, V_H55, V_H56, V_H57, V_H58, V_H59, V_H60, V_H61, V_H62, V_H63, V_H64, V_H65, V_H66, V_H67, V_H68, V_H69, V_H70, V_H71, V_H72, V_H73, V_H74, V_H75, V_H76, V_H77, V_H78, V_H79, V_H80, 81, V_H82, V_H83, V_H84, V_H85, V_H 86, V_H 87, V_H88, V_H89, V_H90, V_H91, V_H92, V_H93, and V_H94, etc. Still another example of such an antigen binding protein comprises (a) one V_L1, and (b) one of V_L2, V_L3, V_L4, V_L5, V_L6, V_L7, V_L8, V_L9, V_L10, V_L11, V_L12, V_L13, V_L14, V_L15, V_L16, V_L17, V_L18, V_L19, V_L20, V_L21, V_L22, V_L23, V_L24, V_L25, V_L26, V_L27, V_L28, V_L29, V_L30, V_L31, V_L32, V_L33, V_L34, V_L35, V_L36, V_L37, V_L38, V_L39, V_L40, V_L41, V_L42, V_L43, V_L44, V_L45, V_L46, V_L47, V_L48, V_L49, V_L50, V_L51, V_L52, V_L53, V_L54, V_L55, V_L56, V_L57, V_L58, V_L59, V_L60, V_L61, V_L62, V_L63, V_L64, V_L65, V_L66, V_L67, V_L68, V_L69, V_L70, V_L71, V_L72, V_L73, V_L74, V_L75, V_L76, V_L77, V_L78, V_L79, V_L80, V_L81, V_L82, V_L83, V_L84, V_L85, V_L86, V_L87, V_L88, V_L89, V_L90, V_L91, V_L92, V_L93, V_L94, V_L95, V_L96, V_L97, V_L98, V_L99 and V_L100. Again another example of such an antigen binding protein comprises (a) one V_L2, and (b) one of V_L1, V_L3, V_L4, V_L5, V_L6, V_L7, V_L8, V_L9, V_L10, V_L11, V_L12, V_L13, V_L14, V_L15, V_L16, V_L17, V_L18, V_L19, V_L20, V_L21, V_L22, V_L23, V_L24, V_L25, V_L26, V_L27, V_L28, V_L29, V_L30, V_L31, V_L32, V_L33, V_L34, V_L35, V_L36, V_L37, V_L38, V_L39, V_L40, V_L41, V_L42, V_L43, V_L44, V_L45, V_L46, V_L47, V_L48, V_L49, V_L50, V_L51, V_L52, V_L53, V_L54, V_L55, V_L56, V_L57, V_L58, V_L59, V_L60, V_L61, V_L62, V_L63, V_L64, V_L65, V_L66, V_L67, V_L68, V_L69, V_L70, V_L71, V_L72, V_L73, V_L74, V_L75, V_L76, V_L77, V_L78, V_L79, V_L80, V_L81, V_L82, V_L83, V_L84, V_L85, V_L86, V_L87, V_L88, V_L89, V_L90, V_L91, V_L92, V_L93, V_L94, V_L95, V_L96, V_L97, V_L98, V_L99 and V_L100. Again another example of such an antigen binding protein comprises (a) one V_L3, and (b) one of V_L1, V_L2, V_L4, V_L5, V_L6, V_L7, V_L8, V_L9, V_L10, V_L11, V_L12, V_L13, V_L14, V_L15, V_L16, V_L17, V_L18, V_L19, V_L20, V_L21, V_L22, V_L23, V_L24, V_L25, V_L26, V_L27, V_L28, V_L29, V_L30, V_L31, V_L32, V_L33, V_L34, V_L35, V_L36, V_L37, V_L38, V_L39, V_L40, V_L41, V_L42, V_L43, V_L44, V_L45, V_L46, V_L47, V_L48, V_L49, V_L50, V_L51, V_L52, V_L53, V_L54, V_L55, V_L56, V_L57, V_L58, V_L59, V_L60, V_L61, V_L62, V_L63, V_L64, V_L65, V_L66, V_L67, V_L68, V_L69, V_L70, V_L71, V_L72, V_L73, V_L74, V_L75, V_L76, V_L77, V_L78, V_L79, V_L80, V_L81, V_L82, V_L83, V_L84, V_L85, V_L86, V_L87, V_L88, V_L89, V_L90, V_L91, V_L92, V_L93, V_L94, V_L95, V_L96, V_L97, V_L98, V_L99 and V_L100, etc.

The various combinations of heavy chain variable regions can be combined with any of the various combinations of light chain variable regions.

In other embodiments, an antigen binding protein comprises two identical light chain variable regions and/or two identical heavy chain variable regions. As an example, the antigen binding protein can be an antibody or immunologically functional fragment thereof that includes two light chain variable regions and two heavy chain variable regions in combinations of pairs of light chain variable regions and pairs of heavy chain variable regions as listed in Tables 2A and 2B.

Some antigen binding proteins that are provided comprise a heavy chain variable domain comprising a sequence of amino acids that differs from the sequence of a heavy chain variable domain selected from V_H1, V_H2, V_H3, V_H4, V_H5, V_H6, V_H7, V_H8, V_H9, V_H10, V_H11, V_H12, V_H13, V_H14, V_H15, V_H16, V_H17, V_H18, V_H19, V_H20, V_H21 V_H22, V_H23, V_H24, V_H25, V_H26, V_H27, V_H28, V_H29, V_H30, V_H31, V_H32, V_H33, V_H34, V_H35, V_H36, V_H37, V_H38, V_H39, V_H40, V_H41, V_H42, V_H43, V_H44, V_H45, V_H46, V_H47, V_H48, V_H49, V_H50, V_H51, V_H52, V_H53, V_H54, V_H55, V_H56, V_H57, V_H58, V_H59, V_H60, V_H61, V_H62, V_H63, V_H64, V_H65, V_H66, V_H67, V_H68, V_H69, V_H70, V_H71, V_H72, V_H73, V_H74, V_H75, V_H76, V_H77, V_H78, V_H79, V_H80, 81, V_H82, V_H83, V_H84, V_H85, V_H 86, V_H 87, V_H88, V_H89, V_H90, V_H91, V_H92, V_H93, and V_H94 at only 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14 or 15 amino acid residues, wherein each such sequence difference is independently either a deletion, insertion or substitution of one amino acid, with the deletions, insertions and/or substitutions resulting in no more than 15 amino acid changes relative to the foregoing variable domain sequences. The heavy chain variable region in some antigen binding proteins comprises a sequence of amino acids that has at least 70%, 75%, 80%, 85%, 90%, 95%, 97% or 99% sequence identity to the amino acid sequences of the heavy chain variable region of V_H1, V_H2, V_H3, V_H4, V_H5, V_H6, V_H7, V_H8, V_H9, V_H10, V_H11, V_H12, V_H13, V_H14, V_H15, V_H16, V_H17, V_H18, V_H19, V_H20, V_H21 V_H22, V_H23, V_H24, V_H25, V_H26, V_H27, V_H28, V_H29, V_H30, V_H31, V_H32, V_H33, V_H34, V_H35, V_H36, V_H37, V_H38, V_H39, V_H40, V_H41, V_H42, V_H43, V_H44, V_H45, V_H46, V_H47, V_H48, V_H49, V_H50, V_H51, V_H52, V_H53, V_H54, V_H55, V_H56, V_H57, V_H58, V_H59, V_H60, V_H61, V_H62, V_H63, V_H64, V_H65, V_H66, V_H67, V_H68, V_H69, V_H70, V_H71, V_H72, V_H73, V_H74, V_H75, V_H76, V_H77, V_H78, V_H79, V_H80, 81, V_H82, V_H83, V_H84, V_H85, V_H 86, V_H 87, V_H88, V_H89, V_H90, V_H91, V_H92, V_H93, and V_H94.

Certain antigen binding proteins comprise a light chain variable domain comprising a sequence of amino acids that differs from the sequence of a light chain variable domain selected from V_L1, V_L2, V_L3, V_L4, V_L5, V_L6, V_L7, V_L8, V_L9, V_L10, V_L11, V_L12, V_L13, V_L14, V_L15, V_L16, V_L17, V_L18, V_L19, VL20, V_L21, V_L22, V_L23, V_L24, V_L25, V_L26, V_L27, V_L28, V_L29, V_L30, V_L31, V_L32, V_L33, V_L34, V_L35, V_L36, V_L37, V_L38, V_L39, V_L40, V_L41, V_L42, V_L43, V_L44, V_L45, V_L46, V_L47, V_L48, V_L49, V_L50, V_L51, V_L52, V_L53, V_L54, V_L55, V_L56, V_L57, V_L58, V_L59, V_L60, V_L61, V_L62, V_L63, V_L64, V_L65, V_L66, V_L67, V_L68, V_L69, V_L70, V_L71, V_L72, V_L73, V_L74, V_L75, V_L76, V_L77, V_L78, V_L79, V_L80, V_L81, V_L82, V_L83, V_L84, V_L85, V_L86, V_L87, V_L88, V_L89, V_L90, V_L91, V_L92, V_L93, V_L94, V_L95, V_L96, V_L97, V_L98, V_L99 and V_L100 at only 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14 or 15 amino acid residues, wherein each such sequence difference is independently either a deletion, insertion or substitution of one amino acid, with the deletions, insertions and/or substitutions resulting in no more than 15 amino acid changes relative to the foregoing variable domain sequences. The light chain variable region in some antigen binding proteins comprises a sequence of amino acids that has at least 70%, 75%, 80%, 85%, 90%, 95%, 97% or 99% sequence identity to the amino acid sequences of the light chain variable region of V_L1, V_L2, V_L3, V_L4, V_L5, V_L6, V_L7, V_L8, V_L9, V_L10, V_L11, V_L12, V_L13, V_L14, V_L15, V_L16, V_L17, V_L18, V_L19, V_L20, V_L21, V_L22, V_L23, V_L24, V_L25, V_L26, V_L27, V_L28, V_L29, V_L30, V_L31, V_L32, V_L33, V_L34, V_L35, V_L36, V_L37, V_L38, V_L39, V_L40, V_L41, V_L42, V_L43, V_L44, V_L45, V_L46, V_L47, V_L48, V_L49, V_L50, V_L51, V_L52, V_L53, V_L54, V_L55, V_L56, V_L57, V_L58, V_L59, V_L60, V_L61, V_L62, V_L63, V_L64, V_L65, V_L66, V_L67, V_L68, V_L69, V_L70, V_L71, V_L72, V_L73, V_L74, V_L75, V_L76, V_L77, V_L78, V_L79, V_L80, V_L81, V_L82, V_L83, V_L84, V_L85, V_L86, V_L87, V_L88, V_L89, V_L90, V_L91, V_L92, V_L93, V_L94, V_L95, V_L96, V_L97, V_L98, V_L99 and V_L100.

In additional instances, antigen binding proteins comprise the following pairings of light chain and heavy chain variable domains: V_L1 with V_H1, V_L2 with V_H1, V_L3 with V_H2 or V_H3, V_L4 with V_H4, V_L5 with V_H5, V_L6 with V_H6, V_L7 with V_H6, V_L8 with V_H7 or V_H8, V_L9 with V_H9, V_L10 with V_H9, V_L11 with V_H 10, V_L12 with V_H11, V_L13 with V_H12, V_L13 with V_H14, V_L14 with V_H13, V_L15 with V_H14, V_L16 with V_H15, V_L17 with V_H16, V_L18 with V_H17, V_L19 with V_H18, V_L20 with V_H19, V_L21 with V_H20, V_L22 with V_H21, V_L23 with V_H22, V_L24 with V_H23, V_L25 with V_H24, V_L26 with V_H25, V_L27 with V_H26, V_L28 with V_H27, V_L29 with V_H28, V_L30 with V_H29, V_L31 with V_H30, V_L32 with V_H31, V_L33 with V_H32, V_L34 with V_H33, V_L35 with V_H34, V_L36 with V_H35, V_L37 with V_H36, V_L38 with V_H37, V_L39 with V_H38, V_L40 with V_H39, V_L41 with V_H40, V_L42 with V_H41, V_L43 with V_H42, V_L44 with V_H43, V_L45 with V_H44, V_L46 with V_H45, V_L47 with V_H46, V_L48 with V_H47, V_L49 with V_H48, V_L50 with V_H49, V_L51 with V_H50, 52 with V_H51, V_L53 with V_H52, V_L54 with V_H53, V_L55 with 54, and V_L56 with V_H54, V_L57 with V_H54, V_L58 with V_H55, V_L59 with V_H56, V_L60 with V_H57, V_L61 with V_H58, V_L62 with V_H59, V_L63 with V_H60, V_L64 with V_H1, V_L65 with V_H62, V_L66 with V_H63, V_L67 with V_H64, V_L68 with V_H65, V_L69 with V_H66, V_L70 with V_H67, V_L71 with V_H68, V_L72 with V_H69, V_L73 with V_H70, V_L74 with V_H70, and V_L75 with V_H70, V_L76 with V_H71, V_L77 with V_H72, V_L78 with V_H73, V_L79 with V_H74, V_L80 with V_H75, V_L81 with V_H76, V_L82 with V_H77, V_L83 with V_H78, V_L84 with V_H79, V_L85 with V_H80, V_L86 with V_H81, V_L87 with V_H82, V_L88 with V_H86, V_L89 with V_H83, V_L90 with V_H84, V_L91 with V_H85, V_L 92 with V_H 87, V_L 93 with V_H 88, V_L 94 with V_H 88, V_L 95 with V_H 89, V_L 96 with V_H 90, V_L 97 with V_H 91, V_L 98 with V_H 92, V_L 99 with V_H 93, and V_L 100 with V_H 94.

In some instances, the antigen binding proteins in the above pairings can comprise amino acid sequences that have 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% sequence identity with the specified variable domains.

Still other antigen binding proteins, e.g., antibodies or immunologically functional fragments, include variant forms of a variant heavy chain and a variant light chain as just described.

Antigen Binding Protein CDRs

In various embodiments, the antigen binding proteins disclosed herein can comprise polypeptides into which one or more CDRs are grafted, inserted and/or joined. An antigen binding protein can have 1, 2, 3, 4, 5 or 6 CDRs. An antigen binding protein thus can have, for example, one heavy chain CDR1 ("CDRH1"), and/or one heavy chain CDR2 ("CDRH2"), and/or one heavy chain CDR3 ("CDRH3"), and/or one light chain CDR1 ("CDRL1"), and/or one light chain CDR2 ("CDRL2"), and/or one light chain CDR3 ("CDRL3"). Some antigen binding proteins include both a CDRH3 and a CDRL3. Specific heavy and light chain CDRs are identified in Tables 3A and 3B, respectively, infra.

Complementarity determining regions (CDRs) and framework regions (FR) of a given antibody can be identified using the system described by Kabat et al., (1991) "Sequences of Proteins of Immunological Interest", 5th Ed., US Dept. of Health and Human Services, PHS, NIH, NIH Publication no. 91-3242. Although presented in the Kabat nomenclature scheme, as desired, the CDRs disclosed herein can also be redefined according an alternative nomenclature scheme, such as that of Chothia (see Chothia & Lesk, (1987) J. Mol. Biol. 196:901-917; Chothia et al., (1989) Nature 342:878-883 or Honegger & Pluckthun, (2001) J. Mol. Biol. 309:657-670). Certain antibodies that are disclosed herein comprise one or more amino acid sequences that are identical or have substantial sequence identity to the amino acid sequences of one or more of the CDRs presented in Table 3A (CDRHs) and Table 3B (CDRLs), infra.



48C9	603	CDRH1-1	GYYWT
49A12
51E2
48F3	604	CDRH1-2	GYYWS
51E5
52C5
55E4
60G5.1
49B11
50H10
53C1
56G1
51C1
48F8	605	CDRH1-3	SYSMN
51G2
56A7
53B9
56B4
57E7
57F11
56E4
55E6
48H11	606	CDRH1-4	GYYKH
48G4	607	CDRH1-5	ELSIH
53C3.1
49A10	608	CDRH1-6	NYGMH
58C2
59G10.2
48D4
49C8	609	CDRH1-7	SYDID
52H1
49G2	610	CDRH1-8	NYGMR
50C12
55G11
49G3	611	CDRH1-9	NPRMGVS
49H12	612	CDRH1-10	SYDIN
54A1
55G9
50G1	613	CDRH1-11	SYGLH
51A8	614	CDRH1-12	SYGMH
52C1
53H5.2
56C11
60D7
64H5
65G4
66G2
68G5
64C8
67G8
68D3v2
51C10.1	615	CDRH1-13	NYAMS
59D10v1
59D10v2
51C10.2	616	CDRH1-14	SGGYYWS
64A6
52A8	617	CDRH1-15	GYYLH
66B4
52B8	618	CDRH1-16	YYYWS
52F8	619	CDRH1-17	GYYTH
52H2	620	CDRH1-18	TYYWS
53F6	621	CDRH1-19	TYGMH
53H5.3	622	CDRH1-20	DYYWN
54H10.1	623	CDRH1-21	SYAMS
60F9
61G5
55D1
48H3
53C11
48B4
52D6
55D3	624	CDRH1-22	SGVYYWN
55E9	625	CDRH1-23	SFGMH
55G5	626	CDRH1-24	SYYWS
65C3
68D5
67F5
55A7
56E7	627	CDRH1-25	SYWIG
67A5
67C10
64H6
56G3.2	628	CDRH1-26	SYYWN
56G3.3	629	CDRH1-27	SSSYYWG
55B10
61H5
52B9
57B12	630	CDRH1-28	SGVYYWS
57D9	631	CDRH1-29	SNSATWN
59A10	632	CDRH1-30	DSYMS
49H4
59C9	633	CDRH1-31	SYSMS
58A5
57A4
57F9
59G10.3	634	CDRH1-32	HYAMS
60G5.2	635	CDRH1-33	NYGIS
63G8	636	CDRH1-34	SYGIH
64A8
67B4
68D3
64E6	637	CDRH1-35	SGDYYWT
65E8
65F11
67G7
63H11
63F5
65C1
66F6
63B6	638	CDRH1-36	SGDYYWS
64D4
65F9
64B10 64B10v2
63E6	639	CDRH1-37	GYYMH
66F7
50G5 v1
50G5 v2
67G10v1	640	CDRH1-38	NAWMS
67G10v2
63A10 65H11
53C3.2	641	CDRH1-39	SGNYYWS
64A7	642	CDRH1-40	SDTSYWG
50D4	643	CDRH1-41	SHDIN
61E1	644	CDRH1-42	SNSAAWN
66D4	645	CDRH1-43	GYYIH
54H10.3
65B1	646	CDRH1-44	GYFMH
67A4	647	CDRH1-45	TYDMH
65B4	648	CDRH1-46	SYDMH
65E3	649	CDRH1-47	NYNMH
65D4	650	CDRH1-48	FYGMH
65D1	651	CDRH1-49	YYYIH
65B7	652	CDRH1-50	SDAYYWS
68C8	653	CDRH1-51	SGDNYWS
63F9	654	CDRH1-52	SGGYYWN
67F6v1	655	CDRH1-53	GYWIG
67F6v2
48C9	656	CDRH2-1	EINHSENTNYNPSLKS
52C5
55E4
56G1
49A12
51E2
60G5.1
49B11
50H10
53C1
51C1
48F3	657	CDRH2-2	EITHTGSSNYNPSLKS
48F8	658	CDRH2-3	SISSSSSYEYYVDSVKG
53B9
56B4
57E7
57F11
48H11	659	CDRH2-4	WINPNSGATKYAQKFQG
48G4	660	CDRH2-5	GFDPEDGETIYAQKFQG
53C3.1
49A10	661	CDRH2-6	IIWYDGSNKNYADSVKG
48D4
49C8	662	CDRH2-7	WMNPNGGNTGYAQKFQG
52H1
49G2	663	CDRH2-8	LIWYDGSNKFYADSVKG
50C12
55G11
49G3	664	CDRH2-9	HIFSNDEKSYSTSLKS
49H12	665	CDRH2-10	WMNPYSGSTGYAQNFQG
50G1	666	CDRH2-11	VIWNDGSNKLYADSVKG
51A8	667	CDRH2-12	VISYDGSNKYYADSVKG
63G8
64A8
67B4
68D3
51C10.1	668	CDRH2-13	GISGSSAGTYYADSVKG
59D10v1
59D10v2
51C10.2	669	CDRH2-14	YIYYNGSPYDNPSLKR
51E5	670	CDRH2-15	ELDHSGSINYNPSLKS
51G2	671	CDRH2-16	SISSSSTYIYYADSVKG
56A7
56E4
52A8	672	CDRH2-17	WINPNSAATNYAPKFQG
52B8	673	CDRH2-18	YIYYSGSTNYNPSLKS
55A7
52C1	674	CDRH2-19	VIWYDGSNNYYADSVKG
52F8	675	CDRH2-20	WINPSSGDTKYAQKFQG
52H2	676	CDRH2-21	YIFYNGNANYSPSLKS
53F6	677	CDRH2-22	VIWYDGSNKYYADSVKG
60D7
65D4
53H5.2	678	CDRH2-23	LISYDGSNKYYADSVKG
53H5.3	679	CDRH2-24	EINHSGTTNYNPSLKS
54A1	680	CDRH2-25	WMNPHSGNTGYAQKFQG
55G9
54H10.1	681	CDRH2-26	AISGSGRTTYSADSVKG
55D1
48H3
53C11
55D3	682	CDRH2-27	YLYYSGSTYYNPSLKS
55E9	683	CDRH2-28	LIWYDGDNKYYADSVKG
55G5	684	CDRH2-29	RIYISGSTNYNPSLEN
56C11	685	CDRH2-30	VIWYDGSYQFYADSVKG
56E7	686	CDRH2-31	IIYPGDSDTRYSPSFQG
67A5
67C10
67F6v1
67F6v2
56G3.2	687	CDRH2-32	RIYTSGSTNYNPSLKS
56G3.3	688	CDRH2-33	MIYYSGTTYYNPSLKS
55B10
56G3.3
57B12	689	CDRH2-34	YIYYSGSTYYNPSLKS
63H11
66F6
65F9
57D9	690	CDRH2-35	RTYYRSKWYNDYAVSVKS
61E1
58C2	691	CDRH2-36	VIWNDGNNKYYADSVKG
59A10	692	CDRH2-37	SISSSGSIVYFADSVKG
49H4
59C9	693	CDRH2-38	SISSSSTYIYYAD SLKG
58A5
57A4
57F9
59G10.2	694	CDRH2-39	ITSYGGSNKNYADSVKG
59G10.3	695	CDRH2-40	AISGSGAGTFYADSMKG
60F9	696	CDRH2-41	VISDSGGSTYYADSVKG
48B4
52D6
60G5.2	697	CDRH2-42	WISAYNGYSNYAQKFQD
61G5	698	CDRH2-43	VISGSGGDTYYADSVKG
64E6	699	CDRH2-44	YIYYTGSTYYNPSLKS
65E8
65F11
67G7
63B6	700	CDRH2-45	YIYYSGTTYYNPSLKS
64D4
65C3	701	CDRH2-46	YIYYTGSTNYNPSLKS
68D5
63E6	702	CDRH2-47	WMNPNSGATKYAQKFQG
66F7
64H5	703	CDRH2-48	VIWDDGSNKYYADSVKG
65G4
67G10v1	704	CDRH2-49	RIKSKTDGGTTEYAAPVKG
67G10v2
63F5	705	CDRH2-50	YIYYSGSAYYNPSLKS
64A7	706	CDRH2-51	NIYYSGTTYFNPSLKS
65C1	707	CDRH2-52	YIFYSGSTYYNPSLKS
65B7
66B4	708	CDRH2-53	WINPNSGGTDYAQKFQG
66G2	709	CDRH2-54	GISYDGSNKNYADSVKG
68G5	710	CDRH2-55	VIWYDGSNKYHADSVKG
66D4	711	CDRH2-56	WINPPSGATNYAQKFRG
65B1	712	CDRH2-57	WINPNSGATNYAQKFHG
67A4	713	CDRH2-58	AIGIAGDTYYSDSVKG
65B4	714	CDRH2-59	TIDTAGDAYYPGSVKG
63A10	715	CDRH2-60	RIKSKTDGGTTDYAAPVKG
67G10v1
67G10v2
65H11	716	CDRH2-61	RIIGKTDGGTTDYAAPVKG
64C8	717	CDRH2-62	VISYDGSNKHYADSVKG
65E3	718	CDRH2-63	VLWYDGNTKYYADSVKG
65D1	719	CDRH2-64	LIWYDGSNKDYADSVKG
67G8	720	CDRH2-65	VIWYDGSNKDYADSVKG
64A6	721	CDRH2-66	YIYYSGGTHYNPSLKS
67F5	722	CDRH2-67	YIYYSGNTNYNPSLKS
64B10	723	CDRH2-68	FIYYSGGTNYNPSLKS
68C8	724	CDRH2-69	FMFYSGSTNYNPSLKS
64H6	725	CDRH2-70	IIYPGDSETRYSPSFQG
63F9	726	CDRH2-71	YIYDSGSTYYNPSLKS
61H5	727	CDRH2-72	SIYYSGTTYYNPSLKS
52B9
50G5v1	728	CDRH2-73	WINPDSGGTNYAQKFQG
50G5v2
54H10.3	729	CDRH2-74	WINPNSGGTNYAQKFRG
50D4	730	CDRH2-75	WMNPYSGSTGLAQRFQD
55E6	731	CDRH2-76	YISSGSSTIYHADSVKG
53C3.2	732	CDRH2-77	YIYHSGSAYYNPSLKS
64B10v2	1868	CDRH2-78	FIYYSGGTNYNPPLKS
68D3v2	1869	CDRH2-79	FISYAGSNKYYADSVKG
48C9	733	CDRH3-1	ESGNFPFDY
49A12
51E2
48F3	734	CDRH3-2	GGILWFGEQAFDI
48F8	735	CDRH3-3	SLSIAVAASDY
53B9
56B4
57E7
57F11
48H11	736	CDRH3-4	EVPDGIWAGSNAFDF
48G4	737	CDRH3-5	HSGSGRFYYYYYGMDV
53C3.1
49A10	738	CDRH3-6	DQDYDFWSGYPYFYYYGMDV
48D4
49C8	739	CDRH3-7	GKEFSRAEFDY
52H1
49G2	740	CDRH3-8	DRYYDFWSGYPYFFYYGLDV
50C12
55G11
49G3	741	CDRH3-9	VDTLNYHYYGMDV
49H12	742	CDRH3-10	YNWNYGAFDF
54A1
55G9
50G1	743	CDRH3-11	DQYYDFWSGYPYYHYYGMDV
51A8	744	CDRH3-12	ADGDYPYYYYYYGMDV
51C10.1	745	CDRH3-13	DWSIAVAGTFDY
59D10v1
59D10v2
51C10.2	746	CDRH3-14	GALYGMDV
51E5	747	CDRH3-15	VLGSTLDY
51G2	748	CDRH3-16	DTYISGWNYGMDV
52A8	749	CDRH3-17	EGGTYNWFDP
52B8	750	CDRH3-18	GTRAFDI
52C1	751	CDRH3-19	DRAGASPGMDV
52C5	752	CDRH3-20	VTGTDAFDF
60G5.1
49B11
50H10
53C1
51C1
55E4
56G1
52F8	753	CDRH3-21	SGWYPSYYYGMDV
52H2	754	CDRH3-22	ETDYGDYARPFEY
53F6	755	CDRH3-23	GHYDSSGPRDY
53H5.2	756	CDRH3-24	EANWGYNYYGMDV
53H5.3	757	CDRH3-25	ILRYFDWLEYYFDY
61E1	758	CDRH3-26	EGSWSSFFDY
54H10	759	CDRH3-27	EQQWLVYFDY
55D1
48H3
53C11
55D3	760	CDRH3-28	DGITMVRGVTHYYGMDV
57B12
55E6	761	CDRH3-29	EGYYDSSGYYYNGMDV
55E9	762	CDRH3-30	NSGWDYFYYYGMDV
55G5	763	CDRH3-31	SGSYSFDY
56A7	764	CDRH3-32	DIYSSGWSYGMDV
56E4
56C11	765	CDRH3-33	DHVWRTYRYIFDY
56E7	766	CDRH3-34	AQLGIFDY
50G5v1	767	CDRH3-35	GGYSYGYEDYYGMDV
50G5v2
56G3.2	768	CDRH3-36	GPLWFDY
56G3.3	769	CDRH3-37	VAAVYWYFDL
55B10
61H5
52B9
55A7	770	CDRH3-38	GITGTIDF
57D9	771	CDRH3-39	IVVVPAVLFDY
58C2	772	CDRH3-40	DQNYDFWNGYPYYFYYGMDV
59A10	773	CDRH3-41	ETFSSGWFDAFDI
49H4
59C9	774	CDRH3-42	DRWSSGWNEGFDY
58A5
57A4
57F9
53C3.2	775	CDRH3-43	TTGASDI
59G10.2	776	CDRH3-44	EAGYSFDY
59G10.3	777	CDRH3-45	DLRIAVAGSFDY
60D7	778	CDRH3-46	DQYFDFWSGYPFFYYYGMDV
60F9	779	CDRH3-47	DHSSGWYYYGMDV
48B4
52D6
60G5.2	780	CDRH3-48	EEKQLVKDYYYYGMDV
61G5	781	CDRH3-49	DHTSGWYYYGMDV
63G8	782	CDRH3-50	TVTKEDYYYYGMDV
64A8
67B4
68D3
66G2
64E6	783	CDRH3-51	MTTPYWYFDL
65E8
65F11
67G7
63H11
63F5
66F6
63B6	784	CDRH3-52	MTTPYWYFGL
64D4
65C3	785	CDRH3-53	EYYYGSGSYYP
68D5
67F5
63E6	786	CDRH3-54	ELGDYPFFDY
66F7
64H5	787	CDRH3-55	EYVAEAGFDY
65G4
67G10v1	788	CDRH3-56	DSSGSYYVEDYFDY
67G10 v2
63A10
65H11
64A7	789	CDRH3-57	LRGVYWYFDL
65C1	790	CDRH3-58	MTSPYWYFDL
66B4	791	CDRH3-59	DAATGRYYFDN
68G5	792	CDRH3-60	DPGYSYGHFDY
66D4	793	CDRH3-61	ETGTWSFFDY
65B1	794	CDRH3-62	ELGIFNWFDP
67A4	795	CDRH3-63	DRSSGRFGDYYGMDV
65B4	796	CDRH3-64	DRSSGRFGDFYGMDV
64C8	797	CDRH3-65	ELLWFGEYGVDHGMDV
65E3	798	CDRH3-66	DVYGDYFAY
65D4	799	CDRH3-67	ALNWNFFDY
65D1	800	CDRH3-68	EGTTRRGFDY
67G8	801	CDRH3-69	SAVALYNWFDP
65B7	802	CDRH3-70	ESRILYFNGYFQH
64A6	803	CDRH3-71	VLHYSDSRGYSYYSDF
65F9	804	CDRH3-72	VLHYYDSSGYSYYFDY
64B10v1	805	CDRH3-73	YSSTWDYYYGVDV
64B10v2
68C8	806	CDRH3-74	YRSDWDYYYGMDV
67A5	807	CDRH3-75	RASRGYRFGLAFAI
67C10	808	CDRH3-76	RASRGYRYGLAFAI
64H6	809	CDRH3-77	VAVSAFNWFDP
63F9	810	CDRH3-78	DVLMVYTKGGYYYYGVDV
67F6v1	811	CDRH3-79	RASRGYSYGHAFDF
67F6v2
50D4	812	CDRH3-80	DLSSGYYYYGLDV
54H10.3	813	CDRH3-81	EEDYSDHHYFDY
66D4	1870	CDRH3-82	ETGTWNFFDY
68D3v2	1871	CDRH3-83	TVTEEDYYYYGMDV



48C9	814		CDRL1-1	RASQNIRTYLN
49A12
51E2
48F3	815		CDRL1-2	RASQRISSYLN
48F8	816		CDRL1-3	RASQDIGNSLH
53B9
56B4
57E7
57F11
48H11	817		CDRL1-4	RASQNIRSYLN
49A10	818		CDRL1-5	RSSQSLLDSDDGNTYLD
48D4
49C8	819		CDRL1-6	QASQDINIYLN
52H1
49G2	820		CDRL1-7	RSSQSLLDSDDGDTYLD
50C12
55G11
60D7
50G1
49G3	821		CDRL1-8	QASQGISNYLN
49H12	822		CDRL1-9	QASQDITKYLN
51A8	823		CDRL1-10	TRSSGSIASDYVQ
51C10.1	824		CDRL1-11	SGDALPKKYAY
51C10.2	825		CDRL1-12	SGDELGDKYAC
51E5	826		CDRL1-13	RASQDIRNDLG
63G8v1
64A8
67B4
68D3
51G2	827		CDRL1-14	RASQGISSWLA
59A10
49H4
52A8	828		CDRL1-15	RASQTISSYLN
52B8	829		CDRL1-16	RASQSVSDILA
52C1	830		CDRL1-17	SGSSSNIGINYVS
52C5	831		CDRL1-18	RASQSISNYLN
55E4
49B11
50H10
53C1
56G1
51C1
60G5.1
52F8	832		CDRL1-19	RSSQSLLHSNGYNYLD
52H2	833		CDRL1-20	RASQSVRSSYLA
53F6	834		CDRL1-21	RSSQSLQHSNGYNYLD
53H5.2	835		CDRL1-22	RASQGIRNDLG
50G5 v1
66G2
53H5.3	836		CDRL1-23	RASQSVSSNVA
54A1	837		CDRL1-24	QASQDISIYLN
55G9
54H10.1	838		CDRL1-25	RASQSFSSSYLA
55D1
48H3
53C11
55D3	839		CDRL1-26	RASQDISNYLA
50D4
55E9	840		CDRL1-27	RSSQSLLHSNGFNYLD
55G5	841		CDRL1-28	SGDNLGDKYAF
56A7	842		CDRL1-29	RASQDISSWLA
56E4
56C11	843		CDRL1-30	GGNDIGSKSVH
56E7	844		CDRL1-31	QASQDIKKFLN
56G3.2	845		CDRL1-32	RARQSVGSNLI
56G3.3	846		CDRL1-33	RASQSVSRDYLA
55B10
61H5
52B9
57B12	847		CDRL1-34	RASHDISNYLA
57D9	848		CDRL1-35	RASPSVSSSYLA
53C3.2	849		CDRL1-36	RASQSISSNLA
59C9	850		CDRL1-37	RASQDIDSWLV
58A5
57A4
57F9
59D10 v1	851		CDRL1-38	SGDAVPKKYAN
59D10 v2	852		CDRL1-39	SGDKLGDKYVC
65D1
59G10.2	853		CDRL1-40	SGDNLGDKYAC
59G10.3	854		CDRL1-41	SGSSSNIGDNYVS
54H10.3	855		CDRL1-84	RASQTISIYLN
60F9	856		CDRL1-43	RASQRVPSSYIV
48B4
52D6
60G5.2	857		CDRL1-44	SGNKLGDKYVC
61G5	858		CDRL1-45	RASQRVPSSYLV
64E6	859		CDRL1-46	RASQSVRNSYLA
65E8
65F11
67G7
63H11
66F6
63B6	860		CDRL1-47	RASQSVSNSYLA
64D4
65C3	861		CDRL1-48	RASQSVSSQLA
68D5
63E6	862		CDRL1-49	RTSQSISSYLN
66F7	863		CDRL1-50	RTSQSISNYLN
64H5	864		CDRL1-51	GGNNIGSKNVH
65G4
65E3
64H6
67G10 v1	865		CDRL1-52	GGNNIGSKAVH
63A10 v1
63A10v2
67G10 v2	866		CDRL1-53	SGDKLGDKYAC
63F5	867		CDRL1-54	RASQTVRNNYLA
64A7	868		CDRL1-55	RASQSVSRNYLA
65C1	869		CDRL1-56	RASQTIRNSYLA
66B4	870		CDRL1-57	RASQGISRWLA
55A7	871		CDRL1-58	RASQSISSYLN
68G5	872		CDRL1-59	GGNNIGSINVH
66D4	873		CDRL1-60	RASQIISRYLN
65B1	874		CDRL1-61	RASQNINNYLN
67A4	875		CDRL1-62	GGNNIGSKSVH
65B4	876		CDRL1-63	GGNNIGSKSVQ
55E6	877		CDRL1-64	RASQSVSRSHLA
65H11	878		CDRL1-65	GGNNIGSKTVH
64C8	879		CDRL1-66	RSSPSLVYSDGNTYLN
65D4	880		CDRL1-67	GGNDIGSKNVH
61E1	881		CDRL1-68	RASQSIGTFLN
67G8	882		CDRL1-69	GGNNIGSYNVF
65B7	883		CDRL1-70	RASQSVSSMYLA
64A6	884		CDRL1-71	RASQSVNSNLA
65F9	885		CDRL1-72	RASQSVSSNLA
67F5
64B10	886		CDRL1-73	SGSSSNIGNNYVA
68C8	887		CDRL1-74	SGSSSNIGNNYVS
67A5 67C10	888		CDRL1-75	RSSQSLLNSDDGNTYLD
63F9	889		CDRL1-76	RASQDIRNDLA
67F6v1	890		CDRL1-77	RSSQSLLNSDAGTTYLD
50G5v2	891		CDRL1-78	RSSQRLVYSDGNTYLN
48G4	892		CDRL1-79	RASQSVASSYLV
53C3.1
58C2	893		CDRL1-81	RSSQSLFDNDDGDTYLD
68G8v2	1872		CDRL1-82	RASQGIRSGLG
68G8v3
65B7v1	1873		CDRL1-83	RASQSVSSIYLA
67F6v2	1874		CDRL1-84	RSSQSLLNSDAGTTYLD
65B7v2	1875		CDRL1-85	RSSQSLVYSDGDTYLN
65H11v2	1876		CDRL1-86	SGDKLGDRYVC
63A10v3	1877		CDRL1-87	SGDKLGNRYTC
48C9	894		CDRL2-1	VASSLES
49A12
51E2
48F3	895		CDRL2-2	AVSSLQS
48F8	896		CDRL2-3	FASQSFS
53B9
56B4
57E7
57F11
48H11	897		CDRL2-4	GASNLQS
49A10	898		CDRL2-5	TLSYRAS
48D4
49G2
50C12
55G11
60D7
67A5
67C10
50G1
58C2		VL91
49C8	899		CDRL2-6	DVSNLET
52H1
54A1
55G9
49G3	900		CDRL2-7	DASNLET
56E7
49H12	901		CDRL2-8	DTFILET
51A8	902		CDRL2-9	EDKERSS
51C10.1	903		CDRL2-10	EDSKRPS
59D10v1
51C10.2	904		CDRL2-11	QDTKRPS
59G10.2
51E5	905		CDRL2-12	AASSLQF
51G2	906		CDRL2-13	DASSLQS
52A8	907		CDRL2-14	AASSLQS
52C5
53H5.2
55D3
56G1
57B12
63E6
66F7
66D4
50G5 v1
51C1
55A7
61E1
60G5.1


52B8	908	CDRL2-15	GASTRAT
53H5.3
65F9
52C1	909	CDRL2-16	DNNKRPS
59G10.3
68C8
52F8	910	CDRL2-17	LGSNRAS
55E9
52H2	911	CDRL2-18	GASRRAT
53F6	912	CDRL2-19	LDSNRAS
54H10.1	913	CDRL2-20	GASSRAT
55D1
48H3
53C11
57D9
61H5
52B9
63F5
64A7
65B7
55E6
55E4	914	CDRL2-21	TASSLQS
49B11
50H10
53C1
50G5v2 65B7v2	915	CDRL2-22	KVSNWDS
55G5	916	CDRL2-23	QDNKRPS
56A7	917	CDRL2-24	DASTLQS
56E4
56C11	918	CDRL2-25	DDSDRPS
67A4
65B4
56G3.2	919	CDRL2-26	GASSRDT
56G3.3	920	CDRL2-27	GASARAT
55B10
59A10	921	CDRL2-28	GASSLQS
49H4
59C9	922	CDRL2-29	AASNLQR
58A5
57A4
57F9
63G8v1
63G8v2
63G8v3
64A8
67B4
68D3
59D10 v2	923	CDRL2-30	QNNKRPS
60F9	924	CDRL2-31	GSSNRAT
48B4
52D6
60G5.2	925	CDRL2-32	QDSKRPS
65D1
65H11v2
61G5	926	CDRL2-33	GASNRAT
64E6	927	CDRL2-34	GAFSRAS
65E8
65F11
67G7
63H11
63B6	928	CDRL2-35	GAFSRAT
64D4
65C1
66F6
48G4
53C3.1
65C3	929	CDRL2-36	GASNRAI
68D5
64H5	930	CDRL2-37	RDSKRPS
65G4
67G8
64H6
67G10 v1	931	CDRL2-38	SDSNRPS
65H11
67G10 v2	932	CDRL2-39	QDNERPS
66B4	933	CDRL2-40	AASSLKS
66G2	934	CDRL2-41	AASNLQS
68G5	935	CDRL2-42	RDRNRPS
65E3
65D4
65B1	936	CDRL2-43	TTSSLQS
53C3.2	937	CDRL2-44	GTSIRAS
63A10v1	938	CDRL2-45	CDSNRPS
63A10v2
54H10.3	939	CDRL2-46	SASSLQS
64C8	940	CDRL2-47	KGSNWDS
64A6	941	CDRL2-48	GTSTRAT
67F5	942	CDRL2-49	GSSNRAI
64B10	943	CDRL2-50	DNDKRPS
63F9	944	CDRL2-51	ASSSLQS
67F6	945	CDRL2-52	TLSFRAS
67F6v2
50D4	946	CDRL2-53	AASTLLS
63A10v3	1878	CDRL2-54	QDSERPS
48C9	947	CDRL3-1	QQSDSIPRT
49A12
51E2
48F3	948	CDRL3-2	QQSYSATFT
48F8	949	CDRL3-3	HQSSDLPLT
53B9
56B4
57E7
57F11
48H11	950	CDRL3-4	QQSYNTPCS
49A10	951	CDRL3-5	MQRIEFPIT
48D4
67C10
67F6v1
67F6v1
49C8	952	CDRL3-6	QQYDNLPFT
52H1
49G2	953	CDRL3-7	MQHIEFPST
50C12
55G11
49G3	954	CDRL3-8	HQYDDLPLT
49H12	955	CDRL3-9	QQYDNLPLT
54A1
55G9
51A8	956	CDRL3-10	QSYDRNNHVV
51C10.1	957	CDRL3-11	YSTDSSVNHVV
51C10.2	958	CDRL3-12	QAWDSGTVV
51E5	959	CDRL3-13	LQHSSYPLT
51G2	960	CDRL3-14	QQTNSFPPWT
56A7
56E4
59A10
49H4
59C9
58A5
57A4
57F9
52A8	961	CDRL3-15	QQSYSTPLT
65B1
52B8	962	CDRL3-16	QQYNNWPLT
56G3.2
52C1	963	CDRL3-17	GTWDSSLSAVV
64B10
68C8
52C5	964	CDRL3-18	QQSSSIPWT
55E4
49B11
50H10
53C1
51C1
60G5.1
52F8	965	CDRL3-19	MQALQTPFT
52H2	966	CDRL3-20	QQYGSSPRS
53F6	967	CDRL3-21	MQGLQTPPT
53H5.2	968	CDRL3-22	LQHKSYPFT
53H5.3	969	CDRL3-23	QQFSNSIT
54H10.1	970	CDRL3-24	QQYGSSRT
55D1
48H3
53C11
55D3	971	CDRL3-25	QQYNIYPRT
55E9	972	CDRL3-26	MQALQTLIT
55G5	973	CDRL3-27	QAWDSATVI
56C11	974	CDRL3-28	QVWDSSSDVV
56E7	975	CDRL3-29	QQYAILPFT
56G1	976	CDRL3-30	QQSSTIPWT
56G3.3	977	CDRL3-31	QQYGRSLFT
55B10
61H5
52B9
57B12	978	CDRL3-32	QQYNTYPRT
57D9	979	CDRL3-33	HQYGTSPCS
59D10 v1	980	CDRL3-34	YSTDSSGNHVV
59D10 v2	981	CDRL3-35	QAWDSSTAV
59G10.2	982	CDRL3-36	QAWDSSTTWV
59G10.3	983	CDRL3-37	GTWDSSLSVMV
60D7	984	CDRL3-38	MQRIEFPLT
50G1
60F9	985	CDRL3-39	QQYGSSPPWT
48B4
52D6
61G5
60G5.2	986	CDRL3-40	QAWDSSTWV
63G8v1	987	CDRL3-41	LQHNSYPLT
63G8v2
64A8
67B4
68D3
64E6	988	CDRL3-42	QQFGSSLT
65E8
65F11
67G7
63H11
63F5
65C1
66F6
63B6	989	CDRL3-43	QQFGRSFT
64D4
65C3	990	CDRL3-44	QQYNNWPWT
68D5
63E6	991	CDRL3-45	QQSYSTSLT
66F7
64H5	992	CDRL3-46	QVWDSSSVV
65G4
67G10 v1	993	CDRL3-47	QVWDSSSDGV
67G10 v2	994	CDRL3-48	QAWDSTTVV
64A10v3
64A7	995	CDRL3-49	QQYGSSSLCS
66B4	996	CDRL3-50	QQANSFPPT
66G2	997	CDRL3-51	LQLNGYPLT
68G5	998	CDRL3-52	QLWDSSTVV
66D4	999	CDRL3-53	QQSYSSPLT
54H10.3
55A7	1000	CDRL3-54	QQTYSAPFT
67A4	1001	CDRL3-55
65B4			QVWDSSSDHVV
63A10	1002	CDRL3-56	HACGSSSSDGV
65H11	1003	CDRL3-57	QVWDSSCDGV
64C8	1004	CDRL3-58	IQDTHWPTCS
65E3	1005	CDRL3-59
67G8			QVWDSSTVV
65D4	1006	CDRL3-60	QVWDSNPVV
65D1	1007	CDRL3-61	QAWDSRV
65B7v1	1008	CDRL3-62	QQYGSSCS
64A6	1009	CDRL3-63
65F9			QQYNTWPWT
67F5	1010	CDRL3-64	QQYEIWPWT
55E6	1011	CDRL3-65	QQYGSSPWT
67A5	1012	CDRL3-66
58C2			MQRLEFPIT
61E1	1013	CDRL3-67	QQSFSTPLT
64H6	1014	CDRL3-68	QVWDSSPVV
63F9	1015	CDRL3-69	LQRNSYPLT
53C3.2	1016	CDRL3-70	HQYTNWPRT
48G4	1017	CDRL3-71	QQYGTSPFT
53C3.1
50G5 v1	1018	CDRL3-72	LQHNSYPRT
50D4	1019	CDRL3-74	QKYYSAPFT
50G5 v2	1020	CDRL3-75	MEGTHWPRD
63G8v3	1879	CDRL3-76	LQHNTYPLT
65B7v2	1880	CDRL3-77	MQGTHWRGWT
65H11v2	1881	CDRL3-78	QAWDSITVV
63A10v1	1882	CDRL3-79	QVWDSSSDGV



48C9	1021	CDRH1-1	GGTTACTACTGGACC
49A12
51E2
48F3	1022	CDRH1-2	GGTTACTACTGGAGC
51E5
52C5
55E4
60G50.1
49B11
50H10
53C1
56G1
51C1
48F8	1023	CDRH1-3	AGCTATAGCATGAAC
51G2
56A7
53B9
56B4
57E7
57F11
56E4
55E6
48H11	1024	CDRH1-4	GGCTACTATAAGCAC
48G4	1025	CDRH1-5	GAATTATCCATACAC
49A10	1026	CDRH1-6	AACTATGGCATGCAC
58C2
59G10.2
48D4
49C8	1027	CDRH1-7	AGTTATGATATCGAC
52H1
49G2	1028	CDRH1-8	AACTATGGCATGCGC
50C12
55G11
49G3	1029	CDRH1-9	AATCCTAGAATGGGTGTGAGC
49H12	1030	CDRH1-10	AGTTACGATATCAAC
54A1
55G9
50G1	1031	CDRH1-11	AGCTATGGCCTGCAC
51A8	1032	CDRH1-12	AGCTATGGCATGCAC
52C1
53H5.2
56C11
60D7
64H5
65G4
66G2
68G5
64C8
67G8
68D3v2
51C10.1	1033	CDRH1-13	AACTATGCCATGAGC
59D10v1
59D10v2
51C10.2	1034	CDRH1-14	AGTGGTGGTTACTACTGGAGC
64A6
52A8	1035	CDRH1-15	GGCTACTATTTGCAC
66B4
52B8	1036	CDRH1-16	TATTATTACTGGAGT
52F8	1037	CDRH1-17	GGCTACTATACACAC
52H2	1038	CDRH1-18	ACTTACTACTGGAGC
53F6	1039	CDRH1-19	ACCTATGGCATGCAC
53H5.3	1040	CDRH1-20	GATTACTACTGGAAC
54H10.1	1041	CDRH1-21	AGCTATGCCATGAGC
60F9
61G5
55D1
48H3
53C11
48B4
52D6
55D3	1042	CDRH1-22	AGTGGTGTTTACTACTGGAAC
55E9	1043	CDRH1-23	AGCTTTGGCATGCAC
55G5	1044	CDRH1-24	AGTTACTACTGGAGC
65C3
68D5
67F5
55A7
56E7	1045	CDRH1-25	AGCTACTGGATCGGC
67A5
67C10
64H6
56G3.2	1046	CDRH1-26	AGTTACTACTGGAAC
56G3.3	1047	CDRH1-27	AGTAGTAGTTACTACTGGGGC
55B10
61H5
52B9
57B12	1048	CDRH1-28	AGTGGTGTTTACTACTGGAGC
57D9	1049	CDRH1-29	AGCAACAGTGCTACTTGGAAC
59A10	1050	CDRH1-30	GACTCCTACATGAGC
49H4
59C9	1051	CDRH1-31	AGCTATAGCATGAGT
58A5
57A4
57F9
59G10.3	1052	CDRH1-32	CACTATGCCATGAGC
60G5.2	1053	CDRH1-33	AACTATGGTATCAGC
63G8	1054	CDRH1-34	AGCTATGGCATACAC
64A8
67B4
68D3
64E6	1055	CDRH1-35	AGTGGTGATTACTACTGGACC
65E8
65F11
67G7
63H11
63F5
65C1
66F6
63B6	1056	CDRH1-36	AGTGGTGATTACTACTGGAGC
64D4
65F9
64B10v1
64B10v1
63E6	1057	CDRH1-37	AGTGGTGATTACTACTGGACC
66F7
50G5 v1
50G5 v2
67G10v1	1058	CDRH1-38	AACGCCTGGATGAGT
67G10v2
63A10
65H11
53C3.2	1059	CDRH1-39	AGTGGTAATTACTACTGGAGC
64A7	1060	CDRH1-40	AGTGATACTTCCTACTGGGGC
50D4	1061	CDRH1-41	AGTCATGATATCAAC
61E1	1062	CDRH1-42	AGCAACAGTGCTGCTTGGAAC
66D4	1063	CDRH1-43	GGCTACTATATACAC
54H10.3		CDRH1-43	GGCTACTATATACAC
65B1	1064	CDRH1-44	GGCTACTTTATGCAC
67A4	1065	CDRH1-45	ACCTACGACATGCAC
65B4	1066	CDRH1-46	AGTTACGACATGCAC
65E3	1067	CDRH1-47	AACTATAACATGCAC
65D4	1068	CDRH1-48	TTCTATGGCATGCAC
65D1	1069	CDRH1-49	TACTATTACATTCAC
65B7	1070	CDRH1-50	AGTGATGCTTACTACTGGAGC
68C8	1071	CDRH1-51	AGTGGTGATAACTACTGGAGC
63F9	1072	CDRH1-52	AGTGGTGGTTACTACTGGAAC
67F6	1073	CDRH1-53	GGCTACTGGATCGGC
48C9	1074	CDRH2-1
52C5
55E4
56G1
49A12
51E2
60G5.1
49B11
50H10
53C1
51C1
48F3	1075	CDRH2-2
48F8	1076	CDRH2-3
53B9
56B4
57E7
57F11
48H11	1077	CDRH2-4
48G4	1078	CDRH2-5
53C3.1	1078	CDRH2-5
49A10	1079	CDRH2-6
48D4
49C8	1080	CDRH2-7
49G2	1081	CDRH2-8
50C12
55G11
49G3	1082	CDRH2-9
49H12	1083	CDRH2-10
50G1	1084	CDRH2-11
51A8	1085	CDRH2-12
63G8
64A8
67B4
68D3
51C10.1	1086	CDRH2-13
59D10v1
59D10v2
51C10.2	1087	CDRH2-14
51E5	1088	CDRH2-15
51G2	1089	CDRH2-16
56A7
56E4
52A8	1090	CDRH2-17
52B8	1091	CDRH2-18
55A7
52C1	1092	CDRH2-19
52F8	1093	CDRH2-20
52H2	1094	CDRH2-21
53F6	1095	CDRH2-22
60D7
65D4
53H5.2	1096	CDRH2-23
53H5.3	1097	CDRH2-24
54A1	1098	CDRH2-25
55G9
54H10.1	1099	CDRH2-26
55D1
48H3
53C11
55D3	1100	CDRH2-27
55E9	1101	CDRH2-28
55G5	1102	CDRH2-29
56C11	1103	CDRH2-30
56E7	1104	CDRH2-31
67A5
67C10
67F6
56G3.2	1105	CDRH2-32
56G3.3	1106	CDRH2-33
57B12	1107	CDRH2-34
63H11
66F6
65F9
57D9	1108	CDRH2-35
61E1
58C2	1109	CDRH2-36
59A10	1110	CDRH2-37
49H4
59C9	1111	CDRH2-38
58A5
57A4
57F9
59G10.2	1112	CDRH2-39
59G10.3	1113	CDRH2-40
60F9	1114	CDRH2-41
48B4
52D6
60G5.2	1115	CDRH2-42
61G5	1116	CDRH2-43
64E6	1117	CDRH2-44
65E8
65F11
67G7
63B6	1118	CDRH2-45
64D4
65C3	1119	CDRH2-46
68D5
63E6	1120	CDRH2-47
66F7
64H5	1121	CDRH2-48
65G4
67G10v1	1122	CDRH2-49
67G10v2
63F5	1123	CDRH2-50
64A7	1124	CDRH2-51
65C1	1125	CDRH2-52
65B7
66B4	1126	CDRH2-53
66G2	1127	CDRH2-54
68G5	1128	CDRH2-55
66D4	1129	CDRH2-56
65B1	1130	CDRH2-57
67A4	1131	CDRH2-58
65B4	1132	CDRH2-59
63A10	1133	CDRH2-60
67G10v1
67G10v2
65H11	1134	CDRH2-61
64C8	1135	CDRH2-62
65E3	1136	CDRH2-63
65D1	1137	CDRH2-64
67G8	1138	CDRH2-65
64A6	1139	CDRH2-66
67F5	1140	CDRH2-67
64B10	1141	CDRH2-68
68C8	1142	CDRH2-69
64H6	1143	CDRH2-70
63F9	1144	CDRH2-71
61H5	1145	CDRH2-72
52B9
50G5v1	1146	CDRH2-73
50G5v2
54H10.3	1147	CDRH2-74
50D4	1148	CDRH2-75
55E6	1149	CDRH2-76
53C3.2	1150	CDRH2-77
64B10v2	1883	CDRH2-78
68D3v2	1884	CDRH2-79
48C9	1151	CDRH3-1	GAGAGTGGGAACTTCCCCTTTGACTAC
49A12
51E2
48F3	1152	CDRH3-2
48F8	1153	CDRH3-3
53B9
56B4
57E7
57F11
48H11	1154	CDRH3-4
48G4	1155	CDRH3-5
53C3.1
49A10	1156	CDRH3-6
48D4
49C8	1157	CDRH3-7
49G2	1158	CDRH3-8
50C12
55G11
49G3	1159	CDRH3-9
49H12	1160	CDRH3-10	TATAATTGGAACTATGGGGCTTTTGATTTC
54A1
55G9
50G1	1161	CDRH3-11
51A8	1162	CDRH3-12
51C10.1	1163	CDRH3-13
59D10 v1
59D10 v2
51C10.2	1164	CDRH3-14	GGGGCCCTCTACGGTATGGACGTC
51E5	1165	CDRH3-15	GTCCTGGGATCTACTCTTGACTAT
51G2	1166	CDRH3-16
52A8	1167	CDRH3-17	GAGGGTGGAACTTACAACTGGTTCGACCCC
52B8	1168	CDRH3-18	GGAACTAGGGCTTTTGATATC
52C1	1169	CDRH3-19
52C5	1170	CDRH3-20	GTAACTGGAACGGATGCTTTTGATTTC
60G5.1
49B11
50H10
53C1
51C1
55E4
56G1
52F8	1171	CDRH3-21
52H2	1172	CDRH3-22
53F6	1173	CDRH3-23
53H5.2	1174	CDRH3-24
53H5.3	1175	CDRH3-25
61E1	1176	CDRH3-26	GAGGGCAGCTGGTCCTCCTTCTTTGACTAC
54H10.1	1177	CDRH3-27	GAACAGCAGTGGCTGGTTTATTTTGACTAC
55D1
48H3
53C11
55D3	1178	CDRH3-28
57B12
55E6	1179	CDRH3-29
55E9	1180	CDRH3-30
55G5	1181	CDRH3-31	AGTGGGAGCTACTCCTTTGACTAC
56A7	1182	CDRH3-32
56E4
56C11	1183	CDRH3-33
56E7	1184	CDRH3-34	GCACAACTGGGGATCTTTGACTAC
50G5v1	1185	CDRH3-35
50G5v2
56G3.2	1186	CDRH3-36	GGCCCTCTTTGGTTTGACTAC
56G3.3	1187	CDRH3-37	GTGGCAGCAGTTTACTGGTATTTCGATCTC
55B10
61H5
52B9
55A7	1188	CDRH3-38	GGGATAACTGGAACTATTGACTTC
57D9	1189	CDRH3-39
58C2	1190	CDRH3-40
59A10	1191	CDRH3-41
49H4
59C9	1192	CDRH3-42
58A5
57A4
57F9
53C3.2	1193	CDRH3-43	ACTACGGGTGCTTCTGATATC
59G10.2	1194	CDRH3-44	GAGGCCGGGTATAGCTTTGACTAC
59G10.3	1195	CDRH3-45
60D7	1196	CDRH3-46
60F9	1197	CDRH3-47
48B4
52D6
60G5.2	1198	CDRH3-48
61G5	1199	CDRH3-49
63G8	1200	CDRH3-50
64A8
67B4
68D3
66G2
64E6	1201	CDRH3-51	ATGACTACCCCTTACTGGTACTTCGATCTC
65E8
65F11
67G7
63H11
63F5
66F6
63B6	1202	CDRH3-52	ATGACTACTCCTTACTGGTACTTCGGTCTC
64D4			ATGACTACTCCTTACTGGTACTTCGGTCTC
65C3	1203	CDRH3-53
68D5
67F5
63E6	1204	CDRH3-54	GAACTCGGTGACTACCCCTTTTTTGACTAC
66F7			GAACTCGGTGACTACCCCTTTTTTGACTAC
64H5	1205	CDRH3-55	GAATACGTAGCAGAAGCTGGTTTTGACTAC
65G4			GAATACGTAGCAGAAGCTGGTTTTGACTAC
67G10v1	1206	CDRH3-56
67G10v2
63A10
65H11
64A7	1207	CDRH3-57	CTCCGAGGGGTCTACTGGTACTTCGATCTC
65C1	1208	CDRH3-58	ATGACTTCCCCTTACTGGTACTTCGATCTC
66B4	1209	CDRH3-59
68G5	1210	CDRH3-60
66D4	1211	CDRH3-61	GAGACTGGAACTTGGAGCTTCTTTGACTAC
65B1	1212	CDRH3-62	GAACTGGGGATCTTCAACTGGTTCGACCCC
67A4	1213	CDRH3-63
65B4	1214	CDRH3-64
64C8	1215	CDRH3-65
65E3	1216	CDRH3-66	GATGTCTACGGTGACTATTTTGCGTAC
65D4	1217	CDRH3-67	GCCCTCAACTGGAACTTTTTTGACTAC
65D1	1218	CDRH3-68	GAAGGGACAACTCGACGGGGATTTGACTAC
67G8	1219	CDRH3-69
65B7	1220	CDRH3-70
64A6	1221	CDRH3-71
65F9	1222	CDRH3-72
64B10	1223	CDRH3-73
68C8	1224	CDRH3-74
67A5	1225	CDRH3-75
67C10	1226	CDRH3-76
64H6	1227	CDRH3-77
63F9	1228	CDRH3-78
67F6	1229	CDRH3-79
50D4	1230	CDRH3-80
54H10.3	1231	CDRH3-81
66D4	1885	CDRH3-82	GAGACTGGAACTTGGAACTTCTTTGACTAC
68D3v2	1886	CDRH3-83



48C9	1232	CDRL1-1
49A12
51E2
48F3	1233	CDRL1-2
48F8	1234	CDRL1-3
53B9
56B4
57E7
57F11
48H11	1235	CDRL1-4
49A10	1236	CDRL1-5
48D4
49C8	1237	CDRL1-6
52H1
49G2	1238	CDRL1-7
50C12
55G11
50G1
60D7
49G3	1239	CDRL1-8
49H12	1240	CDRL1-9
51A8	1241	CDRL1-10
51C10.1	1242	CDRL1-11
51C10.2	1243	CDRL1-12
51E5	1244	CDRL1-13
63G8v1
64A8
67B4
68D3
51G2	1245	CDRL1-14
52A8	1246	CDRL1-15
52B8	1247	CDRL1-16
52C1	1248	CDRL1-17
52C5	1249	CDRL1-18
55E4
49B11
50H10
53C1
56G1
51C1
60G5.1
52F8	1250	CDRL1-19
52H2	1251	CDRL1-20
53F6	1252	CDRL1-21
53H5.2	1253	CDRL1-22
50G5v1
53H5.3	1254	CDRL1-23
54A1	1255	CDRL1-24
55G9
54H10.1	1256	CDRL1-25
55D1
48H3
53C11
55D3	1257	CDRL1-26
50D4
55E9	1258	CDRL1-27
55G5	1259	CDRL1-28
56A7	1260	CDRL1-29
56E4
56C11	1261	CDRL1-30
56E7	1262	CDRL1-31
56G3.2	1263	CDRL1-32
56G3.3	1264	CDRL1-33
55B10
61H5
52B9
57B12	1265	CDRL1-34
57D9	1266	CDRL1-35
53C3.2	1267	CDRL1-36
59C9	1268	CDRL1-37
58A5
57A4
57F9
59D10 v1	1269	CDRL1-38
59D10	1270	CDRL1-39
v2
65D1
59G10.2	1271	CDRL1-40
59G10.3	1272	CDRL1-41
54H10.3	1273	CDRL1-42
60F9	1274	CDRL1-43
48B4
52D6
60G5.2	1275	CDRL1-44
61G5	1276	CDRL1-45
64E6	1277	CDRL1-46
65E8
65F11
67G7
63H11
66F6
63B6	1278	CDRL1-47
64D4
65C3	1279	CDRL1-48
68D5
63E6	1280	CDRL1-49
66F7	1281	CDRL1-50
64H5	1282	CDRL1-51
65G4
65E3
64H6
67G10	1283	CDRL1-52
v1
63A10
63A10v2
67G10	1284	CDRL1-53
v2
63F5	1285	CDRL1-54
64A7	1286	CDRL1-55
65C1	1287	CDRL1-56
66B4	1288	CDRL1-57
55A7	1289	CDRL1-58
68G5	1290	CDRL1-59
66D4	1291	CDRL1-60
65B1	1292	CDRL1-61
67A4	1293	CDRL1-62
65B4	1294	CDRL1-63
55E6	1295	CDRL1-64
65H11	1296	CDRL1-65
64C8	1297	CDRL1-66
65D4	1298	CDRL1-67
61E1	1299	CDRL1-68
67G8	1300	CDRL1-69
65B7	1301	CDRL1-70
64A6	1302	CDRL1-71
65F9	1303	CDRL1-72
67F5
64B10	1304	CDRL1-73
68C8	1305	CDRL1-74
67A5	1306	CDRL1-75
67C10
63F9	1307	CDRL1-76
67F6v1	1308	CDRL1-77
67F6v2
50G5 v2	1309	CDRL1-78
48G4	1310	CDRL1-79
53C3.1
58C2	1311	CDRL1-81
65B7v1	1887	CDRL1-82
65B7v2	1888	CDRL1-83
63G8v3	1889	CDRL1-84
63G8v2
63A10v3	1890	CDRL1-85
65H11v2	1891	CDRL1-86
48C9	1312	CDRL2-1	GTTGCATCCAGTTTGGAAAGT
49A12
51E2
48F3	1313	CDRL2-2	GCTGTATCCAGTTTGCAAAGT
48F8	1314	CDRL2-3	TTTGCTTCCCAGTCCTTCTCA
53B9
56B4
57E7
57F11
48H11	1315	CDRL2-4	GGTGCATCTAATTTACAGAGT
49A10	1316	CDRL2-5	ACGCTTTCCTATCGGGCCTCT
48D4
49G2
50C12
55G11
60D7
67A5
67C10
50G1
60D7
58C2
49C8	1317	CDRL2-6	GATGTATCCAATTTGGAAACA
52H1
54A1
55G9
49G3	1318	CDRL2-7	GATGCATCCAATTTGGAAACA
56E7			GATGCATCCAATTTGGAAACA
49H12	1319	CDRL2-8	GATACATTCATTTTGGAAACA
51A8	1320	CDRL2-9	GAGGATAAAGAAAGATCCTCT
51C10.1	1321	CDRL2-10	GAGGACAGCAAACGACCCTCC
59D10
v1
51C10.2	1322	CDRL2-11	CAAAATAACAAGCGGCCCTCA
59G10.2			CAAAATAACAAGCGGCCCTCA
51E5	1323	CDRL2-12	GCTGCATCCAGTTTGCAATTT
51G2	1324	CDRL2-13	GATGCATCCAGTTTGCAAAGT
52A8	1325	CDRL2-14	GCTGCATCCAGTTTGCAAAGT
52C5
53H5.2
55D3
56G1
57B12
63E6
66F7
66D4
50G5 v1
51C1
55A7
61E1
60G5.1
52B8	1326	CDRL2-15	GGTGCATCCACCAGGGCCACT
53H5.3
65F9
52C1	1327	CDRL2-16	GACAATAATAAGCGACCCTCA
59G10.3
68C8
52F8	1328	CDRL2-17	TTGGGTTCTAATCGGGCCTCC
55E9			TTGGGTTCTAATCGGGCCTCC
52H2	1329	CDRL2-18	GGTGCATCCAGGAGGGCCACT
53F6	1330	CDRL2-19	TTGGATTCTAATCGGGCCTCC
54H10.1	1331	CDRL2-20	GGTGCATCCAGCAGGGCCACT
55D1
48H3
53C11
57D9
61H5
52B9
63F5
64A7
65B7v1
55E6
55E4	1332	CDRL2-21	ACAGCTTCCAGTTTGCAAAGT
49BG11
50H10
53C1
50G5v2	1333	CDRL2-22	AAGGTTTCTAACTGGGACTCT
65B7v2			AAGGTTTCTAACTGGGACTCT
55G5	1334	CDRL2-23	CAAGATACCAAGCGGCCCTCA
56A7	1335	CDRL2-24	GATGCATCCACTTTGCAAAGT
56E4			GATGCATCCACTTTGCAAAGT
56C11	1336	CDRL2-25	GATGATAGCGACCGGCCCTCA
67A4
65B4
56G3.2	1337	CDRL2-26	GGTGCATCCAGCAGGGACACT
56G3.3	1338	CDRL2-27	GGTGCATCCGCCAGGGCCACT
55B10			GGTGCATCCGCCAGGGCCACT
59A10	1339	CDRL2-28	GGTGCATCCAGTTTGCAAAGT
49H4			GGTGCATCCAGTTTGCAAAGT
59C9	1340	CDRL2-29	GCTGCATCCAATTTGCAAAGA
58A5
57A4
57F9
63G8v1
63GBv2
63G8v3
64A8
67B4
68D3
59D10	1341	CDRL2-30	CAAGATACCAAGCGGCCCTCA
v2			CAAGATACCAAGCGGCCCTCA
60F9	1342	CDRL2-31	GGTTCATCCAACAGGGCCACT
48B4
52D6
60G5.2	1343	CDRL2-32	CAAGATAGCAAGCGGCCCTCA
65D1
65H11v2
61G5	1344	CDRL2-33	GGTGCATCCAACAGGGCCACA
64E6	1345	CDRL2-34	GGTGCATTTAGCAGGGCCTCT
65E8
65F11
67G7
63H11
63B6	1346	CDRL2-35	GGTGCATTCAGTAGGGCCACT
64D4
65C1
66F6
48G4
53C3.1
65C3	1347	CDRL2-36	GGTGCCTCCAACAGGGCCATT
68D5
64H5	1348	CDRL2-37	AGGGATAGCAAGCGGCCCTCT
65G4
67G8
64H6
67G10	1349	CDRL2-38	AGCGATAGCAACCGGCCCTCA
v1
65H11
67G10	1350	CDRL2-39	CAAGATAACGAGCGGCCCTCA
v2			CAAGATAACGAGCGGCCCTCA
66B4	1351	CDRL2-40	GCTGCATCCAGTTTGAAAAGT
66G2	1352	CDRL2-41	GCTGCATCCAATTTGCAAAGT
68G5	1353	CDRL2-42	AGGGATAGGAACCGGCCCTCT
65E3
65D4
65B1	1354	CDRL2-43	ACTACATCCAGTTTGCAAAGT
53C3.2	1355	CDRL2-44	GGTACATCCATCAGGGCCAGT
63A10v1	1356	CDRL2-45	TGTGATAGCAACCGGCCCTCA
63A10v2			TGTGATAGCAACCGGCCCTCA
54H10.3	1357	CDRL2-46	TCTGCATCCAGTTTGCAAAGT
64C8	1358	CDRL2-47	AAGGGTTCTAACTGGGACTCA
64A6	1359	CDRL2-48	GGTACATCCACCAGGGCCACT
67F5	1360	CDRL2-49	GGTTCATCCAACAGGGCCATT
64B10	1361	CDRL2-50	GACAATGATAAGCGACCCTCA
63F9	1362	CDRL2-51	GCTTCATCCAGTTTGCAAAGT
67F6v2	1363	CDRL2-52	ACGCTTTCCTTTCGGGCCTCT
50D4	1364	CDRL2-53	GCTGCATCCACTTTGCTATCA
68A10v3	1892	CDRL2-54	CAAGATAGCGAGCGGCCCTCA
48C9	1365	CDRL3-1	CAACAGAGTGACAGTATCCCTCGGACG
49A12
51E2
48F3	1366	CDRL3-2	CAACAGAGTTACAGTGCTACATTCACT
48F8	1367	CDRL3-3	CATCAGAGTAGTGATTTACCGCTCACT
53B9
56B4
57E7
57F11
48H11	1368	CDRL3-4	CAACAGAGTTACAATACCCCGTGCAGT
49A10	1369	CDRL3-5	ATGCAACGTATAGAGTTTCCGATCACC
48D4
67F6v2
49C8	1370	CDRL3-6	CAACAATATGATAATCTCCCATTCACT
52H1
67C10
67F6v1
49G2	1371	CDRL3-7	ATGCAACATATAGAATTTCCTTCGACC
50C12
55G11
49G3	1372	CDRL3-8	CACCAGTATGATGATCTCCCGCTCACT CAACAGTATGACAATTTACCGCTCACC
49H12	1373	CDRL3-9
54A1
55G9
51A8	1374	CDRL3-10	CAGTCTTATGATCGCAACAATCATGTGG TT
51C10.1	1375	CDRL3-11	TACTCAACAGACAGCAGTGTTAATCATG TGGTA
51C10.2	1376	CDRL3-12	CAGGCGTGGGATAGTAGTACTGCGGTA
51E5	1377	CDRL3-13	CTACAACATAGTAGTTACCCGCTCACT
51G2	1378	CDRL3-14
56A7
56E4
59A10
49H4
59C9
58A5
57A4
57F9
52A8	1379	CDRL3-15	CAGCAGAGTTACAGTACCCCGCTCACT
65B1			CAGCAGAGTTACAGTACCCCGCTCACT
52B8	1380	CDRL3-16	CAGCAGTATAATAACTGGCCGCTCACT
56G3.2			CAGCAGTATAATAACTGGCCGCTCACT
52C1	1381	CDRL3-17
64B10
68C8
52C5	1382	CDRL3-18	CAACAGAGTTCCAGTATCCCTTGGACG
55E4
49B11
50H10
53C1
51C1
60G5.1
52F8	1383	CDRL3-19	ATGCAAGCTCTACAAACTCCATTCACT
52H2	1384	CDRL3-20	CAGCAGTATGGTAGTTCACCTCGCAGT
53F6	1385	CDRL3-21	ATGCAAGGTCTACAAACTCCTCCCACT
53H5.2	1386	CDRL3-22	CTACAGCATAAGAGTTACCCATTCACT
53H5.3	1387	CDRL3-23	CAGCAGTTTAGTAACTCAATCACC
54H10.1	1388	CDRL3-24	CAGCAGTATGGTAGCTCACGGACG
55D1
48H3
53C11
55D3	1389	CDRL3-25	CAACAGTATAATATTTACCCTCGGACG
55E9	1390	CDRL3-26	ATGCAAGCTCTACAAACTCTCATCACC
55G5	1391	CDRL3-27	CAGGCGTGGGACAGCGGCACTGTGGTA
56C11	1392	CDRL3-28
56E7	1393	CDRL3-29	CAACAATATGCTATTCTCCCATTCACT
56G1	1394	CDRL3-30	CAACAGAGTTCCACTATCCCTTGGACG
56G3.3	1395	CDRL3-31	CAGCAATATGGTAGATCACTATTCACT
55B10
61H5
52B9
57B12	1396	CDRL3-32	CAACAATATAATACTTACCCTCGGACG
57D9	1397	CDRL3-33	CATCAGTATGGTACCTCACCGTGCAGT
59D10	1398	CDRL3-34
v1
59D10	1399	CDRL3-35
v2
59G10.2	1400	CDRL3-36	CAGGCGTGGGACAGCGCCACTGTGATT
59G10.3	1401	CDRL3-37
60D7	1402	CDRL3-38	ATGCAACGTATAGAGTTTCCGCTCACT
50G1			ATGCAACGTATAGAGTTTCCGCTCACT
60F9	1403	CDRL3-39
48B4
52D6
61G5
60G5.2	1404	CDRL3-40	CAGGCGTGGGACAGCAGCACTTGGGTG
63G8v1	1405	CDRL3-41	CTCCAGCATAATAGTTACCCTCTCACT
63G8v2
64A8
67B4
68D3
64E6	1406	CDRL3-42	CAGCAGTTTGGAAGCTCACTCACT
65E8
65F11
67G7
63H11
63F5
65C1
66F6
63B6	1407	CDRL3-43	CAGCAGTTTGGTAGGTCATTCACT
64D4			CAGCAGTTTGGTAGGTCATTCACT
65C3	1408	CDRL3-44	CAGCAGTATAATAACTGGCCGTGGACG
68D5			CAGCAGTATAATAACTGGCCGTGGACG
63E6	1409	CDRL3-45	CAACAGAGTTACAGTACCTCGCTCACT
66F7			CAACAGAGTTACAGTACCTCGCTCACT
64H5	1410	CDRL3-46	CAGGTGTGGGACAGCAGTAGTGTGGTA
65G4			CAGGTGTGGGACAGCAGTAGTGTGGTA
67G10	1411	CDRL3-47
v1
67G10	1412	CDRL3-48	CAGGCGTGGGACAGCACCACTGTGGTA
v2
63A10v2
63A10v3
64A7	1413	CDRL3-49
66B4	1414	CDRL3-50	CAACAGGCTAACAGTTTCCCTCCGACG
66G2	1415	CDRL3-51	CTACAACTTAATGGTTACCCTCTCACT
68G5	1416	CDRL3-52	CAGTTGTGGGACAGCAGCACTGTGGTT
66D4	1417	CDRL3-53	CAACAGAGTTACAGTTCCCCGCTCACT
54H10.3			CAACAGAGTTACAGTTCCCCGCTCACT
55A7	1418	CDRL3-54	CAACAGACTTACAGTGCCCCATTCACT
67A4	1419	CDRL3-55
65B4
63A10	1420	CDRL3-56
65H11	1421	CDRL3-57
64C8	1422	CDRL3-58
65E3	1423	CDRL3-59	CAGGTGTGGGACAGCAGCACTGTGGTC
67G8			CAGGTGTGGGACAGCAGCACTGTGGTC
65D4	1424	CDRL3-60	CAGGTGTGGGACAGCAACCCTGTGGTA
65D1	1425	CDRL3-61	CAGGCGTGGGACAGCAGGGTA
65B7v1	1426	CDRL3-62	CAGCAGTATGGTAGCTCGTGCAGT
64A6	1427	CDRL3-63	CAGCAATATAATACCTGGCCGTGGACG
65F9			CAGCAATATAATACCTGGCCGTGGACG
67F5	1428	CDRL3-64	CAGCAGTATGAAATTTGGCCGTGGACG
55E6	1429	CDRL3-65	CAGCAGTATGGTAGTTCACCGTGGACG
67A5	1430	CDRL3-66	ATGCAACGTCTAGAGTTTCCTATTACC
58C2			ATGCAACGTCTAGAGTTTCCTATTACC
61E1	1431	CDRL3-67	CAACAGAGTTTCAGTACCCCGCTCACT
64H6	1432	CDRL3-68	CAGGTGTGGGACAGCAGTCCTGTGGTA
63F9	1433	CDRL3-69	CTACAGCGTAATAGTTACCCGCTCACT
53C3.2	1434	CDRL3-70	CACCAGTATACTAACTGGCCTCGGACG
48G4	1435	CDRL3-71	CAGCAGTATGGTACCTCACCATTTACT
53C3.1			CAGCAGTATGGTACCTCACCATTTACT
50G5 v1	1436	CDRL3-72	CTACAGCATAATAGTTACCCTCGGACG
64B10v2	1893	CDRL3-73
50D4	1437	CDRL3-74	CAAAAGTATTACAGTGCCCCTTTCACT
50G5 v2	1438	CDRL3-75	ATGGAAGGTACACACTGGCCTCGGGAC
63G8v3	1894	CDRL3-76	CTCCAACATAATACTTACCCTCTCACT
65B7v2	1895	CDRL3-77
65H11v2	1896	CDRL3-78	CAGGCGTGGGACAGCATCACTGTGGTA
63A10v1	1897	CDRL3-79

The structure and properties of CDRs within a naturally occurring antibody has been described, supra. Briefly, in a traditional antibody, the CDRs are embedded within a framework in the heavy and light chain variable region where they constitute the regions responsible for antigen binding and recognition. A variable region comprises at least three heavy or light chain CDRs, see, e.g., Kabat et al., (1991) "Sequences of Proteins of Immunological Interest", 5th Ed., US Dept. of Health and Human Services, PHS, NIH, NIH Publication no. 91-3242; see also Chothia and Lesk, (1987) J. Mol. Biol. 196:901-917; Chothia et al., (1989) Nature 342: 877-883), within a framework region (designated framework regions 1-4, FR1, FR2, FR3, and FR4, by Kabat et al., (1991); see also Chothia and Lesk, (1987) supra). The CDRs provided herein, however, can not only be used to define the antigen binding domain of a traditional antibody structure, but can be embedded in a variety of other polypeptide structures, as described herein.

In one aspect, the CDRs provided are (a) a CDRH selected from the group consisting of (i) a CDRH1 selected from the group consisting of SEQ ID NOS 603-655; (ii) a CDRH2 selected from the group consisting of SEQ ID NOS 656-732; (iii) a CDRH3 selected from the group consisting of SEQ ID NOS 733-813; and (iv) a CDRH of (i), (ii) and (iii) that contains one or more amino acid substitutions, deletions or insertions of no more than five, four, three, two, or one amino acids; (B) a CDRL selected from the group consisting of (i) a CDRL1 selected from the group consisting of SEQ ID NOS 814-893; (ii) a CDRL2 selected from the group consisting of SEQ ID NOS 894-946; (iii) a CDRL3 selected from the group consisting of SEQ ID NOS 947-1020; and (iv) a CDRL of (i), (ii) and (iii) that contains one or more amino acid substitutions, deletions or insertions of no more than 1, 2, 3, 4, or 5 amino acids amino acids.

In another aspect, an antigen binding protein comprises 1, 2, 3, 4, 5, or 6 variant forms of the CDRs listed in Tables 3A and 3B, infra, each having at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% sequence identity to a CDR sequence listed in Tables 3A and 3B,infra. Some antigen binding proteins comprise 1, 2, 3, 4, 5, or 6 of the CDRs listed in Tables 3A and 3B, infra, each differing by no more than 1, 2, 3, 4 or 5 amino acids from the CDRs listed in these tables.

In still another aspect, an antigen binding protein includes the following associations of CDRL1, CDRL2 and CDRL3, presented for convenience in tabular form and in reference to the clone source of the association:



63G8	CDRL1-13	CDRL2-29	CDRL3-41
64A8	CDRL1-13	CDRL2-29	CDRL3-41
67B4	CDRL1-13	CDRL2-29	CDRL3-41
68D3	CDRL1-13	CDRL2-29	CDRL3-41
64E6	CDRL1-46	CDRL2-34	CDRL3-42
65E8	CDRL1-46	CDRL2-34	CDRL3-42
65F11	CDRL1-46	CDRL2-34	CDRL3-42
67G7	CDRL1-46	CDRL2-34	CDRL3-42
63B6	CDRL1-47	CDRL2-3	CDRL3-43
64D4	CDRL1-47	CDRL2-3	CDRL3-43
65C3	CDRL1-48	CDRL2-36	CDRL3-44
68D5	CDRL1-48	CDRL2-36	CDRL3-44
63E6	CDRL1-49	CDRL2-14	CDRL3-45
66F7	CDRL1-50	CDRL2-14	CDRL3-45
64H5	CDRL1-51	CDRL2-37	CDRL3-46
65G4	CDRL1-51	CDRL2-37	CDRL3-46
67G10v1	CDRL1-52	CDRL2-38	CDRL3-47
67G10v2	CDRL1-53	CDRL2-39	CDRL3-48
66B4	CDRL1-57	CDRL2-40	CDRL3-50
66G2	CDRL1-22	CDRL2-41	CDRL3-51
68G5	CDRL1-59	CDRL2-42	CDRL3-52
63F5	CDRL1-54	CDRL2-20	CDRL3-42
66F6	CDRL1-46	CDRL2-35	CDRL3-42
65C1	CDRL1-56	CDRL2-35	CDRL3-42
64A7	CDRL1-55	CDRL2-20	CDRL3-49
66D4	CDRL1-60	CDRL2-14	CDRL3-53
65B1	CDRL1-61	CDRL2-43	CDRL3-15
67A4	CDRL1-62	CDRL2-25	CDRL3-55
65B4	CDRL1-63	CDRL2-25	CDRL3-55
63A10	CDRL1-52	CDRL2-45	CDRL3-56
65H11	CDRL1-65	CDRL2-38	CDRL3-57
64C8	CDRL1-66	CDRL2-47	CDRL3-58
65E3	CDRL1-51	CDRL2-42	CDRL3-59
65D4	CDRL1-67	CDRL2-42	CDRL3-60
65D1	CDRL1-39	CDRL2-32	CDRL3-61
67G8	CDRL1-69	CDRL2-37	CDRL3-59
65B7	CDRL1-70	CDRL2-20	CDRL3-62
64A6	CDRL1-71	CDRL2-48	CDRL3-63
65F9	CDRL1-72	CDRL2-15	CDRL3-63
67F5	CDRL1-72	CDRL2-49	CDRL3-64
64B10	CDRL1-73	CDRL2-50	CDRL3-17
68C8	CDRL1-74	CDRL2-16	CDRL3-17
67A5	CDRL1-75	CDRL2-5	CDRL3-66
67C10	CDRL1-75	CDRL2-5	CDRL3-5
64H6	CDRL1-51	CDRL2-37	CDRL3-68
63F9	CDRL1-76	CDRL2-51	CDRL3-69
67F6	CDRL1-77	CDRL2-52	CDRL3-5
48H11	CDRL1-4	CDRL2-4	CDRL3-4
52A8	CDRL1-15	CDRL2-14	CDRL3-15
52F8	CDRL1-19	CDRL2-17	CDRL3-19
49H12	CDRL1-9	CDRL2-8	CDRL3-9
54A1	CDRL1-24	CDRL2-6	CDRL3-9
55G9	CDRL1-24	CDRL2-6	CDRL3-9
49C8	CDRL1-6	CDRL2-6	CDRL3-6
52H1	CDRL1-6	CDRL2-6	CDRL3-6
60G5.2	CDRL1-44	CDRL2-32	CDRL3-40
49G3	CDRL1-8	CDRL2-7	CDRL3-8
59A10	CDRL1-14	CDRL2-28	CDRL3-14
49H4	CDRL1-14	CDRL2-28	CDRL3-14
48F8	CDRL1-3	CDRL2-3	CDRL3-3
53B9	CDRL1-3	CDRL2-3	CDRL3-3
56B4	CDRL1-3	CDRL2-3	CDRL3-3
57E7	CDRL1-3	CDRL2-3	CDRL3-3
57F11	CDRL1-3	CDRL2-3	CDRL3-3
59C9	CDRL1-37	CDRL2-29	CDRL3-14
58A5	CDRL1-37	CDRL2-29	CDRL3-14
57A4	CDRL1-37	CDRL2-29	CDRL3-14
57F9	CDRL1-37	CDRL2-29	CDRL3-14
51G2	CDRL1-14	CDRL2-13	CDRL3-14
56A7	CDRL1-29	CDRL2-24	CDRL3-14
56E4	CDRL1-29	CDRL2-24	CDRL3-14
54H10.1	CDRL1-25	CDRL2-20	CDRL3-24
55D1	CDRL1-25	CDRL2-20	CDRL3-24
48H3	CDRL1-25	CDRL2-20	CDRL3-24
53C11	CDRL1-25	CDRL2-20	CDRL3-24
59G10.3	CDRL1-41	CDRL2-16	CDRL3-37
51C10.1	CDRL1-12	CDRL2-10	CDRL3-11
59D10 v1	CDRL1-38	CDRL2-10	CDRL3-34
59D10 v2	CDRL1-39	CDRL2-30	CDRL3-35
60F9	CDRL1-43	CDRL2-31	CDRL3-39
48B4	CDRL1-43	CDRL2-31	CDRL3-39
52D6	CDRL1-43	CDRL2-31	CDRL3-39
61G5	CDRL1-45	CDRL2-33	CDRL3-39
59G10.2	CDRL1-40	CDRL2-11	CDRL3-36
51A8	CDRL1-10	CDRL2-9	CDRL3-10
53H5.2	CDRL1-22	CDRL2-14	CDRL3-22
53F6	CDRL1-21	CDRL2-19	CDRL3-21
56C11	CDRL1-30	CDRL2-25	CDRL3-28
49A10	CDRL1-5	CDRL2-5	CDRL3-5
48D4	CDRL1-5	CDRL2-5	CDRL3-5
49G2	CDRL1-7	CDRL2-5	CDRL3-7
50C12	CDRL1-7	CDRL2-5	CDRL3-7
55G11	CDRL1-7	CDRL2-5	CDRL3-7
52C1	CDRL1-17	CDRL2-16	CDRL3-17
55E9	CDRL1-27	CDRL2-17	CDRL3-26
60D7	CDRL1-1	CDRL2-5	CDRL3-38
51C10.2	CDRL1-12	CDRL2-11	CDRL3-12
55D3	CDRL1-26	CDRL2-14	CDRL3-25
57B12	CDRL1-34	CDRL2-14	CDRL3-32
52C5	CDRL1-18	CDRL2-14	CDRL3-18
55E4	CDRL1-18	CDRL2-21	CDRL3-18
49B11	CDRL1-18	CDRL2-21	CDRL3-18
50H10	CDRL1-18	CDRL2-21	CDRL3-18
53C1	CDRL1-18	CDRL2-21	CDRL3-18
56G1	CDRL1-18	CDRL2-14	CDRL3-30
48F3	CDRL1-2	CDRL2-2	CDRL3-2
48C9	CDRL1-1	CDRL2-1	CDRL3-1
49A12	CDRL1-1	CDRL2-1	CDRL3-1
51E2	CDRL1-1	CDRL2-1	CDRL3-1
51E5	CDRL1-13	CDRL2-12	CDRL3-13
53H5.3	CDRL1-23	CDRL2-15	CDRL3-23
56G3.3	CDRL1-33	CDRL2-27	CDRL3-31
55B10	CDRL1-33	CDRL2-27	CDRL3-31
52B8	CDRL1-16	CDRL2-15	CDRL3-16
55G5	CDRL1-28	CDRL2-23	CDRL3-27
52H2	CDRL1-20	CDRL2-18	CDRL3-20
56G3.2	CDRL1-32	CDRL2-26	CDRL3-16
56E7	CDRL1-31	CDRL2-7	CDRL3-29
57D9	CDRL1-35	CDRL2-20	CDRL3-33
61H5	CDRL1-33	CDRL2-20	CDRL3-31
52B9	CDRL1-33	CDRL2-20	CDRL3-31
48G4	CDRL1-79	CDRL2-35	CDRL3-71
53C3.1	CDRL1-79	CDRL2-35	CDRL3-71
50G1	CDRL1-7	CDRL2-5	CDRL3-38
58C2	CDRL1-81	CDRL2-5	CDRL3-66
60G5.1	CDRL1-18	CDRL2-14	CDRL3-18
54H10.3	CDRL1-42	CDRL2-46	CDRL3-53
50G5 v1	CDRL1-22	CDRL2-14	CDRL3-72
50G5 v2	CDRL1-78	CDRL2-22	CDRL3-75
51C1	CDRL1-18	CDRL2-14	CDRL3-18
53C3.2	CDRL1-36	CDRL2-44	CDRL3-70
50D4	CDRL1-26	CDRL2-53	CDRL3-74
55A7	CDRL1-58	CDRL2-14	CDRL3-54
55E6	CDRL1-64	CDRL2-20	CDRL3-65
61E1	CDRL1-68	CDRL2-14	CDRL3-67
63H11	CDRL1-46	CDRL2-34	CDRL3-42

In an additional aspect, an antigen binding protein includes the following associations of CDRH1, CDRH2 and CDRH3, presented for convenience in tablular form and in reference to the clone source of the association:



63G8	CDRH1-34	CDRH2-12	CDRH3-50
64A8	CDRH1-34	CDRH2-12	CDRH3-50
67B4	CDRH1-34	CDRH2-12	CDRH3-50
68D3	CDRH1-34	CDRH2-12	CDRH3-50
64E6	CDRH1-35	CDRH2-44	CDRH3-51
65E8	CDRH1-35	CDRH2-44	CDRH3-51
65F11	CDRH1-35	CDRH2-44	CDRH3-51
67G7	CDRH1-35	CDRH2-44	CDRH3-51
63B6	CDRH1-36	CDRH2-45	CDRH3-52
64D4	CDRH1-36	CDRH2-45	CDRH3-52
65C3	CDRH1-24	CDRH2-46	CDRH3-53
68D5	CDRH1-24	CDRH2-46	CDRH3-53
63E6	CDRH1-37	CDRH2-47	CDRH3-54
66F7	CDRH1-37	CDRH2-47	CDRH3-54
64H5	CDRH1-12	CDRH2-48	CDRH3-55
65G4	CDRH1-12	CDRH2-48	CDRH3-55
67G10v1	CDRH1-38	CDRH2-49	CDRH3-56
67G10v2	CDRH1-38	CDRH2-49	CDRH3-56
66B4	CDRH1-15	CDRH2-53	CDRH3-59
66G2	CDRH1-12	CDRH2-54	CDRH3-50
68G5	CDRH1-12	CDRH2-55	CDRH3-60
63F5	CDRH1-35	CDRH2-50	CDRH3-51
66F6	CDRH1-35	CDRH2-34	CDRH3-51
65C1	CDRH1-35	CDRH2-52	CDRH3-58
64A7	CDRH1-40	CDRH2-51	CDRH3-57
66D4	CDRH1-43	CDRH2-56	CDRH3-61
65B1	CDRH1-44	CDRH2-57	CDRH3-62
67A4	CDRH1-45	CDRH2-58	CDRH3-63
65B4	CDRH1-46	CDRH2-59	CDRH3-64
63A10	CDRH1-38	CDRH2-60	CDRH3-56
65H11	CDRH1-38	CDRH2-61	CDRH3-56
64C8	CDRH1-12	CDRH2-62	CDRH3-65
65E3	CDRH1-47	CDRH2-63	CDRH3-66
65D4	CDRH1-48	CDRH2-22	CDRH3-67
65D1	CDRH1-49	CDRH2-64	CDRH3-68
67G8	CDRH1-12	CDRH2-65	CDRH3-69
65B7	CDRH1-50	CDRH2-52	CDRH3-70
64A6	CDRH1-14	CDRH2-66	CDRH3-71
65F9	CDRH1-36	CDRH2-34	CDRH3-72
67F5	CDRH1-24	CDRH2-67	CDRH3-53
64B10	CDRH1-36	CDRH2-68	CDRH3-73
68C8	CDRH1-51	CDRH2-69	CDRH3-74
67A5	CDRH1-25	CDRH2-31	CDRH3-75
67C10	CDRH1-25	CDRH2-31	CDRH3-76
64H6	CDRH1-25	CDRH2-70	CDRH3-77
63F9	CDRH1-52	CDRH2-71	CDRH3-78
67F6	CDRH1-53	CDRH2-31	CDRH3-79
48H11	CDRH1-4	CDRH2-4	CDRH3-4
52A8	CDRH1-15	CDRH2-17	CDRH3-17
52F8	CDRH1-17	CDRH2-20	CDRH3-21
49H12	CDRH1-10	CDRH2-10	CDRH3-10
54A1	CDRH1-10	CDRH2-25	CDRH3-10
55G9	CDRH1-10	CDRH2-25	CDRH3-10
49C8	CDRH1-7	CDRH2-7	CDRH3-7
52H1	CDRH1-7	CDRH2-7	CDRH3-7
60G5.2	CDRH1-33	CDRH2-42	CDRH3-48
49G3	CDRH1-9	CDRH2-9	CDRH3-9
59A10	CDRH1-30	CDRH2-37	CDRH3-41
49H4	CDRH1-30	CDRH2-37	CDRH3-41
48F8	CDRH1-3	CDRH2-3	CDRH3-3
53B9	CDRH1-3	CDRH2-3	CDRH3-3
56B4	CDRH1-3	CDRH2-3	CDRH3-3
57E7	CDRH1-3	CDRH2-3	CDRH3-3
57F11	CDRH1-3	CDRH2-3	CDRH3-3
59C9	CDRH1-31	CDRH2-38	CDRH3-42
58A5	CDRH1-31	CDRH2-38	CDRH3-42
57A4	CDRH1-31	CDRH2-38	CDRH3-42
57F9	CDRH1-31	CDRH2-38	CDRH3-42
51G2	CDRH1-3	CDRH2-16	CDRH3-16
56A7	CDRH1-3	CDRH2-16	CDRH3-32
56E4	CDRH1-3	CDRH2-16	CDRH3-32
54H10.1	CDRH1-21	CDRH2-26	CDRH3-27
55D1	CDRH1-21	CDRH2-26	CDRH3-27
48H3	CDRH1-21	CDRH2-26	CDRH3-27
53C11	CDRH1-21	CDRH2-26	CDRH3-27
59G10.3	CDRH1-32	CDRH2-40	CDRH3-45
51C10.1	CDRH1-13	CDRH2-13	CDRH3-13
59D10 v1	CDRH1-13	CDRH2-13	CDRH3-13
59D10 v2	CDRH1-13	CDRH2-13	CDRH3-13
60F9	CDRH1-21	CDRH2-41	CDRH3-47
48B4	CDRH1-21	CDRH2-41	CDRH3-47
52D6	CDRH1-21	CDRH2-41	CDRH3-47
61G5	CDRH1-21	CDRH2-43	CDRH3-49
59G10.2	CDRH1-6	CDRH2-39	CDRH3-44
51A8	CDRH1-12	CDRH2-12	CDRH3-12
53H5.2	CDRH1-12	CDRH2-23	CDRH3-24
53F6	CDRH1-19	CDRH2-22	CDRH3-23
56C11	CDRH1-12	CDRH2-30	CDRH3-33
49A10	CDRH1-6	CDRH2-6	CDRH3-6
48D4	CDRH1-6	CDRH2-6	CDRH3-6
49G2	CDRH1-8	CDRH2-8	CDRH3-8
50C12	CDRH1-8	CDRH2-8	CDRH3-8
55G11	CDRH1-8	CDRH2-8	CDRH3-8
52C1	CDRH1-12	CDRH2-19	CDRH3-19
55E9	CDRH1-23	CDRH2-28	CDRH3-30
60D7	CDRH1-12	CDRH2-22	CDRH3-46
51C10.2	CDRH1-14	CDRH2-14	CDRH3-14
55D3	CDRH1-22	CDRH2-27	CDRH3-28
57B12	CDRH1-28	CDRH2-34	CDRH3-28
52C5	CDRH1-2	CDRH2-1	CDRH3-20
60G5.1	CDRH1-2	CDRH2-1	CDRH3-20
55E4	CDRH1-2	CDRH2-1	CDRH3-20
49B11	CDRH1-2	CDRH2-1	CDRH3-20
50H10	CDRH1-2	CDRH2-1	CDRH3-20
53C1	CDRH1-2	CDRH2-1	CDRH3-20
56G1	CDRH1-2	CDRH2-1	CDRH3-20
48F3	CDRH1-2	CDRH2-2	CDRH3-2
48C9	CDRH1-1	CDRH2-1	CDRH3-1
49A12	CDRH1-1	CDRH2-1	CDRH3-1
51E2	CDRH1-1	CDRH2-1	CDRH3-1
51E5	CDRH1-2	CDRH2-15	CDRH3-15
53H5.3	CDRH1-20	CDRH2-24	CDRH3-25
56G3.3	CDRH1-27	CDRH2-33	CDRH3-37
55B10	CDRH1-27	CDRH2-33	CDRH3-37
52B8	CDRH1-16	CDRH2-18	CDRH3-18
55G5	CDRH1-24	CDRH2-29	CDRH3-31
52H2	CDRH1-18	CDRH2-21	CDRH3-22
56G3.2	CDRH1-26	CDRH2-32	CDRH3-36
56E7	CDRH1-25	CDRH2-31	CDRH3-34
57D9	CDRH1-29	CDRH2-35	CDRH3-39
48G4	CDRH1-5	CDRH2-5	CDRH3-5
53C3.1	CDRH1-5	CDRH2-5	CDRH3-5
50G1	CDRH1-11	CDRH2-11	CDRH3-11
58C2	CDRH1-6	CDRH2-36	CDRH3-40
63H11	CDRH1-35	CDRH2-34	CDRH3-51
61H5	CDRH1-27	CDRH2-72	CDRH3-37
52B9	CDRH1-27	CDRH2-72	CDRH3-37
54H10.3	CDRH1-43	CDRH2-74	CDRH3-81
50G5 v1	CDRH1-37	CDRH2-73	CDRH3-35
50G5 v2	CDRH1-37	CDRH2-73	CDRH3-35
51C1	CDRH1-2	CDRH2-1	CDRH3-20
53C3.2	CDRH1-39	CDRH2-77	CDRH3-43
50D4	CDRH1-41	CDRH2-75	CDRH3-80
55A7	CDRH1-24	CDRH2-18	CDRH3-38
55E6	CDRH1-3	CDRH2-76	CDRH3-29
61E1	CDRH1-42	CDRH2-35	CDRH3-26

In an additional aspect, an antigen binding protein includes the following associations of CDRH1, CDRH2, CDRH3, CDRL1, CDRL2 and CDRL3 presented for convenience in tablular form and in reference to the clone source of the association:



63G8	CDRL1-13	CDRL2-29	CDRL3-41	CDRH1-34	CDRH2-12	CDRH3-50
64A8	CDRL1-13	CDRL2-29	CDRL3-41	CDRH1-34	CDRH2-12	CDRH3-50
67B4	CDRL1-13	CDRL2-29	CDRL3-41	CDRH1-34	CDRH2-12	CDRH3-50
68D3	CDRL1-13	CDRL2-29	CDRL3-41	CDRH1-34	CDRH2-12	CDRH3-50
64E6	CDRL1-46	CDRL2-34	CDRL3-42	CDRH1-35	CDRH2-44	CDRH3-51
65E8	CDRL1-46	CDRL2-34	CDRL3-42	CDRH1-35	CDRH2-44	CDRH3-51
65F11	CDRL1-46	CDRL2-34	CDRL3-42	CDRH1-35	CDRH2-44	CDRH3-51
67G7	CDRL1-46	CDRL2-34	CDRL3-42	CDRH1-35	CDRH2-44	CDRH3-51
63B6	CDRL1-47	CDRL2-3	CDRL3-43	CDRH1-36	CDRH2-45	CDRH3-52
64D4	CDRL1-47	CDRL2-3	CDRL3-43	CDRH1-36	CDRH2-45	CDRH3-52
65C3	CDRL1-48	CDRL2-36	CDRL3-44	CDRH1-24	CDRH2-46	CDRH3-53
68D5	CDRL1-48	CDRL2-36	CDRL3-44	CDRH1-24	CDRH2-46	CDRH3-53
63E6	CDRL1-49	CDRL2-14	CDRL3-45	CDRH1-37	CDRH2-47	CDRH3-54
66F7	CDRL1-50	CDRL2-14	CDRL3-45	CDRH1-37	CDRH2-47	CDRH3-54
64H5	CDRL1-51	CDRL2-37	CDRL3-46	CDRH1-12	CDRH2-48	CDRH3-55
65G4	CDRL1-51	CDRL2-37	CDRL3-46	CDRH1-12	CDRH2-48	CDRH3-55
67G10v1	CDRL1-52	CDRL2-38	CDRL3-47	CDRH1-38	CDRH2-49	CDRH3-56
67G10v2	CDRL1-53	CDRL2-39	CDRL3-48	CDRH1-38	CDRH2-49	CDRH3-56
66B4	CDRL1-57	CDRL2-40	CDRL3-50	CDRH1-15	CDRH2-53	CDRH3-59
66G2	CDRL1-22	CDRL2-41	CDRL3-51	CDRH1-12	CDRH2-54	CDRH3-50
68G5	CDRL1-59	CDRL2-42	CDRL3-52	CDRH1-12	CDRH2-55	CDRH3-60
63F5	CDRL1-54	CDRL2-20	CDRL3-42	CDRH1-35	CDRH2-50	CDRH3-51
66F6	CDRL1-46	CDRL2-35	CDRL3-42	CDRH1-35	CDRH2-34	CDRH3-51
65C1	CDRL1-56	CDRL2-35	CDRL3-42	CDRH1-35	CDRH2-52	CDRH3-58
64A7	CDRL1-55	CDRL2-20	CDRL3-49	CDRH1-40	CDRH2-51	CDRH3-57
66D4	CDRL1-60	CDRL2-14	CDRL3-53	CDRH1-43	CDRH2-56	CDRH3-61
65B1	CDRL1-61	CDRL2-43	CDRL3-15	CDRH1-44	CDRH2-57	CDRH3-62
67A4	CDRL1-62	CDRL2-25	CDRL3-55	CDRH1-45	CDRH2-58	CDRH3-63
65B4	CDRL1-63	CDRL2-25	CDRL3-55	CDRH1-46	CDRH2-59	CDRH3-64
63A10	CDRL1-52	CDRL2-45	CDRL3-56	CDRH1-38	CDRH2-60	CDRH3-56
65H11	CDRL1-65	CDRL2-38	CDRL3-57	CDRH1-38	CDRH2-61	CDRH3-56
64C8	CDRL1-66	CDRL2-47	CDRL3-58	CDRH1-12	CDRH2-62	CDRH3-65
65E3	CDRL1-51	CDRL2-42	CDRL3-59	CDRH1-47	CDRH2-63	CDRH3-66
65D4	CDRL1-67	CDRL2-42	CDRL3-60	CDRH1-48	CDRH2-22	CDRH3-67
65D1	CDRL1-39	CDRL2-32	CDRL3-61	CDRH1-49	CDRH2-64	CDRH3-68
67G8	CDRL1-69	CDRL2-37	CDRL3-59	CDRH1-12	CDRH2-65	CDRH3-69
65B7	CDRL1-70	CDRL2-20	CDRL3-62	CDRH1-50	CDRH2-52	CDRH3-70
64A6	CDRL1-71	CDRL2-48	CDRL3-63	CDRH1-14	CDRH2-66	CDRH3-71
65F9	CDRL1-72	CDRL2-15	CDRL3-63	CDRH1-36	CDRH2-34	CDRH3-72
67F5	CDRL1-72	CDRL2-49	CDRL3-64	CDRH1-24	CDRH2-67	CDRH3-53
64B10	CDRL1-73	CDRL2-50	CDRL3-17	CDRH1-36	CDRH2-68	CDRH3-73
68C8	CDRL1-74	CDRL2-16	CDRL3-17	CDRH1-51	CDRH2-69	CDRH3-74
67A5	CDRL1-75	CDRL2-5	CDRL3-66	CDRH1-25	CDRH2-31	CDRH3-75
67C10	CDRL1-75	CDRL2-5	CDRL3-5	CDRH1-25	CDRH2-31	CDRH3-76
64H6	CDRL1-51	CDRL2-37	CDRL3-68	CDRH1-25	CDRH2-70	CDRH3-77
63F9	CDRL1-76	CDRL2-51	CDRL3-69	CDRH1-52	CDRH2-71	CDRH3-78
67F6	CDRL1-77	CDRL2-52	CDRL3-5	CDRH1-53	CDRH2-31	CDRH3-79
48H11	CDRL1-4	CDRL2-4	CDRL3-4	CDRH1-4	CDRH2-4	CDRH3-4
52A8	CDRL1-15	CDRL2-14	CDRL3-15	CDRH1-15	CDRH2-17	CDRH3-17
52F8	CDRL1-19	CDRL2-17	CDRL3-19	CDRH1-17	CDRH2-20	CDRH3-21
49H12	CDRL1-9	CDRL2-8	CDRL3-9	CDRH1-10	CDRH2-10	CDRH3-10
54A1	CDRL1-24	CDRL2-6	CDRL3-9	CDRH1-10	CDRH2-25	CDRH3-10
55G9	CDRL1-24	CDRL2-6	CDRL3-9	CDRH1-10	CDRH2-25	CDRH3-10
49C8	CDRL1-6	CDRL2-6	CDRL3-6	CDRH1-7	CDRH2-7	CDRH3-7
52H1	CDRL1-6	CDRL2-6	CDRL3-6	CDRH1-7	CDRH2-7	CDRH3-7
60G5.2	CDRL1-44	CDRL2-32	CDRL3-40	CDRH1-33	CDRH2-42	CDRH3-48
49G3	CDRL1-8	CDRL2-7	CDRL3-8	CDRH1-9	CDRH2-9	CDRH3-9
59A10	CDRL1-14	CDRL2-28	CDRL3-14	CDRH1-30	CDRH2-37	CDRH3-41
49H4	CDRL1-14	CDRL2-28	CDRL3-14	CDRH1-30	CDRH2-37	CDRH3-41
48F8	CDRL1-3	CDRL2-3	CDRL3-3	CDRH1-3	CDRH2-3	CDRH3-3
53B9	CDRL1-3	CDRL2-3	CDRL3-3	CDRH1-3	CDRH2-3	CDRH3-3
56B4	CDRL1-3	CDRL2-3	CDRL3-3	CDRH1-3	CDRH2-3	CDRH3-3
57E7	CDRL1-3	CDRL2-3	CDRL3-3	CDRH1-3	CDRH2-3	CDRH3-3
57F11	CDRL1-3	CDRL2-3	CDRL3-3	CDRH1-3	CDRH2-3	CDRH3-3
59C9	CDRL1-37	CDRL2-29	CDRL3-14	CDRH1-31	CDRH2-38	CDRH3-42
58A5	CDRL1-37	CDRL2-29	CDRL3-14	CDRH1-31	CDRH2-38	CDRH3-42
57A4	CDRL1-37	CDRL2-29	CDRL3-14	CDRH1-31	CDRH2-38	CDRH3-42
57F9	CDRL1-37	CDRL2-29	CDRL3-14	CDRH1-31	CDRH2-38	CDRH3-42
51G2	CDRL1-14	CDRL2-13	CDRL3-14	CDRH1-3	CDRH2-16	CDRH3-16
56A7	CDRL1-29	CDRL2-24	CDRL3-14	CDRH1-3	CDRH2-16	CDRH3-32
56E4	CDRL1-29	CDRL2-24	CDRL3-14	CDRH1-3	CDRH2-16	CDRH3-32
54H10.1	CDRL1-25	CDRL2-20	CDRL3-24	CDRH1-21	CDRH2-26	CDRH3-27
55D1	CDRL1-25	CDRL2-20	CDRL3-24	CDRH1-21	CDRH2-26	CDRH3-27
48H3	CDRL1-25	CDRL2-20	CDRL3-24	CDRH1-21	CDRH2-26	CDRH3-27
53C11	CDRL1-25	CDRL2-20	CDRL3-24	CDRH1-21	CDRH2-26	CDRH3-27
59G10.3	CDRL1-41	CDRL2-16	CDRL3-37	CDRH1-32	CDRH2-40	CDRH3-45
51C10.1	CDRL1-12	CDRL2-10	CDRL3-11	CDRH1-13	CDRH2-13	CDRH3-13
59D10 v1	CDRL1-38	CDRL2-10	CDRL3-34	CDRH1-13	CDRH2-13	CDRH3-13
59D10 v2	CDRL1-39	CDRL2-30	CDRL3-35	CDRH1-13	CDRH2-13	CDRH3-13
60F9	CDRL1-43	CDRL2-31	CDRL3-39	CDRH1-21	CDRH2-41	CDRH3-47
48B4	CDRL1-43	CDRL2-31	CDRL3-39	CDRH1-21	CDRH2-41	CDRH3-47
52D6	CDRL1-43	CDRL2-31	CDRL3-39	CDRH1-21	CDRH2-41	CDRH3-47
61G5	CDRL1-45	CDRL2-33	CDRL3-39	CDRH1-21	CDRH2-43	CDRH3-49
59G10.2	CDRL1-40	CDRL2-11	CDRL3-36	CDRH1-6	CDRH2-39	CDRH3-44
51A8	CDRL1-10	CDRL2-9	CDRL3-10	CDRH1-12	CDRH2-12	CDRH3-12
53H5.2	CDRL1-22	CDRL2-14	CDRL3-22	CDRH1-12	CDRH2-23	CDRH3-24
53F6	CDRL1-21	CDRL2-19	CDRL3-21	CDRH1-19	CDRH2-22	CDRH3-23
56C11	CDRL1-30	CDRL2-25	CDRL3-28	CDRH1-12	CDRH2-30	CDRH3-33
49A10	CDRL1-5	CDRL2-5	CDRL3-5	CDRH1-6	CDRH2-6	CDRH3-6
48D4	CDRL1-5	CDRL2-5	CDRL3-5	CDRH1-6	CDRH2-6	CDRH3-6
49G2	CDRL1-7	CDRL2-5	CDRL3-7	CDRH1-8	CDRH2-8	CDRH3-8
50C12	CDRL1-7	CDRL2-5	CDRL3-7	CDRH1-8	CDRH2-8	CDRH3-8
55G11	CDRL1-7	CDRL2-5	CDRL3-7	CDRH1-8	CDRH2-8	CDRH3-8
52C1	CDRL1-17	CDRL2-16	CDRL3-17	CDRH1-12	CDRH2-19	CDRH3-19
55E9	CDRL1-27	CDRL2-17	CDRL3-26	CDRH1-23	CDRH2-28	CDRH3-30
60D7	CDRL1-1	CDRL2-5	CDRL3-38	CDRH1-12	CDRH2-22	CDRH3-46
51C10.2	CDRL1-12	CDRL2-11	CDRL3-12	CDRH1-14	CDRH2-14	CDRH3-14
55D3	CDRL1-26	CDRL2-14	CDRL3-25	CDRH1-22	CDRH2-27	CDRH3-28
57B12	CDRL1-34	CDRL2-14	CDRL3-32	CDRH1-28	CDRH2-34	CDRH3-28
52C5	CDRL1-18	CDRL2-14	CDRL3-18	CDRH1-2	CDRH2-1	CDRH3-20
60G5.1	CDRL1-18	CDRL2-14	CDRL3-18	CDRH1-2	CDRH2-1	CDRH3-20
55E4	CDRL1-18	CDRL2-21	CDRL3-18	CDRH1-2	CDRH2-1	CDRH3-20
49B11	CDRL1-18	CDRL2-21	CDRL3-18	CDRH1-2	CDRH2-1	CDRH3-20
50H10	CDRL1-18	CDRL2-21	CDRL3-18	CDRH1-2	CDRH2-1	CDRH3-20
53C1	CDRL1-18	CDRL2-21	CDRL3-18	CDRH1-2	CDRH2-1	CDRH3-20
56G1	CDRL1-18	CDRL2-14	CDRL3-30	CDRH1-2	CDRH2-1	CDRH3-20
48F3	CDRL1-2	CDRL2-2	CDRL3-2	CDRH1-2	CDRH2-2	CDRH3-2
48C9	CDRL1-1	CDRL2-1	CDRL3-1	CDRH1-1	CDRH2-1	CDRH3-1
49A12	CDRL1-1	CDRL2-1	CDRL3-1	CDRH1-1	CDRH2-1	CDRH3-1
51E2	CDRL1-1	CDRL2-1	CDRL3-1	CDRH1-1	CDRH2-1	CDRH3-1
51E5	CDRL1-13	CDRL2-12	CDRL3-13	CDRH1-2	CDRH2-15	CDRH3-15
53H5.3	CDRL1-23	CDRL2-15	CDRL3-23	CDRH1-20	CDRH2-24	CDRH3-25
56G3.3	CDRL1-33	CDRL2-27	CDRL3-31	CDRH1-27	CDRH2-33	CDRH3-37
55B10	CDRL1-33	CDRL2-27	CDRL3-31	CDRH1-27	CDRH2-33	CDRH3-37
52B8	CDRL1-16	CDRL2-15	CDRL3-16	CDRH1-16	CDRH2-18	CDRH3-18
55G5	CDRL1-28	CDRL2-23	CDRL3-27	CDRH1-24	CDRH2-29	CDRH3-31
52H2	CDRL1-20	CDRL2-18	CDRL3-20	CDRH1-18	CDRH2-21	CDRH3-22
56G3.2	CDRL1-32	CDRL2-26	CDRL3-16	CDRH1-26	CDRH2-32	CDRH3-36
56E7	CDRL1-31	CDRL2-7	CDRL3-29	CDRH1-25	CDRH2-31	CDRH3-34
57D9	CDRL1-35	CDRL2-20	CDRL3-33	CDRH1-29	CDRH2-35	CDRH3-39
61H5	CDRL1-33	CDRL2-20	CDRL3-31	CDRH1-27	CDRH2-72	CDRH3-37
52B9	CDRL1-33	CDRL2-20	CDRL3-31	CDRH1-27	CDRH2-72	CDRH3-37
48G4	CDRL1-79	CDRL2-35	CDRL3-71	CDRH1-5	CDRH2-5	CDRH3-5
53C3.1	CDRL1-79	CDRL2-35	CDRL3-71	CDRH1-5	CDRH2-5	CDRH3-5
50G1	CDRL1-7	CDRL2-5	CDRL3-38	CDRH1-11	CDRH2-11	CDRH3-11
58C2	CDRL1-81	CDRL2-5	CDRL3-66	CDRH1-6	CDRH2-36	CDRH3-40
54H10.3	CDRL1-42	CDRL2-46	CDRL3-53	CDRH1-43	CDRH2-74	CDRH3-81
50G5 v1	CDRL1-22	CDRL2-14	CDRL3-72	CDRH1-37	CDRH2-73	CDRH3-35
50G5 v2	CDRL1-78	CDRL2-22	CDRL3-75	CDRH1-37	CDRH2-73	CDRH3-35
51C1	CDRL1-18	CDRL2-14	CDRL3-18	CDRH1-2	CDRH2-1	CDRH3-20
53C3.2	CDRL1-36	CDRL2-44	CDRL3-70	CDRH1-39	CDRH2-77	CDRH3-43
50D4	CDRL1-26	CDRL2-53	CDRL3-74	CDRH1-41	CDRH2-75	CDRH3-80
55A7	CDRL1-58	CDRL2-14	CDRL3-54	CDRH1-24	CDRH2-18	CDRH3-38
55E6	CDRL1-64	CDRL2-20	CDRL3-65	CDRH1-3	CDRH2-76	CDRH3-29
61E1	CDRL1-68	CDRL2-14	CDRL3-67	CDRH1-42	CDRH2-35	CDRH3-26
63H11	CDRL1-46	CDRL2-34	CDRL3-42	CDRH1-35	CDRH2-34	CDRH3-51

Consensus Sequences

In yet another aspect, the CDRs disclosed herein include consensus sequences derived from groups of related monoclonal antibodies. As described herein, a "consensus sequence" refers to amino acid sequences having conserved amino acids common among a number of sequences and variable amino acids that vary within a given amino acid sequences. The CDR consensus sequences provided include CDRs corresponding to each of CDRH1, CDRH2, CDRH3, CDRL1, CDRL2 and CDRL3.

Consensus sequences were determined using standard analyses of the CDRs corresponding to the V_H and V_L of the disclosed antigen binding proteins shown in Tables 3A and 3B, some of which specifically bind to a complex comprising β-Klotho and at least one of (i) FGFR1c, (ii) FGFR2c and (iii) FGFR3c. The consensus sequences can be determined by keeping the CDRs contiguous within the same sequence corresponding to a V_H or V_L.

Light Chain CDR3

Group 1 QQFGSSLT (SEQ ID NO: 1439) Group 2

QQS	Y	S	T	S	LT (SEQ ID NO: 1440)
QQS	Y	S	S	P	LT (SEQ ID NO: 1441)
QQS	F	S	T	P	LT (SEQ ID NO: 1442)

wherein X₁ is Y or F; X₂ is T or S; and X₃ is P or S. Group 3

LQ	R	N	SYP	L	T (SEQ ID NO: 1444)
LQ	H	N	SYP	R	T (SEQIDNO: 1445)
LQ	H	S	SYP	L	T (SEQ ID NO: 1446)

wherein X₄ is H or R; X₅ is N or S; and X₆ is L or R. Group 4

MQR	I	EFP	L	T (SEQ ID NO: 1448)
MQR	I	EFP	I	T (SEQ ID NO: 1449)
MQR	L	EFP	I	T (SEQ ID NO: 1450)

wherein X₇ is I or L; and X₈ us I or L. Group 5

Q	V	WDS	N	P	VV (SEQ ID NO: 1452)
Q	L	WDS	S	T	VV (SEQ ID NO: 1453)
Q	V	WDS	S	S	VV (SEQ ID NO: 1454)
Q	V	WDS	S	P	VV (SEQ ID NO: 1455)
Q	V	WDS	S	T	VV (SEQ ID NO: 1456)

wherein X₉ is V or L; X₁₀ is S or N; and X₁₁ is T, P or S. Group 6

QQYN	N	WP	L	T (SEQ ID NO: 1458)
QQYN	N	WP	W	T (SEQ ID NO: 1459)
QQYN	T	WP	W	T (SEQ ID NO: 1460)

wherein X₁₂ is N or T; and X₁₃ is W or L. Group 7

QVWDSS	S	D	H	V	V	(SEQ ID NO: 1462)
QVWDSS	S	D	V	V	(SEQ ID NO: 1463)
QVWDSS	C	D	G	V	(SEQ ID NO: 1464)
QVWDSS	S	D	G	V	(SEQ ID NO: 1465)
(SEQ ID NO: 1466)

wherein X₁₄ is S or C; X₁₅ is H, V or G; and X₁₆ is V or absent. Group 8

QQSS	S	IPWT (SEQ ID NO: 1467)
QQSS	T	IPWT (SEQ ID NO: 1468)

wherein X₁₇ is S or T. Group 9 QQTNSFPPWT (SEQ ID NO: 1470) Group 10

GTWDSSLS	A	V	V (SEQ ID NO: 1471)
GTWDSSLS	V	V	V (SEQ ID NO: 1472)
GTWDSSLS	A	M	V (SEQ ID NO: 1473)

wherein X₁₈ is A or V; and X₁₉ is V or M. Group 11

QQYDNLP	L	T (SEQ ID NO: 1475)
QQYDNLP	F	T (SEQ ID NO: 1476)

wherein X₂₀ is L or F. Group 12

QQYGSS	P	PWT (SEQ ID NO: 1478)
QQYGSS	PWT (SEQ ID NO: 1479)

wherein X₂₁ is P or absent. Group 13

QQYG	R	S	L	FT (SEQ ID NO: 1481)
QQYG	T	S	P	FT (SEQ ID NO: 1482)

wherein X₂₂ is R or T; and X₂₃ is L or P. Group 14

QQYGSS	R	S (SEQ ID NO: 1484)
QQYGSS	P	R	S (SEQ ID NO: 1485)
QQYGSS	R	T (SEQ ID NO: 1486)
QQYGSS	C	S (SEQ ID NO: 1487)

wherein X₂₄ is P or absent; X₂₅ is R or C and X₂₆ is S or T. Group 15

T	T	W	V (SEQ ID NO: 1489)
QADWSS	T	A	V (SEQ ID NO: 1490)
QADWSS	T	W	V (SEQ ID NO: 1491)

wherein X₂₇ is T or absent; and X₂₈ is W or A. Group 16

QADWS	G	TV	V (SEQ ID NO: 1493)
QADWS	T	TV	V (SEQ ID NO: 1494)
QAWDS	A	TV	I (SEQ ID NO: 1495)

wherein X₂₉ is G, T, A or absent; and X₃₀ is V or I. Group 17

QQ	S	YSA	T	FT (SEQ ID NO: 1497)
QQ	T	YSA	P	FT (SEQ ID NO: 1498)

wherein X₃₁ is S or T; and X₃₂ is T or P. Group 18

QQYN	I	YPRT (SEQ ID NO: 1500)
QQYN	T	YPRT (SEQ ID NO: 1501)

wherein X₃₃ is I or T. Group 19

HQ	S	S	DLPLT (SEQ ID NO: 1503)
HQ	Y	D	DLPLT (SEQ ID NO: 1504)

wherein X₃₄ is S or Y; and X₃₅ is S or D. Group 20

MQALQT	P	F	T (SEQ ID NO: 1506)
MQALQT	L	I	T (SEQ ID NO: 1507)

wherein X₃₆ is P or L; and X₃₇ is F or I. Group 21 QQFGRSFT (SEQ ID NO: 1509) Group 22

YSTDSS	V	NHVV (SEQ ID NO: 1510)
YSTDSS	G	(SEQ ID NO: 1511)

wherein X₃₈ is V or G.

Light Chain CDR2

Group 1

A	ASSL	Q	S (SEQ ID NO: 1513)
S	ASSL	Q	S (SEQ ID NO: 1514)
A	ASSL	Q	F (SEQ ID NO: 1515)
A	ASSL	K	S (SEQ ID NO: 1516)

wherein X₃₉ is A or S; X₄₀ is Q or K; and X₄₁ is S or F. Group 2

G	A	S	S	R	A	T (SEQ ID NO: 1518)
G	A	S	S	R	D	T (SEQ ID NO: 1519)
G	T	S	T	R	A	T (SEQ ID NO: 1520)
G	A	S	T	R	A	T (SEQ ID NO: 1521)
G	A	S	A	R	A	T (SEQ ID NO: 1522)
G	A	S	R	R	A	T (SEQ ID NO: 1523)
G	A	S	N	R	A	T (SEQ ID NO: 1524)

wherein X₄₂ is A or T; X₄₃ is S, T, A, R or N; and X₄₄ is A or D. Group 3

GAFSRA	S (SEQ ID NO: 1526)
GAFSRA	T (SEQ ID NO: 1527)

wherein X₄₅ is S or T. Group 4

Q	D	T	KRPS (SEQ ID NO: 1529)
R	D	S	KRPS (SEQ ID NO: 1530)
E	D	S	KRPS (SEQ ID NO: 1531)
Q	D	S	KRPS (SEQ ID NO: 1532)

wherein X₄₆ is Q, R or E; and X₄₇ isT or S. Group 5

TLS	Y	RAS (SEQ ID NO: 1534)
TLS	F	RAS (SEQ ID NO: 1535)

wherein X₄₈ is Y or F. Group 6

AASNLQ	R (SEQ ID NO: 1537)
AASNLQ	S (SEQ ID NO: 1538)

wherein X49 is R or S. Group 7

G	A	SNRA I (SEQ ID NO: 1540)
G	S	SNRA I (SEQ ID NO: 1541)
G	S	SNRA T (SEQ ID NO: 1542)

wherein X₅₀ is A or S; and X₅₁ is I or T. Group 8

D	A	S	S	LQS (SEQ ID NO: 1544)
D	A	S	T	LQS (SEQ ID NO: 1545)
G	A	S	S	LQS (SEQ ID NO: 1546)
G	A	S	N	LQS (SEQ ID NO: 1547)

wherein X₅₂ is D or G; and X₅₃ is S, T or N. Group 9

DN	N	KRPS (SEQ ID NO: 1549)
DN	D	KRPS (SEQ ID NO: 1550)

wherein X₅₃ is N or D. Group 10

D	A	SNLET (SEQ ID NO: 1552)
D	V	SNLET (SEQ ID NO: 1553)

wherein X₅₄ is A or V. Group 11

L	G	SNRAS (SEQ ID NO: 1555)
L	D	SNRAS (SEQ ID NO: 1556)

wherein X₅₅ is G or D. Group 12

Q	D	N	K	RPS (SEQ ID NO: 1558)
Q	N	N	K	RPS (SEQ ID NO: 1559)
Q	D	N	E	RPS (SEQ ID NO: 1560)

wherein X₅₆ is D or N; and X₅₇ is K or E. Group 13 RDRNRPS (SEQ ID NO: 1562) Group 14

S	DSNRPS (SEQ ID NO: 1563)
C	DSNRPS (SEQ ID NO: 1564)

wherein X₅₈ is S or C. Group 15 DDSDRPS (SEQ ID NO: 1566) Group 16

A	V	SSLQS (SEQ ID NO: 1567)
A	S	SSLQS (SEQ ID NO: 1568)

wherein X₅₉ is S or V. Group 17

T	A	SSLQS (SEQ ID NO: 1570)
T	T	SSLQS (SEQ ID NO: 1571)

wherein X₆₀ is A or T. Group 18

K	V	SNWDS (SEQ ID NO: 1573)
K	G	SNWDS (SEQ ID NO: 1574)

wherein X₆₁ is V or G.

Light Chain CDR1

Group 1

RAS	Q	S	V	S	D	I	L	A (SEQ ID NO: 1576)
RAS	P	S	V	S	S	S	Y	L	A (SEQ ID NO: 1577)
RAS	Q	S	F	S	S	S	Y	L	A (SEQ ID NO: 1578)
RAS	Q	S	V	S	R	S	H	L	A (SEQ ID NO: 1579)
RAS	Q	S	V	S	R	D	Y	L	A (SEQ ID NO: 1580)
RAS	Q	S	V	S	R	N	Y	L	A (SEQ ID NO: 1581)
RAS	Q	S	V	S	S	M	Y	L	A (SEQ ID NO: 1582)
RAS	Q	S	V	S	S	Q	L	A (SEQ ID NO: 1583)
RAS	Q	S	I	S	S	N	L	A (SEQ ID NO: 1584)
RAS	Q	S	V	S	S	N	L	A (SEQ ID NO: 1585)
RAS	Q	S	V	S	S	N	V	A (SEQ ID NO: 1586)
RAS	Q	S	V	N	S	N	L	A (SEQ ID NO: 1587)
RAS	Q	S	V	R	S	S	S	L	A (SEQ ID NO: 1588)
RAS	Q	S	V	S	N	S	S	L	A (SEQ ID NO: 1589)
RAS	Q	S	V	R	N	S	S	L	A (SEQ ID NO: 1590)

wherein X₆₂ is P or Q; X₆₃ is V, I or F; X₆₄ is S, R or absent; X₆₅ is S, R or N; X₆₆ is D, S, N or M; X₆₇ is I, Y, H, Q, N or S; and X₆₈ is L or V. Group 2

R	A	SQ	I	I	S	R	YLN (SEQ ID NO: 1592)
R	T	SQ	S	I	S	S	YLN (SEQ ID NO: 1593)
R	A	SQ	S	I	S	N	YLN (SEQ ID NO: 1594)
R	T	SQ	S	I	S	S	YLN (SEQ ID NO: 1595)
R	A	SQ	T	I	S	YLN (SEQ ID NO: 1596)
R	A	SQ	R	I	S	S	YLN (SEQ ID NO: 1597)
R	A	SQ	S	I	S	S	YLN (SEQ ID NO: 1598)
R	A	SQ	N	I	R	T	YLN (SEQ ID NO: 1599)
R	A	SQ	N	I	R	S	YLN (SEQ ID NO: 1600)
R	A	SQ	N	I	N	N	YLN (SEQ ID NO: 1601)

wherein X₆₉ is A or T; X₇₀ is I, S, T or N; X₇₁ is R, S or N; and X₇₂ is R, S, N, or I. Group 3

GGN	N	IGS	Y	N	V	H (SEQ ID NO: 1603)
GGN	N	IGS	I	N	V	H (SEQ ID NO: 1604)
GGN	N	IGS	K	S	V	Q (SEQ ID NO: 1605)
GGN	D	IGS	K	S	V	H (SEQ ID NO: 1606)
GGN	N	IGS	K	S	V	H (SEQ ID NO: 1607)
GGN	N	IGS	K	T	V	H (SEQ ID NO: 1608)
GGN	N	IGS	K	A	V	H (SEQ ID NO: 1609)
GGN	N	IGS	K	N	V	H (SEQ ID NO: 1610)
GGN	D	IGS	K	N	V	H (SEQ ID NO: 1611)

wherein X₇₃ is N, or D; X₇₄ is Y, I or K; X₇₅ is N, S, T or A; and X₇₆ is H or Q. Group 4

RASQ	D	IRNDL	G (SEQ ID NO: 1613)
RASQ	D	IRNDL	A (SEQ ID NO: 1614)
RASQ	G	IRNDL	G (SEQ ID NO: 1615)

wherein X₇₇ is D or G; and X₇₈ is G or A. Group 5

RSSQSL	L	N	S	D	A	G	T	TYLD (SEQ ID NO: 1617)
RSSQSL	F	D	N	D	D	G	D	TYLD (SEQ ID NO: 1618)
RSSQSL	L	N	S	D	D	G	N	TYLD (SEQ ID NO: 1619)
RSSQSL	L	D	S	D	D	G	D	TYLD (SEQ ID NO: 1620)
RSSQSL	L	D	S	D	D	G	N	TYLD (SEQ ID NO: 1621)
G

wherein X₇₉ is L or F; X₈₀ is N or D; X₈₁ is S or N; X₈₂ is A or D; and X₈₃ is T, D or N. Group 6

SG	N	K	LGDKY	V	C (SEQ ID NO: 1623)
SG	D	K	LGDKY	V	C (SEQ ID NO: 1624)
SG	D	K	LGDKY	A	C (SEQ ID NO: 1625)
SG	D	E	LGDKY	A	C (SEQ ID NO: 1626)
SG	D	N	LGDKY	A	F (SEQ ID NO: 1627)
SG	D	N	LGDKY	A	C (SEQ ID NO: 1628)

wherein X₈₄ is N or D; X₈₅ is K, E or N; X₈₆ is V or A; and X₈₇ is C or F. Group 7

QASQ G	I	S	N	Y	LN (SEQ ID NO: 1630)
QASQ D	I	K	K	F	LN (SEQ ID NO: 1631)
QASQ D	I	N	I	Y	LN (SEQ ID NO: 1632)
QASQ D	I	S	I	Y	LN (SEQ ID NO: 1633)
QASQ D	I	T	K	Y	LN (SEQ ID NO: 1634)

wherein X₈₈ isG or D; X₈₉ is S, K N or T; X₉₀ is N, K or I; and X₉₁ is Y or F. Group 8

RASQ D	I	D	S	WL	V (SEQ ID NO: 1636)
RASQ G	I	S	R	WL	A (SEQ ID NO: 1637)
RASQ D	I	S	S	WL	A (SEQ ID NO: 1638)
RASQ G	I	S	S	WL	A (SEQ ID NO: 1639)

wherein X₉₂ is D or G; X₉₃ isD or S; X₉₄ is R or S; and X₉₅ is V or A. Group 9

SGSSSNIG	N	NYV	A (SEQ ID NO: 1641)
SGSSSNIG	I	NYV	S (SEQ ID NO: 1642)
SGSSSNIG	D	NYV	S (SEQ ID NO: 1643)
SGSSSNIG	N	NYV	S (SEQ ID NO: 1644)

wherein X₉₆ is N, I or D; and X₉₇ is A or S. Group 10

RAS	Q	DISNYLA (SEQ ID NO: 1646)
RAS	H	DISNYLA (SEQ ID NO: 1647)

wherein X₉₈ is Q or H. Group 11

RASQ	R	V	P	SSY	V (SEQ ID NO: 1649)
RASQ	R	V	P	SSY	V (SEQ ID NO: 1650)
RASQ	S	V	A	SSY	V (SEQ ID NO: 1651)

wherein X₉₉ is R or S; X₁₀₀ is P or A; and X₁₀₁ is I or L. Group 12

RSSQSL	L	HSNG	Y	NYLD (SEQ ID NO: 1653)
RSSQSL	L	HSNG	F	NYLD (SEQ ID NO: 1654)
RSSQSL	Q	HSNG	Y	NYLD (SEQ ID NO: 1655)

wherein X₁₀₂ is L or Q; and X₁₀₃ is Y or F. Group 13

RASQT	V	RN	N	YLA (SEQ ID NO: 1657)
RASQT	I	RN	S	YLA (SEQ ID NO: 1658)

wherein X₁₀₄ is V or I; and X₁₀₅ is N or S. Group 14

RSS	Q	R	LVYSDGNTYLN (SEQ ID NO: 1660)
RSS	P	S	LVYSDGNTYLN (SEQ ID NO: 1661)

wherein X₁₀₆ is Q or P; and X₁₀₇ is R or S. Group 15

SGDA	L	PKKYA	Y (SEQ ID NO: 1663)
SGDA	V	PKKYA	N (SEQ ID NO: 1664)

wherein X₁₀₈ is L or V; and X₁₀₉ is Y or N.

Heavy Chain CDR3

Group 1

MT	T	PYWYF	D	L (SEQ ID NO: 1666)
MT	S	PYWYF	D	L (SEQ ID NO: 1667)
MT	T	PYWYF	G	L (SEQ ID NO: 1668)

wherein X₁₁₀ is T or S; and X₁₁₁ is D or G. Group 2

D R Y (SEQ ID NO: 1670)	Y	DFW	S	GYP	Y	F	R	YYG	L	DV
D Q Y (SEQ ID NO: 1671)	F	DFW	S	GYP	F	F	Y	YYG	M	DV
D Q D (SEQ ID NO: 1672)	Y	DFW	S	GYP	Y	F	Y	YYG	M	DV
D Q N (SEQ ID NO: 1673)	Y	DFW	N	GYP	Y	Y	F	YYG	M	DV
D Q Y (SEQ ID NO: 1674)	Y	DFW	S	GYP	Y	Y	H	YYG	M	DV

wherein X₁₁₂ is R or Q; X₁₁₃ is Y, D or N; X₁₁₄ is Y or F; X₁₁₅ is S or N; X₁₁₆ is Y or F; X₁₁₇ is F or Y; X₁₁₈ is R, Y, F or H; and X₁₁₉ is L or M. Group 3 VTGTDAFDF (SEQ ID NO: 1676) Group 4 TVTKEDYYYYGMDV (SEQ ID NO: 1677) Group 5 DSSGSYYVEDYFDY (SEQ ID NO: 1678) Group 6

D	W	S	IAVAG	T	FDY (SEQ ID NO: 1679)
D	L	R	IAVAG	S	FDY (SEQ ID NO: 1680)

wherein X₁₁₉ is W or L; X₁₂₀ is S or R; and X₁₂₁ is T or S. Group 7 EYYYGSGSYYP (SEQ ID NO: 1682) Group 8 ELGDYPFFDY (SEQ ID NO: 1683) Group 9 EYVAEAGFDY (SEQ ID NO: 1684) Group 10 VAAVYWYFDL (SEQ ID NO: 1685) Group 11 YNWNYGAFDF (SEQ ID NO: 1686) Group 12

RASRGYR	F	GLAFAI (SEQ ID NO: 1687)
RASRGYR	Y	GLAFAI (SEQ ID NO: 1688)

wherein X₁₂₂ is F or Y. Group 13 DGITMVRGVTHYYGMDV (SEQ ID NO: 1690) Group 14

DH	S	SGWYYYGMDV (SEQ ID NO: 1691)
DH	T	SCWYYYGMDV (SEQ ID NO: 1692)

wherein X₁₂₃ is S or T. Group 15

Y	S	T	WDYYYG	V	DV (SEQ ID NO: 1694)
Y	R	D	WDYYYG	M	DV (SEQ ID NO: 1695)

wherein X₁₂₄ is S or R; X₁₂₅ is T or D; and X₁₂₆ is V or M. Group 16

VLHY	S	DS	R	GYSYY	S	D	F (SEQ ID NO: 1697)
VLHY	Y	DS	S	GYSYY	F	D	Y (SEQ ID NO: 1698)

wherein X₁₂₇ is S or Y; X₁₂₈ is R or S; X₁₂₉ is S or F; and X₁₃₀ is F or Y.

Heavy Chain CDR2

Group 1

N	I	Y	Y	S	G	T	T	Y	F	NPSLKS (SEQ ID NO: 1700)
F	I	Y	Y	S	G	G	T	N	Y	NPSLKS (SEQ ID NO: 1701)
Y	I	Y	Y	S	G	G	T	H	Y	NPSLKS (SEQ ID NO: 1702)
Y	I	Y	H	S	G	S	A	Y	Y	NPSLKS (SEQ ID NO: 1703)
Y	I	Y	D	S	G	S	T	Y	Y	NPSLKS (SEQ ID NO: 1704)
S	I	Y	Y	S	G	T	T	Y	Y	NPSLKS (SEQ ID NO: 1705)
M	I	Y	Y	S	G	T	T	Y	Y	NPSLKS (SEQ ID NO: 1706)
Y	I	Y	Y	S	G	T	T	Y	Y	NPSLKS (SEQ ID NO: 1707)
Y	I	Y	Y	S	G	S	A	Y	Y	NPSLKS (SEQ ID NO: 1708)
Y	I	F	Y	S	G	S	T	Y	Y	NPSLKS (SEQ ID NO: 1709)
Y	L	Y	Y	S	G	S	T	Y	Y	NPSLKS (SEQ ID NO: 1710)
Y	I	Y	Y	S	G	S	T	Y	Y	NPSLKS (SEQ ID NO: 1711)
Y	I	Y	Y	T	G	S	T	Y	Y	NPSLKS (SEQ ID NO: 1712)
Y	I	Y	Y	T	G	S	T	N	Y	NPSLKS (SEQ ID NO: 1713)
Y	I	Y	Y	S	G	N	T	N	Y	NPSLKS (SEQ ID NO: 1714)
Y	I	Y	Y	S	G	S	T	N	Y	NPSLKS (SEQ ID NO: 1715)
G

wherein X₁₃₁ is N, F, Y, S or M; X₁₃₂ is I or L; X₁₃₃ is Y or F; X₁₃₄ is Y, H or D; X₁₃₅ is S or T; X₁₃₆ is T, G, S or T; X₁₃₇ is T or A; X₁₃₈ is Y, N or H; and X₁₃₉ is F or Y. Group 2

L	I	W	Y	DG	D	N	K	Y	Y	ADSVKG (SEQ ID NO: 1717)
G	I	S	Y	DG	S	N	K	N	Y	ADSVKG (SEQ ID NO: 1718)
I	I	W	Y	DG	S	N	K	N	Y	ADSVKG (SEQ ID NO: 1719)
L	I	W	Y	DG	S	N	K	N	Y	ADSVKG (SEQ ID NO: 1720)
L	I	W	Y	DG	S	N	K	D	Y	ADSVKG (SEQ ID NO: 1721)
V	I	W	Y	DG	S	N	K	D	Y	ADSVKG (SEQ ID NO: 1722)
L	I	S	Y	DG	S	N	K	Y	Y	ADSVKG (SEQ ID NO: 1723)
V	I	S	Y	DG	S	N	K	H	Y	ADSVKG (SEQ ID NO: 1724)
V	I	S	Y	DG	S	N	K	Y	Y	ADSVKG (SEQ ID NO: 1725)
V	I	W	D	DG	S	N	K	Y	Y	ADSVKG (SEQ ID NO: 1726)
V	I	W	D	DG	S	N	N	Y	Y	ADSVKG (SEQ ID NO: 1727)
V	I	W	Y	DG	S	N	K	Y	H	ADSVKG (SEQ ID NO: 1728)
V	I	W	Y	DG	S	N	K	Y	Y	ADSVKG (SEQ ID NO: 1729)
V	I	W	N	DG	N	N	K	Y	Y	ADSVKG (SEQ ID NO: 1730)
V	I	W	N	DG	S	N	K	N	Y	ADSVKG (SEQ ID NO: 1731)
N

wherein X₁₄₀ is L, G, I or V;X₁₄₁ is W or S; X₁₄₂ is Y, D or N; X₁₄₃ is S or D; X₁₄₄ is K or N; X₁₄₅ is Y, N, D, or H; and X₁₄₆ is Y or H. Group 3

W	I	NP	P	SG	A	T	N	YAQKF	R	G (SEQ ID NO: 1733)
W	I	NP	N	SG	G	T	N	YAQKF	R	G (SEQ ID NO: 1734)
W	I	NP	N	SG	A	T	N	YAQKF	H	G (SEQ ID NO: 1735)
W	I	NP	S	SG	D	T	K	YAQKF	Q	G (SEQ ID NO: 1736)
W	M	NP	N	SG	A	T	K	YAQKF	Q	G (SEQ ID NO: 1737)

W	I	NP	N	SG	A	T	K	YAQKF	Q	G (SEQ ID NO: 1738)
W	I	NP	D	SG	G	T	N	YAQKF	Q	G (SEQ ID NO: 1739)
W	I	NP	N	SG	G	T	D	YAQKF	Q	G (SEQ ID NO: 1740)

wherein X₁₄₇ is I or M; X₁₄₈ is P, N, S or D; X₁₄₉ is A, G or D; X₁₅₀ isN, K, or D; X₁₅₁ is R, H or Q. Group 4

EINHS	E	N	TNYNPSLKS (SEQ ID NO: 1742)
EINHS	G	T	TNYNPSLKS (SEQ ID NO: 1743)

wherein X₁₅₂ is E or G; and X₁₅₃ is N or T. Group 5

IIYPGDS	D	TRYSPSFQG (SEQ ID NO: 1745)
IIYPGDS	E	TRYSPSFQG (SEQ ID NO: 1746)

wherein X₁₅₄ is D or E. Group 6

SISSSS	T	Y	I	YY	A	DS	V	KG (SEQ ID NO: 1748)
SISSSS	T	Y	I	YY	A	DS	L	KG (SEQ ID NO: 1749)
SISSSS	S	Y	E	YY	V	DS	V	KG (SEQ ID NO: 1750)

wherein X₁₅₅ is T or S; X₁₅₆ is I or E; X₁₅₇ is A or V; and X₁₅₈ is V or L. Group 7

RI	K	S	KTDGGTT	D	YAAPVKG (SEQ ID NO: 1752)
RI	K	S	KTDGGTT	E	YAAPVKG (SEQ ID NO: 1753)
RI	I	G	KTDGGTT	D	YAAPVKG (SEQ ID NO: 1754)

wherein X₁₅₉ is K or I; X₁₆₀ is S or G; and X₁₆₁ is D or E. Group 8 GISGSSAGTYYADSVGK (SEQ ID NO: 1756) Group 9

VIS	D	SGG	S	TYYADSVKG (SEQ ID NO: 1757)
VIS	G	SGG	D	TYYADSVKG (SEQ ID NO: 1758)

wherein X₁₆₂ is D or G; and X₁₆₃ is S or D. Group 10 RTYYRSKWYNDYAVSVKS (SEQ ID NO: 1760) Group 11

RIY	I	SGSTNYNPSL	E	N (SEQ ID NO: 1761)
RIY	T	SGSTNYNPSL	K	S (SEQ ID NO: 1762)

wherein X₁₆₄ is I or T; X₁₆₅ is E or K; and X₁₆₆ is N or S. Group 12

WMNPYSGSTG	Y	AQ	N	FQ	G (SEQ ID NO: 1764)
WMNPYSGSTG	L	AQ	R	FQ	D (SEQ ID NO: 1765)

wherein X₁₆₇ is Y or L; X₁₆₈ is N or R; and X₁₆₉ is G or D.

Heavy Chain CDR1

Group 1

SG	V	Y	YW	N (SEQ ID NO: 1767)
SG	V	Y	YW	S (SEQ ID NO: 1768)
SG	G	Y	YW	N (SEQ ID NO: 1769)
SG	G	Y	YW	S (SEQ ID NO: 1770)
SG	D	N	TW	S (SEQ ID NO: 1771)
SG	N	Y	TW	S (SEQ ID NO: 1772)
SG	D	Y	TW	T (SEQ ID NO: 1773)
SG	D	Y	TW	S (SEQ ID NO: 1774)

wherein X₁₇₀ is V,G, N or D; X₁₇₁ is Y or N; and X₁₇₂ is N, S or T. Group 2

T	YYW	S (SEQ ID NO: 1776)
Y	YYW	S (SEQ ID NO: 1777)
S	YYW	S (SEQ ID NO: 1778)
G	YYW	S (SEQ ID NO: 1779)
G	YYW	T (SEQ ID NO: 1780)

wherein X₁₇₃ isT, S or G; and X₁₇₄ is S or T. Group 3

S	Y	GMH (SEQ ID NO: 1782)
S	F	GMH (SEQ ID NO: 1783)
T	Y	GMH (SEQ ID NO: 1784)
F	Y	GMH (SEQ ID NO: 1785)

wherein X₁₇₅ is S, T or F; and X₁₇₆ is Y or F. Group 4

SY	A	M	S (SEQ ID NO: 1787)
SY	S	M	N (SEQ ID NO: 1788)
SY	S	M	S (SEQ ID NO: 1789)

wherein X₁₇₇ is A or S; and X₁₇₈ is S, N or M. Group 5

Y	YY	I	H (SEQ ID NO: 1791)
G	YY	L	H (SEQ ID NO: 1792)
G	YY	K	H (SEQ ID NO: 1793)
G	YY	T	H (SEQ ID NO: 1794)
G	YY	I	H (SEQ ID NO: 1795)

wherein X₁₇₉ is Y or G; and X₁₈₀ is I, L, K or T. Group 6

SYG	I	H (SEQ ID NO: 1797)
SYG	L	H (SEQ ID NO: 1798)

wherein X₁₈₁ is L or I. Group 7

NY	G	M	H (SEQ ID NO: 1800)
NY	G	M	R (SEQ ID NO: 1801)
NY	N	M	H (SEQ ID NO: 1802)

wherein X₁₈₂ is G or N; and X₁₈₃ is H, R or M. Group 8

S	YWIG (SEQ ID NO: 1804)
G	YWIG (SEQ ID NO: 1805)

wherein X₁₈₄ is S or G. Group 9

GY	Y	MH (SEQ ID NO: 1807)
GY	F	MH (SEQ ID NO: 1808)

wherein X₁₈₅ is Y or F. Group 10

S	Y	DI	N (SEQ ID NO: 1810)
S	H	DI	N (SEQ ID NO: 1811)
S	Y	DI	D (SEQ ID NO: 1812)

wherein X₁₈₆ is Y or H; and X₁₈₇ is N or D. Group 11

N	YAMS (SEQ ID NO: 1814)
H	YAMS (SEQ ID NO: 1815)

wherein X₁₈₈ is N or H. Group 12 NAWMS (SEQ ID NO: 1817) Group 13 SSSYYWG (SEQ ID NO: 1818) Group 14

D	YYWN (SEQ ID NO: 1819)
S	YYWN (SEQ ID NO: 1820)

wherein X₁₈₉ is D or S. Group 15

SNSA	T	WN (SEQ ID NO: 1822)
SNSA	A	WN (SEQ ID NO: 1823)

wherein X₁₉₀ is T or A. Group 16

S	YDMH (SEQ ID NO: 1825)
T	YDMH (SEQ ID NO: 1826)

wherein X₁₉₁ is S or T.

In some cases an antigen binding protein comprises at least one heavy chain CDR1, CDR2, or CDR3 having one of the above consensus sequences. In some cases, an antigen binding protein comprises at least one light chain CDR1, CDR2, or CDR3 having one of the above consensus sequences. In other cases, the antigen binding protein comprises at least two heavy chain CDRs according to the determined consensus sequences, and/or at least two light chain CDRs according to the determined consensus sequences. In still other cases, the antigen binding protein comprises at least three heavy chain CDRs according to the determined consensus sequences, and/or at least three light chain CDRs according to the determined consensus sequences.

Exemplary Antigen Binding Proteins

According to one aspect, an isolated antigen binding protein comprising (a) one or more heavy chain complementary determining regions (CDRHs) comprising one or more of: (i) a CDRH1 selected from the group consisting of SEQ ID NOS 603-655; (ii) a CDRH2 selected from the group consisting of SEQ ID NOS 656-732; (iii) a CDRH3 selected from the group consisting of SEQ ID NOS 733-813; and (iv) a CDRH of (i), (ii) and (iii) that comprises ten, nine, eight, seven, six, five, four, three, two or one amino acid substitutions, deletions, insertions and combinations thereof; (b) one or more light chain complementary determining regions (CDRLs) comprising one or more of: (i) a CDRL1 selected from the group consisting of SEQ ID NOS 814-893; (ii) a CDRL2 comprising one or more of SEQ ID NOS 894-946; (iii) a CDRL3 comprising one or more of SEQ ID NOS 947-1020; and (iv) a CDRL of (i), (ii) and (iii) that comprises ten, nine, eight, seven, six, five, four, three, four, two or one amino acid substitutions, deletions or insertions and combinations thereof; or (c) one or more heavy chain CDRHs of (a) and one or more light chain CDRLs of (b).

In another embodiment, the CDRHs have at least 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% sequence identity with an amino acid sequence selected from the group consisting of SEQ ID NOS 603-813, and/or the CDRLs have at least 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% sequence identity with an amino acid sequence selected from the group consisting of SEQ ID NOS 814-1020. In a further embodiment, the VH is selected from the group consisting of SEQ ID NOS 316-409, and/or the V_L is selected from the group consisting of SEQ ID NOS 217-315.

According to one aspect, an isolated antigen binding protein comprising (a) one or more variable heavy chains (V_Hs) comprising one or more of: (i) SEQ ID NOS 316-409; and (ii) a V_H of (i) that comprises ten, nine, eight, seven, six, five, four, three, two or one amino acid substitutions, deletions, insertions and combinations thereof; (b) one or more variable light chains (V_Ls) selected from the group consisting of: (i) SEQ ID NOS 217-315, and (ii) a V_L of (i) that comprises ten, nine, eight, seven, six, five, four, three, two or one amino acid substitutions, deletions, insertions and combinations thereof; or (c) one or more variable heavy chains of (a) and one or more variable light chains of (b).

In another embodiment, the variable heavy chain (V_H) has at least 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% sequence identity with an amino acid sequence selected from the group consisting of SEQ ID NOS 36-409, and/or the variable light chain (V_L) has at least 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%. 98% or 99% sequence identity with an amino acid sequence selected from the group consisting of SEQ ID NOS 217-315.

In one aspect, also provided is an antigen binding protein that specifically binds to a linear or three-dimensional epitope comprising one or more amino acid residues from FGFR1c, FGRF2c and FGFR3c.

In one aspect, also provided is an antigen binding protein that specifically binds to a linear or three-dimensional epitope comprising one or more amino acid residues from β-Klotho.

In another aspect, also provided is an isolated antigen binding protein that specifically binds to a linear or three-dimensional epitope comprising one or more amino acid residues from both β-Klotho and one or more amino acid residues from FGFR1c, FGFR2c and FGFR3c.

In yet another embodiment, the isolated antigen binding protein described hereinabove comprises a first amino acid sequence comprising at least one of the CDRH consensus sequences disclosed herein, and a second amino acid sequence comprising at least one of the CDRL consensus sequences disclosed herein.

In one aspect, the first amino acid sequence comprises at least two of the CDRH consensus sequences, and/or the second amino acid sequence comprises at least two of the CDRL consensus sequences. In certain embodiments, the first and the second amino acid sequence are covalently bonded to each other.

In a further embodiment, the first amino acid sequence of the isolated antigen binding protein comprises the CDRH3, the CDRH2 and the CDRH1 parings shown in Table 5 for each clone, and/or the second amino acid sequence of the isolated antigen binding protein comprises the CDRL3, the CDRL2 and the CDRL1 pairings shown in Table 4 or each clone.

In a further embodiment, the antigen binding protein comprises at least two CDRH sequences of heavy chain sequences H1, H2, H3, H4, H5, H6, H7, H8, H9, H10, H11, H12, H13, H14, H15, H16, H17 or H18, H19, H20, H21, H22, H23, H24, H25, H26, H27, H28, H29, H30, H31, H32, H33, H34, H35, H36, H37, H38, H39, H40, H41, H42, H43, H44, H45, H146, H46, H48, H49, H50, H51, H52, H53, H54, H55, H56, H57, H58, H59, H60, H61, H62, H63, H64, H65, H66, H67, H68, H69, H70, H71, H72, H73, H74, H75, H76, H77, H78, H79, H80, H81, H82, H83, H84, H85, H86, H87, H88, H89, H90, H91, H92, H93 and H94, as shown in Tables 3A and 4A.

In again a further embodiment, the antigen binding protein comprises at least two CDRL sequences of light chain sequences L1, L2, L3, L4, L5, L6, L7, L8, L9, L10, L11, L12, L13, L14, L15, L16, L17, L18, L19, L20, L21, L22, L23, L24, L25, L26, L27, L28, L29, L30, L31, L32, L33, L34, L35, L36, L37, L38, L39, L40, L41, L42, L43, L44, L45, L46, L47, L48, L49, L50, L51, L52, L53, L54, L55, L56, L57, L58, L59, L60, L61, L62, L63, L64, L65, L66, L67, L68, L69, L70, L71, L72, L73, L74, L75, L76, L77, L78, L79, L80, L81, L82, L83, L84, L85, L86, L87, L88, L89, L90, L91, L92, L93, L94, L95, L96, L97, L98, L99 and L100, as shown in Tables 3B and 4B.

In still a further embodiment, the antigen binding protein comprises at least two CDRH sequences of heavy chain sequences H1, H2, H3, H4, H5, H6, H7, H8, H9, H10, H11, H12, H13, H14, H15, H16, H17 or H18, H19, H20, H21, H22, H23, H24, H25, H26, H27, H28, H29, H30, H31, H32, H33, H34, H35, H36, H37, H38, H39, H40, H41, H42, H43, H44, H45, H146, H46, H48, H49, H50, H51, H52, H53, H54, H55, H56, H57, H58, H59, H60, H61, H62, H63, H64, H65, H66, H67, H68, H69, H70, H71, H72, H73, H74, H75, H76, H77, H78, H79, H80, H81, H82, H83, H84, H85, H86, H87, H88, H89, H90, H91, H92, H93 and H94, as shown in Tables 3A and 4A, and at least two CDRLs of light chain sequences L1, L2, L3, L4, L5, L6, L7, L8, L9, L10, L11, L12, L13, L14, L15, L16, L17, L18, L19, L20, L21, L22, L23, L24, L25, L26, L27, L28, L29, L30, L31, L32, L33, L34, L35, L36, L37, L38, L39, L40, L41, L42, L43, L44, L45, L46, L47, L48, L49, L50, L51, L52, L53, L54, L55, L56, L57, L58, L59, L60, L61, L62, L63, L64, L65, L66, L67, L68, L69, L70, L71, L72, L73, L74, L75, L76, L77, L78, L79, L80, L81, L82, L83, L84, L85, L86, L87, L88, L89, L90, L91, L92, L93, L94, L95, L96, L97, L98, L99 and L100, as shown in Tables 3B and 4B.

In again another embodiment, the antigen binding protein comprises the CDRH1, CDRH2, and CDRH3 sequences of heavy chain sequences H1, H2, H3, H4, H5, H6, H7, H8, H9, H10, H11, H12, H13, H14, H15, H16, H17 or H18, H19, H20, H21, H22, H23, H24, H25, H26, H27, H28, H29, H30, H31, H32, H33, H34, H35, H36, H37, H38, H39, H40, H41, H42, H43, H44, H45, H146, H46, H48, H49, H50, H51, H52, H53, H54, H55, H56, H57, H58, H59, H60, H61, H62, H63, H64, H65, H66, H67, H68, H69, H70, H71, H72, H73, H74, H75, H76, H77, H78, H79, H80, H81, H82, H83, H84, H85, H86, H87, H88, H89, H90, H91, H92, H93 and H94, as shown in Tables 3A and 4A.

In yet another embodiment, the antigen binding protein comprises the CDRL1, CDRL2, and CDRL3 sequences of light chain sequences L1, L2, L3, L4, L5, L6, L7, L8, L9, L10, L11, L12, L13, L14, L15, L16, L17, L18, L19, L20, L21, L22, L23, L24, L25, L26, L27, L28, L29, L30, L31, L32, L33, L34, L35, L36, L37, L38, L39, L40, L41, L42, L43, L44, L45, L46, L47, L48, L49, L50, L51, L52, L53, L54, L55, L56, L57, L58, L59, L60, L61, L62, L63, L64, L65, L66, L67, L68, L69, L70, L71, L72, L73, L74, L75, L76, L77, L78, L79, L80, L81, L82, L83, L84, L85, L86, L87, L88, L89, L90, L91, L92, L93, L94, L95, L96, L97, L98, L99 and L100, as shown in Tables 3B and 4B.

In yet another embodiment, the antigen binding protein comprises all six CDRs of an antigen binding protein comprising the following V_H and V_L pairs: V_L1 with V_L1; V_L2 with V_H1; V_L3 with V_H2 or V_H3; V_L4 with V_H4; V_L5 with V_H5; V_L6 with V_H6; V_L7 with V_H6; V_L8 with V_H7 or V_H8; V_L9 with V_H9; V_L10 with V_H9; V_L11 with V_H 10; V_L12 with V_H11; V_L13 with V_H12; V_L13 with V_H14; V_L14 with V_H13; V_L15 with V_H14; V_L16 with V_H15; V_L17 with V_H16; V_L18 with V_H17; V_L19 with V_H18; V_L20 with V_H19; V_L21 with V_H20; V_L22 with V_H21; V_L23 with V_H22; V_L24 with V_H23; V_L25 with V_H24; V_L26 with V_H25; V_L27 with V_H26; V_L28 with V_H27; V_L29 with V_H28; V_L30 with V_H29; V_L31 with V_H30; V_L32 with V_H31; V_L33 with V_H32;, V_L34 with V_H33; V_L35 with V_H34; V_L36 with V_H35; V_L37 with V_H36; V_L38 with V_H37; V_L39 with V_H38; V_L40 with V_H39; V_L41 with V_H40; V_L42 with V_H41; V_L43 with V_H42; V_L44 with V_H43; V_L45 with V_H44; V_L46 with V_H45; V_L47 with V_H46; V_L48 with V_H47; V_L49 with V_H48; V_L50 with V_H49; V_L51 with V_H50; V_L52 with V_H51; V_L53 with V_H52; V_L54 with V_H53; V_L55 with V_H54; V_L56 with V_H54; V_L57 with V_H54; V_L58 with V_H55; V_L59 with V_H56; V_L60 with V_H57; V_L61 with V_H58; V_L62 with V_H59; V_L63 with V_H60; V_L64 with V_L1; V_L65 with V_H62; V_L66 with V_H63; V_L67 with V_H64; V_L68 with V_H65; V_L69 with V_H66; V_L70 with V_H67; V_L71 with V_H68; V_L72 with V_H69; V_L73 with V_H70; V_L74 with V_H70; V_L75 with V_H70; V_L76 with V_H71; V_L77 with V_H72; V_L78 with V_H73; V_L79 with V_H74; V_L80 with V_H75; V_L81 with V_H76; V_L82 with V_H77; V_L83 with V_H78; V_L84 with V_H79; V_L85 with V_H80; V_L86 with V_H81; V_L87 with V_H82; V_L88 with V_H86; V_L89 with V_H83; V_L90 with V_H84; V_L91 with V_H85; V_L 92 with V_H 87; V_L 93 with V_H 88; V_L 94 with V_H 88; V_L 95 with V_H 89; V_L 96 with V_H 90; V_L 97 with V_H 91; V_L 98 with V_H 92; V_L 99 with V_H 93; and V_L 100 with V_H 94; as shown in Tables 2A and 2B and Tables 4A and 4B.

In one aspect, the isolated antigen binding proteins that specifically bind to a complex comprising β-Klotho and at least one of (i) FGFR1c, (ii) FGFR2c and (iii) FGFR3c provided herein can be a monoclonal antibody, a polyclonal antibody, a recombinant antibody, a human antibody, a humanized antibody, a chimeric antibody, a multispecific antibody, or an antibody fragment thereof.

In another embodiment, the antibody fragment of the isolated antigen-binding proteins provided herein can be a Fab fragment, a Fab' fragment, an F(ab')₂ fragment, an Fv fragment, a diabody, or a single chain antibody molecule.

In a further embodiment, an isolated antigen binding protein that specifically binds to a complex comprising β-Klotho and at least one of (i) FGFR1c, (ii) FGFR2c and (iii) FGFR3c provided herein is a human antibody and can be of the IgG1-, IgG2- IgG3- or IgG4-type.

In another embodiment, an isolated antigen binding protein that specifically binds to a complex comprising β-Klotho and at least one of (i) FGFR1c, (ii) FGFR2c and (iii) FGFR3c comprises a light or a heavy chain polypeptide as set forth in Tables 1A-1B. In some embodiments, an antigen binding protein that specifically binds to a complex comprising β-Klotho and at least one of (i) FGFR1c, (ii) FGFR2c and (iii) FGFR3c comprises a variable light or variable heavy domain such as those listed in Tables 2A-2B. In still other embodiments, an antigen binding protein that specifically binds to a complex comprising β-Klotho and at least one of (i) FGFR1c, (ii) FGFR2c and (iii) FGFR3c comprises one, two or three CDRHs or one, two or three CDRLs as set forth in Tables 3A-3B, 4A-4B, infra. Such antigen binding proteins, and indeed any of the antigen binding proteins disclosed herein, can be PEGylated with one or more PEG molecules, for examples PEG molecules having a molecular weight selected from the group consisting of 5K, 10K, 20K, 40K, 50K, 60K, 80K, 100K or greater than 100K.

In yet another aspect, any antigen binding protein that specifically binds to a complex comprising β-Klotho and at least one of (i) FGFR1c, (ii) FGFR2c and (iii) FGFR3c provided herein can be coupled to a labeling group and can compete for binding to the extracellular portion of the individual protein components of a complex comprising β-Klotho and at least one of (i) FGFR1c, (ii) FGFR2c and (iii) FGFR3c with an antigen binding protein of one of the isolated antigen binding proteins provided herein. In one embodiment, the isolated antigen binding protein provided herein can reduce blood glucose levels, decrease triglyceride and cholesterol levels or improve other glycemic parameters and cardiovascular risk factors when administered to a patient.

As will be appreciated, for any antigen binding protein comprising more than one CDR provided in Tables 3A-3B, and 4A-4B, any combination of CDRs independently selected from the depicted sequences may be useful. Thus, antigen binding proteins with one, two, three, four, five or six of independently selected CDRs can be generated. However, as will be appreciated by those in the art, specific embodiments generally utilize combinations of CDRs that are non-repetitive, e.g., antigen binding proteins are generally not made with two CDRH2 regions, etc.

Some of the antigen binding proteins that specifically bind to a complex comprising β-Klotho and at least one of (i) FGFR1c, (ii) FGFR2c and (iii) FGFR3c that are provided herein are discussed in more detail below.

Antigen Binding Proteins and Binding Epitopes and Binding Domains

When an antigen binding protein is said to bind an epitope on a complex β-Klotho and at least one of (i) FGFR1c, (ii) FGFR2c and (iii) FGFR3c, or the extracellular domain of a protein component of such a complex, what is meant is that the antigen binding protein specifically binds to a specified portion of the complex comprising β-Klotho and an FGFR (e.g., FGFR1c, FGFR2c or FGFR3c) or to the extracellular domain of such a complex. In some embodiments, e.g., in certain cases where the antigen binding protein binds only β-Klotho, the antigen binding protein can specifically bind to a polypeptide consisting of specified residues (e.g., a specified segment of β-Klotho). In other embodiments, e.g., in certain cases where an antigen binding protein interacts with both β-Klotho and at least one of (i) FGFR1c, (ii) FGFR2c and (iii) FGFR3c, the antigen binding protein can bind residues, sequences of residues, or regions in both β-Klotho and at least one of (i) FGFR1c, (ii) FGFR2c and (iii) FGFR3c, depending on which receptor the antigen binding protein recognizes. In still other embodiments the antigen binding protein will bind residues, sequences or residues or regions of a complex comprising β-Klotho and at least one of (i) FGFR1c, (ii) FGFR2c and (iii) FGFR3c, for example FGFR1c.

In any of the foregoing embodiments, such an antigen binding protein does not need to contact every residue of β-Klotho or a complex comprising β-Klotho and at least one of (i) FGFRl c, (ii) FGFR2c and (iii) FGFR3c, or the extracellular domain of the recited proteins or complexes. Nor does every single amino acid substitution or deletion within β-Klotho or a complex comprising β-Klotho and at least one of (i) FGFR1c, (ii) FGFR2c and (iii) FGFR3c, or the extracellular domain of the recited proteins or complexes, necessarily significantly affect binding affinity.

Epitope specificity and the binding domain(s) of an antigen binding protein can be determined by a variety of methods. Some methods, for example, can use truncated portions of an antigen. Other methods utilize antigen mutated at one or more specific residues, such as by employing an alanine scanning or arginine scanning-type approach or by the generation and study of chimeric proteins in which various domains, regions or amino acids are swapped between two proteins (e.g., mouse and human forms of one or more of the antigens or target proteins), or by protease protection assays.

Competing Antigen Binding Proteins

In another aspect, antigen binding proteins are provided that compete with one of the exemplified antibodies or functional fragments for binding to a complex comprising β-Klotho and at least one of (i) FGFR1c, (ii) FGFR2c and (iii) FGFR3c. Such antigen binding proteins can also bind to the same epitope as one of the herein exemplified antigen binding proteins, or an overlapping epitope. Antigen binding proteins and fragments that compete with or bind to the same epitope as the exemplified antigen binding proteins are expected to show similar functional properties. The exemplified antigen binding proteins and fragments include those with the heavy and light chains H1-H94 and L1-L100, variable region domains V_L1-V_L100 and V_H1-V_H94, and CDRs provided herein, including those in Tables 1, 2, 3, and 4. Thus, as a specific example, the antigen binding proteins that are provided include those that compete with an antibody comprising:

(a) 1, 2, 3, 4, 5 or all 6 of the CDRs listed for an antigen binding protein listed in Tables 3A and 3B, and 4A and 4B, infra;
(b) a V_H and a V_L selected from V_L1-V_L100 and V_H1-V_H94 and listed for an antigen binding protein listed in Tables 2A and 2B; or
(c) two light chains and two heavy chains as specified for an antigen binding protein listed in Tables 1A and 1B, infra.

Thus, in one embodiment, the present disclosure provides antigen binding proteins, including human antibodies, that competes for binding to a complex comprising β-Klotho and at least one of (i) FGFR1c, (ii) FGFR2c and (iii) FGFR3c with a reference antibody, wherein the reference antibody comprises a combination of light chain and heavy chain variable domain sequences selected from the group consisting of V_L1 with V_H1, V_L2 with V_H1, V_L3 with V_H2 or V_H3, V_L4 with V_H4, V_L5 with V_H5, V_L6 with V_H6, V_L7 with V_H6, V_L8 with V_H7 or V_H8, V_L9 with V_H9, V_L10 with V_H9, V_L11 with V_H 10, V_L12 with V_H11, V_L13 with V_H12, V_L13 with V_H14, V_L14 with V_H13, V_L15 with V_H14, V_L16 with V_H15, V_L17 with V_H16, V_L18 with V_H17, V_L19 with V_H18, V_L20 with V_H19, V_L21 with V_H20, V_L22 with V_H21, V_L23 with V_H22, V_L24 with V_H23, V_L25 with V_H24, V_L26 with V_H25, V_L27 with V_H26, V_L28 with V_H27, V_L29 with V_H28, V_L30 with V_H29, V_L31 with V_H30, V_L32 with V_H31, V_L33 with V_H32, V_L34 with V_H33, V_L35 with V_H34, V_L36 with V_H35, V_L37 with V_H36, V_L38 with V_H37, V_L39 with V_H38, V_L40 with V_H39, V_L41 with V_H40, V_L42 with V_H41, V_L43 with V_H42, V_L44 with V_H43, V_L45 with V_H44, V_L46 with V_H45, V_L47 with V_H46, V_L48 with V_H47, V_L49 with V_H48, V_L50 with V_H49, V_L51 with V_H50, 52 with V_H51, V_L53 with V_H52, V_L54 with V_H53, V_L55 with 54, and V_L56 with V_H54, V_L57 with V_H54, V_L58 with V_H55, V_L59 with V_H56, V_L60 with V_H57, V_L61 with V_H58, V_L62 with V_H59, V_L63 with V_H60, V_L64 with V_H1, V_L65 with V_H62, V_L66 with V_H63, V_L67 with V_H64, V_L68 with V_H65, V_L69 with V_H66, V_L70 with V_H67, V_L71 with V_H68, V_L72 with V_H69, V_L73 with V_H70, V_L74 with V_H70, and V_L75 with V_H70, V_L76 with V_H71, V_L77 with V_H72, V_L78 with V_H73, V_L79 with V_H74, V_L80 with V_H75, V_L81 with V_H76, V_L82 with V_H77, V_L83 with V_H78, V_L84 with V_H79, V_L85 with V_H80, V_L86 with V_H81, V_L87 with V_H82, V_L88 with V_H86, V_L89 with V_H83, V_L90 with V_H84, V_L91 with V_H85, V_L 92 with V_H 87, V_L 93 with V_H 88, V_L 94 with V_H 88, V_L 95 with V_H 89, V_L 96 with V_H 90, V_L 97 with V_H 91, V_L 98 with V_H 92, V_L 99 with V_H 93, and V_L 100 with V_H 94.

In another embodiment, the present disclosure provides antigen binding proteins, including human antibodies, that compete for binding to a complex comprising β-Klotho and at least one of (i) FGFR1c, (ii) FGFR2c and (iii) FGFR3c with a reference antibody, wherein the reference antibody is 63G8, 64A8, 67B4, 68D3, 64E6, 65E8, 65F11, 67G7, 63B6, 64D4, 65C3, 68D5, 63E6, 66F7, 64H5, 65G4, 67G10v1, 67G10v2, 66B4, 66G2, 68G5, 63F5, 66F6, 65C1, 64A7, 66D4, 65B1, 67A4, 65B4, 63A10, 65H11, 64C8, 65E3, 65D4, 65D1, 67G8, 65B7, 64A6, 65F9, 67F5, 64B10, 68C8, 67A5, 67C10, 64H6, 63F9, 67F6, 48H11, 52A8, 52F8, 49H12, 54A1, 55G9, 49C8, 52H1, 60G5.2, 49G3, 59A10, 48F8, 53B9, 56B4, 57E7, 57F11, 59C9, 58A5, 57A4, 57F9, 51G2, 56A7, 56E4, 54H10, 55D1, 48H3, 53C11, 59G10.3, 51C10.1, 59D10v1, 59D10v2, 60F9, 48B4, 52D6, 61G5, 59G10.2, 51A8, 53H5.2, 53F6, 56C11, 49A10, 48D4, 49G2, 50C12, 55G11, 52C1, 55E9, 60D7, 51C10.2, 55D3, 57B12, 52C5, 60G5.1, 55E4, 49B11, 50H10, 53C1, 56G1, 48F3, 48C9, 49A12, 51E2, 51E5, 53H5.3, 56G3.3, 55B10, 52B8, 55G5, 52H2, 56G3.2, 6E7, 57D9, 61H5, 48G4, 50G1, 58C2, 50D4, 50G5v1, 50G5v2, 51C1, 53C3.2, 54H10.3, 55A7, 55E6, 61E1, 53C3.1, 49H4, and 51E2.

In a further embodiment, an isolated antigen binding protein, such as a human antibody, is provided that binds to a complex comprising β-Klotho and at least one of (i) FGFR1c, (ii) FGFR2c and (iii) FGFR3c with substantially the same Kd as a reference antibody; initiates FGF21-like signaling in an in vitro ELK-Luciferase assay to the same degree as a reference antibody; lowers blood glucose; lowers serum lipid levels; and/or competes for binding with said reference antibody to a complex comprising β-Klotho and at least one of (i) FGFR1c, (ii) FGFR2c and (iii) FGFR3c, wherein the reference antibody is selected from the group consisting of 63G8, 64A8, 67B4, 68D3, 64E6, 65E8, 65F11, 67G7, 63B6, 64D4, 65C3, 68D5, 63E6, 66F7, 64H5, 65G4, 67G10v1, 67G10v2, 66B4, 66G2, 68G5, 63F5, 66F6, 65C1, 64A7, 66D4, 65B1, 67A4, 65B4, 63A10, 65H11, 64C8, 65E3, 65D4, 65D1, 67G8, 65B7, 64A6, 65F9, 67F5, 64B10, 68C8, 67A5, 67C10, 64H6, 63F9, 67F6, 48H11, 52A8, 52F8, 49H12, 54A1, 55G9, 49C8, 52H1, 60G5.2, 49G3, 59A10, 48F8, 53B9, 56B4, 57E7, 57F11, 59C9, 58A5, 57A4, 57F9, 51G2, 56A7, 56E4, 54H10, 55D1, 48H3, 53C11, 59G10.3, 51C10.1, 59D10v1, 59D10v2, 60F9, 48B4, 52D6, 61G5, 59G10.2, 51A8, 53H5.2, 53F6, 56C11, 49A10, 48D4, 49G2, 50C12, 55G11, 52C1, 55E9, 60D7, 51C10.2, 55D3, 57B12, 52C5, 60G5.1, 55E4, 49B11, 50H10, 53C1, 56G1, 48F3, 48C9, 49A12, 51E2, 51E5, 53H5.3, 56G3.3, 55B10, 52B8, 55G5, 52H2, 56G3.2, 6E7, 57D9, 61H5, 48G4, 50G1, 58C2, 50D4, 50G5v1, 50G5v2, 51C1, 53C3.2, 54H10.3, 55A7, 55E6, 61E1, 53C3.1, 49H4, and 51E2.

The ability to compete with an antibody can be determined using any suitable assay, such as those described herein, in which antigen binding proteins 63G8, 64A8, 67B4, 68D3, 64E6, 65E8, 65F11, 67G7, 63B6, 64D4, 65C3, 68D5, 63E6, 66F7, 64H5, 65G4, 67G10v1, 67G10v2, 66B4, 66G2, 68G5, 63F5, 66F6, 65C1, 64A7, 66D4, 65B1, 67A4, 65B4, 63A10, 65H11, 64C8, 65E3, 65D4, 65D1, 67G8, 65B7, 64A6, 65F9, 67F5, 64B10, 68C8, 67A5, 67C10, 64H6, 63F9, 67F6, 48H11, 52A8, 52F8, 49H12, 54A1, 55G9, 49C8, 52H1, 60G5.2, 49G3, 59A10, 48F8, 53B9, 56B4, 57E7, 57F11, 59C9, 58A5, 57A4, 57F9, 51G2, 56A7, 56E4, 54H10, 55D1, 48H3, 53C11, 59G10.3, 51C10.1, 59D10v1, 59D10v2, 60F9, 48B4, 52D6, 61G5, 59G10.2, 51A8, 53H5.2, 53F6, 56C11, 49A10, 48D4, 49G2, 50C12, 55G11, 52C1, 55E9, 60D7, 51C10.2, 55D3, 57B12, 52C5, 60G5.1, 55E4, 49B11, 50H10, 53C1, 56G1, 48F3, 48C9, 49A12, 51E2, 51E5, 53H5.3, 56G3.3, 55B10, 52B8, 55G5, 52H2, 56G3.2, 6E7, 57D9, 61H5, 48G4, 50G1, 58C2, 50D4, 50G5v1, 50G5v2, 51C1, 53C3.2, 54H10.3, 55A7, 55E6, 61E1, 53C3.1, 49H4, and 51E2 can be used as the reference antibody.

Monoclonal Antibodies

The antigen binding proteins that are provided include monoclonal antibodies that bind to a complex comprising β-Klotho and at least one of (i) FGFR1c, (ii) FGFR2c and (iii) FGFR3c, and induce FGF21-like signaling to various degrees. Monoclonal antibodies can be produced using any technique known in the art, e.g., by immortalizing spleen cells harvested from the transgenic animal after completion of the immunization schedule. The spleen cells can be immortalized using any technique known in the art, e.g., by fusing them with myeloma cells to produce hybridomas. Myeloma cells for use in hybridoma-producing fusion procedures preferably are non-antibody-producing, have high fusion efficiency, and enzyme deficiencies that render them incapable of growing in certain selective media which support the growth of only the desired fused cells (hybridomas). Examples of suitable cell lines for use in mouse fusions include Sp-20, P3-X63/Ag8, P3-X63-Ag8.653, NS1/1.Ag 4 1, Sp210-Agl4, FO, NSO/U, MPC-11, MPC11-X45-GTG 1.7 and S194/5XXO Bul; examples of cell lines used in rat fusions include R210.RCY3, Y3-Ag 1.2.3, IR983F and 4B210. Other cell lines useful for cell fusions are U-266, GM1500-GRG2, LICR-LON-HMy2 and UC729-6.

In some instances, a hybridoma cell line is produced by immunizing an animal (e.g., a transgenic animal having human immunoglobulin sequences) with an immunogen comprising (1) cell-bound receptor of CHO transfectants expressing full length human FGFR1c and β-Klotho at the cell surface, obtained by transfecting CHO cells with cDNA encoding a human full length FGFR1c polypeptide of SEQ ID NO: 4 and cDNA encoding a human β-Klotho polypeptide of SEQ ID NO: 7 with cells incubated with FGF21 prior to freezing (as shown in Example 2); or (2) cell-bound receptor of 293T transfectants expressing full length human β-Klotho and an N-terminal truncated form of human FGFR1c encompassing amino acid residue #141 to #822 polypeptide of SEQ ID NO: 4 (as shown in Example 2); harvesting spleen cells from the immunized animal; fusing the harvested spleen cells to a myeloma cell line, thereby generating hybridoma cells; establishing hybridoma cell lines from the hybridoma cells, and identifying a hybridoma cell line that produces an antibody that binds to a complex comprising β-Klotho and at least one of (i) FGFR1c, (ii) FGFR2c and (iii) FGFR3c and can induce FGF21-like signaling (e.g., as described in Example 4). Such hybridoma cell lines, and the monoclonal antibodies produced by them, form aspects of the present disclosure.

Monoclonal antibodies secreted by a hybridoma cell line can be purified using any technique known in the art. Hybridomas or mAbs can be further screened to identify mAbs with particular properties, such as the ability to induce FGF21-like signaling. Examples of such screens are provided herein.

Chimeric and Humanized Antibodies

Chimeric and humanized antibodies based upon the foregoing sequences can readily be generated. One example is a chimeric antibody, which is an antibody composed of protein segments from different antibodies that are covalently joined to produce functional immunoglobulin light or heavy chains or immunologically functional portions thereof. Generally, a portion of the heavy chain and/or light chain is identical with or homologous to a corresponding sequence in antibodies derived from a particular species or belonging to a particular antibody class or subclass, while the remainder of the chain(s) is/are identical with or homologous to a corresponding sequence in antibodies derived from another species or belonging to another antibody class or subclass. For methods relating to chimeric antibodies, see, for example, United States Patent No. 4,816,567 ; and Morrison et al., (1985) Proc. Natl. Acad. Sci. USA 81:6851-6855, which are hereby incorporated by reference. CDR grafting is described, for example, in United States Patent No. 6,180,370 , No. 5,693,762 , No. 5,693,761 , No. 5,585,089 , and No. 5,530,101 .

Generally, a goal of making a chimeric antibody is to create a chimera in which the number of amino acids from the intended patient/recipient species is maximized. One example is the "CDR-grafted" antibody, in which the antibody comprises one or more complementarity determining regions (CDRs) from a particular species or belonging to a particular antibody class or subclass, while the remainder of the antibody chain(s) is/are identical with or homologous to a corresponding sequence in antibodies derived from another species or belonging to another antibody class or subclass. For use in humans, the variable region or selected CDRs from a rodent antibody often are grafted into a human antibody, replacing the naturally-occurring variable regions or CDRs of the human antibody.

One useful type of chimeric antibody is a "humanized" antibody. Generally, a humanized antibody is produced from a monoclonal antibody raised initially in a non-human animal. Certain amino acid residues in this monoclonal antibody, typically from non-antigen recognizing portions of the antibody, are modified to be homologous to corresponding residues in a human antibody of corresponding isotype. Humanization can be performed, for example, using various methods by substituting at least a portion of a rodent variable region for the corresponding regions of a human antibody (see, e.g., United States Patent No. 5,585,089 , and No. 5,693,762 ; Jones et al., (1986) Nature 321:522-525; Riechmann et al., (1988) Nature 332:323-27; Verhoeyen et al., (1988) Science 239:1534-1536).

In one aspect, the CDRs of the light and heavy chain variable regions of the antibodies provided herein (e.g., in Tables 3-4 and 21-23) are grafted to framework regions (FRs) from antibodies from the same, or a different, phylogenetic species. For example, the CDRs of the heavy and light chain variable regions V_H1, V_H2, V_H3, V_H4, V_H5, V_H6, V_H7, V_H8, V_H9, V_H10, V_H11, V_H12, V_H13, V_H14, V_L15, V_L16, V_L17, V_L18, V_L19, V_H20, V_H21 V_H22, V_H23, V_H24, V_H25, V_H26, V_H27, V_H28, V_H29, V_H30, V_H31, V_H32, V_H33, V_H34, V_H35, V_H36, V_H37, V_H38, V_H39, V_H40, V_H41, V_H42, V_H43, V_H44, V_H45, V_H46, V_H47, V_H48, V_H49, V_H50, V_H51, V_H52, V_H53, V_H54, V_H55, V_H56, V_H57, V_H58, V_H59, V_H60, V_H61, V_H62, V_H63, V_H64, V_H65, V_H66, V_H67, V_H68, V_H69, V_H70, V_H71, V_H72, V_H73, V_H74, V_H75, V_H76, V_H77, V_H78, V_H79, V_H80, 81, V_H82, V_H83, V_H84, V_H85, V_H 86, V_H 87, V_H88, V_H89, V_H90, V_H91, V_H92, V_H93, and V_H94 and/or V_L1, V_L2, V_L3, V_L4, V_L5, V_L6, V_L7, V_L8, V_L9, V_L10, V_L11, V_L12, V_L13, V_L14, V_L15, V_L16, V_L17, V_L18, V_L19, V_L20, V_L21, V_L22, V_L23, V_L24, V_L25, V_L26, V_L27, V_L28, V_L29, V_L30, V_L31, V_L32, V_L33, V_L34, V_L35, V_L36, V_L37, V_L38, V_L39, V_L40, V_L41, V_L42, V_L43, V_L44, V_L45, V_L46, V_L47, V_L48, V_L49, V_L50, V_L51, V_L52, V_L53, V_L54, V_L55, V_L56, V_L57, V_L58, V_L59, V_L60, V_L61, V_L62, V_L63, V_L64, V_L65, V_L66, V_L67, V_L68, V_L69, V_L70, V_L71, V_L72, V_L73, V_L74, V_L75, V_L76, V_L77, V_L78, V_L79, V_L80, V_L81, V_L82, V_L83, V_L84, V_L85, V_L86, V_L87, V_L88, V_L89, V_L90, V_L91, V_L92, V_L93, V_L94, V_L95, V_L96, V_L97, V_L98, V_L99 and V_L100 can be grafted to consensus human FRs. To create consensus human FRs, FRs from several human heavy chain or light chain amino acid sequences can be aligned to identify a consensus amino acid sequence. In other embodiments, the FRs of a heavy chain or light chain disclosed herein are replaced with the FRs from a different heavy chain or light chain. In one aspect, rare amino acids in the FRs of the heavy and light chains of an antigen binding protein (e.g., an antibody) that specifically binds to a complex comprising β-Klotho and at least one of (i) FGFR1c, (ii) FGFR2c and (iii) FGFR3c are not replaced, while the rest of the FR amino acids are replaced. A "rare amino acid" is a specific amino acid that is in a position in which this particular amino acid is not usually found in an FR. Alternatively, the grafted variable regions from the one heavy or light chain can be used with a constant region that is different from the constant region of that particular heavy or light chain as disclosed herein. In other embodiments, the grafted variable regions are part of a single chain Fv antibody.

In certain embodiments, constant regions from species other than human can be used along with the human variable region(s) to produce hybrid antibodies.

Fully Human Antibodies

Fully human antibodies are provided by the instant disclosure. Methods are available for making fully human antibodies specific for a given antigen without exposing human beings to the antigen ("fully human antibodies"). One specific means provided for implementing the production of fully human antibodies is the "humanization" of the mouse humoral immune system. Introduction of human immunoglobulin (Ig) loci into mice in which the endogenous Ig genes have been inactivated is one means of producing fully human monoclonal antibodies (mAbs) in mouse, an animal that can be immunized with any desirable antigen. Using fully human antibodies can minimize the immunogenic and allergic responses that can sometimes be caused by administering mouse or mouse-derived mAbs to humans as therapeutic agents.

Fully human antibodies can be produced by immunizing transgenic animals (typically mice) that are capable of producing a repertoire of human antibodies in the absence of endogenous immunoglobulin production. Antigens for this purpose typically have six or more contiguous amino acids, and optionally are conjugated to a carrier, such as a hapten. See, e.g., Jakobovits et al., (1993) Proc. Natl. Acad. Sci. USA 90:2551-2555; Jakobovits et al., (1993) Nature 362:255-258; and Bruggermann et al., (1993) Year in Immunol. 7:33. In one example of such a method, transgenic animals are produced by incapacitating the endogenous mouse immunoglobulin loci encoding the mouse heavy and light immunoglobulin chains therein, and inserting into the mouse genome large fragments of human genome DNA containing loci that encode human heavy and light chain proteins. Partially modified animals, which have less than the full complement of human immunoglobulin loci, are then cross-bred to obtain an animal having all of the desired immune system modifications. When administered an immunogen, these transgenic animals produce antibodies that are immunospecific for the immunogen but have human rather than murine amino acid sequences, including the variable regions. For further details of such methods, see, e.g., WO96/33735 and WO94/02602 . Additional methods relating to transgenic mice for making human antibodies are described in United States Patent No. 5,545,807 ; No. 6,713,610 ; No. 6,673,986 ; No. 6,162,963 ; No. 5,545,807 ; No. 6,300,129 ; No. 6,255,458 ; No. 5,877,397 ; No. 5,874,299 and No. 5,545,806 ; in PCT publications WO91/10741 , WO90/04036 , and in EP 546073 and EP 546073 .

According to certain embodiments, antibodies of the invention can be prepared through the utilization of a transgenic mouse that has a substantial portion of the human antibody producing genome inserted but that is rendered deficient in the production of endogenous, murine antibodies. Such mice, then, are capable of producing human immunoglobulin molecules and antibodies and are deficient in the production of murine immunoglobulin molecules and antibodies. Technologies utilized for achieving this result are disclosed in the patents, applications and references disclosed in the specification, herein. In certain embodiments, one can employ methods such as those disclosed in PCT Published Application No. WO 98/24893 or in Mendez et al., (1997) Nature Genetics, 15:146-156, which are hereby incorporated by reference for any purpose.

Generally, fully human monoclonal antibodies specific for a complex comprising β-Klotho and at least one of (i) FGFR1c, (ii) FGFR2c and (iii) FGFR1c can be produced as follows. Transgenic mice containing human immunoglobulin genes are immunized with the antigen of interest, e.g. those described herein, lymphatic cells (such as B-cells) from the mice that express antibodies are obtained. Such recovered cells are fused with a myeloid-type cell line to prepare immortal hybridoma cell lines, and such hybridoma cell lines are screened and selected to identify hybridoma cell lines that produce antibodies specific to the antigen of interest. In certain embodiments, the production of a hybridoma cell line that produces antibodies specific to a complex comprising β-Klotho and at least one of (i) FGFR1c, (ii) FGFR2c and (iii) FGFR1c is provided.

In certain embodiments, fully human antibodies can be produced by exposing human splenocytes (B or T cells) to an antigen in vitro, and then reconstituting the exposed cells in an immunocompromised mouse, e.g. SCID or nod/SCID. See, e.g., Brams et al., J.Immunol. 160: 2051-2058 (1998); Carballido et al., Nat. Med., 6: 103-106 (2000). In certain such approaches, engraftment of human fetal tissue into SCID mice (SCID-hu) results in long-term hematopoiesis and human T-cell development. See, e.g., McCune et al., Science, 241:1532-1639 (1988); Ifversen et al., Sem. Immunol., 8:243-248 (1996). In certain instances, humoral immune response in such chimeric mice is dependent on co-development of human T-cells in the animals. See, e.g., Martensson et al., Immunol., 83:1271-179 (1994). In certain approaches, human peripheral blood lymphocytes are transplanted into SCID mice. See, e.g., Mosier et al., Nature, 335:256-259 (1988). In certain such embodiments, when such transplanted cells are treated either with a priming agent, such as Staphylococcal Enterotoxin A (SEA), or with anti-human CD40 monoclonal antibodies, higher levels of B cell production is detected. See, e.g., Martensson et al., Immunol., 84: 224-230 (1995); Murphy et al., Blood, 86:1946-1953 (1995).

Thus, in certain embodiments, fully human antibodies can be produced by the expression of recombinant DNA in host cells or by expression in hybridoma cells. In other embodiments, antibodies can be produced using the phage display techniques described herein.

The antibodies described herein were prepared through the utilization of the XENOMOUSE® technology, as described herein. Such mice, then, are capable of producing human immunoglobulin molecules and antibodies and are deficient in the production of murine immunoglobulin molecules and antibodies. Technologies utilized for achieving the same are disclosed in the patents, applications, and references disclosed in the background section herein. In particular, however, a preferred embodiment of transgenic production of mice and antibodies therefrom is disclosed in U.S. Patent Application Serial No. 08/759,620, filed December 3, 1996 and International Patent Application Nos. WO 98/24893, published June 11, 1998 and WO 00/76310, published December 21, 2000 , the disclosures of which are hereby incorporated by reference. See also Mendez et al., Nature Genetics, 15:146-156 (1997), the disclosure of which is hereby incorporated by reference.

Through the use of such technology, fully human monoclonal antibodies to a variety of antigens have been produced. Essentially, XENOMOUSE® lines of mice are immunized with an antigen of interest (e.g. an antigen provided herein), lymphatic cells (such as B-cells) are recovered from the hyper-immunized mice, and the recovered lymphocytes are fused with a myeloid-type cell line to prepare immortal hybridoma cell lines. These hybridoma cell lines are screened and selected to identify hybridoma cell lines that produced antibodies specific to the antigen of interest. Provided herein are methods for the production of multiple hybridoma cell lines that produce antibodies specific to a complex comprising β-Klotho and at least one of (i) FGFR1c, (ii) FGFR2c and (iii) FGFRlc. Further, provided herein are characterization of the antibodies produced by such cell lines, including nucleotide and amino acid sequence analyses of the heavy and light chains of such antibodies.

The production of the XENOMOUSE® strains of mice is further discussed and delineated in U.S. Patent Application Serial Nos. 07/466,008, filed January 12, 1990 , 07/610,515, filed November 8, 1990 , 07/919,297, filed July 24, 1992 , 07/922,649, filed July 30, 1992 , 08/031,801, filed March 15, 1993 , 08/112,848, filed August 27, 1993 , 08/234,145, filed April 28, 1994 , 08/376,279, filed January 20, 1995 , 08/430, 938, filed April 27, 1995 , 08/464,584, filed June 5, 1995 , 08/464,582, filed June 5, 1995 , 08/463,191, filed June 5, 1995 , 08/462,837, filed June 5, 1995 , 08/486,853, filed June 5, 1995 , 08/486,857, filed June 5, 1995 , 08/486,859, filed June 5, 1995 , 08/462,513, filed June 5, 1995 , 08/724,752, filed October 2, 1996 , 08/759,620, filed December 3, 1996 , U.S. Publication 2003/0093820, filed November 30, 2001 and U.S. Patent Nos. 6,162,963 , 6,150,584 , 6,114,598 , 6,075,181 , and 5,939,598 and Japanese Patent Nos. 3 068 180 B2 , 3 068 506 B2 , and 3 068 507 B2 . See also European Patent No., EP 0 463 151 B1, grant published June 12, 1996 , International Patent Application No., WO 94/02602, published February 3, 1994 , International Patent Application No., WO 96/34096, published October 31, 1996 , WO 98/24893, published June 11, 1998 , WO 00/76310, published December 21, 2000 . The disclosures of each of the above-cited patents, applications, and references are hereby incorporated by reference in their entirety.

Using hybridoma technology, antigen-specific human mAbs with the desired specificity can be produced and selected from the transgenic mice such as those described herein. Such antibodies can be cloned and expressed using a suitable vector and host cell, or the antibodies can be harvested from cultured hybridoma cells.

Fully human antibodies can also be derived from phage-display libraries (as described in Hoogenboom et al., (1991) J. Mol. Biol. 227:381 ; and Marks et al., (1991) J. Mol. Biol. 222:581). Phage display techniques mimic immune selection through the display of antibody repertoires on the surface of filamentous bacteriophage, and subsequent selection of phage by their binding to an antigen of choice. One such technique is described in PCT Publication No. WO 99/10494 (hereby incorporated by reference), which describes the isolation of high affinity and functional agonistic antibodies for MPL- and msk-receptors using such an approach.

Bispecific or Bifunctional Antigen Binding Proteins

Also provided are bispecific and bifunctional antibodies that include one or more CDRs or one or more variable regions as described above. A bispecific or bifunctional antibody in some instances can be an artificial hybrid antibody having two different heavy/light chain pairs and two different binding sites. Bispecific antibodies can be produced by a variety of methods including, but not limited to, fusion of hybridomas or linking of Fab' fragments. See, e.g., Songsivilai & Lachmann, (1990) Clin. Exp. Immunol. 79:315-321; Kostelny et al., (1992) J. Immunol. 148:1547-1553. When an antigen binding protein of the instant disclosure binds to a complex comprising β-Klotho and at least one of (i) FGFR1c, (ii) FGFR2c and (iii) FGFR3c, the binding may lead to the activation of FGF21-like activity as measured by the FGF21-like functional and signaling assays described in Examples 4-6; when such an antigen binding protein is an antibody it is referred to as an agonistic antibody.

Various Other Forms

Some of the antigen binding proteins that specifically bind to a complex comprising β-Klotho and at least one of (i) FGFR1c, (ii) FGFR2c and (iii) FGFR3c that are provided in the present disclosure include variant forms of the antigen binding proteins disclosed herein (e.g., those having the sequences listed in Tables 1-4 and 6-23).

In various embodiments, the antigen binding proteins disclosed herein can comprise one or more non-naturally occurring/encoded amino acids. For instance, some of the antigen binding proteins have one or more non-naturally occurring/encoded amino acid substitutions in one or more of the heavy or light chains, variable regions or CDRs listed in Tables 1-23. Examples of non-naturally occurring/encoded amino acids (which can be substituted for any naturally-occurring amino acid found in any sequence disclosed herein, as desired) include: 4-hydroxyproline, γ-carboxyglutamate, ε-N,N,N-trimethyllysine, ε-N-acetyllysine, O-phosphoserine, N-acetylserine, N-formylmethionine, 3-methylhistidine, 5-hydroxylysine, σ-N-methylarginine, and other similar amino acids and imino acids (e.g., 4-hydroxyproline). In the polypeptide notation used herein, the left-hand direction is the amino terminal direction and the right-hand direction is the carboxyl-terminal direction, in accordance with standard usage and convention. A non-limiting lists of examples of non-naturally occurring/encoded amino acids that can be inserted into an antigen binding protein sequence or substituted for a wild-type residue in an antigen binding sequence include β-amino acids, homoamino acids, cyclic amino acids and amino acids with derivatized side chains. Examples include (in the L-form or D-form; abbreviated as in parentheses): citrulline (Cit), homocitrulline (hCit), Nα-methylcitrulline (NMeCit), Nα-methylhomocitrulline (Nα-MeHoCit), ornithine (Orn), Nα-Methylornithine (Nα-MeOrn or NMeOrn), sarcosine (Sar), homolysine (hLys or hK), homoarginine (hArg or hR), homoglutamine (hQ), Nα-methylarginine (NMeR), Nα-methylleucine (Nα-MeL or NMeL), N-methylhomolysine (NMeHoK), Nα-methylglutamine (NMeQ), norleucine (Nle), norvaline (Nva), 1,2,3,4-tetrahydroisoquinoline (Tic), Octahydroindole-2-carboxylic acid (Oic), 3-(1-naphthyl)alanine (1-Nal), 3-(2-naphthyl)alanine (2-Nal), 1,2,3,4-tetrahydroisoquinoline (Tic), 2-indanylglycine (IgI), para-iodophenylalanine (pI-Phe), para-aminophenylalanine (4AmP or 4-Amino-Phe), 4-guanidino phenylalanine (Guf), glycyllysine (abbreviated "K(Nε-glycyl)" or "K(glycyl)" or "K(gly)"), nitrophenylalanine (nitrophe), aminophenylalanine (aminophe or Amino-Phe), benzylphenylalanine (benzylphe), γ-carboxyglutamic acid (γ-carboxyglu), hydroxyproline (hydroxypro), p-carboxyl-phenylalanine (Cpa), α-aminoadipic acid (Aad), Nα-methyl valine (NMeVal), N-α-methyl leucine (NMeLeu), Nα-methylnorleucine (NMeNle), cyclopentylglycine (Cpg), cyclohexylglycine (Chg), acetylarginine (acetylarg), α, β-diaminopropionoic acid (Dpr), α, γ-diaminobutyric acid (Dab), diaminopropionic acid (Dap), cyclohexylalanine (Cha), 4-methyl-phenylalanine (MePhe), β, β-diphenyl-alanine (BiPhA), aminobutyric acid (Abu), 4-phenyl-phenylalanine (or biphenylalanine; 4Bip), α-amino-isobutyric acid (Aib), beta-alanine, beta-aminopropionic acid, piperidinic acid, aminocaprioic acid, aminoheptanoic acid, aminopimelic acid, desmosine, diaminopimelic acid, N-ethylglycine, N-ethylaspargine, hydroxylysine, allo-hydroxylysine, isodesmosine, allo-isoleucine, N-methylglycine, N-methylisoleucine, N-methylvaline, 4-hydroxyproline (Hyp), γ-carboxyglutamate, ε-N,N,N-trimethyllysine, ε-N-acetyllysine, O-phosphoserine, N-acetylserine, N-formylmethionine, 3-methylhistidine, 5-hydroxylysine, ω-methylarginine, 4-Amino-O-Phthalic Acid (4APA), and other similar amino acids, and derivatized forms of any of those specifically listed.

Additionally, the antigen binding proteins can have one or more conservative amino acid substitutions in one or more of the heavy or light chains, variable regions or CDRs listed in Tables 1-4 and 6-23. Naturally-occurring amino acids can be divided into classes based on common side chain properties:

1) hydrophobic: norleucine, Met, Ala, Val, Leu, Ile;
2) neutral hydrophilic: Cys, Ser, Thr, Asn, Gln;
3) acidic: Asp, Glu;
4) basic: His, Lys, Arg;
5) residues that influence chain orientation: Gly, Pro; and
6) aromatic: Trp, Tyr, Phe.

Conservative amino acid substitutions can involve exchange of a member of one of these classes with another member of the same class. Conservative amino acid substitutions can encompass non-naturally occurring/encoded amino acid residues, which are typically incorporated by chemical peptide synthesis rather than by synthesis in biological systems. See Table 8, infra. These include peptidomimetics and other reversed or inverted forms of amino acid moieties.

Non-conservative substitutions can involve the exchange of a member of one of the above classes for a member from another class. Such substituted residues can be introduced into regions of the antibody that are homologous with human antibodies, or into the non-homologous regions of the molecule.

In making such changes, according to certain embodiments, the hydropathic index of amino acids can be considered. The hydropathic profile of a protein is calculated by assigning each amino acid a numerical value ("hydropathy index") and then repetitively averaging these values along the peptide chain. Each amino acid has been assigned a hydropathic index on the basis of its hydrophobicity and charge characteristics. They are: isoleucine (+4.5); valine (+4.2); leucine (+3.8); phenylalanine (+2.8); cysteine/cystine (+2.5); methionine (+1.9); alanine (+1.8); glycine (-0.4); threonine (-0.7); serine (-0.8); tryptophan (-0.9); tyrosine (-1.3); proline (-1.6); histidine (-3.2); glutamate (-3.5); glutamine (-3.5); aspartate (-3.5); asparagine (-3.5); lysine (-3.9); and arginine (-4.5).

The importance of the hydropathic profile in conferring interactive biological function on a protein is understood in the art (see, e.g., Kyte et al., 1982, J. Mol. Biol. 157:105-131). It is known that certain amino acids can be substituted for other amino acids having a similar hydropathic index or score and still retain a similar biological activity. In making changes based upon the hydropathic index, in certain embodiments, the substitution of amino acids whose hydropathic indices are within ±2 is included. In some aspects, those which are within ±1 are included, and in other aspects, those within ±0.5 are included.

It is also understood in the art that the substitution of like amino acids can be made effectively on the basis of hydrophilicity, particularly where the biologically functional protein or peptide thereby created is intended for use in immunological embodiments, as in the present case. In certain embodiments, the greatest local average hydrophilicity of a protein, as governed by the hydrophilicity of its adjacent amino acids, correlates with its immunogenicity and antigen-binding or immunogenicity, that is, with a biological property of the protein.

The following hydrophilicity values have been assigned to these amino acid residues: arginine (+3.0); lysine (+3.0); aspartate (+3.0±1); glutamate (+3.0±1); serine (+0.3); asparagine (+0.2); glutamine (+0.2); glycine (0); threonine (-0.4); proline (-0.5±1); alanine (-0.5); histidine (-0.5); cysteine (-1.0); methionine (-1.3); valine (-1.5); leucine (-1.8); isoleucine (-1.8); tyrosine (-2.3); phenylalanine (-2.5) and tryptophan (-3.4). In making changes based upon similar hydrophilicity values, in certain embodiments, the substitution of amino acids whose hydrophilicity values are within ±2 is included, in other embodiments, those which are within ±1 are included, and in still other embodiments, those within ±0.5 are included. In some instances, one can also identify epitopes from primary amino acid sequences on the basis of hydrophilicity. These regions are also referred to as "epitopic core regions."

Exemplary conservative amino acid substitutions are set forth in Table 8.



Ala	Ser
Arg	Lys
Asn	Gln, His
Asp	Glu
Cys	Ser
Gln	Asn
Glu	Asp
Gly	Pro
His	Asn, Gln
Ile	Leu, Val
Leu	Ile, Val
Lys	Arg, Gln, Glu
Met	Leu, Ile
Phe	Met, Leu, Tyr
Ser	Thr
Thr	Ser
Trp	Tyr
Tyr	Trp, Phe
Val	Ile, Leu

A skilled artisan will be able to determine suitable variants of polypeptides as set forth herein using well-known techniques coupled with the information provided herein. One skilled in the art can identify suitable areas of the molecule that can be changed without destroying activity by targeting regions not believed to be important for activity. The skilled artisan also will be able to identify residues and portions of the molecules that are conserved among similar polypeptides. In further embodiments, even areas that can be important for biological activity or for structure can be subject to conservative amino acid substitutions without destroying the biological activity or without adversely affecting the polypeptide structure.

Additionally, one skilled in the art can review structure-function studies identifying residues in similar polypeptides that are important for activity or structure. In view of such a comparison, one can predict the importance of amino acid residues in a protein that correspond to amino acid residues important for activity or structure in similar proteins. One skilled in the art can opt for chemically similar amino acid substitutions for such predicted important amino acid residues.

One skilled in the art can also analyze the three-dimensional structure and amino acid sequence in relation to that structure in similar polypeptides. In view of such information, one skilled in the art can predict the alignment of amino acid residues of an antibody with respect to its three dimensional structure. One skilled in the art can choose not to make radical changes to amino acid residues predicted to be on the surface of the protein, since such residues can be involved in important interactions with other molecules. Moreover, one skilled in the art can generate test variants containing a single amino acid substitution at each desired amino acid residue. These variants can then be screened using assays for FGF21-like signaling, (including those described in the Examples provided herein) thus yielding information regarding which amino acids can be changed and which must not be changed. In other words, based on information gathered from such routine experiments, one skilled in the art can readily determine the amino acid positions where further substitutions should be avoided either alone or in combination with other mutations.

A number of scientific publications have been devoted to the prediction of secondary structure. See, Moult, (1996) Curr. Op. in Biotech. 7:422-427; Chou et al., (1974) Biochem. 13:222-245; Chou et al., (1974) Biochemistry 113:211-222; Chou et al., (1978) Adv. Enzymol. Relat. Areas Mol. Biol. 47:45-148; Chou et al., (1979) Ann. Rev. Biochem. 47:251-276; and Chou et al., (1979) Biophys. J. 26:367-384. Moreover, computer programs are currently available to assist with predicting secondary structure. One method of predicting secondary structure is based upon homology modeling. For example, two polypeptides or proteins that have a sequence identity of greater than 30%, or similarity greater than 40% can have similar structural topologies. The growth of the protein structural database (PDB) has provided enhanced predictability of secondary structure, including the potential number of folds within a polypeptide's or protein's structure. See, Holm et al., (1999) Nucl. Acid. Res. 27:244-247. It has been suggested (Brenner et al., (1997) Curr. Op. Struct. Biol. 7:369-376) that there are a limited number of folds in a given polypeptide or protein and that once a critical number of structures have been resolved, structural prediction will become dramatically more accurate.

Additional methods of predicting secondary structure include "threading" (Jones, (1997) Curr. Opin. Struct. Biol. 7:377-387; Sippl et al., (1996) Structure 4:15-19), "profile analysis" (Bowie et al., (1991) Science 253:164-170; Gribskov et al., (1990) Meth. Enzym. 183:146-159; Gribskov et al., (1987) Proc. Nat. Acad. Sci. 84:4355-4358), and "evolutionary linkage" (See, Holm, (1999) supra; and Brenner, (1997) supra).

In some embodiments, amino acid substitutions are made that: (1) reduce susceptibility to proteolysis, (2) reduce susceptibility to oxidation, (3) alter binding affinity for forming protein complexes, (4) alter ligand or antigen binding affinities, and/or (4) confer or modify other physicochemical or functional properties on such polypeptides. For example, single or multiple amino acid substitutions (in some embodiments, conservative amino acid substitutions) can be made in the naturally-occurring sequence. Substitutions can be made in that portion of the antibody that lies outside the domain(s) forming intermolecular contacts. In such embodiments, conservative amino acid substitutions can be used that do not substantially change the structural characteristics of the parent sequence (e.g., one or more replacement amino acids that do not disrupt the secondary structure that characterizes the parent or native antigen binding protein). Examples of art-recognized polypeptide secondary and tertiary structures are described in Creighton, Proteins: Structures and Molecular Properties 2nd edition, 1992, W. H. Freeman & Company; Creighton, Proteins: Structures and Molecular Principles, 1984, W. H. Freeman & Company; Introduction to Protein Structure (Branden and Tooze, eds.), 2nd edition, 1999, Garland Publishing; Petsko & Ringe, Protein Structure and Function, 2004, New Science Press Ltd; and Thornton et al., (1991) Nature 354:105, which are each incorporated herein by reference.

Additional preferred antibody variants include cysteine variants wherein one or more cysteine residues in the parent or native amino acid sequence are deleted from or substituted with another amino acid (e.g., serine). Cysteine variants are useful, inter alia when antibodies must be refolded into a biologically active conformation. Cysteine variants can have fewer cysteine residues than the native antibody, and typically have an even number to minimize interactions resulting from unpaired cysteines.

The heavy and light chains, variable regions domains and CDRs that are disclosed can be used to prepare polypeptides that contain an antigen binding region that can specifically bind to a complex comprising β-Klotho and at least one of (i) FGFR1c, (ii) FGFR2c and (iii) FGFR3c and may induce FGF21-like signaling. For example, one or more of the CDRs listed in Tables 3-4 and 21-23 can be incorporated into a molecule (e.g., a polypeptide) covalently or noncovalently to make an immunoadhesion. An immunoadhesion can incorporate the CDR(s) as part of a larger polypeptide chain, can covalently link the CDR(s) to another polypeptide chain, or can incorporate the CDR(s) noncovalently. The CDR(s) enable the immunoadhesion to bind specifically to a particular antigen of interest (e.g., to a complex comprising β-Klotho and at least one of (i) FGFR1c, (ii) FGFR2c and (iii) FGFR3c or an epitope thereon).

The heavy and light chains, variable regions domains and CDRs that are disclosed can be used to prepare polypeptides that contain an antigen binding region that can specifically bind to a complex comprising β-Klotho and at least one of (i) FGFR1c, (ii) FGFR2c and (iii) FGFR3c and may induce FGF21-like signaling. For example, one or more of the CDRs listed in Tables 3-4 and 21-23 can be incorporated into a molecule (e.g., a polypeptide) that is structurally similar to a "half" antibody comprising the heavy chain, the light chain of an antigen binding protein paired with a Fc fragment so that the antigen binding region is monovalent (like a Fab fragment) but with a dimeric Fc moiety.

Mimetics (e.g., "peptide mimetics" or "peptidomimetics") based upon the variable region domains and CDRs that are described herein are also provided. These analogs can be peptides, non-peptides or combinations of peptide and non-peptide regions. Fauchere, (1986) Adv. Drug Res. 15:29; Veber and Freidinger, (1985) TINS p. 392; and Evans et al., (1987) J. Med. Chem. 30:1229, which are incorporated herein by reference for any purpose. Peptide mimetics that are structurally similar to therapeutically useful peptides can be used to produce a similar therapeutic or prophylactic effect. Such compounds are often developed with the aid of computerized molecular modeling. Generally, peptidomimetics are proteins that are structurally similar to an antibody displaying a desired biological activity, such as here the ability to specifically bind to a complex comprising β-Klotho and at least one of (i) FGFR1c, (ii) FGFR2c and (iii) FGFR3c, but have one or more peptide linkages optionally replaced by a linkage selected from: -CH₂NH-, -CH₂S-, -CH₂-CH₂-, -CH-CH-(cis and trans), -COCH₂-, -CH(OH)CH₂-, and -CH₂SO-, by methods well known in the art. Systematic substitution of one or more amino acids of a consensus sequence with a D-amino acid of the same type (e.g., D-lysine in place of L-lysine) can be used in certain embodiments to generate more stable proteins. In addition, constrained peptides comprising a consensus sequence or a substantially identical consensus sequence variation can be generated by methods known in the art (Rizo and Gierasch, (1992) Ann. Rev. Biochem. 61:387), incorporated herein by reference), for example, by adding internal cysteine residues capable of forming intramolecular disulfide bridges which cyclize the peptide.

Derivatives of the antigen binding proteins that specifically bind to a complex comprising β-Klotho and at least one of (i) FGFR1c, (ii) FGFR2c and (iii) FGFR3c that are described herein are also provided. The derivatized antigen binding proteins can comprise any molecule or substance that imparts a desired property to the antibody or fragment, such as increased half-life in a particular use. The derivatized antigen binding protein can comprise, for example, a detectable (or labeling) moiety (e.g., a radioactive, colorimetric, antigenic or enzymatic molecule, a detectable bead (such as a magnetic or electrodense (e.g., gold) bead), or a molecule that binds to another molecule (e.g., biotin or streptavidin), a therapeutic or diagnostic moiety (e.g., a radioactive, cytotoxic, or pharmaceutically active moiety), or a molecule that increases the suitability of the antigen binding protein for a particular use (e.g., administration to a subject, such as a human subject, or other in vivo or in vitro uses). Examples of molecules that can be used to derivatize an antigen binding protein include albumin (e.g., human serum albumin) and polyethylene glycol (PEG). Albumin-linked and PEGylated derivatives of antigen binding proteins can be prepared using techniques well known in the art. Certain antigen binding proteins include a PEGylated single chain polypeptide as described herein. In one embodiment, the antigen binding protein is conjugated or otherwise linked to transthyretin ("TTR") or a TTR variant. The TTR or TTR variant can be chemically modified with, for example, a chemical selected from the group consisting of dextran, poly(n-vinyl pyrrolidone), polyethylene glycols, propropylene glycol homopolymers, polypropylene oxide/ethylene oxide co-polymers, polyoxyethylated polyols and polyvinyl alcohols.

Other derivatives include covalent or aggregative conjugates of the antigen binding proteins that specifically bind to a complex comprising β-Klotho and at least one of (i) FGFR1c, (ii) FGFR2c and (iii) FGFR3c that are disclosed herein with other proteins or polypeptides, such as by expression of recombinant fusion proteins comprising heterologous polypeptides fused to the N-terminus or C-terminus of an antigen binding protein that induces FGF21-like signaling. For example, the conjugated peptide can be a heterologous signal (or leader) polypeptide, e.g., the yeast alpha-factor leader, or a peptide such as an epitope tag. An antigen binding protein-containing fusion protein of the present disclosure can comprise peptides added to facilitate purification or identification of an antigen binding protein that specifically binds to a complex comprising β-Klotho and at least one of (i) FGFR1c, (ii) FGFR2c and (iii) FGFR3c (e.g., a poly-His tag) and that can induce FGF21-like signaling. An antigen binding protein that specifically binds to a complex comprising β-Klotho and at least one of (i) FGFR1c, (ii) FGFR2c and (iii) FGFR3c also can be linked to the FLAG peptide as described in Hopp et al., (1988) Bio/Technology 6:1204; and United States Patent No. 5,011,912 . The FLAG peptide is highly antigenic and provides an epitope reversibly bound by a specific monoclonal antibody (mAb), enabling rapid assay and facile purification of expressed recombinant protein. Reagents useful for preparing fusion proteins in which the FLAG peptide is fused to a given polypeptide are commercially available (Sigma, St. Louis, MO).

Multimers that comprise one or more antigen binding proteins that specifically bind to a complex comprising β-Klotho and at least one of (i) FGFR1c, (ii) FGFR2c and (iii) FGFR3c form another aspect of the present disclosure. Multimers can take the form of covalently-linked or non-covalently-linked dimers, trimers, or higher multimers. Multimers comprising two or more antigen binding proteins that bind to a complex comprising β-Klotho and at least one of (i) FGFR1c, (ii) FGFR2c and (iii) FGFR3c and which may induce FGF21-like signaling are contemplated for use as therapeutics, diagnostics and for other uses as well, with one example of such a multimer being a homodimer. Other exemplary multimers include heterodimers, homotrimers, heterotrimers, homotetramers, heterotetramers, etc.

One embodiment is directed to multimers comprising multiple antigen binding proteins that specifically bind to a complex comprising β-Klotho and at least one of (i) FGFR1c, (ii) FGFR2c and (iii) FGFR3c joined via covalent or non-covalent interactions between peptide moieties fused to an antigen binding protein that specifically binds to a complex comprising β-Klotho and at least one of (i) FGFR1c, (ii) FGFR2c and (iii) FGFR3c. Such peptides can be peptide linkers (spacers), or peptides that have the property of promoting multimerization. Leucine zippers and certain polypeptides derived from antibodies are among the peptides that can promote multimerization of antigen binding proteins attached thereto, as described in more detail herein.

In particular embodiments, the multimers comprise from two to four antigen binding proteins that bind to a complex comprising β-Klotho and at least one of (i) FGFR1c, (ii) FGFR2c and (iii) FGFR3c. The antigen binding protein moieties of the multimer can be in any of the forms described above, e.g., variants or fragments. Preferably, the multimers comprise antigen binding proteins that have the ability to specifically bind to a complex comprising β-Klotho and at least one of (i) FGFR1c, (ii) FGFR2c and (iii) FGFR3c.

In one embodiment, an oligomer is prepared using polypeptides derived from immunoglobulins. Preparation of fusion proteins comprising certain heterologous polypeptides fused to various portions of antibody-derived polypeptides (including the Fc domain) has been described, e.g., by Ashkenazi et al., (1991) Proc. Natl. Acad. Sci. USA 88:10535; Byrn et al., (1990) Nature 344:677; and Hollenbaugh et al., (1992) Current Protocols in Immunology, Suppl. 4, pages 10.19.1-10.19.11.

One embodiment comprises a dimer comprising two fusion proteins created by fusing an antigen binding protein that specifically binds to a complex comprising β-Klotho and at least one of (i) FGFR1c, (ii) FGFR2c and (iii) FGFR3c to the Fc region of an antibody. The dimer can be made by, for example, inserting a gene fusion encoding the fusion protein into an appropriate expression vector, expressing the gene fusion in host cells transformed with the recombinant expression vector, and allowing the expressed fusion protein to assemble much like antibody molecules, whereupon interchain disulfide bonds form between the Fc moieties to yield the dimer.

The term "Fc polypeptide" as used herein includes native and mutein forms of polypeptides derived from the Fc region of an antibody. Truncated forms of such polypeptides containing the hinge region that promotes dimerization also are included. Fusion proteins comprising Fc moieties (and oligomers formed therefrom) offer the advantage of facile purification by affinity chromatography over Protein A or Protein G columns.

One suitable Fc polypeptide, described in PCT application WO 93/10151 and United States Patent No. 5,426,048 and No. 5,262,522 , is a single chain polypeptide extending from the N-terminal hinge region to the native C-terminus of the Fc region of a human IgG1 antibody. Another useful Fc polypeptide is the Fc mutein described in United States Patent No. 5,457,035 , and in Baum et al., (1994) EMBO J. 13:3992-4001. The amino acid sequence of this mutein is identical to that of the native Fc sequence presented in WO 93/10151 , except that amino acid 19 has been changed from Leu to Ala, amino acid 20 has been changed from Leu to Glu, and amino acid 22 has been changed from Gly to Ala. The mutein exhibits reduced affinity for Fc receptors.

In other embodiments, the variable portion of the heavy and/or light chains of a antigen binding protein such as disclosed herein can be substituted for the variable portion of an antibody heavy and/or light chain.

Alternatively, the oligomer is a fusion protein comprising multiple antigen binding proteins that specifically bind to a complex comprising β-Klotho and at least one of (i) FGFR1c, (ii) FGFR2c and (iii) FGFR3c with or without peptide linkers (spacer peptides). Among the suitable peptide linkers are those described in United States Patent. No. 4,751,180 and No. 4,935,233 .

Another method for preparing oligomeric derivatives comprising that antigen binding proteins that specifically bind to a complex comprising β-Klotho and at least one of (i) FGFR1c, (ii) FGFR2c and (iii) FGFR3c involves use of a leucine zipper. Leucine zipper domains are peptides that promote oligomerization of the proteins in which they are found. Leucine zippers were originally identified in several DNA-binding proteins (Landschultz et al., (1988) Science 240:1759-64), and have since been found in a variety of different proteins. Among the known leucine zippers are naturally occurring peptides and derivatives thereof that dimerize or trimerize. Examples of leucine zipper domains suitable for producing soluble oligomeric proteins are described in PCT application WO 94/10308 , and the leucine zipper derived from lung surfactant protein D (SPD) described in Hoppe et al., (1994) FEBS Letters 344:191, hereby incorporated by reference. The use of a modified leucine zipper that allows for stable trimerization of a heterologous protein fused thereto is described in Fanslow et al., (1994) Semin. Immunol. 6:267-278. In one approach, recombinant fusion proteins comprising an antigen binding protein fragment or derivative that specifically binds to a complex comprising β-Klotho and an FGFR (e.g., FGFR1c, FGFR2c or FGFR3c) is fused to a leucine zipper peptide are expressed in suitable host cells, and the soluble oligomeric antigen binding protein fragments or derivatives that form are recovered from the culture supernatant.

In certain embodiments, the antigen binding protein has a K_D (equilibrium binding affinity) of less than 1 pM, 10 pM, 100 pM, 1 nM, 2 nM, 5 nM, 10 nM, 25 nM or 50 nM.

In another aspect the instant disclosure provides an antigen binding protein having a half-life of at least one day in vitro or in vivo (e.g., when administered to a human subject). In one embodiment, the antigen binding protein has a half-life of at least three days. In another embodiment, the antibody or portion thereof has a half-life of four days or longer. In another embodiment, the antibody or portion thereof has a half-life of eight days or longer. In another embodiment, the antibody or portion thereof has a half-life of ten days or longer. In another embodiment, the antibody or portion thereof has a half-life of eleven days or longer. In another embodiment, the antibody or portion thereof has a half-life of fifteen days or longer. In another embodiment, the antibody or antigen-binding portion thereof is derivatized or modified such that it has a longer half-life as compared to the underivatized or unmodified antibody. In another embodiment, an antigen binding protein that specifically binds to a complex comprising β-Klotho and at least one of (i) FGFR1c, (ii) FGFR2c and (iii) FGFR3c contains point mutations to increase serum half life, such as described in WO 00/09560, published Feb. 24, 2000 , incorporated by reference.

Glycosylation

An antigen binding protein that specifically binds to a complex comprising β-Klotho and at least one of (i) FGFR1c, (ii) FGFR2c and (iii) FGFR3c can have a glycosylation pattern that is different or altered from that found in the native species. As is known in the art, glycosylation patterns can depend on both the sequence of the protein (e.g., the presence or absence of particular glycosylation amino acid residues, discussed below), or the host cell or organism in which the protein is produced. Particular expression systems are discussed below.

Glycosylation of polypeptides is typically either N-linked or O-linked. N-linked refers to the attachment of the carbohydrate moiety to the side chain of an asparagine residue. The tri-peptide sequences asparagine-X-serine and asparagine-X-threonine, where X is any amino acid except proline, are the recognition sequences for enzymatic attachment of the carbohydrate moiety to the asparagine side chain. Thus, the presence of either of these tri-peptide sequences in a polypeptide creates a potential glycosylation site. O-linked glycosylation refers to the attachment of one of the sugars N-acetylgalactosamine, galactose, or xylose, to a hydroxyamino acid, most commonly serine or threonine, although 5-hydroxyproline or 5-hydroxylysine can also be used.

Addition of glycosylation sites to the antigen binding protein is conveniently accomplished by altering the amino acid sequence such that it contains one or more of the above-described tri-peptide sequences (for N-linked glycosylation sites). The alteration can also be made by the addition of, or substitution by, one or more serine or threonine residues to the starting sequence (for O-linked glycosylation sites). For ease, the antigen binding protein amino acid sequence can be altered through changes at the DNA level, particularly by mutating the DNA encoding the target polypeptide at preselected bases such that codons are generated that will translate into the desired amino acids.

Another means of increasing the number of carbohydrate moieties on the antigen binding protein is by chemical or enzymatic coupling of glycosides to the protein. These procedures are advantageous in that they do not require production of the protein in a host cell that has glycosylation capabilities for N- and O-linked glycosylation. Depending on the coupling mode used, the sugar(s) can be attached to (a) arginine and histidine; (b) free carboxyl groups; (c) free sulfhydryl groups such as those of cysteine; (d) free hydroxyl groups such as those of serine, threonine, or hydroxyproline; (e) aromatic residues such as those of phenylalanine, tyrosine, or tryptophan; or (f) the amide group of glutamine. These methods are described in WO 87/05330 and in Aplin & Wriston, (1981) CRC Crit. Rev. Biochem. 10:259-306.

Removal of carbohydrate moieties present on the starting antigen binding protein can be accomplished chemically or enzymatically. Chemical deglycosylation requires exposure of the protein to the compound trifluoromethanesulfonic acid, or an equivalent compound. This treatment results in the cleavage of most or all sugars except the linking sugar (N-acetylglucosamine or N-acetylgalactosamine), while leaving the polypeptide intact. Chemical deglycosylation is described by Hakimuddin et al., (1987) Arch. Biochem. Biophys. 259:52-57 and by Edge et al., (1981) Anal. Biochem. 118:131-37. Enzymatic cleavage of carbohydrate moieties on polypeptides can be achieved by the use of a variety of endo- and exo-glycosidases as described by Thotakura et al., (1987) Meth. Enzymol. 138:350-59. Glycosylation at potential glycosylation sites can be prevented by the use of the compound tunicamycin as described by Duskin et al., (1982) J. Biol. Chem. 257:3105-09. Tunicamycin blocks the formation of protein-N-glycoside linkages.

Hence, aspects of the present disclosure include glycosylation variants of antigen binding proteins that specifically bind to a complex comprising β-Klotho and at least one of (i) FGFR1c, (ii) FGFR2c and (iii) FGFR3c wherein the number and/or type of glycosylation site(s) has been altered compared to the amino acid sequences of the parent polypeptide. In certain embodiments, antibody protein variants comprise a greater or a lesser number of N-linked glycosylation sites than the native antibody. An N-linked glycosylation site is characterized by the sequence: Asn-X-Ser or Asn-X-Thr, wherein the amino acid residue designated as X can be any amino acid residue except proline. The substitution of amino acid residues to create this sequence provides a potential new site for the addition of an N-linked carbohydrate chain. Alternatively, substitutions that eliminate or alter this sequence will prevent addition of an N-linked carbohydrate chain present in the native polypeptide. For example, the glycosylation can be reduced by the deletion of an Asn or by substituting the Asn with a different amino acid. In other embodiments, one or more new N-linked sites are created. Antibodies typically have a N-linked glycosylation site in the Fc region.

Labels and Effector Groups

In some embodiments, an antigen binding protein that specifically binds to a complex comprising β-Klotho and at least one of (i) FGFR1c, (ii) FGFR2c and (iii) FGFR3c comprises one or more labels. The term "labeling group" or "label" means any detectable label. Examples of suitable labeling groups include, but are not limited to, the following: radioisotopes or radionuclides (e.g., ³H, ¹⁴C, ¹⁵N, ³⁵S , ⁹⁰Y, ⁹⁹Tc, ¹¹¹In, ¹²⁵I, ¹³¹I), fluorescent groups (e.g., FITC, rhodamine, lanthanide phosphors), enzymatic groups (e.g., horseradish peroxidase, β-galactosidase, luciferase, alkaline phosphatase), chemiluminescent groups, biotinyl groups, or predetermined polypeptide epitopes recognized by a secondary reporter (e.g., leucine zipper pair sequences, binding sites for secondary antibodies, metal binding domains, epitope tags). In some embodiments, the labeling group is coupled to the antigen binding protein via spacer arms of various lengths to reduce potential steric hindrance. Various methods for labeling proteins are known in the art and can be used as is seen fit.

The term "effector group" means any group coupled to an antigen binding protein that specifically binds to a complex comprising β-Klotho and at least one of (i) FGFR1c, (ii) FGFR2c and (iii) FGFR3c and that acts as a cytotoxic agent. Examples for suitable effector groups are radioisotopes or radionuclides (e.g., ³H, ¹⁴C, ¹⁵N, ³⁵S, ⁹⁰Y, ⁹⁹Tc, ¹¹¹In, ¹²⁵I, ¹³¹I). Other suitable groups include toxins, therapeutic groups, or chemotherapeutic groups. Examples of suitable groups include calicheamicin, auristatins, geldanamycin and cantansine. In some embodiments, the effector group is coupled to the antigen binding protein via spacer arms of various lengths to reduce potential steric hindrance.

In general, labels fall into a variety of classes, depending on the assay in which they are to be detected: a) isotopic labels, which can be radioactive or heavy isotopes; b) magnetic labels (e.g., magnetic particles); c) redox active moieties; d) optical dyes; enzymatic groups (e.g. horseradish peroxidase, β-galactosidase, luciferase, alkaline phosphatase); e) biotinylated groups; and f) predetermined polypeptide epitopes recognized by a secondary reporter (e.g., leucine zipper pair sequences, binding sites for secondary antibodies, metal binding domains, epitope tags, etc.). In some embodiments, the labeling group is coupled to the antigen binding protein via spacer arms of various lengths to reduce potential steric hindrance. Various methods for labeling proteins are known in the art.

Specific labels include optical dyes, including, but not limited to, chromophores, phosphors and fluorophores, with the latter being specific in many instances. Fluorophores can be either "small molecule" fluores, or proteinaceous fluores.

By "fluorescent label" is meant any molecule that can be detected via its inherent fluorescent properties. Suitable fluorescent labels include, but are not limited to, fluorescein, rhodamine, tetramethylrhodamine, eosin, erythrosin, coumarin, methyl-coumarins, pyrene, Malacite green, stilbene, Lucifer Yellow, Cascade Blue, Texas Red, IAEDANS, EDANS, BODIPY FL, LC Red 640, Cy 5, Cy 5.5, LC Red 705, Oregon green, the Alexa-Fluor dyes (Alexa Fluor 350, Alexa Fluor 430, Alexa Fluor 488, Alexa Fluor 546, Alexa Fluor 568, Alexa Fluor 594, Alexa Fluor 633, Alexa Fluor 647, Alexa Fluor 660, Alexa Fluor 680), Cascade Blue, Cascade Yellow and R-phycoerythrin (PE) (Molecular Probes, Eugene, OR), FITC, Rhodamine, and Texas Red (Pierce, Rockford, IL), Cy5, Cy5.5, Cy7 (Amersham Life Science, Pittsburgh, PA). Suitable optical dyes, including fluorophores, are described in Molecular Probes Handbook by Richard P. Haugland and in subsequent editions, including Molecular Probes Handbook, A Guide to Fluorescent Probes and Labeling Technologies, 11th edition, Johnson and Spence (eds), hereby expressly incorporated by reference.

Suitable proteinaceous fluorescent labels also include, but are not limited to, green fluorescent protein, including a Renilla, Ptilosarcus, or Aequorea species of GFP (Chalfie et al., (1994) Science 263:802-805), eGFP (Clontech Labs., Inc., Genbank Accession Number U55762), blue fluorescent protein (BFP, Quantum Biotechnologies, Inc., Quebec, Canada; Stauber, (1998) Biotechniques 24:462-71; Heim et al., (1996) Curr. Biol. 6:178-82), enhanced yellow fluorescent protein (EYFP, Clontech Labs., Inc.), luciferase (Ichiki et al., (1993) J. Immunol. 150:5408-17), β-galactosidase (Nolan et al., (1988) Proc. Natl. Acad. Sci. U.S.A. 85:2603-07) and Renilla (WO92/15673 , WO95/07463 , WO98/14605 , WO98/26277 , WO99/49019 , United States Patent Nos. 5292658 , 5418155 , 5683888 , 5741668 , 5777079 , 5804387 , 5874304 , 5876995 and 5925558 ).

Preparing Of Antigen Binding Proteins

Non-human antibodies that are provided can be, for example, derived from any antibody-producing animal, such as a mouse, rat, rabbit, goat, donkey, or non-human primate (such as a monkey, (e.g., cynomolgus or rhesus monkey) or an ape (e.g., chimpanzee)). Non-human antibodies can be used, for instance, in in vitro cell culture and cell-culture based applications, or any other application where an immune response to the antibody does not occur or is insignificant, can be prevented, is not a concern, or is desired. In certain embodiments, the antibodies can be produced by immunizing with cell bound receptor from CHO transfectants expressing full length human FGFR1c and β-Klotho at the cell surface following incubated with FGF21; or with cell bound receptor of 293T transfectants expressing full length human β-Klotho and an N-terminal truncated version of human FGFR1c encompassing amino acid residues 141 to 822 of the polypeptide of SEQ ID NO: 4; or with full-length β-Klotho, FGFR1c, FGFR2c or FGFR3c; or with the extracellular domain of β-Klotho, FGFR1c, FGFR2c or FGFR3c; or with two of β-Klotho, FGFR1c, FGFR2c, and FGFR3c; or with whole cells expressing FGFR1c, β-Klotho or both FGFR1c and β-Klotho; or with membranes prepared from cells expressing FGFR1c, β-Klotho or both FGFR1c and β-Klotho; or with fusion proteins, e.g., Fc fusions comprising FGFR1c, β-Klotho or FGFR1c and β-Klotho (or extracellular domains thereof) fused to Fc, and other methods known in the art, e.g., as described in the Examples presented herein. Alternatively, the certain non-human antibodies can be raised by immunizing with amino acids which are segments of one or more of β-Klotho, FGFR1c, FGFR2c or FGFR3c that form part of the epitope to which certain antibodies provided herein bind. The antibodies can be polyclonal, monoclonal, or can be synthesized in host cells by expressing recombinant DNA.

Fully human antibodies can be prepared as described above by immunizing transgenic animals containing human immunoglobulin loci or by selecting a phage display library that is expressing a repertoire of human antibodies.

The monoclonal antibodies (mAbs) can be produced by a variety of techniques, including conventional monoclonal antibody methodology, e.g., the standard somatic cell hybridization technique of Kohler & Milstein, (1975) Nature 256:495-97. Alternatively, other techniques for producing monoclonal antibodies can be employed, for example, the viral or oncogenic transformation of B-lymphocytes. One suitable animal system for preparing hybridomas is the murine system, which is a very well established procedure. Immunization protocols and techniques for isolation of immunized splenocytes for fusion are known in the art. For such procedures, B cells from immunized mice are fused with a suitable immortalized fusion partner, such as a murine myeloma cell line. If desired, rats or other mammals besides can be immunized instead of mice and B cells from such animals can be fused with the murine myeloma cell line to form hybridomas. Alternatively, a myeloma cell line from a source other than mouse can be used. Fusion procedures for making hybridomas also are well known. SLAM technology can also be employed in the production of antibodies.

The single chain antibodies that are provided can be formed by linking heavy and light chain variable domain (Fv region) fragments via an amino acid bridge (short peptide linker), resulting in a single polypeptide chain. Such single-chain Fvs (scFvs) can be prepared by fusing DNA encoding a peptide linker between DNAs encoding the two variable domain polypeptides (V_L and V_H). The resulting polypeptides can fold back on themselves to form antigen-binding monomers, or they can form multimers (e.g., dimers, trimers, or tetramers), depending on the length of a flexible linker between the two variable domains (Kortt et al., (1997) Prot. Eng. 10:423; Kortt et al., (2001) Biomol. Eng. 18:95-108). By combining different V_L and V_H-comprising polypeptides, one can form multimeric scFvs that bind to different epitopes (Kriangkum et al., (2001) Biomol. Eng. 18:31-40). Techniques developed for the production of single chain antibodies include those described in U.S. Pat. No. 4,946,778 ; Bird et al., (1988) Science 242:423-26; Huston et al., (1988) Proc. Natl. Acad. Sci. U.S.A. 85:5879-83; Ward et al., (1989) Nature 334:544-46, de Graaf et al., (2002) Methods Mol Biol. 178:379-387. Single chain antibodies derived from antibodies provided herein include, but are not limited to scFvs comprising the variable domain combinations of the heavy and light chain variable regions depicted in Table 2, or combinations of light and heavy chain variable domains which include the CDRs depicted in Tables 3-4 and 6-23.

Antibodies provided herein that are of one subclass can be changed to antibodies from a different subclass using subclass switching methods. Thus, IgG antibodies can be derived from an IgM antibody, for example, and vice versa. Such techniques allow the preparation of new antibodies that possess the antigen binding properties of a given antibody (the parent antibody), but also exhibit biological properties associated with an antibody isotype or subclass different from that of the parent antibody. Recombinant DNA techniques can be employed. Cloned DNA encoding particular antibody polypeptides can be employed in such procedures, e.g., DNA encoding the constant domain of an antibody of the desired isotype. See, e.g., Lantto et al., (2002) Methods Mol. Biol. 178:303-16.

Accordingly, the antibodies that are provided include those comprising, for example, the variable domain combinations described, supra., having a desired isotype (for example, IgA, IgG1, IgG2, IgG3, IgG4, IgE, and IgD) as well as Fab or F(ab')₂ fragments thereof. Moreover, if an IgG4 is desired, it can also be desired to introduce a point mutation (e.g., a mutation from CPSCP to CPPCP (SEQ ID NOs 1828 and 1829, respectively, in order of appearance) in the hinge region as described in Bloom et al., (1997) Protein Science 6:407-15, incorporated by reference herein) to alleviate a tendency to form intra-H chain disulfide bonds that can lead to heterogeneity in the IgG4 antibodies.

Moreover, techniques for deriving antibodies having different properties (i.e., varying affinities for the antigen to which they bind) are also known. One such technique, referred to as chain shuffling, involves displaying immunoglobulin variable domain gene repertoires on the surface of filamentous bacteriophage, often referred to as phage display. Chain shuffling has been used to prepare high affinity antibodies to the hapten 2-phenyloxazol-5-one, as described by Marks et al., (1992) Nature Biotechnology 10:779-83.

Conservative modifications can be made to the heavy and light chain variable regions described in Table 2, or the CDRs described in Tables 3A and 3B, 4A and 4B, and Tables 6-23 (and corresponding modifications to the encoding nucleic acids) to produce an antigen binding protein having functional and biochemical characteristics. Methods for achieving such modifications are described herein.

Antigen binding proteins that specifically bind to a complex comprising β-Klotho and at least one of (i) FGFR1c, (ii) FGFR2c and (iii) FGFR3c can be further modified in various ways. For example, if they are to be used for therapeutic purposes, they can be conjugated with polyethylene glycol (PEGylated) to prolong the serum half-life or to enhance protein delivery. PEG can be attached directely to the antigen binding protein or it can be attached via a linker, such as a glycosidic linkage.

Alternatively, the V region of the subject antibodies or fragments thereof can be fused with the Fc region of a different antibody molecule. The Fc region used for this purpose can be modified so that it does not bind complement, thus reducing the likelihood of inducing cell lysis in the patient when the fusion protein is used as a therapeutic agent. In addition, the subject antibodies or functional fragments thereof can be conjugated with human serum albumin to enhance the serum half-life of the antibody or fragment thereof. Another useful fusion partner for the antigen binding proteins or fragments thereof is transthyretin (TTR). TTR has the capacity to form a tetramer, thus an antibody-TTR fusion protein can form a multivalent antibody which can increase its binding avidity.

Alternatively, substantial modifications in the functional and/or biochemical characteristics of the antigen binding proteins described herein can be achieved by creating substitutions in the amino acid sequence of the heavy and light chains that differ significantly in their effect on maintaining (a) the structure of the molecular backbone in the area of the substitution, for example, as a sheet or helical conformation, (b) the charge or hydrophobicity of the molecule at the target site, or (c) the bulkiness of the side chain. A "conservative amino acid substitution" can involve a substitution of a native amino acid residue with a nonnative residue that has little or no effect on the polarity or charge of the amino acid residue at that position. See, Table 8, supra. Furthermore, any native residue in the polypeptide can also be substituted with alanine, as has been previously described for alanine scanning mutagenesis.

Amino acid substitutions (whether conservative or non-conservative) of the subject antibodies can be implemented by those skilled in the art by applying routine techniques. Amino acid substitutions can be used to identify important residues of the antibodies provided herein, or to increase or decrease the affinity of these antibodies for a complex comprising β-Klotho and at least one of (i) FGFR1c, (ii) FGFR2c and (iii) FGFR3c or for modifying the binding affinity of other antigen-binding proteins described herein.

Methods of Expressing Antigen Binding Proteins

Expression systems and constructs in the form of plasmids, expression vectors, transcription or expression cassettes that comprise at least one polynucleotide as described above are also provided herein, as well host cells comprising such expression systems or constructs.

The antigen binding proteins provided herein can be prepared by any of a number of conventional techniques. For example, antigen binding proteins that specifically bind to a complex comprising β-Klotho and at least one of (i) FGFR1c, (ii) FGFR2c and (iii) FGFR3c can be produced by recombinant expression systems, using any technique known in the art. See, e.g., Monoclonal Antibodies, Hybridomas: A New Dimension in Biological Analyses, (Kennet et al., eds.) Plenum Press (1980) and subsequent editions; and Harlow & Lane, (1988) supra.

Antigen binding proteins can be expressed in hybridoma cell lines (e.g., in particular antibodies can be expressed in hybridomas) or in cell lines other than hybridomas. Expression constructs encoding the antibodies can be used to transform a mammalian, insect or microbial host cell. Transformation can be performed using any known method for introducing polynucleotides into a host cell, including, for example packaging the polynucleotide in a virus or bacteriophage and transducing a host cell with the construct by transfection procedures known in the art, as exemplified by United States Patent Nos. 4,399,216 ; 4,912,040 ; 4,740,461 ; and 4,959,455 . The optimal transformation procedure used will depend upon which type of host cell is being transformed. Methods for introduction of heterologous polynucleotides into mammalian cells are well known in the art and include, but are not limited to, dextran-mediated transfection, calcium phosphate precipitation, polybrene mediated transfection, protoplast fusion, electroporation, encapsulation of the polynucleotide(s) in liposomes, mixing nucleic acid with positively-charged lipids, and direct microinjection of the DNA into nuclei.

Recombinant expression constructs typically comprise a nucleic acid molecule encoding a polypeptide comprising one or more of the following: one or more CDRs provided herein; a light chain constant region; a light chain variable region; a heavy chain constant region (e.g., C_H1, C_H2 and/or C_H3); and/or another scaffold portion of an antigen binding protein. These nucleic acid sequences are inserted into an appropriate expression vector using standard ligation techniques. In one embodiment, the heavy or light chain constant region is appended to the C-terminus of the anti-β-Klotho/FGFR (e.g., FGFR1c, FGFR2c or FGFR3c) complex-specific heavy or light chain variable region and is ligated into an expression vector. The vector is typically selected to be functional in the particular host cell employed (i.e., the vector is compatible with the host cell machinery, permitting amplification and/or expression of the gene can occur). In some embodiments, vectors are used that employ protein-fragment complementation assays using protein reporters, such as dihydrofolate reductase (see, for example, U.S. Patent No. 6,270,964 , which is hereby incorporated by reference). Suitable expression vectors can be purchased, for example, from Invitrogen Life Technologies or BD Biosciences. Other useful vectors for cloning and expressing the antibodies and fragments include those described in Bianchi and McGrew, (2003) Biotech. Biotechnol. Bioeng. 84:439-44, which is hereby incorporated by reference. Additional suitable expression vectors are discussed, for example, in "Gene Expression Technology," Methods Enzymol., vol. 185, (Goeddel et al., ed.), (1990), Academic Press.

Typically, expression vectors used in any of the host cells will contain sequences for plasmid maintenance and for cloning and expression of exogenous nucleotide sequences. Such sequences, collectively referred to as "flanking sequences" in certain embodiments will typically include one or more of the following nucleotide sequences: a promoter, one or more enhancer sequences, an origin of replication, a transcriptional termination sequence, a complete intron sequence containing a donor and acceptor splice site, a sequence encoding a leader sequence for polypeptide secretion, a ribosome binding site, a polyadenylation sequence, a polylinker region for inserting the nucleic acid encoding the polypeptide to be expressed, and a selectable marker element.

Optionally, an expression vector can contain a "tag"-encoding sequence, i.e., an oligonucleotide molecule located at the 5' or 3' end of an antigen binding protein coding sequence; the oligonucleotide sequence encodes polyHis (such as hexaHis, HHHHHH (SEQ ID NO: 1830)), or another "tag" such as FLAG, HA (hemaglutinin influenza virus), or myc, for which commercially available antibodies exist. This tag is typically fused to the polypeptide upon expression of the polypeptide, and can serve as a means for affinity purification or detection of the antigen binding protein from the host cell. Affinity purification can be accomplished, for example, by column chromatography using antibodies against the tag as an affinity matrix. Optionally, the tag can subsequently be removed from the purified antigen binding protein by various means such as using certain peptidases for cleavage.

Flanking sequences can be homologous (i.e., from the same species and/or strain as the host cell), heterologous (i.e., from a species other than the host cell species or strain), hybrid (i.e., a combination of flanking sequences from more than one source), synthetic or native. As such, the source of a flanking sequence can be any prokaryotic or eukaryotic organism, any vertebrate or invertebrate organism, or any plant, provided that the flanking sequence is functional in, and can be activated by, the host cell machinery.

Flanking sequences useful in the vectors can be obtained by any of several methods well known in the art. Typically, flanking sequences useful herein will have been previously identified by mapping and/or by restriction endonuclease digestion and can thus be isolated from the proper tissue source using the appropriate restriction endonucleases. In some cases, the full nucleotide sequence of a flanking sequence can be known. Here, the flanking sequence can be synthesized using the methods described herein for nucleic acid synthesis or cloning.

Whether all or only a portion of the flanking sequence is known, it can be obtained using polymerase chain reaction (PCR) and/or by screening a genomic library with a suitable probe such as an oligonucleotide and/or flanking sequence fragment from the same or another species. Where the flanking sequence is not known, a fragment of DNA containing a flanking sequence can be isolated from a larger piece of DNA that can contain, for example, a coding sequence or even another gene or genes. Isolation can be accomplished by restriction endonuclease digestion to produce the proper DNA fragment followed by isolation using agarose gel purification, column chromatography or other methods known to the skilled artisan. The selection of suitable enzymes to accomplish this purpose will be readily apparent to one of ordinary skill in the art.

An origin of replication is typically a part of those prokaryotic expression vectors purchased commercially, and the origin aids in the amplification of the vector in a host cell. If the vector of choice does not contain an origin of replication site, one can be chemically synthesized based on a known sequence, and ligated into the vector. For example, the origin of replication from the plasmid pBR322 (GenBank Accession # J01749, New England Biolabs, Beverly, MA) is suitable for most gram-negative bacteria, and various viral origins (e.g., SV40, polyoma, adenovirus, vesicular stomatitus virus (VSV), or papillomaviruses such as HPV or BPV) are useful for cloning vectors in mammalian cells. Generally, the origin of replication component is not needed for mammalian expression vectors (for example, the SV40 origin is often used only because it also contains the virus early promoter).

A transcription termination sequence is typically located 3' to the end of a polypeptide coding region and serves to terminate transcription. Usually, a transcription termination sequence in prokaryotic cells is a G-C rich fragment followed by a poly-T sequence. While the sequence is easily cloned from a library or even purchased commercially as part of a vector, it can also be readily synthesized using methods for nucleic acid synthesis such as those described herein.

A selectable marker gene encodes a protein necessary for the survival and growth of a host cell grown in a selective culture medium. Typical selection marker genes encode proteins that (a) confer resistance to antibiotics or other toxins, e.g., ampicillin, tetracycline, or kanamycin for prokaryotic host cells; (b) complement auxotrophic deficiencies of the cell; or (c) supply critical nutrients not available from complex or defined media. Specific selectable markers are the kanamycin resistance gene, the ampicillin resistance gene, and the tetracycline resistance gene. Advantageously, a neomycin resistance gene can also be used for selection in both prokaryotic and eukaryotic host cells.

Other selectable genes can be used to amplify the gene that will be expressed. Amplification is the process wherein genes that are required for production of a protein critical for growth or cell survival are reiterated in tandem within the chromosomes of successive generations of recombinant cells. Examples of suitable selectable markers for mammalian cells include dihydrofolate reductase (DHFR) and promoterless thymidine kinase genes. Mammalian cell transformants are placed under selection pressure wherein only the transformants are uniquely adapted to survive by virtue of the selectable gene present in the vector. Selection pressure is imposed by culturing the transformed cells under conditions in which the concentration of selection agent in the medium is successively increased, thereby leading to the amplification of both the selectable gene and the DNA that encodes another gene, such as an antigen binding protein that binds to a complex comprising β-Klotho and at least one of (i) FGFR1c, (ii) FGFR2c and (iii) FGFR3c. As a result, increased quantities of a polypeptide such as an antigen binding protein are synthesized from the amplified DNA.

A ribosome-binding site is usually necessary for translation initiation of mRNA and is characterized by a Shine-Dalgarno sequence (prokaryotes) or a Kozak sequence (eukaryotes). The element is typically located 3' to the promoter and 5' to the coding sequence of the polypeptide to be expressed.

In some cases, such as where glycosylation is desired in a eukaryotic host cell expression system, one can manipulate the various pre- or pro-sequences to improve glycosylation or yield. For example, one can alter the peptidase cleavage site of a particular signal peptide, or add prosequences, which also can affect glycosylation. The final protein product can have, in the -1 position (relative to the first amino acid of the mature protein), one or more additional amino acids incident to expression, which may not have been totally removed. For example, the final protein product can have one or two amino acid residues found in the peptidase cleavage site, attached to the amino-terminus. Alternatively, use of some enzyme cleavage sites can result in a slightly truncated form of the desired polypeptide, if the enzyme cuts at such area within the mature polypeptide.

Expression and cloning will typically contain a promoter that is recognized by the host organism and operably linked to the molecule encoding an antigen binding protein that specifically binds to a complex comprising β-Klotho and at least one of (i) FGFR1c, (ii) FGFR2c and (iii) FGFR3c. Promoters are untranscribed sequences located upstream (i.e., 5') to the start codon of a structural gene (generally within about 100 to 1000 bp) that control transcription of the structural gene. Promoters are conventionally grouped into one of two classes: inducible promoters and constitutive promoters. Inducible promoters initiate increased levels of transcription from DNA under their control in response to some change in culture conditions, such as the presence or absence of a nutrient or a change in temperature. Constitutive promoters, on the other hand, uniformly transcribe a gene to which they are operably linked, that is, with little or no control over gene expression. A large number of promoters, recognized by a variety of potential host cells, are well known. A suitable promoter is operably linked to the DNA encoding heavy chain or light chain comprising an antigen binding protein by removing the promoter from the source DNA by restriction enzyme digestion and inserting the desired promoter sequence into the vector.

Suitable promoters for use with yeast hosts are also well known in the art. Yeast enhancers are advantageously used with yeast promoters. Suitable promoters for use with mammalian host cells are well known and include, but are not limited to, those obtained from the genomes of viruses such as polyoma virus, fowlpox virus, adenovirus (such as Adenovirus 2), bovine papilloma virus, avian sarcoma virus, cytomegalovirus, retroviruses, hepatitis-B virus, and Simian Virus 40 (SV40). Other suitable mammalian promoters include heterologous mammalian promoters, for example, heat-shock promoters and the actin promoter.

Additional promoters which can be of interest include, but are not limited to: SV40 early promoter (Benoist & Chambon, (1981) Nature 290:304-310); CMV promoter (Thornsen et al., (1984) Proc. Natl. Acad. U.S.A. 81:659-663); the promoter contained in the 3' long terminal repeat of Rous sarcoma virus (Yamamoto et al., (1980) Cell 22:787-97); herpes thymidine kinase promoter (Wagner et al., (1981) Proc. Natl. Acad. Sci. U.S.A. 78:1444-45); promoter and regulatory sequences from the metallothionine gene (Prinster et al., (1982) Nature 296:39-42); and prokaryotic promoters such as the beta-lactamase promoter (Villa-Kamaroff et al., (1978) Proc. Natl. Acad. Sci. U.S.A. 75:3727-31); or the tac promoter (DeBoer et al., (1983) Proc. Natl. Acad. Sci. U.S.A. 80:21-25). Also of interest are the following animal transcriptional control regions, which exhibit tissue specificity and have been utilized in transgenic animals: the elastase I gene control region that is active in pancreatic acinar cells (Swift et al., (1984) Cell 38:639-46; Ornitz et al., (1986) Cold Spring Harbor Symp. Quant. Biol. 50:399-409; MacDonald, (1987) Hepatology 7:425-515); the insulin gene control region that is active in pancreatic beta cells (Hanahan, (1985) Nature 315:115-22); the immunoglobulin gene control region that is active in lymphoid cells (Grosschedl et al., (1984) Cell 38:647-58; Adames et al., (1985) Nature 318:533-38; Alexander et al., (1987) Mol. Cell. Biol. 7:1436-44); the mouse mammary tumor virus control region that is active in testicular, breast, lymphoid and mast cells (Leder et al., (1986) Cell 45:485-95); the albumin gene control region that is active in liver (Pinkert et al., (1987) Genes and Devel. 1:268-76); the alpha-feto-protein gene control region that is active in liver (Krumlauf et al., (1985) Mol. Cell. Biol. 5:1639-48; Hammer et al., (1987) Science 253:53-58); the alpha 1-antitrypsin gene control region that is active in liver (Kelsey et al., (1987) Genes and Devel. 1:161-71); the beta-globin gene control region that is active in myeloid cells (Mogram et al., (1985) Nature 315:338-40; Kollias et al., (1986) Cell 46:89-94); the myelin basic protein gene control region that is active in oligodendrocyte cells in the brain (Readhead et al., (1987) Cell 48:703-12); the myosin light chain-2 gene control region that is active in skeletal muscle (Sani, (1985) Nature 314:283-86); and the gonadotropic releasing hormone gene control region that is active in the hypothalamus (Mason et al., (1986) Science 234:1372-78).

An enhancer sequence can be inserted into the vector to increase transcription of DNA encoding light chain or heavy chain comprising an antigen binding protein that specifically binds to a complex comprising β-Klotho and at least one of (i) FGFR1c, (ii) FGFR2c and (iii) FGFR3c by higher eukaryotes, e.g., a human antigen binding protein that specifically binds to a complex comprising β-Klotho and at least one of (i) FGFR1c, (ii) FGFR2c and (iii) FGFR3c. Enhancers are cis-acting elements of DNA, usually about 10-300 bp in length, that act on the promoter to increase transcription. Enhancers are relatively orientation and position independent, having been found at positions both 5' and 3' to the transcription unit. Several enhancer sequences available from mammalian genes are known (e.g., globin, elastase, albumin, alpha-feto-protein and insulin). Typically, however, an enhancer from a virus is used. The SV40 enhancer, the cytomegalovirus early promoter enhancer, the polyoma enhancer, and adenovirus enhancers known in the art are exemplary enhancing elements for the activation of eukaryotic promoters. While an enhancer can be positioned in the vector either 5' or 3' to a coding sequence, it is typically located at a site 5' from the promoter. A sequence encoding an appropriate native or heterologous signal sequence (leader sequence or signal peptide) can be incorporated into an expression vector, to promote extracellular secretion of the antibody. The choice of signal peptide or leader depends on the type of host cells in which the antibody is to be produced, and a heterologous signal sequence can replace the native signal sequence. Examples of signal peptides that are functional in mammalian host cells include the following: the signal sequence for interleukin-7 (IL-7) described in US Patent No. 4,965,195 ; the signal sequence for interleukin-2 receptor described in Cosman et al., (1984) Nature 312:768-71; the interleukin-4 receptor signal peptide described in EP Patent No. 0367 566 ; the type I interleukin-1 receptor signal peptide described in U.S. Patent No. 4,968,607 ; the type II interleukin-1 receptor signal peptide described in EP Patent No. 0 460 846 .

Expression vectors can be constructed from a starting vector such as a commercially available vector. Such vectors can but need not contain all of the desired flanking sequences. Where one or more of the flanking sequences are not already present in the vector, they can be individually obtained and ligated into the vector. Methods used for obtaining each of the flanking sequences are well known to one skilled in the art.

After the vector has been constructed and a nucleic acid molecule encoding light chain, a heavy chain, or a light chain and a heavy chain comprising an antigen binding protein that specifically binds to a complex comprising β-Klotho and at least one of (i) FGFR1c, (ii) FGFR2c and (iii) FGFR3c has been inserted into the proper site of the vector, the completed vector can be inserted into a suitable host cell for amplification and/or polypeptide expression. The transformation of an expression vector for an antigen binding protein into a selected host cell can be accomplished by well known methods including transfection, infection, calcium phosphate co-precipitation, electroporation, microinjection, lipofection, DEAE-dextran mediated transfection, or other known techniques. The method selected will in part be a function of the type of host cell to be used. These methods and other suitable methods are well known to the skilled artisan, and are set forth, for example, in Sambrook et al., (2001), supra.

A host cell, when cultured under appropriate conditions, synthesizes an antigen binding protein that can subsequently be collected from the culture medium (if the host cell secretes it into the medium) or directly from the host cell producing it (if it is not secreted). The selection of an appropriate host cell will depend upon various factors, such as desired expression levels, polypeptide modifications that are desirable or necessary for activity (such as glycosylation or phosphorylation) and ease of folding into a biologically active molecule.

Mammalian cell lines available as hosts for expression are well known in the art and include, but are not limited to, immortalized cell lines available from the American Type Culture Collection (ATCC), including but not limited to HeLa cells, Human Embryonic Kidney 293 cells (HEK293 cells), Chinese hamster ovary (CHO) cells, HeLa cells, baby hamster kidney (BHK) cells, monkey kidney cells (COS), human hepatocellular carcinoma cells (e.g., Hep G2), and a number of other cell lines. In certain embodiments, cell lines can be selected through determining which cell lines have high expression levels and constitutively produce antigen binding proteins with desirable binding properties (e.g., the ability to bind to a complex comprising β-Klotho and at least one of (i) FGFR1c, (ii) FGFR2c and (iii) FGFR3c). In another embodiment, a cell line from the B cell lineage that does not make its own antibody but has a capacity to make and secrete a heterologous antibody can be selected. The ability to induce FGF21-like signaling can also form a selection criterion.

Uses of Antigen Binding Proteins for Diagnostic and Therapeutic Purposes

The antigen binding proteins disclosed herein are useful for detecting to a complex comprising β-Klotho and at least one of (i) FGFR1c, (ii) FGFR2c and (iii) FGFR3c in biological samples and identification of cells or tissues that produce one or more of β-Klotho and at least one of (i) FGFR1c, (ii) FGFR2c and (iii) FGFR3c. For instance, the antigen binding proteins disclosed herein can be used in diagnostic assays, e.g., binding assays to detect and/or quantify a complex comprising β-Klotho and at least one of (i) FGFR1c, (ii) FGFR2c and (iii) FGFR3c expressed in a tissue or cell.

Antigen binding proteins that specifically bind to a complex comprising β-Klotho and at least one of (i) FGFR1c, (ii) FGFR2c and (iii) FGFR3c can also be used in treatment of diseases related to FGF21-like signaling in a patient in need thereof, such as type 2 diabetes, obesity, dyslipidemia, NASH, cardiovascular disease, and metabolic syndrome. By forming a signaling complex comprising an antigen binding protein and a complex comprising β-Klotho and at least one of (i) FGFR1c, (ii) FGFR2c and (iii) FGFR3c, the natural in vivo activity of FGF21, which associates with a complex comprising β-Klotho and at least one of (i) FGFR1c, (ii) FGFR2c and (iii) FGFR3c in vivo to initiate signaling, can be mimicked and/or enchanced, leading to therapeutic effects.

Indications

A disease or condition associated with human FGF21 includes any disease or condition whose onset in a patient is influenced by, at least in part, the lack of or therapeutically insufficient induction of FGF21-like signaling, which is initiated in vivo by the formation of a complex comprising β-Klotho and at least one of (i) FGFR1c, (ii) FGFR2c and (iii) FGFR3c. The severity of the disease or condition can also be decreased by the induction of FGF21-like signaling. Examples of diseases and conditions that can be treated with the antigen binding proteins provided herein include type 2 diabetes, obesity, dyslipidemia, NASH, cardiovascular disease, and metabolic syndrome.

The antigen binding proteins described herein can be used to treat type 2 diabetes, obesity, dyslipidemia, NASH, cardiovascular disease, and metabolic syndrome, or can be employed as a prophylactic treatment administered, e.g., daily, weekly, biweekly, monthly, bimonthly, biannually, etc to prevent or reduce the frequency and/or severity of symptoms, e.g., elevated plasma glucose levels, elevated triglycerides and/or cholesterol levels, thereby providing an improved glycemic and cardiovascular risk factor profile.

Diagnostic Methods

The antigen binding proteins described herein can be used for diagnostic purposes to detect, diagnose, or monitor diseases and/or conditions associated with FGFR1c, FGFR2c, FGFR3c, β-Klotho, FGF21 and/or complexes comprising combinations thereof. Also provided are methods for the detection of the presence of to a complex comprising β-Klotho and at least one of (i) FGFR1c, (ii) FGFR2c and (iii) FGFR3c in a sample using classical immunohistological methods known to those of skill in the art (e.g., Tijssen, (1985) "Practice and Theory of Enzyme Immunoassays" in Laboratory Techniques in Biochemistry and Molecular Biology, 15 (Burdon & van Knippenberg, eds.), Elsevier Biomedical); Zola, (1987) Monoclonal Antibodies: A Manual of Techniques, pp. 147-58 (CRC Press, Inc.); Jalkanen et al., (1985) J. Cell. Biol. 101:976-85; Jalkanen et al., (1987) J. Cell Biol. 105:3087-96). The detection of a complex comprising β-Klotho and at least one of (i) FGFR1c, (ii) FGFR2c and (iii) FGFR3c can be performed in vivo or in vitro.

Diagnostic applications provided herein include use of the antigen binding proteins to detect expression/formation of a complex comprising β-Klotho and at least one of (i) FGFR1c, (ii) FGFR2c and (iii) FGFR3c, and/or binding to a complex comprising β-Klotho and at least one of (i) FGFR1c, (ii) FGFR2c and (iii) FGFR3c. Examples of methods useful in the detection of the presence of a complex comprising β-Klotho and at least one of (i) FGFR1c, (ii) FGFR2c and (iii) FGFR3c include immunoassays, such as the enzyme linked immunosorbent assay (ELISA) and the radioimmunoassay (RIA).

For diagnostic applications, the antigen binding protein typically will be labeled with a detectable labeling group. Suitable labeling groups include, but are not limited to, the following: radioisotopes or radionuclides (e.g., ³H, ¹⁴C, ¹⁵N, ³⁵S, ⁹⁰Y, ⁹⁹Tc, ¹¹¹In, ¹²⁵I, ¹³¹I), fluorescent groups (e.g., FITC, rhodamine, lanthanide phosphors), enzymatic groups (e.g., horseradish peroxidase, β-galactosidase, luciferase, alkaline phosphatase), chemiluminescent groups, biotinyl groups, or predetermined polypeptide epitopes recognized by a secondary reporter (e.g., leucine zipper pair sequences, binding sites for secondary antibodies, metal binding domains, epitope tags). In some embodiments, the labeling group is coupled to the antigen binding protein via spacer arms of various lengths to reduce potential steric hindrance. Various methods for labeling proteins are known in the art and can be used.

In another aspect, an antigen binding protein can be used to identify a cell or cells that express a complex comprising β-Klotho and at least one of (i) FGFR1c, (ii) FGFR2c and (iii) FGFR3c. In a specific embodiment, the antigen binding protein is labeled with a labeling group and the binding of the labeled antigen binding protein to a complex comprising β-Klotho and at least one of (i) FGFR1c, (ii) FGFR2c and (iii) FGFR3c is detected. In a further specific embodiment, the binding of the antigen binding protein to a complex comprising β-Klotho and at least one of (i) FGFR1c, (ii) FGFR2c and (iii) FGFR3c detected in vivo. In a further specific embodiment, the antigen binding protein is isolated and measured using techniques known in the art. See, for example, Harlow & Lane, (1988) supra; Current Protocols In Immunology (John E. Coligan, ed), John Wiley & Sons (1993 ed., and supplements and/or updates).

Another aspect provides for detecting the presence of a test molecule that competes for binding to a complex comprising β-Klotho and at least one of (i) FGFR1c, (ii) FGFR2c and (iii) FGFR3c with the antigen binding proteins provided, as disclosed herein. An example of one such assay could involve detecting the amount of free antigen binding protein in a solution containing an amount of a complex comprising β-Klotho and at least one of (i) FGFR1c, (ii) FGFR2c and (iii) FGFR3c in the presence or absence of the test molecule. An increase in the amount of free antigen binding protein (i.e., the antigen binding protein not bound to a complex comprising β-Klotho and at least one of (i) FGFR1c, (ii) FGFR2c and (iii) FGFR3c) would indicate that the test molecule is capable of competing for binding to a complex comprising β-Klotho and at least one of (i) FGFR1c, (ii) FGFR2c and (iii) FGFR3c with the antigen binding protein. In one embodiment, the antigen binding protein is labeled with a labeling group. Alternatively, the test molecule is labeled and the amount of free test molecule is monitored in the presence and absence of an antigen binding protein.

Methods of Treatment: Pharmaceutical Formulations and Routes of Administration

Methods of using the disclosed antigen binding proteins are also provided. In some methods, an antigen binding protein is provided to a patient, which induces FGF21-like signaling.

Pharmaceutical compositions that comprise a therapeutically effective amount of one or a plurality of the antigen binding proteins and a pharmaceutically acceptable diluent, carrier, solubilizer, emulsifier, preservative, and/or adjuvant are also provided. In addition, methods of treating a patient by administering such pharmaceutical composition are included. The term "patient" includes human patients.

Acceptable formulation materials are nontoxic to recipients at the dosages and concentrations employed. In specific embodiments, pharmaceutical compositions comprising a therapeutically effective amount of human antigen binding proteins that specifically bind to a complex comprising β-Klotho and at least one of (i) FGFR1c, (ii) FGFR2c and (iii) FGFR3c are provided.

In certain embodiments, acceptable formulation materials preferably are nontoxic to recipients at the dosages and concentrations employed. In certain embodiments, the pharmaceutical composition can contain formulation materials for modifying, maintaining or preserving, for example, the pH, osmolarity, viscosity, clarity, color, isotonicity, odor, sterility, stability, rate of dissolution or release, adsorption or penetration of the composition. In such embodiments, suitable formulation materials include, but are not limited to, amino acids (such as glycine, glutamine, asparagine, arginine or lysine); antimicrobials; antioxidants (such as ascorbic acid, sodium sulfite or sodium hydrogen-sulfite); buffers (such as borate, bicarbonate, Tris-HCl, citrates, phosphates or other organic acids); bulking agents (such as mannitol or glycine); chelating agents (such as ethylenediamine tetraacetic acid (EDTA)); complexing agents (such as caffeine, polyvinylpyrrolidone, beta-cyclodextrin or hydroxypropyl-beta-cyclodextrin); fillers; monosaccharides; disaccharides; and other carbohydrates (such as glucose, mannose or dextrins); proteins (such as serum albumin, gelatin or immunoglobulins); coloring, flavoring and diluting agents; emulsifying agents; hydrophilic polymers (such as polyvinylpyrrolidone); low molecular weight polypeptides; salt-forming counterions (such as sodium); preservatives (such as benzalkonium chloride, benzoic acid, salicylic acid, thimerosal, phenethyl alcohol, methylparaben, propylparaben, chlorhexidine, sorbic acid or hydrogen peroxide); solvents (such as glycerin, propylene glycol or polyethylene glycol); sugar alcohols (such as mannitol or sorbitol); suspending agents; surfactants or wetting agents (such as Pluronics, PEG, sorbitan esters, polysorbates such as polysorbate 20, polysorbate, triton, tromethamine, lecithin, cholesterol, tyloxapal); stability enhancing agents (such as sucrose or sorbitol); tonicity enhancing agents (such as alkali metal halides, preferably sodium or potassium chloride, mannitol sorbitol); delivery vehicles; diluents; excipients and/or pharmaceutical adjuvants. See, e.g., Remington's Pharmaceutical Sciences, 18th Edition, (A.R. Gennaro, ed.), 1990, Mack Publishing Company, and subsequent editions.

In certain embodiments, the optimal pharmaceutical composition will be determined by one skilled in the art depending upon, for example, the intended route of administration, delivery format and desired dosage. See, for example, Remington's Pharmaceutical Sciences, supra. In certain embodiments, such compositions can influence the physical state, stability, rate of in vivo release and rate of in vivo clearance of the antigen binding proteins disclosed. In certain embodiments, the primary vehicle or carrier in a pharmaceutical composition can be either aqueous or non-aqueous in nature. For example, a suitable vehicle or carrier can be water for injection, physiological saline solution or artificial cerebrospinal fluid, possibly supplemented with other materials common in compositions for parenteral administration. Neutral buffered saline or saline mixed with serum albumin are further exemplary vehicles. In specific embodiments, pharmaceutical compositions comprise Tris buffer of about pH 7.0-8.5, or acetate buffer of about pH 4.0-5.5, and can further include sorbitol or a suitable substitute. In certain embodiments, compositions comprising antigen binding proteins that specifically bind to a complex comprising β-Klotho and at least one of (i) FGFR1c, (ii) FGFR2c and (iii) FGFR3c can be prepared for storage by mixing the selected composition having the desired degree of purity with optional formulation agents (see, Remington's Pharmaceutical Sciences, supra for examples of suitable formulation agents) in the form of a lyophilized cake or an aqueous solution. Further, in certain embodiments, antigen binding proteins that bind to a complex comprising β-Klotho and at least one of (i) FGFR1c, (ii) FGFR2c and (iii) FGFR3c can be formulated as a lyophilizate using appropriate excipients such as sucrose.

The pharmaceutical compositions can be selected for parenteral delivery. Alternatively, the compositions can be selected for inhalation or for delivery through the digestive tract, such as orally. Preparation of such pharmaceutically acceptable compositions is within the skill of the art.

The formulation components are present preferably in concentrations that are acceptable to the site of administration. In certain embodiments, buffers are used to maintain the composition at physiological pH or at a slightly lower pH, typically within a pH range of from about 5 to about 8.

When parenteral administration is contemplated, the therapeutic compositions can be provided in the form of a pyrogen-free, parenterally acceptable aqueous solution comprising the desired antigen binding protein in a pharmaceutically acceptable vehicle. A particularly suitable vehicle for parenteral injection is sterile distilled water in which the antigen binding protein is formulated as a sterile, isotonic solution, properly preserved. In certain embodiments, the preparation can involve the formulation of the desired molecule with an agent, such as injectable microspheres, bio-erodible particles, polymeric compounds (such as polylactic acid or polyglycolic acid), beads or liposomes, that can provide controlled or sustained release of the product which can be delivered via depot injection. In certain embodiments, hyaluronic acid can also be used, which can have the effect of promoting sustained duration in the circulation. In certain embodiments, implantable drug delivery devices can be used to introduce the desired antigen binding protein.

Certain pharmaceutical compositions are formulated for inhalation. In some embodiments, antigen binding proteins that bind to a complex comprising β-Klotho and at least one of (i) FGFR1c, (ii) FGFR2c and (iii) FGFR3c are formulated as a dry, inhalable powder. In specific embodiments, antigen binding protein inhalation solutions can also be formulated with a propellant for aerosol delivery. In certain embodiments, solutions can be nebulized. Pulmonary administration and formulation methods therefore are further described in International Patent Application No. PCT/US94/001875 , which is incorporated by reference and describes pulmonary delivery of chemically modified proteins. Some formulations can be administered orally. Antigen binding proteins that specifically bind to a complex comprising β-Klotho and at least one of (i) FGFR1c, (ii) FGFR2c and (iii) FGFR3c that are administered in this fashion can be formulated with or without carriers customarily used in the compounding of solid dosage forms such as tablets and capsules. In certain embodiments, a capsule can be designed to release the active portion of the formulation at the point in the gastrointestinal tract when bioavailability is maximized and pre-systemic degradation is minimized. Additional agents can be included to facilitate absorption of an antigen binding protein. Diluents, flavorings, low melting point waxes, vegetable oils, lubricants, suspending agents, tablet disintegrating agents, and binders can also be employed.

Some pharmaceutical compositions comprise an effective quantity of one or a plurality of human antigen binding proteins that specifically bind to a complex comprising β-Klotho and at least one of (i) FGFR1c, (ii) FGFR2c and (iii) FGFR3c in a mixture with nontoxic excipients that are suitable for the manufacture of tablets. By dissolving the tablets in sterile water, or another appropriate vehicle, solutions can be prepared in unit-dose form. Suitable excipients include, but are not limited to, inert diluents, such as calcium carbonate, sodium carbonate or bicarbonate, lactose, or calcium phosphate; or binding agents, such as starch, gelatin, or acacia; or lubricating agents such as magnesium stearate, stearic acid, or talc.

Additional pharmaceutical compositions will be evident to those skilled in the art, including formulations involving antigen binding proteins that specifically bind to a complex comprising β-Klotho and at least one of (i) FGFR1c, (ii) FGFR2c and (iii) FGFR3c in sustained- or controlled-delivery formulations. Techniques for formulating a variety of other sustained- or controlled-delivery means, such as liposome carriers, bio-erodible microparticles or porous beads and depot injections, are also known to those skilled in the art. See, for example, International Patent Application No. PCT/US93/00829 , which is incorporated by reference and describes controlled release of porous polymeric microparticles for delivery of pharmaceutical compositions. Sustained-release preparations can include semipermeable polymer matrices in the form of shaped articles, e.g., films, or microcapsules. Sustained release matrices can include polyesters, hydrogels, polylactides (as disclosed in U.S. Patent No. 3,773,919 and European Patent Application Publication No. EP 058481 , each of which is incorporated by reference), copolymers of L-glutamic acid and gamma ethyl-L-glutamate (Sidman et al., (1983) Biopolymers 2:547-556), poly (2-hydroxyethyl-inethacrylate) (Langer et al., (1981) J. Biomed. Mater. Res. 15:167-277 and Langer, (1982) Chem. Tech. 12:98-105), ethylene vinyl acetate (Langer et al., (1981) supra) or poly-D(-)-3-hydroxybutyric acid (European Patent Application Publication No. EP 133988 ). Sustained release compositions can also include liposomes that can be prepared by any of several methods known in the art. See, e.g., Eppstein et al., (1985) Proc. Natl. Acad. Sci. U.S.A. 82:3688-3692; European Patent Application Publication Nos. EP 036676 ; EP 088046 and EP 143949 , incorporated by reference.

Pharmaceutical compositions used for in vivo administration are typically provided as sterile preparations. Sterilization can be accomplished by filtration through sterile filtration membranes. When the composition is lyophilized, sterilization using this method can be conducted either prior to or following lyophilization and reconstitution. Compositions for parenteral administration can be stored in lyophilized form or in a solution. Parenteral compositions generally are placed into a container having a sterile access port, for example, an intravenous solution bag or vial having a stopper pierceable by a hypodermic injection needle.

In certain embodiments, cells expressing a recombinant antigen binding protein as disclosed herein are encapsulated for delivery (see, Tao et al., Invest. Ophthalmol Vis Sci (2002) 43:3292-3298 and Sieving et al., Proc. Natl. Acad. Sciences USA (2006) 103:3896-3901).

In certain formulations, an antigen binding protein has a concentration of between 10 mg/ml and 150 mg/ml. Some formulations contain a buffer, sucrose and polysorbate. An example of a formulation is one containing 50-100 mg/ml of antigen binding protein, 5-20 mM sodium acetate, 5-10% w/v sucrose, and 0.002 - 0.008% w/v polysorbate. Certain, formulations, for instance, contain 1-100 mg/ml of an antigen binding protein in 9-11 mM sodium acetate buffer, 8-10% w/v sucrose, and 0.005-0.006% w/v polysorbate. The pH of certain such formulations is in the range of 4.5-6. Other formulations can have a pH of 5.0-5.5.

Once the pharmaceutical composition has been formulated, it can be stored in sterile vials as a solution, suspension, gel, emulsion, solid, crystal, or as a dehydrated or lyophilized powder. Such formulations can be stored either in a ready-to-use form or in a form (e.g., lyophilized) that is reconstituted prior to administration. Kits for producing a single-dose administration unit are also provided. Certain kits contain a first container having a dried protein and a second container having an aqueous formulation. In certain embodiments, kits containing single and multi-chambered pre-filled syringes (e.g., liquid syringes and lyosyringes) are provided. The therapeutically effective amount of an antigen binding protein-containing pharmaceutical composition to be employed will depend, for example, upon the therapeutic context and objectives. One skilled in the art will appreciate that the appropriate dosage levels for treatment will vary depending, in part, upon the molecule delivered, the indication for which the antigen binding protein is being used, the route of administration, and the size (body weight, body surface or organ size) and/or condition (the age and general health) of the patient. In certain embodiments, the clinician can titer the dosage and modify the route of administration to obtain the optimal therapeutic effect.

A typical dosage can range from about 1 µg/kg to up to about 30 mg/kg or more, depending on the factors mentioned above. In specific embodiments, the dosage can range from 10 µg/kg up to about 35 mg/kg, optionally from 0.1 mg/kg up to about 35 mg/kg, alternatively from 0.3 mg/kg up to about 20 mg/kg. In some applications, the dosage is from 0.5 mg/kg to 20 mg/kg and in other applications the dosage is from 21-100 mg/kg. In some instances, an antigen binding protein is dosed at 0.3-20 mg/kg. The dosage schedule in some treatment regimes is at a dose of 0.3 mg/kg qW-20 mg/kg qW.

Dosing frequency will depend upon the pharmacokinetic parameters of the particular antigen binding protein in the formulation used. Typically, a clinician administers the composition until a dosage is reached that achieves the desired effect. The composition can therefore be administered as a single dose, or as two or more doses (which can but need not contain the same amount of the desired molecule) over time, or as a continuous infusion via an implantation device or catheter. Appropriate dosages can be ascertained through use of appropriate dose-response data. In certain embodiments, the antigen binding proteins can be administered to patients throughout an extended time period. Chronic administration of an antigen binding protein minimizes the adverse immune or allergic response commonly associated with antigen binding proteins that are not fully human, for example an antibody raised against a human antigen in a non-human animal, for example, a non-fully human antibody or non-human antibody produced in a non-human species.

The route of administration of the pharmaceutical composition is in accord with known methods, e.g., orally, through injection by intravenous, intraperitoneal, intracerebral (intra-parenchymal), intracerebroventricular, intramuscular, intra-ocular, intraarterial, intraportal, or intralesional routes; by sustained release systems or by implantation devices. In certain embodiments, the compositions can be administered by bolus injection or continuously by infusion, or by implantation device.

The composition also can be administered locally via implantation of a membrane, sponge or another appropriate material onto which the desired molecule has been absorbed or encapsulated. In certain embodiments, where an implantation device is used, the device can be implanted into any suitable tissue or organ, and delivery of the desired molecule can be via diffusion, timed-release bolus, or continuous administration.

It also can be desirable to use antigen binding protein pharmaceutical compositions ex vivo. In such instances, cells, tissues or organs that have been removed from the patient are exposed to antigen binding protein pharmaceutical compositions after which the cells, tissues and/or organs are subsequently implanted back into the patient.

In particular, antigen binding proteins that specifically bind to a complex comprising β-Klotho and at least one of (i) FGFR1c, (ii) FGFR2c and (iii) FGFR3c can be delivered by implanting certain cells that have been genetically engineered, using methods such as those described herein and known in the art, to express and secrete the polypeptide. In certain embodiments, such cells can be animal or human cells, and can be autologous, heterologous, or xenogeneic. In certain embodiments, the cells can be immortalized. In other embodiments, in order to decrease the chance of an immunological response, the cells can be encapsulated to avoid infiltration of surrounding tissues. In further embodiments, the encapsulation materials are typically biocompatible, semi-permeable polymeric enclosures or membranes that allow the release of the protein product(s) but prevent the destruction of the cells by the patient's immune system or by other detrimental factors from the surrounding tissues.

Combination Therapies

In another aspect, the present disclosure provides a method of treating a subject for diabetes with a therapeutic antigen binding protein of the present disclosure, such as the fully human therapeutic antibodies described herein, together with one or more other treatments. In one embodiment, such a combination therapy achieves an additive or synergistic effect. The antigen binding proteins can be administered in combination with one or more of the type 2 diabetes or obesity treatments currently available. These treatments for diabetes include biguanide (metaformin), and sulfonylureas (such as glyburide, glipizide). Additional treatments directed at maintaining glucose homeostasis include PPAR gamma agonists (pioglitazone, rosiglitazone); glinides (meglitinide, repaglinide, and nateglinide); DPP-4 inhibitors (Januvia® and Onglyza®) and alpha glucosidase inhibitors (acarbose, voglibose).

Additional combination treatments for diabetes include injectable treatments such as insulin and incretin mimetics (Byetta®, Exenatide®), other GLP-1 (glucagon-like peptide) analogs such as Victoza® (liraglutide), other GLP-1R agonists and Symlin® (pramlintide).

Additional combination treatments directed at weight loss include Meridia® and Xenical®.

Embodiments of the present invention are disclosed in the following (Embodiments 1 to 23). The definitions given herein above apply accordingly.

1. An isolated antigen binding protein that induces FGF21-mediated signaling.
2. An isolated antigen binding protein that specifically binds to a complex comprising β-Klotho and at least one of (i) FGFR1c, (ii) FGFR2c and (iii) FGFR3c.
3. The antigen binding protein of embodiment 1, wherein the antigen binding protein comprises one or more of:
1. (a) a light chain CDR3 comprising one or more of:
  1. (i) a light chain CDR3 sequence that differs by three , two or one amino acid additions, substitutions, deletions and combinations thereof from a CDR3 of one or more of CDRL3-1 to CDRL3-75 of Table 3B (SEQ ID NOs 947-1020, respectively, in order of appearance);
  2. (ii) QQFGSSLT (SEQ ID NO: 988);
  3. (iii) QQSX₁SX₂X₃ LT (SEQ ID NO: 1443);
  4. (iv) LQX₄X₅SYPX₆T (SEQ ID NO: 1447);
  5. (v) MQRX₇EFPX₈T (SEQ ID NO: 1451);
  6. (vi) QX₉WDSX₁₀ X₁₁VV (SEQ ID NO: 1457);
  7. (vii) QQYNX₁₂WP X₁₃T (SEQ ID NO: 1461);
  8. (viii) QVWDSSX₁₄ DX₁₅VX₁₆ (SEQ ID NO: 1466);
  9. (ix) QQSS X₁₇IPWT (SEQ ID NO: 1469);
  10. (x) QQTNSFPPWT (SEQ ID NO: 1470);
  11. (xi) GTWDSSLSX₁₈X₁₉V (SEQ ID NO: 1474);
  12. (xii) QQYDNLPX₂₀T (SEQ ID NO: 1477);
  13. (xiii) QQYGSSX₂₁PWT (SEQ ID NO: 1480);
  14. (xiv) QQYGX₂₂SX₂₃FT (SEQ ID NO: 1483);
  15. (xv) QQYGSSX₂₄X₂₅X₂₆ (SEQ ID NO: 1488);
  16. (xvi) QAWDSSX₂₇TX₂₈V (SEQ ID NO: 1492);
  17. (xvii) QAWDSX₂₉TVX₃₀ (SEQ ID NO: 1496);
  18. (xviii) QQX₃₁YSAX₃₂FT (SEQ ID NO: 1499);
  19. (xix) QQYNX₃₃YPRT (SEQ ID NO: 1502);
  20. (xx) HQX₃₄X₃₅DLPLT (SEQ ID NO: 1505);
  21. (xxi) MQALQTX₃₆X₃₇T (SEQ ID NO: 1508);
  22. (xxii) QQFGRSFT (SEQ ID NO: 1509);
  23. (xxiii) YSTDSSX₃₈NHVV (SEQ ID NO: 1512);
2. (b) a heavy chain CDR3 sequence comprising one or more of:
  1. (i) a heavy chain CDR3 that differs by eight, seven, six, five, four, three, two or one amino acid additions, substitutions, deletions and combinations thereof from a CDR3 of one or more of CDRH3-1 to CDRH3-81 of Table 3A (SEQ ID NOs 733-813, respectively, in order of appearance);
  2. (ii) MTX₁₁₀PYWYFX₁₁₁ L (SEQ ID NO: 1669);
  3. (iii) DX₁₁₂X₁₁₃X₁₁₄DFWX₁₁₅GYPX₁₁₆X₁₁₇X₁₁₈YYGX₁₁₉DV (SEQ ID NO: 1675);
  4. (iv) VTGTDAFDF (SEQ ID NO: 1676);
  5. (v) TVTKEDYYYYGMDV (SEQ ID NO: 1677);
  6. (vi) DSSGSYYVEDYFDY (SEQ ID NO: 1678);
  7. (vii) DX₁₁₉X₁₂₀IAVAGX₁₂₁FDY (SEQ ID NO: 1681);
  8. (viii) EYYYGSGSYYP (SEQ ID NO: 1682);
  9. (ix) ELGDYPFFDY (SEQ ID NO: 1683);
  10. (x) EYVAEAGFDY (SEQ ID NO: 1684);
  11. (xi) VAAVYWYFDL (SEQ ID NO: 1685);
  12. (xii) YNWNYGAFDF (SEQ ID NO: 1686);
  13. (xiii) RASRGYRX₁₂₂GLAFAI (SEQ ID NO: 1689);
  14. (xiv) DGITMVRGVTHYYGMDV (SEQ ID NO: 1690);
  15. (xv) DHX₁₂₃SCWYYYGMDV (SEQ ID NO: 1693);
  16. (xvi) YX₁₂₄X₁₂₅WDYYYGX₁₂₆DV (SEQ ID NO: 1696);
  17. (xvii) VLHY X₁₂₇DSX₁₂₈GYSYYX₁₂₉DX₁₃₀ (SEQ ID NO: 1699); or
3. (c) the light chain CDR3 sequence of (a) and the heavy chain CDR3 sequence of (b); wherein,
  - X₁ is Y or F;
  - X₂ is T or S;
  - X₃ is P or S;
  - X₄ is H or R;
  - X₅ is N or S;
  - X₆ is L or R;
  - X₇ is I or L;
  - X₈ is I or L;
  - X₉ is V or L;
  - X₁₀ is S or N;
  - X₁₁ is T, P or S;
  - X₁₂ is N or T;
  - X₁₃ is W or L;
  - X₁₄ is S or C;
  - X₁₅ is H, V or G;
  - X₁₆ is V or absent;
  - X₁₇ is S or T;
  - X₁₈ is A or V;
  - X₁₉ is V or M;
  - X₂₀ is L or F;
  - X₂₁ is P or absent;
  - X₂₂ is R or T;
  - X₂₃ is L or P;
  - X₂₄ is P or absent;
  - X₂₅ is R or C;
  - X₂₆ is S or T;
  - X₂₇ is T or absent;
  - X₂₈ is W or A;
  - X₂₉ is G, T, A or absent;
  - X₃₀ is V or I;
  - X₃₁ is S or T;
  - X₃₂ is T or P;
  - X₃₃ is I or T;
  - X₃₄ is S or Y;
  - X₃₅ is S or D;
  - X₃₆ is P or L;
  - X₃₇ is F or I;
  - X₃₈ is V or G;
  - X₁₁₀ is T or S;
  - X₁₁₁ is D or G;
  - X₁₁₂ is R or Q;
  - X₁₁₃ is Y, D or N;
  - X₁₁₄ is Y or F;
  - X₁₁₅ is S or N;
  - X₁₁₆ is Y or F;
  - X₁₁₇ is F or Y;
  - X₁₁₈ is R, Y, F or H;
  - X₁₁₉ is L or M;
  - X₁₁₉ is W or L;
  - X₁₂₀ is S or R;
  - X₁₂₁ is T or S;
  - X₁₂₂ is F or Y;
  - X₁₂₃ is S or T;
  - X₁₂₄ is S or R;
  - X₁₂₅ is T or D;
  - X₁₂₆ is V or M;
  - X₁₂₇ is S or Y;
  - X₁₂₈ is R or S;
  - X₁₂₉ is S or F; and
  - X₁₃₀ is F or Y.
4. The antigen binding protein of embodiment 1, comprising one or more of:
1. (a) a light chain CDR1 sequence comprising one or more of:
  - (i) a light chain CDR1 that differs by seven, six, five, four, three, two or one amino acid additions, substitutions, deletions and combinations thereof from a CDR1 sequence of one or more of CDRL1-1 to CDRL1-81 of Table 3B (SEQ ID NOs 814-893, respectively, in order of appearance);
  - (ii) RASX₆₂SX₆₃X₆₄X₆₅X₆₆X₆₇X₆₈A (SEQ ID NO: 1591);
  - (iii) RX₆₉SQX₇₀IX₇₁X₇₂YLN (SEQ ID NO: 1602);
  - (iv) GGNX₇₃IGSX₇₄X₇₅VX₇₆ (SEQ ID NO: 1612);
  - (v) RASQX₇₇IRNDLX₇₈ (SEQ ID NO: 1616);
  - (vi) RSSQSLX₇₉X₈₀X₈₁DX₈₂GX₈₃TYLD (SEQ ID NO: 1622);
  - (vii) SGX₈₄ X₈₅LGDKYX₈₆X₈₇ (SEQ ID NO: 1629);
  - viii) QASQX₈₈IX₈₉X₉₀X₉₁LN (SEQ ID NO: 1635);
  - (ix) RASQX₉₂IX₉₃ X₉₄WLX₉₅ (SEQ ID NO: 1640);
  - (x) SGSSSNIGX₉₆NYVX₉₇ (SEQ ID NO: 1645);
  - (xi) RASX₉₈DISNYLA (SEQ ID NO: 1648);
  - (xii) RASQX₉₉VX₁₀₀SSYX₁₀₁V (SEQ ID NO: 1652);
  - (xiii) RSSQSLX₁₀₂HSNGX₁₀₃NYLD (SEQ ID NO: 1656);
  - (xiv) RASQTX₁₀₄RNX₁₀₅YLA (SEQ ID NO: 1659);
  - (xv) RSSX₁₀₆X₁₀₇LVYSDGNTYLN (SEQ ID NO: 1662); and
  - (xvi) SGDAX₁₀₈PKKYAX₁₀₉ (SEQ ID NO: 1665);
2. (b) a light chain CDR2 sequence comprising one or more of:
  1. (i) a light chain CDR2 that differs by three, two or one amino acid additions, substitutions, deletions and combinations thereof from a CDR2 sequence of one or more of of CDRL2-1 to CDRL2-53 of Table 3B (SEQ ID NOs 894-946, respectively, in order of appearance);
  2. (ii) X₃₉ASSLX₄₀X₄₁ (SEQ ID NO: 1517);
  3. (iii) GX₄₂S X₄₃RX₄₄T (SEQ ID NO: 1525);
  4. (iv) GAFSRAX₄₅ (SEQ ID NO: 1528);
  5. (v) X₄₆DX₄₇KRPS (SEQ ID NO: 1533);
  6. (vi) TLSX₄₈RAS (SEQ ID NO: 1536);
  7. (vii) AASNLQX₄₉ (SEQ ID NO: 1539);
  8. (viii) GX₅₀SNRAX₅₁ (SEQ ID NO: 1543);
  9. (ix) X₅₂ASX₅₃LQS (SEQ ID NO: 1548);
  10. (x) DNX₅₃KRPS (SEQ ID NO: 1551);
  11. (xi) DX₅₄SNLET (SEQ ID NO: 1554);
  12. (xii) LX₅₅SNRAS (SEQ ID NO: 1557);
  13. (xiii) QX₅₆NX₅₇RPS (SEQ ID NO: 1561);
  14. (xiv) RDRNRPS (SEQ ID NO: 1562);
  15. (xv) X₅₈DSNRPS (SEQ ID NO: 1565);
  16. (xvi) DDSDRPS (SEQ ID NO: 1566);
  17. (xvii) AX₅₉SSLQS (SEQ ID NO: 1569);
  18. (xviii) TX₆₀SSLQS (SEQ ID NO: 1572); and
  19. (xix) KX₆₁SNWDS (SEQ ID NO: 1575);
3. (c) a heavy chain CDR1 sequence comprising one or more of:
  1. (i) a heavy chain CDR1 that differs by three, two or one amino acid additions, substitutions, deletions and combinations thereof from a CDR1 sequence of one or more of CDRH1-1 to CDRH1-53 of Table 3A (SEQ ID NOs 603-655, respectively, in order of appearance);
  2. (ii) SGX₁₇₀X₁₇₁TWX₁₇₂ (SEQ ID NO: 1775);
  3. (iii) X₁₇₃YYWX₁₇₄ (SEQ ID NO: 1781);
  4. (iv) X₁₇₅X₁₇₆GMS (SEQ ID NO: 1786);
  5. (v) SYX₁₇₇MX₁₇₈ (SEQ ID NO: 1790);
  6. (vi) X₁₇₉YYX₁₈₀H (SEQ ID NO: 1796);
  7. (vii) SYGX₁₈₁H (SEQ ID NO: 1799);
  8. (viii) NYX₁₈₂MX₁₈₃ (SEQ ID NO: 1803);
  9. (ix) X₁₈₄YWIG (SEQ ID NO: 1806);
  10. (x) GYX₁₈₅MH (SEQ ID NO: 1809);
  11. (xi) SX₁₈₆DIX₁₈₇ (SEQ ID NO: 1813);
  12. (xii) X₁₈₈YAMS (SEQ ID NO: 1816);
  13. (xiii) NAWMS (SEQ ID NO: 1817);
  14. (xiv) SSSYYWG (SEQ ID NO: 1818);
  15. (xv) X₁₈₉YYWN (SEQ ID NO: 1821);
  16. (xvi) SNSAX₁₉₀WN (SEQ ID NO: 1824); and
  17. (xvii) X₁₉₁YDMH (SEQ ID NO: 1827);
4. (d) a heavy chain CDR2 sequence comprising one or more of
  1. (i) a heavy chain CDR2 that differs by nine, eight, seven, six, five, four, three, two or one amino acid additions, substitutions, deletions and combinations thereof from a CDR2 sequence of one or more of CDRH2-1 to CDRH2-77 of Table 3A (SEQ ID NOs 656-732, respectively, in order of appearance);
  2. (ii) X₁₃₁X₁₃₂X₁₃₃X₁₃₄X₁₃₅GX₁₃₆X₁₃₇X₁₃₈X₁₃₉NPSLKS (SEQ ID NO: 1716);
  3. (iii) X₁₄₀IX₁₄₁X₁₄₂DGX₁₄₃NX₁₄₄X₁₄₅X₁₄₆ADSVKG (SEQ ID NO: 1732);
  4. (iv) WX₁₄₇NPX₁₄₈SGX₁₄₉TX₁₅₀YAQKF X₁₅₁G (SEQ ID NO: 1741);
  5. (v) EINHSX₁₅₂X₁₅₃TNYNPSLKS (SEQ ID NO: 1744);
  6. (vi) IIYPGDSX₁₅₄TRYSPSFQG (SEQ ID NO: 1747);
  7. (vii) SISSSSX₁₅₅YX_I56YY X₁₅₇DSX₁₅₈KG (SEQ ID NO: 1751);
  8. (viii) RIX₁₅₉ X₁₆₀KTDGGTTX₁₆₁YAAPVKG (SEQ ID NO: 1755);
  9. (ix) GISGSSAGTYYADSVGK (SEQ ID NO: 1756);
  10. (x) VISX₁₆₂SGGX₁₆₃TYYADSVKG (SEQ ID NO: 1759);
  11. (xi) RTYYRSKWYNDYAVSVKS (SEQ ID NO: 1760);
  12. (xii) RIYX₁₆₄SGSTNYNPSLX₁₆₅ X₁₆₆ (SEQ ID NO: 1763); and
  13. (xiii) WMNPYSGSTGX₁₆₇ AQX₁₆₈FQX₁₆₉ (SEQ ID NO: 1766);
  wherein
  - X₃₉ is A or S;
  - X₄₀ is Q or K;
  - X₄₁ is S or F;
  - X₄₂ is A or T;
  - X₄₃ is S, T, A, R or N;
  - X₄₄ is A or D;
  - X₄₅ is S or T;
  - X₄₆ is Q, R or E;
  - X₄₇ is T or S;
  - X₄₈ is Y or F;
  - X₄₉ is R or S;
  - X₅₀ is A or S;
  - X₅₁ is I or T;
  - X₅₂ is D or G;
  - X₅₃ is S, T or N;
  - X₅₃ is N or D;
  - X₅₄ is A or V;
  - X₅₅ is G or D;
  - X₅₆ is D or N;
  - X₅₇ is K or E;
  - X₅₈ is S or C;
  - X₅₉ is S or V;
  - X₆₀ is A or T;
  - X₆₁ is V or G;
  - X₆₂ is P or Q;
  - X₆₃ is V, I or F;
  - X₆₄ is S, R or absent;
  - X₆₅ is S, R or N;
  - X₆₆ is D, S, N or M;
  - X₆₇ is I, Y, H, Q, N or S;
  - X₆₈ is L or V;
  - X₆₉ is A or T;
  - X₇₀ is I, S, T or N;
  - X₇₁ is R, S or N;
  - X₇₂ is R, S, N, or I;
  - X₇₃ is N, or D;
  - X₇₄ is Y, I or K;
  - X₇₅ is N, S, T or A;
  - X₇₆ is H or Q;
  - X₇₇ is D or G;
  - X₇₈ is G or A;
  - X₇₉ is L or F;
  - X₈₀ is N or D;
  - X₈₁ is S or N;
  - X₈₂ is A or D;
  - X₈₃ is T, D or N;
  - X₈₄ is N or D;
  - X₈₅ is K, E or N;
  - X₈₆ is V or A;
  - X₈₇ is C or F;
  - X₈₈ isG or D;
  - X₈₉ is S, K N or T;
  - X₉₀ is N, K or I;
  - X₉₁ is Y or F;
  - X₉₂ is D or G;
  - X₉₃ isD or S;
  - X₉₄ is R or S;
  - X₉₅ is V or A;
  - X₉₆ is N, I or D;
  - X₉₇ is A or S;
  - X₉₈ is Q or H.;
  - X₉₉ is R or S;
  - X₁₀₀ is P or A;
  - X₁₀₁ is I or L;
  - X₁₀₂ is L or Q;
  - X₁₀₃ is Y or F.
  - X₁₀₄ is V or I;
  - X₁₀₅ is N or S;
  - X₁₀₆ is Q or P;
  - X₁₀₇ is R or S;
  - X₁₀₈ is L or V;
  - X₁₀₉ is Y or N;
  - X₁₃₁ is N, F, Y, S or M;
  - X₁₃₂ is I or L;
  - X₁₃₃ is Y or F;
  - X₁₃₄ is Y, H or D;
  - X₁₃₅ is S or T;
  - X₁₃₆ is T, G, S or T;
  - X₁₃₇ is T or A;
  - X₁₃₈ is Y, N or H;
  - X₁₃₉ is F or Y;
  - X₁₄₀ is L, G, I or V;
  - X₁₄₁ is W or S;
  - X₁₄₂ is Y, D or N;
  - X₁₄₃ is S or D;
  - X₁₄₄ is K or N;
  - X₁₄₅ is Y, N, D, or H;
  - X₁₄₆ is Y or H;
  - X₁₄₇ is I or M;
  - X₁₄₈ is P, N, S or D;
  - X₁₄₉ is A, G or D;
  - X₁₅₀ isN, K, or D;
  - X₁₅₁ is R, H or Q;
  - X₁₅₂ is E or G;
  - X₁₅₃ is N or T;
  - X₁₅₄ is D or E;
  - X₁₅₅ is T or S;
  - X₁₅₆ is I or E;
  - X₁₅₇ is A or V;
  - X₁₅₈ is V or L.
  - X₁₅₉ is K or I;
  - X₁₆₀ is S or G;
  - X₁₆₁ is D or E;
  - X₁₆₂ is D or G;
  - X₁₆₃ is S or D;
  - X₁₆₄ is I or T;
  - X₁₆₅ is E or K;
  - X₁₆₆ is N or S;
  - X₁₆₇ is Y or L;
  - X₁₆₈ is N or R;
  - X₁₆₉ is G or D;
  - X₁₇₀ is V, G, N or D;
  - X₁₇₁ is Y or N;
  - X₁₇₂ is N, S or T;
  - X₁₇₃ is T, S or G;
  - X₁₇₄ is S or T;
  - X₁₇₅ is S, T or F;
  - X₁₇₆ is Y or F;
  - X₁₇₇ is A or S;
  - X₁₇₈ is S, N or M;
  - X₁₇₉ is Y or G;
  - X₁₈₀ is I, L, K or T;
  - X₁₈₁ is L or I;
  - X₁₈₂ is G or N;
  - X₁₈₃ is H, R or M;
  - X₁₈₄ is S or G;
  - X₁₈₅ is Y or F;
  - X₁₈₆ is Y or H;
  - X₁₈₇ is N or D;
  - X₁₈₈ is N or H;
  - X₁₈₉ is D or S;
  - X₁₉₀ is T or A;
  - X₁₉₁ is S or T;
5. (e) the light chain CDR1 of (a) and the light chain CDR2 of (b);
6. (f) the light chain CDR1 of (a) and the heavy chain CDR1 of (c);
7. (g) the light chain CDR1 of (a) and the heavy chain CDR2 of (d);
8. (h) the light chain CDR1 (a) and the heavy chain CDR1 of (c);
9. (i) the heavy chain CDR1 of (c) and the heavy chain CDR2 of (d);
10. (j) the light chain CDR2 of (b) and the heavy chain CDR2 of (d);
11. (k) the light chain CDR1 of (a), the light chain CDR2 of (b), and the heavy chain CDR1 of (c);
12. (l) the light chain CDR2 of (b), the heavy CDR1 of (c), and the heavy chain CDR2 of (d);
13. (m) the light chain CDR1 of (a), the heavy chain CDR1 of (c), and the heavy chain CDR2 of (d); and
14. (n) the light chain CDR1 of (a), the light chain CDR2 of (b), the heavy chain CDR2 of (c), and the heavy chain CDR2 of (d).
5. The antigen binding protein of embodiment 1, comprising one or more of:
1. (a) a light chain variable domain comprising one or more of;
  1. (i) a light chain CDR1 sequence selected from CDRL1-1 to CDRL1-81 of Table 3B (SEQ ID NOs 814-893, respectively, in order of appearance);
  2. (ii) a light chain CDR2 sequence selected from CDRL2-1 to CDRL2-53 of Table 3B (SEQ ID NOs 894-946, respectively, in order of appearance);
  3. (iii) a light chain CDR3 sequence selected from CDRL3-1 to CDRL3-75 of Table 3B (SEQ ID NOs 947-1020, respectively, in order of appearance);
2. (b) a heavy chain variable domain comprising one or more of:
  1. (i) a heavy chain CDR1 sequence selected from CDRH1-1 to CDRH1-53 of Table 3A (SEQ ID NOs 603-655, respectively, in order of appearance);
  2. (ii) a heavy chain CDR2 sequence selected from CDRH2-1 to CDRH2-77 of Table 3A (SEQ ID NOs 656-732, respectively, in order of appearance);
  3. (iii) a heavy chain CDR3 sequence selected from CDRH3-1 to CDRH3-81 of Table 3A (SEQ ID NOs 733-813, respectively, in order of appearance); and
3. (c) a combination comprising a light chain variable domain of (a) and a heavy chain variable domain of (b).
6. The antigen binding protein of embodiment 1, wherein the antigen binding protein comprises one or more of:
1. (a) a light chain variable domain sequence comprising one or more of:
  1. (i) amino acids having a sequence at least 80% identical to a light chain variable domain sequence comprising one or more of V_L1-V_L100 of Table 2A (SEQ ID NOs 217-315, respectively, in order of appearance);
  2. (ii) a sequence of amino acids encoded by a polynucleotide sequence that is at least 80% identical to a polynucleotide sequence encoding the light chain variable domain sequence comprising one or more of V_L1-V_L100 of Table 2A(SEQ ID NOs 217-315, respectively, in order of appearance);
2. (b) a heavy chain variable domain sequence comprising one or more of:
  1. (i) a sequence of amino acids that is at least 80% identical to a heavy chain variable domain sequence comprising one or more of V_H1-V_H94 of Table 2B 9SEQ ID NOs 316-409, respectively, in order of appearance);
  2. (ii) a sequence of amino acids encoded by a polynucleotide sequence that is at least 80% identical to a polynucleotide sequence encoding the heavy chain variable domain sequence of V_H1-V_H94 of Table 2B (SEQ ID NOs 316-409, respectively, in order of appearance); and (c) a combination comprising a light chain variable domain of (a) and a heavy chain variable domain of (b).
7. The antigen binding protein of embodiment 6, comprising one or more of:
1. (a) a light chain variable domain sequence comprising one or more of: V_L1-V_L100 of Table 2A (SEQ ID NOs 217-315, respectively, in order of appearance);
2. (b) a heavy chain variable domain sequence comprising one or more of: V_H1-V_H94 of Table 2B (SEQ ID NOs 316-409, respectively, in order of appearance); and
3. (c) a combination comprising a light chain variable domain of (a) and a heavy chain variable domain of (b).
8. The antigen binding protein of embodiment 7, wherein the light chain variable domain and the heavy chain variable domain comprise one or more of: V_L1 and V_H1; V_L2 and V_H1; V_L3 and V_H2 or V_H3; V_L4 and V_H4; V_L5 and V_H5; V_L6 and V_H6; V_L7 and V_H6; V_L8 and V_H7 or V_H8; V_L9 and V_H9; V_L10 and V_H9; V_L11 and V_H 10; V_L12 and V_H11; V_L13 and V_H12; V_L13 and V_H14; V_L14 and V_H13; V_L15 and V_H14; V_L16 and V_H15; V_L17 and V_H16; V_L18 and V_H17; V_L19 and V_H18; V_L20 and V_H19; V_L21 and V_H20; V_L22 and V_H21; V_L23 and V_H22; V_L24 and V_H23; V_L25 and V_H24; V_L26 and V_H25; V_L27 and V_H26; V_L28 and V_H27; V_L29 and V_H28; V_L30 and V_H29; V_L31 and V_H30; V_L32 and V_H31; V_L33 and V_H32; V_L34 and V_H33; V_L35 and V_H34; V_L36 and V_H35; V_L37 and V_H36; V_L38 and V_H37; V_L39 and V_H38; V_L40 and V_H39; V_L41 and V_H40; V_L42 and V_H41; V_L43 and V_H42; V_L44 and V_H43; V_L45 and V_H44; V_L46 and V_H45; V_L47 and V_H46; V_L48 and V_H47; V_L49 and V_H48; V_L50 and V_H49; V_L51 and V_H50; V_L 52 and V_H51; V_L53 and V_H52; V_L54 and V_H53; V_L55 and V_H54; V_L56 and V_H54; V_L57 and V_H54; V_L58 and V_H55; V_L59 and V_H56; V_L60 and V_H57; V_L61 and V_H58; V_L62 and V_H59; V_L63 and V_H60; V_L64 and V_H1; V_L65 and V_H62; V_L66 and V_H63; V_L67 and V_H64; V_L68 and V_H65; V_L69 and V_H66; V_L70 and V_H67; V_L71 and V_H68; V_L72 and V_H69; V_L73 and V_H70; V_L74 and V_H70; V_L75 and V_H70; V_L76 and V_H71; V_L77 and V_H72; V_L78 and V_H73; V_L79 and V_H74; V_L80 and V_H75; V_L81 and V_H76; V_L82 and V_H77; V_L83 and V_H78; V_L84 and V_H79; V_L85 and V_H80; V_L86 and V_H81; V_L87 and V_H82; V_L88 and V_H86; V_L89 and V_H83; V_L90 and V_H84; V_L91 and V_H85; V_L92 and V_H87; V_L93 and V_H88; V_L94 and V_H88; V_L95 and V_H89; V_L96 and V_H90; V_L97 and V_H91; V_L98 and V_H92; V_L99 and V_H93; and V_L100 and V_H94.
9. The antigen binding protein of embodiment 8, further comprising:
1. (a) the kappa light chain constant sequence of SEQ ID NO: 12
2. (b) the lambda light chain constant sequence of SEQ ID NO: 13
3. (c) the heavy chain constant sequence of SEQ ID NO: 11; or
4. (d)
  1. (i) the kappa light chain constant sequence of SEQ ID NO: 12 or the lambda light chain constant sequence of SEQ ID NO: 13, and
  2. (ii) the heavy chain constant sequence of SEQ ID NO: 11.
10. The antigen binding protein of embodiment 1, wherein the antigen binding protein is a human antibody, a humanized antibody, chimeric antibody, a monoclonal antibody, a polyclonal antibody, a recombinant antibody, an antigen-binding antibody fragment, a single chain antibody, a diabody, a triabody, a tetrabody, a Fab fragment, an F(fab')₂ fragment, a domain antibody, an IgD antibody, an IgE antibody, an IgM antibody, an IgG1 antibody, an IgG2 antibody, an IgG3 antibody, an IgG4 antibody, or an IgG4 antibody having at least one mutation in the hinge region.
11. The antigen binding protein of embodiment 1, that, when bound to to a complex comprising β-Klotho and at least one of (i) FGFR1c, (ii) FGFR2c and (iii) FGFR3c:
1. (a) binds to (i) β-Klotho; (ii) FGFR1c, FGFR2c, FGFR3c or FGFR4; or (iii) a complex comprising β-Klotho and one of FGFR1c, FGFR2c, FGFR3c, and FGFR4, with substantially the same Kd as a reference antibody;
2. (b) induces FGF21-like signaling of 10% or greater than the signaling induced by a wild-type FGF21 standard comprising the mature form of SEQ ID NO: 2 as measured in an ELK-luciferase reporter assay;
3. (c) exhibits an EC₅₀ of 10nM or less of FGF21-like signaling in an assay comprising one of:
  1. (i) a FGFR1c/β-Klotho-mediated in vitro recombinant cell-based assay; and
  2. (ii) an in vitro human adipocyte functional assay;
4. (d) exhibits an EC₅₀ of less than 10nM of agonistic activity on FGFR1c in the presence of β--Klotho in an in vitro recombinant FGFR1c receptor mediated reporter assay;
5. (e) exhibits an EC₅₀ of greater than 1µM of agonistic activity on FGFR1c in the absence of β-Klotho in an in vitro recombinant FGFR1c receptor mediated reporter assay;
6. (f) competes for binding with a reference antibody to (i) β-Klotho; (ii) FGFR1c, FGFR2c, FGFR3c or FGFR4; or (iii) a complex comprising β-Klotho and one of FGFR1c, FGFR2c, FGFR3c, and FGFR4, wherein the reference antibody comprises a combination of light chain and heavy chain variable domain sequences selected from the group consisting of V_L1 and V_H1; V_L2 and V_H1; V_L3 and V_H2 or V_H3; V_L4 and V_H4; V_L5 and V_H5; V_L6 and V_H6; V_L7 and V_H6; V_L8 and V_H7 or V_H8; V_L9 and V_H9; V_L10 and V_H9; V_L11 and V_H 10; V_L12 and V_H11; V_L13 and V_H12; V_L13 and V_H14; V_L14 and V_H13; V_L15 and V_H14; V_L16 and V_H15; V_L17 and V_H16; V_L18 and V_H17; V_L19 and V_H18; V_L20 and V_H19; V_L21 and V_H20; V_L22 and V_H21; V_L23 and V_H22; V_L24 and V_H23; V_L25 and V_H24; V_L26 and V_H25; V_L27 and V_H26; V_L28 and V_H27; V_L29 and V_H28; V_L30 and V_H29; V_L31 and V_H30; V_L32 and V_H31; V_L33 and V_H32; V_L34 and V_H33; V_L35 and V_H34; V_L36 and V_H35; V_L37 and V_H36; V_L38 and V_H37; V_L39 and V_H38; V_L40 and V_H39; V_L41 and V_H40; V_L42 and V_H41; V_L43 and V_H42; V_L44 and V_H43; V_L45 and V_H44; V_L46 and V_H45; V_L47 and V_H46; V_L48 and V_H47; V_L49 and V_H48; V_L50 and V_H49; V_L51 and V_H50; V_L52 and V_H51; V_L53 and V_H52; V_L54 and V_H53; V_L55 and V_H54; V_L56 and V_H54; V_L57 and V_H54; V_L58 and V_H55; V_L59 and V_H56; V_L60 and V_H57; V_L61 and V_H58; V_L62 and V_H59; V_L63 and V_H60; V_L64 and V_H1; V_L65 and V_H62; V_L66 and V_H63; V_L67 and V_H64; V_L68 and V_H65; V_L69 and V_H66; V_L70 and V_H67; V_L71 and V_H68; V_L72 and V_H69; V_L73 and V_H70; V_L74 and V_H70; V_L75 and V_H70; V_L76 and V_H71; V_L77 and V_H72; V_L78 and V_H73; V_L79 and V_H74; V_L80 and V_H75; V_L81 and V_H76; V_L82 and V_H77; V_L83 and V_H78; V_L84 and V_H79; V_L85 and V_H80; V_L86 and V_H81; V_L87 and V_H82; V_L88 and V_H86; V_L89 and V_H83; V_L90 and V_H84; V_L91 and V_H85; V_L92 and V_H87; V_L93 and V_H88; V_L94 and V_H88; V_L95 and V_H89; V_L96 and V_H90; V_L97 and V_H91; V_L98 and V_H92; V_L99 and V_H93; and V_L100 and V_H94; and
7. (g) two or more of (a) - (f).
12. The antigen binding protein of embodiment 11, that, when bound to a complex comprising β-Klotho and at least one of (i) FGFR1c, (ii) FGFR2c and (iii) FGFR3c and:
1. (a) lowers blood glucose in an animal model;
2. (b) lowers serum lipid levels in an animal model;
3. (c) lowers insulin levels in an animal model; or
4. (d) two or more of (a) and (b) and (c).
13. The antigen binding protein of embodiment 1, wherein the antigen binding protein comprises one or more of:
1. (a) a heavy chain comprising one of H1-H94;
2. (b) a light chain comprising one of L1-L100; and
3. (c) a combination comprising a heavy chain of (a) and a light chain of (b).
14. A pharmaceutical composition comprising one or more antigen binding proteins of embodiments 1-13 in admixture with a pharmaceutically acceptable carrier thereof.
15. An isolated nucleic acid comprising a polynucleotide sequence encoding the light chain variable domain amino acid sequence, the heavy chain variable domain amino acid sequence, or both amino acid sequences, of an antigen binding protein of embodiments 1-13.
16. The isolated nucleic acid of embodiment 15, wherein the encoded amino acid sequence comprises one or more of:
1. (a) V_L1-V_L100;
2. (b) V_H1-V_H94; and
3. (c) a combination comprising one or more sequences of (a) and one or more sequences of (b).
17. An expression vector comprising the nucleic acid of embodiment 16.
18. An isolated cell comprising the nucleic acid of embodiment 17.
19. An isolated cell comprising the expression vector of embodiment 18.
20. A method of producing an antigen binding protein comprising incubating the host cell of embodiment 19 under conditions that allow it to express the antigen binding protein.
21. A method of preventing or treating a condition in a subject in need of such treatment comprising administering a therapeutically effective amount of the composition of embodiment 14 to the subject, wherein the condition is treatable by lowering one or more of blood glucose, insulin or serum lipid levels.
22. The method of embodiment 21, wherein the condition is type 2 diabetes, obesity, dyslipidemia, NASH, cardiovascular disease or metabolic syndrome.
23. The antigen binding protein of embodiment 1, wherein the antigen binding protein comprises one or more non-naturally occurring or encoded amino acids.

EXAMPLES

The following examples, including the experiments conducted and the results achieved, are provided for illustrative purposes only and are not to be construed as limiting.

EXAMPLE 1 PREPARATION OF FGFR1c AND β-KLOTHO OVER EXPRESSING CELLS FOR USE AS AN ANTIGEN

Nucleic acid sequences encoding the full length human FGFR1c polypepetide (SEQ ID NO: 4; Figures 1A-1B) and a separate sequence encoding the full length human β-Klotho polypeptide (SEQ ID NO: 7; Figures 2A-2C) were subcloned into suitable mammalian cell expression vectors (e.g., pcDNA3.1 Zeo, pcDNA3.1 Hyg (Invitrogen, Carlsbad, CA) or pDSRα24. The pDSRα24 vector contains SV40 early promoter/enhancer for expressing the gene of interest and a mouse DHFR expression cassette for selection in CHO DHFR (-) host cells such as AM1/D CHO (a derivative of DG44, CHO DHFR (-)).

AM-1/D CHO cells were transfected with linearized DNAs of huFGFR1c and huβ-Klotho in standard mammalian cell expression vectors e.g. pcDNA3.1 puro and pcDNA3.1 Hyg with Lipofectamine 2000 (Invitrogen, Carlsbad CA). The transfected cells were trypsinized 2 days after transfection and seeded into media containing the corresponding selection drugs i.e. puromycin and hygromycin. After 2 weeks, the resulting transfected colonies were trypsinized and pooled. Single cell clones from the pools were isolated and screened with antibodies to huFGFR1c and huβKlotho in FACS and Clone 16 was selected due to the high level and balanced expression of the two receptor components.

2x10e9 fresh cells from Clone 16 were harvested from roller bottles into a smaller volume in PBS and incubated with 10µg/ml recombinant FGF21 (Amgen, Thousand Oaks CA) at 4C for 1 hours to form complex with the cell surface receptors. The cells were washed twice with cold PBS, pelleted by centrifugation and frozen in individual vials at 2x10e8 cells for immunization.

HEK 293T cells were transfected with DNA expressing a truncated version of huFGFR1c (a signal peptide VH21 was joined to the remaining FGFR1c from amino acid residue #141 to #822 (in SEQ ID NO: 4) with a deletion that removed both the D1 domain and the acidic box (AB) and DNA expressing the full length huβ-Klotho in pcDNA3.1 series or pTT5 (an expression vector developed by Durocher, NRCC, with CMV promoter and EBV ori) based vector for transient expression. The removal of the D1-AB on FGFR1c was designed to expose epitopes on FGFR1c (e.g., in the D2 and D3 domains) that may be masked by this auto-inhibitory domain (see Mohammadi et al., (2005) Cytokine Growth Factor Reviews, 16, 107-137; Gupte et al., (2011) J. Mol. Biol. 408:491-502).

The expression of β-Klotho and truncated FGFR1c in the transfected 293T cells was verified by the respective specific antibodies in FACS and cells were harvested on day 3 post-transfection and frozen as cell pellet into aliquots for immunization.

Stable CHO or transiently transfected HEK 293T cells expressing FGFR1c and β -Klotho individually or together were also generated and used for titering mouse sera by FACS after immunization and for binding screens of the hybridoma supernatants by FMAT (see Example 3).

EXAMPLE 2 PREPARATION OF MONOCLONAL ANTIBODIES

Immunizations were conducted using one or more suitable forms of FGF21 receptor antigen, including: (1) cell-bound receptor of CHO transfectants expressing full length human FGFR1c and β-Klotho at the cell surface, obtained by transfecting CHO cells with cDNA encoding a human full length FGFR1c polypeptide of SEQ ID NO: 4 (see also Figuresla-b) and cDNA encoding a human β-Klotho polypeptide of SEQ ID NO: 7 (see also Figures 2a-c) in a balanced ratio with cells and incubated with FGF21 prior to freezing; (2) cell-bound receptor of 293T transfectants expressing full length human β-Klotho and an N-terminal truncated form of human FGFR1c encompassing amino acid residues 141-822 polypeptide of SEQ ID NO: 4 (D1 domain of FGFR1c deleted).

A suitable amount of immunogen (i.e., 3 - 4 x 10⁶ cells/mouse of stably transfected CHO cells or transiently transfected 293T cells mentioned above was used for initial immunization in XENOMOUSE® according to the methods disclosed in U.S. Patent Application Serial No. 08/759,620, filed December 3, 1996 and International Patent Application Nos. WO 98/24893 , and WO 00/76310 , the disclosures of which are incorporated by reference. Following the initial immunization, subsequent boost immunizations of immunogen (1.7 x 10⁶ FGF21R transfected cells/mouse) were administered on a schedule and for the duration necessary to induce a suitable anti-FGF21R titer in the mice. Titers were determined by a suitable method, for example, by enzyme immunoassay, fluorescence activated cell sorting (FACS), or by other methods (including combinations of enzyme immunoassays and FACS).

Animals exhibiting suitable titers were identified, and lymphocytes were obtained from draining lymph nodes and, if necessary, pooled for each cohort. Lymphocytes were dissociated from lymphoid tissue by grinding in a suitable medium (for example, Dulbecco's Modified Eagle Medium; DMEM; obtainable from Invitrogen, Carlsbad, CA) to release the cells from the tissues, and suspended in DMEM. B cells were selected and/or expanded using standard methods, and fused with suitable fusion partner, for example, nonsecretory myeloma P3X63Ag8.653 cells (American Type Culture Collection CRL 1580; Kearney et al, (1979) J. Immunol. 123:1548-1550), using techniques that were known in the art.

In one suitable fusion method, lymphocytes were mixed with fusion partner cells at a ratio of 1:4. The cell mixture was gently pelleted by centrifugation at 400 x g for 4 minutes, the supernatant decanted, and the cell mixture gently mixed (for example, by using a 1 ml pipette). Fusion was induced with PEG/DMSO (polyethylene glycol/dimethyl sulfoxide; obtained from Sigma-Aldrich, St. Louis MO; 1 ml per million of lymphocytes). PEG/DMSO was slowly added with gentle agitation over one minute followed, by one minute of mixing. IDMEM (DMEM without glutamine; 2 ml per million of B cells), was then added over 2 minutes with gentle agitation, followed by additional IDMEM (8 ml per million B-cells) which was added over 3 minutes.

The fused cells were pelleted (400 x g 6 minutes) and resuspended in 20 ml Selection media (for example, DMEM containing Azaserine and Hypoxanthine [HA] and other supplemental materials as necessary) per million B-cells. Cells were incubated for 20-30 minutes at 37°C and then resuspended in 200 ml selection media and cultured for three to four days in T175 flasks prior to 96 well plating.

Cells were distributed into 96-well plates using standard techniques to maximize clonality of the resulting colonies. An alternative method was also employed and the fused cells were directly plated clonally into 384-well plates to ensure monoclonality from the start. After several days of culture, supernatants were collected and subjected to screening assays as detailed in the examples below, including confirmation of binding to human FGF21 receptor, specificity and/or cross-species reactivity. Positive cells were further selected and subjected to standard cloning and subcloning techniques. Clonal lines were expanded in vitro, and the secreted human antibodies obtained for analysis.

In this manner, mice were immunized with cells expressing full length FGF21R cells mixed with FGF21, or cells expressing a truncated FGFR1c and full length β-Klotho, with a range of 11-17 immunizations over a period of approximately one to three and one-half months. Several cell lines secreting FGF21R-specific antibodies were obtained, and the antibodies were further characterized. The sequences thereof are presented herein and in the Sequence Listing, and results of various tests using these antibodies are provided.

EXAMPLE 3 SELECTION OF BINDING ANTIBODIES BY FMAT

After 14 days of culture, hybridoma supernatants were screened for FGF21R-specific monoclonal antibodies by Fluorometric Microvolume Assay Technology (FMAT) by screening against either the CHO AM1/D/huFGF21R cell line or recombinant HEK293 cells that were transfected with human FGF21R and counter-screening against parental CHO or HEK293 cells. Briefly the cells in Freestyle media (Invitrogen) were seeded into 384-well FMAT plates in a volume of 50 µL/well at a density of 4,000 cells/well for the stable transfectants, and at a density of 16,000 cells/well for the parental cells, and cells were incubated overnight at 37°C. 10 µL/well of supernatant was then added, and the plates were incubated for approximately one hour at 4°C, after which 10 µL/well of anti-human IgG-Cy5 secondary antibody was added at a concentration of 2.8 µg/ml (400 ng/ml final concentration). Plates were then incubated for one hour at 4°C, and fluorescence was read using an FMAT Cellular Detection System (Applied Biosystems).

In total, over 1,500 hybridoma supernatants were identified as binding to the FGF21 receptor expressing cells but not to parental cells by the FMAT method. These supernatants were then tested in the FGF21 functional assays as described below.

EXAMPLE 4 SELECTION OF ANTIBODIES THAT INDUCE FGF21-LIKE SIGNALING

Experiments were performed to identify functional antibodies that mimic wild-type FGF21 activity (e.g., the ability to induce FGF21-like signaling) using a suitable FGF21 reporter assay. The disclosed FGF21 reporter assay measures activation of FGFR signaling via a MAPK pathway readout. β-Klotho is a co-receptor for FGF21 signaling, and although it is believed not to have any inherent signaling capability due to its very short cytoplasmic domain, it is required for FGF21 to induce signaling through FGFRs.

Example 4.1 ELK-Luciferase Reporter Assay

ELK-luciferase assays were performed using a recombinant human 293T kidney cell or CHO cell system. Specifically, the host cells were engineered to over-express β-Klotho and luciferase reporter constructs. The reporter constructs contain sequences encoding GAL4-ELK1 and 5xUAS-Luc, a luciferase reporter driven by a promoter containing five tandem copies of the Gal4 binding site. Activation of the FGF21 receptor complex in these recombinant reporter cell lines induces intracellular signal transduction, which in turn leads to ERK and ELK phosphorylation. Luciferase activity is regulated by the level of phosphorylated ELK, and is used to indirectly monitor and quantify FGF21 activity.

In one example, CHO cells were transfected sequentially using the Lipofectamine 2000 transfection reagent (Invitrogen) according to the manufacturer's protocol with the receptor constructs expressing β-Klotho, FGFR1c and the reporter plasmids: 5x Gal4-Luciferase (minimal TK promoter with 5xGal4 binding sites upstream of luciferase) and Gal4-ELK1. Gal4-ELK1 binds to the Gal4 binding sites and activates transcription when it is phosphorylated by ERK. Luciferase transcription, and thereby the corresponding enzymatic activity in this context is regulated by the level of phosphorylated ELK1, and is used to indirectly monitor and quantify FGF21 activity.

Clone 16 was selected as the FGF21 luciferase reporter cell line based on the optimal assay window of 10-20 fold with native FGF21 exhibiting an EC50 in the single nM range.

For the assay, the ELK-luciferase reporter cells were plated in 96 well assay plates, and serum starved overnight. FGF21 or test samples were added for 6 hours at 37 degrees. The plates were then allowed to cool to room temperature and the luciferase activity in the cell lysates was measured with Bright-Glo (Promega).

Example 4.2 ERK-Phosphorylation Assay

Alternative host cell lines specifically L6 (a rat myoblastic cell line) was developed and applied to identify antibodies with FGF21-like signaling activity. The rat L6 cell line is a desirable host cell line for the activity assay because it is known to express minimal levels of endogeneous FGF receptors. The L6 cells do not respond to FGF21 even when transfected with β-Klotho expression vector and therefore provides a cleaner background. (Kurosu et al., (2007) J. Biol. Chem. 282, 26687-26695).

Human primary preadipocytes isolated from subcutaneous adipose tissues of multiple healthy nondiabetic donors were purchased from Zen-Bio, Inc. The preadipocytes were plated in 24-well plates and differentiated for 18 days into mature adipocytes. After a 3-hour starvation period, adipocytes were treated with different concentrations of test molecules for 10 minutes. Following treatment, the media was aspirated and cells were snap-frozen in liquid nitrogen. Cell lysates were prepared and ERK phosphorylation was measured using the Phospho-ERK1/2(Thr202/Tyr204;Thr185/Tyr187) /Total ERK1/2 Assay Whole Cell Lysate Kit from Meso Scale Discovery.

L6 cells were maintained in Dulbecco's modified Eagle's medium supplemented with 10% fetal bovine serum and penicillin/streptomycin. Cells were transfected with plasmids expressing β-Klotho and individual FGFR using the Lipofectamine 2000 transfection reagent (Invitrogen) according to the manufacturer's protocol.

Analysis of FGF signaling in L6 cells was performed as described in the literature (Kurosu et al., (2007) J. Biol. Chem. 282, 26687-26695). Cell cultures were collected 10 min after the treatment of FGF21 or test molecules and snap frozen in liquid nitrogen, homogenized in the lysis buffer and ERK phosphorylation was measured using the Phospho-ERK1/2(Thr202/Tyr204;Thr185/Tyr187) /Total ERK1/2 Assay Whole Cell Lysate Kit from Meso Scale Discovery."

In addition, the factor-dependent mouse BaF3 cell-based proliferation assay used frequently for cytokine receptors can also be developed and applied.

Among the hybridoma supernatants tested in the CHO cell (Clone 16) based human FGF21 ELK-luciferase reporter assay, over 140 were identified as positive (> 20% of the activity of FGF21) when compared to 20nM FGF21 as the positive control (Figures 3 and 4). Antibodies can be purified from the conditioned media of the hybridoma cultures of these positives and tested again in the CHO cell based ELK-luciferase reporter assay to assess the potency of the representative antibodies in the dose-responsive assay and determine the EC50. The activities and potency can be confirmed in the L6 cell based ERK1/2-phosphrylation assay. The EC50 is expected to be consistent to the ELK-luciferase assay in the CHO stable cell line Clone 16.

EXAMPLE 5 DETERMINING THAT INDUCTION OF FGF21-LIKE SIGNALING IS SPECIFIC TO THE FGFR/βKLOTHO COMPLEX

FGF21 has been reported to signal through multiple receptor complexes including FGFR1c, 2c, 3c, and 4 when paired with β-Klotho. The selectivity of the FGF21 agonistic antibodies can be determined in the rat myoblastic L6 cells transfected with vectors expressing the respective FGFRs and β-Klotho as described in Example 4.2.

Observed selectivity would be strongly suggestive that the action of these antibodies is β-Klotho-dependent yet it must also involve the FGFR component of the signaling complex. The results are set forth in Table 6 below.



+	+	+	-
+	+	+	+
+	-	-	-
+	-	-	-
+	-	-	-
+	-	-	-
+	-	-	-
+	-	-	-
+	-	-	-
+	-	-	-
+	-	-	-
+	-	-	-
+	-	-	-
+	-	-	-
+	-	-	-
+	-	-	-
+	-	-	-
+	-	-	-
+	-	-	-

EXAMPLE 5.1 BINDING SPECIFICITY IS EXCLUSIVELY β-KLOTHO DEPENDENT

The binding specificity of the reporter assay positive antibodies in the hybridoma supernatants was determined by FACS using 293T cells transiently transfected to express full length FGFR1c alone, β-Klotho alone or FGFR1c and β-Klotho together. Over 98% (141 out of 143 hybridomas) bind β-Klotho alone whereas none bind FGFR1c alone.

EXAMPLE 6 ACTIVITY IN PRIMARY HUMAN ADIPOCYTES

FGF21 stimulates glucose uptake and lipolysis in cultured adipocytes, and adipocytes are considered to be more physiologically relevant than the recombinant reporter cell system.

A panel of the antibodies were tested in the human adipocyte assay for Erk-phosphorylation activity as described in Example 4.2 and compared with FGF21 for their EC₅₀. The results are set forth below in Table 10 below.



Fgf21	0.623
16H7	0.280
49H12.1	0.254
51A8.1	0.213
51E5.1	3.221
54A1.1	0.206
60D7.1	0.496
63A10.1	0.435
64B10.1	0.955
65C3.1	6.387
66G2.1	3.529
67F5.1	1.438
67C10.1	5.789
68C8.1	1.216
49C8.1	0.243
49G3.3	1.424
56E7.3	0.916
58C2.1	0.317

EXAMPLE 7 COMPETITION BINDING AND EPITOPE BINNING

To compare the similarity of the binding sites of the antibodies on the FGF21 receptor, a series of competition binding experiments can be performed and measured by Biacore. In one example, representative agonistic FGF21 receptor antibodies (and any controls) can be immobilized on the sensor chip surface. Soluble human FGFR1c/β--Klotho ECD-Fc complex or β-Klotho can then be captured on the immobilized antibody surfaces. Finally, several of the test FGF21 receptor antibodies can be injected individually over the captured soluble human FGF21 receptor or β-Klotho. If the injected antibody recognizes a distinct binding site relative to that recognized by the immobilized antibody, a second binding event will be observed. If the antibodies recognize very similar binding site, no more binding will be observed.

Alternatively or additionally, a Biacore analysis can be carried out with biotinylated-FGF21 immobilized on the sensor ship. 10 nM soluble β-Klotho is then passed over the chip alone or mixed with the individual test antibodies at 100nM. The results are set forth below in Table 11 below.

EXAMPLE 8 RECOGNITION OF NATIVE AND DENATURED STRUCTURES

The ability of disclosed antigen binding proteins to recognize denatured and native structures was investigated. The procedure and results were as follows.

Example 8.1 FGF21 Receptor Agonistic Antibodies do not Recognize Denatured Structures

Cell lysates from CHO cells stably expressing FGF21 receptor (FGFR1c and β-Klotho) or CHO parental cells were diluted with sample buffer without beta-mercaptoethanol (non-reducing conditions). 20µl of cell lysate were loaded per lane on adjacent lanes separated with a molecular weight marker lane on 4-20% SDS-PAGE gels. Following electrophoresis, the gels were blotted onto 0.2µ nitrocellulose filters. The blots were treated with Tris-buffered saline/Triton-X (TBST) plus 5% non-fat milk (blocking buffer) for 30 minutes. The blots were then cut along the molecular weight marker lanes. The strips were probed with commercial goat anti-murine βKlotho or mouse anti-huFGFRl (R&D Diagnostics) in TBST/5% milk. Blots were incubated with the antibodies for one hour at room temperature, followed by three washes with TBST + 1% milk. The blots were then probed with anti-human or anti-goat IgG-HRP secondary antibodies for 20 min. Blots were given three 15 minute washes with TBST followed by treatment with Pierce Supersignal West Dura developing reagent (1 minute) and exposure to Kodak Biomax X-ray film.

The commercial anti-β-Klotho and anti-FGFRl antibodies detected the corresponding receptor proteins in the SDS-PAGE indicating they bind to denatured receptor proteins.

Example 8.2 FGF21 Receptor Agonistic Antibodies Bind To Native Receptor Structure

A FACS binding assay was performed with several commercially available FGFR1c and β-Klotho antibodies, and several of the disclosed FGF21 receptor agonistic antibodies. The experiments were performed as follows.

CHO cells stably expressing FGF21 receptor were treated with R&D Systems mouse anti-huFGFR1, goat anti-mu β-Klotho (1µg per 1x10⁶ cells in 100µl PBS/0.5% BSA). Cells were incubated with the antibodies at 4°C followed by two washes with PBS/BSA. Cells were then treated with FITC-labeled secondary antibodies at 4°C followed by two washes. The cells were resuspended in 1ml PBS/BSA and antibody binding was analyzed using a FACSCalibur™ instrument.

None of the commercial anti-β-Klotho or anti-FGFRl antibodies tested bind well to cell surface FGF21 receptor, as determined by FACS. This observation further confirmed that the commercial antibodies to the receptor components bind to denatured and non-native structure whereas all of the agonistic antibodies described herein bind receptors on cell surface as shown by FACS or FMAT which were the initial screens.

EXAMPLE 9 ARGININE SCANNING

As described above, antigen binding proteins that bind a complex comprising b-Klotho and one of FGFR1c, FGFR2c and FGFR3c can be created and characterized. To determine the neutralizing determinants on human FGFR1c and/or β-Klotho that these various antigen binding proteins bound, a number of mutant FGFR1c and/or β-Klotho proteins can be constructed having arginine substitutions at select amino acid residues of human FGFR1c and/or β-Klotho. Arginine scanning is an art-recognized method of evaluating where antibodies, or other proteins, bind to another protein, see, e.g., Nanevicz et al., (1995) J. Biol. Chem., 270:37, 21619-25 and Zupnick et al., (2006) J. Biol. Chem., 281:29, 20464-73. In general, the arginine sidechain is positively charged and relatively bulky as compared to other amino acids, which can disrupt antibody binding to a region of the antigen where the mutation is introduced. Arginine scanning is a method that determines if a residue is part of a neutralizing determinant and/or an epitope.

Various amino acids distributed throughout the human FGFR1c and/or β-Klotho extracellular domains can be selected for mutation to arginine. The selection can be biased towards charged or polar amino acids to maximize the possibility of the residue being on the surface and reduce the likelihood of the mutation resulting in misfolded protein. Using standard techniques known in the art, sense and anti-sense oligonucleotides containing the mutated residues can be designed based on criteria provided by Stratagene Quickchange® II protocol kit (Stratagene/Agilent, Santa Clara, CA). Mutagenesis of the wild-type (WT) FGFR1c and/or β-Klotho sequences can be performed using a Quickchange® II kit (Stratagene). Chimeric constructs can be engineered to encode a FLAG-histidine tag (six histidines (SEQ ID NO: 1830)) on the carboxy terminus of the extracellular domain to facilitate purification via the poly-His tag.

Multiplex analysis using the Bio-Plex Workstation and software (BioRad, Hercules, CA) can be performed to determine neutralizing determinants on human FGFR1c and/or β-Klotho by analyzing exemplary human FGFR1c and/or β-Klotho mAbs differential binding to arginine mutants versus wild-type FGFR1c and/or β-Klotho proteins. Any number of bead codes of pentaHis-coated beads ("penta-His" disclosed as SEQ ID NO: 1831) (Qiagen, Valencia, CA) can be used to capture histidine-tagged protein. The bead codes can allow the multiplexing of FGFR1c and/or β-Klotho arginine mutants and wild-type human FGFR1c and/or β-Klotho.

To prepare the beads, 100ul of wild-type FGFR1c and/or β-Klotho and FGFR1c and/or β-Klotho arginine mutant supernatants from transient expression culture are bound to penta-His-coated beads ("penta-His" disclosed as SEQ ID NO: 1831) overnight at 4°C or 2 hours at room temperature with vigorous shaking. The beads are then washed as per the manufacturer's protocol and the bead set pooled and aliquoted into 2 or 3 columns of a 96-well filter plate (Millipore, Bellerica, MA, product #MSBVN1250) for duplicate or triplicate assay points, respectively. 100ul anti-FGFR1c and/or anti-β-Klotho antibodies in 4-fold dilutions are added to the wells, incubated for 1 hour at room temperature, and washed. 100µl of a 1:100 dilution of PE-conjugated anti-human IgG Fc (Jackson Labs., Bar Harbor, ME) is added to each well, incubated for 1 hour at room temperature and washed. Beads are resuspended in 1% BSA, shaken for 3 minutes, and read on the Bio-Plex workstation. Antibody binding to FGFR1c and/or β-Klotho arginine mutant protein is compared to antibody binding to the human FGFR1c and/or β-Klotho wild-type from the same pool. A titration of antibody over approximately a 5 log scale can be performed. Median Fluorescence Intensity (MFI) of FGFR1c and/or β-Klotho arginine mutant proteins can be graphed as a percent of maximum wild-type human FGFR1c and/or β-Klotho signal. Those mutants for which signal from all the antibodies are below a cut-off value, e.g., 30% of wild-type FGFR1c and/or β-Klotho can be deemed to be either of too low a protein concentration on the bead due to poor expression in the transient culture or possibly misfolded and can be excluded from analysis. Mutations (i.e., arginine substitutions) that increase the EC50 for the FGFR1c and/or β-Klotho mAb by a cut-off value, e.g., 3-fold or greater (as calculated by, e.g., GraphPad Prism®) can be considered to have negatively affected FGFR1c and/or β-Klotho mAb binding. Through these methods, neutralizing determinants and epitopes for various FGFR1c and/or β-Klotho antibodies are elucidated.

EXAMPLE 10 PROTEASE PROTECTION ANALYSIS

Regions of the human FGF21 receptor bound by the antigen binding proteins that bind human FGF21 receptor, e.g., FGFR1c, β-Klotho or FGFR1c and β-Klotho complex can be identified by fragmenting human FGF21 receptor into peptides with specific proteases, e.g., AspN, Lys-C, chymotrypsin or trypsin. The sequence of the resulting human FGF21 receptor peptides (i.e., both disulfide- and non-disulfide-containing peptide fragments from FGFR1c and β-Klotho portions) can then be determined. In one example, soluble forms of a human FGF21 receptor, e.g., a complex comprising the FGFR1c ECD-Fc and β-Klotho ECD-Fc heterodimer described herein can be digested with AspN (which cleaves after aspartic acid and some glutamic acid residues at the amino end) by incubating about 100 µg of soluble FGF21 receptor at 1.0 mg/ml in 0.1M sodium phosphate (pH 6.5) for 20 hrs at 37°C with 2 µg of AspN.

A peptide profile of the AspN digests can then be generated on HPLC chromatography while a control digestion with a similar amount of antibody is expected to be essentially resistant to AspN endoprotease. A protease protection assay can then be performed to determine the proteolytic digestion of human FGF21 receptor in the presence of the antigen binding proteins. The general principle of this assay is that binding of an antigen binding protein to the FGF21 receptor can result in protection of certain specific protease cleavage sites and this information can be used to determine the region or portion of FGF21 receptor where the antigen binding protein binds.

Briefly, the peptide digests can be subjected to HPLC peptide mapping; the individual peaks are collected, and the peptides are identified and mapped by on-line electrospray ionization LC-MS (ESI-LC-MS) analyses and/or by N-terminal sequencing. HPLC analyses for these studies can be performed using a narrow bore reverse-phase C18 column (Agilent Technologies) for off-line analysis and using a capillary reverse phase C18 column (The Separation Group) for LC-MS. HPLC peptide mapping can be performed with a linear gradient from 0.05% trifluoroacetic acid (mobile phase A) to 90% acetonitrile in 0.05% trifluoroacetic acid. Columns can be developed at desirable flow rate for narrow bore HPLC for off-line or on-line LC-MS analyses, and for capillary HPLC for on-line LC-MS analyses.

Sequence analyses can be conducted by on-line LC-MS/MS and by Edman sequencing on the peptide peaks recovered from HPLC. On-line ESI LC-MS analyses of the peptide digest can be performed to determine the precise mass and sequence of the peptides that are separated by HPLC. The identities of selected peptides present in the peptide peaks from the protease digestion can thus be determined.

EXAMPLE 11 CONSTRUCTION OF CHIMERIC RECEPTORS

An additional method of determining activation determinants on which these various antigen binding proteins bind is as follows. Specific chimeric FGFR1c and/or β-Klotho proteins between human and mouse species can be constructed, expressed in transient or stable 293 or CHO cells (as described herein) and tested. For example, a chimeric FGF21 receptor can be constructed comprising native human FGFR1c, FGFR2c, FGFR3c or FGFR4 receptors. By way of example, FGFR1c can be paired with chimeric human/mouse β-Klotho in which selected regions or sequences on the human β-Klotho are systematically replaced by the corresponding mouse-specific residues (see, e.g., Figure 2A-2C). Similarly, native human β-Klotho can be paired with chimeric human/mouse FGFR1c, FGFR2c, FGFR3c or FGFR4. Here, selected regions or sequences on the human FGFR1c are systematically replaced by the corresponding mouse-specific residues (see, e.g., the alignments of Figures1A-1B). The critical sequences involved in the binding and/or activity of the antigen binding proteins can be derived through binding assay or activity measurements described in previous Examples 4, 5, 6, and 7 based on the chimeric FGF21 receptors.

Example 11.1 Construction of Specific Chimeras

Human-mouse β-Klotho chimeras were constructed using the methodology described above. A schematic of the chimeras constructed is presented in Figure 4. In summary, the chimeras generated comprised (from N- to C- terminus) a fusion of a human β-Klotho sequence fused to a murine β-Klotho sequence fused to a human β-Klotho sequence. Human β-Klotho (SEQ ID NO:8) was used as a framework into which regions of murine β-Klotho (full length sequence shown in SEQ ID NO:468) were inserted. The regions of murine β-Klotho that were inserted were as follows:

Murine Residues 82P-520P
Murine Residues 506F-1043S
Murine Residues 1M-193L
Murine Residues 82P-302S
Murine Residues 194Y-416G
Murine Residues 302S-506F
Murine Residues 416G-519P
Murine Residues 507P-632G
Murine Residues 520P-735A
Murine Residues 632G-849Q
Murine Residues 735A-963S
Murine Residues 1M-81F
Murine Residues 82P-193L

The chimeras that were generated using the murine β-Klotho sequences comprised the following:


1	huBeta_Klotho (1-81, 523-1044) (muBetaKLOTHO 82-520)	1-81	82-520	523-1044
2	huBeta_Klotho (1-507) (muBetaKLOTHO 506F-1045S)	1-507	506-1043
3	huBeta_Klotho (194-1044) (muBetaKLOTHO 1-L193)		1-193	194-1044
4	huBeta_Klotho (1-81, 303-1044) (muBetaKLOTHO 82P-302S)	1-81	82-302	303-1044
5	huBeta_Klotho (1-193, 419-1044) (muBetaKLOTHO Y194-416G)	1-193	194-416	419-1044
6	huBeta_Klotho(1-301, 509-1044) (muBetaKLOTHO S302-F506)	1-301	302-506	509-1044
7	huBeta_Klotho(1-417, 522-1044) (muBetaKLOTHO G416-F519)	1-417	416-519	522-1044
8	huBeta Klotho (1-507, 635-1044) (muBeta KLOTHO F06-G632)	1-508	507-632	635-1044
9	huBeta Klotho (1-521, 738-1044) (muBeta KLOTHO 520P-735A)	1-521	520-735	738-1044
10	huBeta_Klotho (1-633, 852-1044) (muBeta KLOTHO 632G-849Q)	1-633	632-849	852-1044
11	huBeta_Klotho (1-736, 967-1044) (muBeta KLOTHO 735A-963S)	1-736	735-963	967-1044
12	huBeta_Klotho (82-1044) (muBeta KLOTHO 1-81F)		1-81	82-1044
13	huBeta_Klotho (1-81, 194-1044) (muBeta KLOTHO 82P-193L)	1-81	82-193	194-1044
14	huBeta_Klotho (1-301, 509-743, 967-1044) (muBeta KLOTHO 302-506,742-964)	1-301	302-506, 742-964	967-1044

The generated chimeras comprised the following amino acid sequences:

(i) huBeta_Klotho(1-8L 523-1044)(muBetaKLOTHO 82-520)
(ii) huBeta_Klotho(-507)muBetaKLOTHO 506F-1045S)
(iii) huBeta_Klotho(194-1044)(muBetaKLOTHO 1-L193)
(iv) huBeta_Klotho(1-81, 303-1044)(muBetaKLOTHO 82P-302S)
(v) huBeta_Klotho(1-193, 419-1044)(muBetaKLOTHO Y194-416G)
(vi) huBeta_Klotho(1-301, 509-1044)(muBetaKLOTHO S302-F506)
(vii) huBeta_Klotho(1-417, 522-1044)(muBetaKLOTHO G416-F519)
(viii) huBeta_Klotho(1-507, 635-1044)(muBeta KLOTHO F06-G632)
(ix) huBeta_Klotho(1-521, 738-1044)(muBeta KLOTHO 520P-735A)
(x) huBeta_Klotho(1-633, 852-1044)(muBeta KLOTHO 632G-849Q)
(xi) huBeta_Klotho(1-736, 967-1044)(muBeta KLOTHO 735A-963S)
(xii) huBeta_Klotho(82-1044)(muBeta KLOTHO 1-81F)
(xiii) huBeta_Klotho (1-81,194-1044)(muBeta KLOTHO 82P-193L)
(xiv) huBeta_Klotho (1-301, 509-743, 967-1044) (muBetaKLOTHO 302-506, 742-964)

Various antigen binding proteins provided herein, as well as human FGF21, were tested for the ability to activate chimeras in L6 cells. Figure 5 shows the observed results with each tested molecule.

These data indicate that while human FGF21 was able to activate FGFR1c combined with all of the human/mouse β-Klotho chimeras (the "+" sign indicates activity on the receptor), the substitutions of mouse sequences into human β-Klotho affected the activities of 16H7, 37D3, and 39F7 (See Figure 5). These results suggest that β-Klotho sequences 1-81, 302-522, and 849-1044 are important for the activities of agonistic antigen binding proteins and may represent an important epitope for their function.

In addition, various antigen binding proteins were also tested for binding to the various human/mouse β-Klotho chimeras transiently expressed on the surface of HEK-293T cells by flow cytometry. Transfection and flow-cytometry was performed as described in Example 12. It will be appreciated that antibodies which do not have the ability to cross-bind full length murine β-Klotho are unable to bind the human/mouse β-Klotho chimera if the chimera spans a region of the antibody's binding site. In this manner, the binding profile of each antibody on the panel of chimeras reveals epitope information for the antibody. Data is shown below in Table 10. The anti-β-Klotho antibody 2G10 (which binds both human and mouse β-Klotho) was used as the positive control for expression of each human/mouse chimera. Using this positive control it was determine the expression level of chimeras 7 and 8 were not high enough to provide robust data and therefore they were eliminated from the analysis. One antibody, 26H11, was found to bind to full-length mouse β-Klotho and therefore could not be assigned an epitope in this analysis. Other antibodies which did not cross-bind to mouse β-Klotho could be group into epitope clusters. The first cluster included antibodies 16H7, 46D11, and 49G3.3, which antibodies did not bind to chimera #3 and chimera #12, indicating that the epitope includes the 1-81 region. Additionally, this group of antibodies also lacked observed binding to chimeras 1, 5, 6 and 14, which indicates that the epitope also includes the 294-506 region. Taken together, this data suggests that these antibodies have a complex non-linear type of epitope.

A second cluster included only antibody 65C3.1. This antibody lacked binding to chimeras #2, #11, and #14, indicating an epitope in the region of 849-936. A third cluster, including antibodies 49H12.1, 54A1.1, 49C8.1, 51A8.1, 63A10.1, 64B10.1, 68C8.1 and 39F7, lacked binding to chimera #1, #5, and #6, indicating that their epitope is in the 302-416 region. The forth cluster included antibodies 67C10.1, 51E5.1, 52A8.1, 66G2, 167F5.1, which lacked binding on chimeras #2, #8, #9, #10, #11, and #14 indicating that the epitope for these antibodies lies within region 506-1045. A "+" or "-" symbol in the chart below indicates binding of the respective antibody ("+"), or lack of binding ("-") to the chimera and/or the respective ortholog of β-Klotho, or Mock Cells (negative control).




2G10	+	+	+	+	+	+	+	+	+	+	+	+	+	+	-
26H11	+	+	+	+	+	+	+	+	+	+	+	+	+	+	-
16H7	-	+	-	+	-	-	+	+	+	-	+	-	+	-	-	1-81 & 302-416
49G3.3	-	+	-	+	-	-	+	+	+	-	+	-	+	-	-
46D11	-	+	-	+	-	-	+	+	+	-	+	-	+	-	-
49H12.1	-	+	+	+	-	-	+	+	+	+	+	-	+	-	-	302-416
54A1.1	-	+	+	+	-	-	+	+	+	+	+	-	+	-	-
49C8.1	-	+	+	+	-	-	+	+	+	+	+	-	+	-	-
51A8.1	-	+	+	+	-	-	+	+	+	+	+	-	+	-	-
63A10.1	-	+	+	+	-	-	+	+	+	+	+	-	+	-	-
64B10.1	-	+	+	+	-	-	+	+	+	+	+	-	+	-	-
68C8.1	-	+	+	+	-	-	+	+	+	+	+	-	+	-	-
39F7	-	+	+	+	-	-	+	+	+	+	+	-	+	-	-
65C3.1	+	-	+	+	+	+	+	+	-	+	+	-	+	-	-	849 - 936
67C10.1	+	-	+	+	+	+	-	-	-	+	+	-	+	-	-	506-1045
51E5.1	+	-	+	+	+	+	-	-	-	+	+	-	+	-	-
52A8.1	+	-	+	+	+	+	-	-	-	+	+	-	+	-	-
66G2.1	+	-	+	+	+	+	-	-	-	+	+	-	+	-	-
67F5.1	+	-	+	+	+	+	-	-	-	+	+	-	+	-	-
IgG2/K Control	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-
IgG4/K Control	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-
Secondar y Only	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-

EXAMPLE 12 FGF21 Receptor Agonistic Antibodies Binding Selectivity

A panel of FGF21 receptor agonistic antibodies were assayed using flow cytometry for the binding to human FGFR1/human β-klotho transiently co-transfected HEK293T cells, human FGFR1c transiently transfected HEK293T cells and β-klotho transiently transfected HEK293T cells. In addition, binding was also tested on HEK-293T cells transiently transfected with cynomologous monkey orthologs of FGFR1c and β-klotho. Cells were transfected by preparing 10ug plasmid DNA in 500ul OptiMEM™ media (Invitrogen™) and mixing this with 10ul of 293fectin™ in 500ul OptiMEM™ media, and then incubating the solution for 5 minutes at room temperature. This solution was then added dropwise to 10million HEK293T cells in 10ml of media. 24 hours following transfection, the cells were washed and 50,000 cells were stained with each primary antibody, 50ul of unpurified hybridoma supernatant was diluted 1:2 and used for staining cells. After a one hour incubation at 4°c, the cells were washed and an anti-Human Fc-specific secondary was added. Stained cells were then analyzed on a flow cytometer. The panel of hybridoma supernatants tested all bound specifically to human β-Klotho/human FGFR1c co-transfected cells as well as human β-Klotho transfected alone. Data is shown below in Table 11. No staining was detected for any of the antibodies on cells transfected with FGFR1c alone. All antibodies except 64B10.1 and 68C8.1 specifically detected cynomologous β-Klotho/cynoFGFR1c co-transfected cells.



Antibody	GeoMean	GeoMean	GeoMean	GeoMean	GeoMean
49G3.3	648	706	14891	17919	25947
49H12.1	581	719	16213	21731	20870
51E5.1	723	747	16900	20951	36536
51A8.1	728	795	17799	22826	18476
54A1.1	709	770	14317	18701	11106
59G10.3	686	780	15669	21105	33464
63A10.1	648	834	17442	20432	32558
64B10.1	624	691	14939	19850	701
65C3.1	705	719	13720	18835	24564
66G2.1	695	780	12671	16715	21566
67F5.1	632	757	13482	13948	15784
67C10.1	688	780	15114	18896	4063
68C8.1	592	798	15905	20622	750
16H7 @ 5ug/ml	723	869	16335	20686	31319

EXAMPLE 13 Hotspot/Covariant mutants

A total of 17 antibodies were analyzed for potential hotspots and covariance violations. The designed variants (shown below) outline amino acid substitutions capable of reducing and/or avoiding isomerization, deamidation, oxidation, covariance violations, and the like. In the data below, "02 49C8.1_VK:[F21I]" refers to a variant of the parental antibody 49C8.1 that has a mutation at position 21, from F (Phe) to I (Isoleucine). Note that a structure-based numbering scheme is followed for designating amino acid positions. It will be appreciated that these single point mutations can be combined in any combinatorial manner in order to arrive at a final desired molecule. The data are shown below in Table 15 and Table 16.


02 49C8.1_VK: [F21I]
03 49C8.1_VK: [F91L]
04 49C8.1_VK: [I101F]
05 49C8.1_VK: [I101V]
06 49C8.1_VK: [P141Q]
07 49C8.1_VK: [P141G]
08 49C8.1_VH: [T48P]
09 49C8.1_VH: [N61Q]
10 49C8.1_VH: [G65T]


01 49H12_N83D_VK: [F91L]
02 49H12_N83D_VK: [I101F]
03 49H12_N83D_VK: [I101V]
04 49H12_N83D_VH: [M24K]
05 49H12_N83D_VH: [I30T]
06 49H12_N83D_VH: [T48P]
07 49H12_N83D_VH: [W57Y]
08 49H12_N83D_VH: [W111Y]


01 49G3.3_VK: [F91L]
02 49G3.3_VK: [I101F]
03 49G3.3_VK: [I101V]
04 49G3.3_VK: [G141Q]
05 49G3.3_VH: [E17Q]
06 49G3.3_VH: [V25F]
07 49G3.3_VH: [T56A]
08 49G3.3_VH: [T56G]
09 49G3.3_VH: [T144L]
10 49G3.3_VH: [T144M]


01 51A8.1_VL: [I98T]
02 51A8.1_VL: [I98A]
03 51A8.1_VH: [R17G]
04 51A8.1_VH: [D61E]
05 51A8.1_VH: [D72E]
06 51A8.1_VH: [D110E]


01 51E5.1_VK: [N53K]
02 51E5.1_VK: [R54L]
03 51E5.1_VK: [R54S]
04 51E5.1_VK: [G141Q]
05 51E5.1_VH: [D59E]
06 51E5.1_VH: [H60T]

Antibody 52A8.1
01 52A8.1_VK: [F10S]
02 52A8.1_VK: [H44Y]
03 52A8.1_VK: [H44F]
04 52A8.1_VK: [G141Q]
05 52A8.1_VH: [W57Y]
06 52A8.1_VH: [R95S]
07 52A8.1_VH: [W135Y]


01 54A1.1_N83D_VK: [A5T]
02 54A1.1_N83D_VK: [L46Q]
03 54A1.1_N83D_VK: [G81S]
04 54A1.1_N83D_VK: [F91L]
05 54A1.1_N83D_VK: [I101F]
06 54A1.1_N83D_VK: [I101V]
07 54A1.1_N83D_VK: [P141G]
08 54A1.1_N83D_VK: [P141Q]
09 54A1.1_N83D_VH: [T48P]
10 54A1.1_N83D_VH: [W57Y]
11 54A1.1_N83D_VH: [W111Y]


01 56E7.3_VK: [N53K]
02 56E7.3_VK: [F91L]
03 56E7.3_VK: [I101F]
04 56E7.3_VK: [P141Q]
05 56E7.3_VK: [P141G]
06 56E7.3_VK: [T144K]
07 56E7.3_VK: [T144R]
08 56E7.3_VH: [L31F]
09 56E7.3_VH: [D65E]
10 56E7.3_VH: [T84K]
11 56E7.3_VH: [R95S]


01 58C2.1_VK: [D36E]
02 58C2.1_VH: [R17G]
03 58C2.1_VH: [D61E]
04 58C2.1_VH: [D72E]
05 58C2.1_VH: [N116Q]


01 60D7.1_N30T_VK: [D33E]
02 60D7.1_N30T_VK: [D36E]
03 60D7.1_N30T_VH: [R17G]
04 60D7.1_N30T_VH: [D61E]
05 60D7.1_N30T_VH: [D72E]
06 60D7.1_N30T_VH: [W115Y]


01 63A10.1_C58S_VL: [H9L]
02 63A10.1_C58S_VL: [H9P]
03 63A10.1_C58S_VL: [T15L]
04 63A10.1_C58S_VL: [T15P]
05 63A10.1_C58S_VL: [A16G]
06 63A10.1_C58S_VL: [M18T]
07 63A10.1_C58S_VL: [D51A]
08 63A10.1_C58S_VL: [D51S]
09 63A10.1_C58S_VL: [D51F]
10 63A10.1_C58S_VL: [D67E]
11 63A10.1_C58S_VL: [P83S]
12 63A10.1_C58S_VL: [E97Q]
13 63A10.1_C58S_VL: [D110E]
14 63A10.1_C58S_VL: [D136E]
15 63A10.1_C58S_VH: [D11G]
16 63A10.1_C58S_VH: [K14Q]
17 63A10.1_C58S_VH: [I29F]
18 63A10.1_C58S_VH: [G56S]
19 63A10.1 C58S_VH: [D64E]
20 63A10.1_C58S_VH: [G84D]
21 63A10.1_C58S_VH: [G84N]
22 63A10.1_C58S_VH: [T98A]
23 63A10.1_C58S_VH: [T107A]
24 63A10.1_C58S_VH: [T108R]
25 63A10.1_C58S_VH: [D109E]
26 63A10.1_C58S_VL: [W109Y]


01 63A10.3_ N20R_C42S_VL: [W109Y]
02 63A10.3_ _N20R_C42S_VL: [D67E]
03 63A10.3_ N20R_C42S_VL: [D110E]
04 63A10.3_N20R_C42S_VH: [D11G]
05 63A10.3_N20R_C42S_VH: [K14Q]
06 63A10.3_N20R_C42S_VH: [I29F]
07 63A10.3_N20R_C42S_VH: [G56S]
08 63A10.3_N20R_C42S_VH: [D64E]
09 63A10.3_N20R_C42S_VH: [G84N]
10 63A10.3_N20R_C42S_VH: [T98A]
11 63A10.3_N20R_C42S_VH: [T107A]
12 63A10.3_N20R_C42S_VH: [T108R]
13 63A10.3_N20R_C42S_VH: [D109E]
14 63A10.3_N20R_C42S_VL: [W109Y]


01 64B10.1_VL: [G92A]
02 64B10.1_VL: [G99E]
03 64B10.1_VL: [D110E]
04 64B10.1_VH: [L5Q]
05 64B10.1_VH: [T144L]
06 64B10.1_VH: [T144M]
07 64B10.1_VL: [W109Y]
08 64B10.1_VH: [W113Y]


01 66G2_VK: [R54L]
02 66G2_VK: [K88E]
03 66G2_VK: [K88D]
04 66G2_VK: [N110Q]
05 66G2_VH: [R17G]
06 66G2_VH: [D61E]
07 66G2_VH: [D72E]
08 66G2_VH: [I78F]
09 66G2_VH: [T108K]
10 66G2_VH: [T108R]


1.01 67F5_VK: [H57Y]
2.02 67F5_VK: [Q97E]
3.03 67F5_VK: [S98P]
4.04 67F5_VK: [A99E]
5.05 67F5_VK: [N105Y]
6.06 67F5_VH: [K5Q]
7.07 67F5_VK: [W135Y]
8.08 67F5_VK: [W137Y]


1.01 67C10_VK: [F2I]
2.02 67C10_VK: [D36E]
3.03 67C10_VH: [Q24K]
4.04 67C10_VH: [D65E]


1.01 68C8_VL: [G92A]
2.02 68C8_VL: [G99E]
3.03 68C8_VL: [D110E]
4.04 68C8_VH: [D29G]
5.05 68C8_VH: [H83D]
6.06 68C8_VH: [G107A]
7.07 68C8_VH: [T144L]
8.08 68C8_VH: [T144M]
9.09 68C8_VL: [W109Y]
10.10 68C8_VH: [W113Y]


>49G3.3_VH.07
>49G3.3_VK
>49G3.3 VH.08
>49G3.3_VK
>49G3.3 VH.09
>49G3.3_VK
>49G3.3_VH.10
>51A8.1_VL.01
>51A8.1_VH
>51A8.1_VL.02
>51A8.1_VH
>51A8.1_VL
>51A8.1_VH.03
>51A8.1_VL
>51A8.1_VH.04
>51A8.1_VL
>51A8.1_VH.05
>51A8.1_VL
>51A8.1_VH.06
>51E5.1_VK.01
>51E5.1_VH
>51E5.1_VK.02
>51E5.1_VH
>51E5.1_VK.03
>51E5.1_VH
>51E5.1_VK.04
>51E5.1_VH
>51E5.1_VK
>51E5.1_VH.05
>51E5.1_VK
>51E5.1_VH.06
>52A8.1_VK.01
>52A8.1_VH
>52A8.1_VK.02
>52A8.1_VH
>52A8.1_VK.03
>52A8.1_VH
>52A8.1_VK.04
>52A8.1_VH
>52A8.1_VK
>52A8.1_VH.05
>52A8.1_VK
>52A8.1_VH.06
>52A8.1_VK
>52A8.1 VH.07
>54A1.1_N83D_VK.01
>54A1.1_N83D_VH
>54A1.1_N83D_VK.02
>54A1.1_N83D_VH
>54A1.1_N83D_VK.03
>54A1.1_N83D_VH
>54A1.1_N83D_VK.04
>54A1.1_N83D_VH
>54A1.1_N83D_VK.05
>54A1.1_N83D_VH
>54A1.1_N83D_VK.06
>54A1.1_N83D_VH
>54A1.1_N83D_VK.07
>54A1.1_N83D_VH
>54A1.1_N83D_VK.08
>54A1.1_N83D_VH
>54A1.1_N83D_VK
>54A1.1_N83D_VH.09
>54A1.1_N83D_VK
>54A1.1_N83D_VH.10
>54A1.1_N83D_VK
>54A1.1_N83D_VH.11
>58C2.1_VK.01
>58C2.1_VH
>58C2.1_VK
>58C2.1_VH.02
>58C2.1_VK
>58C2.1_VH.03
>58C2.1_VK
>58C2.1_VH.04
>58C2.1_VK
>58C2.1_VH.05
>56E7.3_VK.01
>56E7.3_VH
>56E7.3_VK.02
>56E7.3 VH
>56E7.3_VK.03
>56E7.3_VH
>56E7.3_VK.04
>56E7.3_VH
>56E7.3_VK.05
>56E7.3_VH
>56E7.3_VK.06
>56E7.3_VH
>56E7.3_VK.07
>56E7.3_VH
>56E7.3_VK
>56E7.3_VH.08
>56E7.3_VK
>56E7.3_VH.09
>56E7.3_VK
>56E7.3_VH.10
>56E7.3_VK
>56E7.3_VH.11
>60D7.1_N30T_VK.01
>60D7.1_N30T_VH
>60D7.1_N30TVK.02
>60D7.1_N30T_VH
>60D7.1_N30T_VK
>60D7.1_N30T_VH.03
>60D7.1_N30T_VK
>60D7.1_N30T_VH.04
>60D7.1_N30T_VK
>60D7.1_N30T_VH.05
>60D7.1_N30T_VK
>60D7.1_N30T_VH.06
>63A10.1_C58S_VL.01
>63A10.1_C58S_VH
>63A10.1_C58S_VL.02
>63A10.1_C58S_VH
>63A10.1_C58S_VL.03
>63A10.1_C58S_VH
>63A10.1_C58S_VL.04
>63A10.1_C58S_VH
>63A10.1_C58S_VL.05
>63A10.1_C58S_VH
>63A10.1_C58S_VL.06
>63A10.1_C58S_VH
>63A10.1_C58S_VL.07
>63A10.1_C58S_VH
>63A10.1_C58S_VL.08
>63A10.1_C58S_VH
>63A10.1_C58S_VL.09
>63A10.1_C58S_VH
>63A10.1_C58S_VL.10
>63A10.1_C58S_VH
>63A10.1_C58S_VL.11
>63A10.1_C58S_VH
>63A10.1_C58S_VL.12
>63A10.1_C58S_VH
>63A10.1_C58S_VL.13
>63A10.1_C58S_VH
>63A10.1_C58S_VL.14
>63A10.1_C58S_VH
>63A10.1_C58S_VL
>63A10.1_C58S_VH.15
>63A10.1_C58S_VL
>63A10.1_C58S_VH.16
>63A10.1_C58S_VL
>63A10.1_C58S_VH.17
>63A10.1_C58S_VL
>63A10.1_C58S_VH.18
>63A10.1_C58S_VL
>63A10.1_C58S_VH.19
>63A10.1_C58S_VL
>63A10.1_C58S_VH.20
>63A10.1_C58S_VL
>63A10.1_C58S_VH.21
>63A10.1_C58S_VL
>63A10.1_C58S_VH.22
>63A10.1_C58S_VL
>63A10.1_C58S_VH.23
>63A10.1_C58S_VL
>63A10.1_C58S_VH.24
>63A10.1_C58S_VL
>63A10.1_C58S_VH.25
>63A10.1_C58S_VL.26
>63A10.1_C58S_VH
>63A10.3_N20R_C42S_VL.01
>63A10.3_N20R_C42S_VH
>63A10.3_N20R_C42S_VL.02
>63A10.3_N20R_C42S_VH
>63A10.3_N20R_C42S_VL.03
>63A10.3_N20R_C42S_VH
>64B10.1_VL.01
>64B10.1_VH
>64B10.1_VL.02
>64B10.1_VH
>64B10.1_VL.03
>64B10.1_VH
>64B10.1_VL
>64B10.1_VH.04
>64B10.1_VL
>64B10.1_VH.05
>64B10.1_VL

>64B10.1_VH.06
>64B10.1_VL.07
>64B10.1_VH
>64B10.1_VL
>64B10.1_VH.08
>66G2_VK.01
>66G2_VH
>66G2_VK.02
>66G2_VH
>66G2_VK.03
>66G2_VH
>66G2_VK.04
>66G2_VH
>66G2_VK
>66G2_VH.05
>66G2_VK
>66G2_VH.06
>66G2_VK
>66G2_VH.07
>66G2_VK
>66G2_VH.08
>66G2_VK
>66G2_VH.09
>66G2_VK
>66G2_VH.10
>67F5_VK.01
>67F5_VH
>67F5_VK.02
>67F5_VH
>67F5_VK.03
>67F5_VH
>67F5_VK.04
>67F5_VH
>67F5_VK.05
>67F5_VH
>67F5_VK
>67F5_VH.06
>67F5_VK.07
>67F5_VH
>67F5_VK.08
>67F5_VH
>67C10_VK.01
>67C10_VH
>67C10VK.02
>67C10_VH
>67C10_VK
>67C10_VH.03
>67C10_VK
>67C10_VH.04
>68C8_VL.01
>68C8_VH
>68C8_VL.02
>68C8_VH
>68C8_VL.03
>68C8_VH
>68C8_VL
>68C8_VH.04
>68C8_VL
>68C8_VH.05
>68C8_VL
>68C8_VH.06
>68C8_VL
>68C8_VH.07
>68C8_VL
>68C8_VH.08
>68C8_VL.09
>68C8_VH
>68C8_VL
>68C8_VH.10

EXAMPLE 14 Immunogenicity Prediction

Immune responses against proteins are enhanced by antigen processing and presentation in the major histocompatability complex (MHC) class II binding site. This interaction is required for T cell help in maturation of antibodies that recognize the protein. Since the binding sites of MHC class II molecules have been characterized, it is possible to predict whether proteins have specific sequences that can bind to a series of common human alleles. Computer algorithms have been created based on literature references and MHC class II crystal structures to determine whether linear 9 amino acid peptide sequences have the potential to break immune tolerance. We used the TEPITOPE™ program called to determine if point mutations of FGF21 are predicted to increase antigen specific T-cells in a majority of humans. Based on the linear protein sequence, none of the mutations examined are expected enhance immunogenicity. Results are shown in Table 17A and Table 17B below.


Met-FGF21	Low
Met-hFGF21(N106D)	Low
Met-FGF21 (N122D)	Low
hFc(R4).L15.hFGF21(G170E)	Low
hFc(R4).L15.hFGF21(P171A)	Low
hFc(R4).L15.hFGF21(S172L)	Low
p30.hFc.L15.hFGF21(A45K, G170E)	Low
p30.hFc.L15.hFGF21 (L98R, P171G)	Low

Clone	Predicted immunogenicity	LC Total Agretopes	LC Tolerant Agretopes	LC Non-Tolerant Agretopes	LC Non-Tolerant HLA DRB1	HC Total Agretopes	HC Tolerant Agretopes	HC Non-tolerant Agretopes	HC Non-tolerant HLA DRB1
	Tier 1	3	3	0	NA	12	12	0	NA
	Tier 1	1	1	0	NA	12	12	0	NA
	Tier 2	3	2	1	0101	16	16	0	NA
					0701
	Tier 2	6	6	0	NA	11	10	1	0401
									0701
	Tier 2	2	2	0	NA	12	11	1	0801
	Tier 3	7	5	2	0101	13	13	0	NA
					0701
					0801
					1301
					1501
	Tier 3	6	4	2	0301	14	14	0	NA
					0801
					1501
	Tier 3	6	5	1	0701	13	12	1	1301
	Tier 4	8	7	1	0301	16	14	2	0401
					0401				1501
					1101
	Tier 4	7	5	2	0801	14	13	1	0401
					1501
	Tier 4	8	7	1	0301	14	12	2	0101
					0401				701
					1101

Each reference cited herein is incorporated by reference in its entirety for all that it teaches and for all purposes.

The present disclosure is not to be limited in scope by the specific embodiments described herein, which are intended as illustrations of individual aspects of the disclosure, and functionally equivalent methods and components form aspects of the disclosure. Indeed, various modifications of the disclosure, in addition to those shown and described herein will become apparent to those skilled in the art from the foregoing description and accompanying drawings. Such modifications are intended to fall within the scope of the appended claims.

Claims

An isolated antigen binding protein that induces FGF21-mediated signaling.
An isolated antigen binding protein that specifically binds to a complex comprising β-Klotho and at least one of (i) FGFR1c, (ii) FGFR2c and (iii) FGFR3c.
The antigen binding protein of claim 1, wherein the antigen binding protein comprises one or more of:
(a) a light chain CDR3 comprising one or more of:
(i) a light chain CDR3 sequence that differs by three , two or one amino acid additions, substitutions, deletions and combinations thereof from a CDR3 of one or more of CDRL3-1 to CDRL3-75 of Table 3B (SEQ ID NOs 947-1020, respectively, in order of appearance);

(ii) QQFGSSLT (SEQ ID NO: 988);

(iii)QQSX₁SX₂X₃ LT (SEQ ID NO: 1443);

(iv)LQX₄X₅SYPX₆T (SEQ ID NO: 1447);

(v) MQRX₇EFPX₈T (SEQ ID NO: 1451);

(vi)QX₉WDSX₁₀X₁₁VV (SEQ ID NO: 1457);

(vii) QQYNX₁₂WPX₁₃T (SEQ ID NO: 1461);

(viii) QVWDSSX₁₄DX₁₅VX₁₆ (SEQ ID NO: 1466);

(ix)QQSSX₁₇IPWT (SEQ ID NO: 1469);

(x) QQTNSFPPWT (SEQ ID NO: 1470);

(xi) GTWDSSLSX₁₈X₁₉V (SEQ ID NO: 1474);

(xii) QQYDNLPX₂₀T (SEQ ID NO: 1477);

(xiii) QQYGSSX₂₁PWT (SEQ ID NO: 1480);

(xiv) QQYGX₂₂SX₂₃FT (SEQ ID NO: 1483);

(xv) QQYGSSX₂₄X₂₅X₂₆ (SEQ ID NO: 1488);

(xvi) QAWDSSX₂₇TX₂₈V (SEQ ID NO: 1492);

(xvii) QAWDSX₂₉TVX₃₀ (SEQ ID NO: 1496);

(xviii) QQX₃₁YSAX₃₂FT (SEQ ID NO: 1499);

(xix) QQYNX₃₃YPRT (SEQ ID NO: 1502);

(xx) HQX₃₄X₃₅DLPLT (SEQ ID NO: 1505);

(xxi) MQALQTX₃₆X₃₇T (SEQ ID NO: 1508);

(xxii) QQFGRSFT (SEQ ID NO: 1509);

(xxiii) YSTDSSX₃₈NHVV (SEQ ID NO: 1512);

(b) a heavy chain CDR3 sequence comprising one or more of:
(i) a heavy chain CDR3 that differs by eight, seven, six, five, four, three, two or one amino acid additions, substitutions, deletions and combinations thereof from a CDR3 of one or more of CDRH3-1 to CDRH3-81 of Table 3A (SEQ ID NOs 733-813, respectively, in order of appearance);

(ii) MTX₁₁₀PYWYFX₁₁₁L (SEQ ID NO: 1669);

(iii)DX₁₁₂X₁₁₃X₁₁₄DFWX₁₁₅GYPX₁₁₆X₁₁₇X₁₁₈YYGX₁₁₉DV (SEQ ID NO: 1675);

(iv)VTGTDAFDF (SEQ ID NO: 1676);

(v) TVTKEDYYYYGMDV (SEQ ID NO: 1677);

(vi)DSSGSYYVEDYFDY (SEQ ID NO: 1678);

(vii) DX₁₁₉X₁₂₀IAVAGX₁₂₁FDY (SEQ ID NO: 1681);

(viii) EYYYGSGSYYP (SEQ ID NO: 1682);

(ix)ELGDYPFFDY (SEQ ID NO: 1683);

(x) EYVAEAGFDY (SEQ ID NO: 1684);

(xi) VAAVYWYFDL (SEQ ID NO: 1685);

(xii) YNWNYGAFDF (SEQ ID NO: 1686);

(xiii) RASRGYRX₁₂₂GLAFAI (SEQ ID NO: 1689);

(xiv) DGITMVRGVTHYYGMDV (SEQ ID NO: 1690);

(xv) DHX₁₂₃SCWYYYGMDV (SEQ ID NO: 1693);

(xvi) YX₁₂₄X₁₂₅WDYYYGX₁₂₆DV (SEQ ID NO: 1696);

(xvii) VLHYX₁₂₇DSX₁₂₈GYSYYX₁₂₉DX₁₃₀ (SEQ ID NO: 1699); or

(c) the light chain CDR3 sequence of (a) and the heavy chain CDR3 sequence of (b); wherein,
X₁ is Y or F;

X₂ is T or S;

X₃ is P or S;

X₄ is H or R;

X₅ is N or S;

X₆ is L or R;

X₇ is I or L;

X₈ is I or L;

X₉ is V or L;

X₁₀ is S or N;

X₁₁ is T, P or S;

X₁₂ is N or T;

X₁₃ is W or L;

X₁₄ is S or C;

X₁₅ is H, V or G;

X₁₆ is V or absent;

X₁₇ is S or T;

X₁₈ is A or V;

X₁₉ is V or M;

X₂₀ is L or F;

X₂₁ is P or absent;

X₂₂ is R or T;

X₂₃ is L or P;

X₂₄ is P or absent;

X₂₅ is R or C;

X₂₆ is S or T;

X₂₇ is T or absent;

X₂₈ is W or A;

X₂₉ is G, T, A or absent;

X₃₀ is V or I;

X₃₁ is S or T;

X₃₂ is T or P;

X₃₃ is I or T;

X₃₄ is S or Y;

X₃₅ is S or D;

X₃₆ is P or L;

X₃₇ is F or I;

X₃₈ is V or G;

X₁₀₀ is T or S;

X₁₁₁ is D or G;

X₁₁₂ is R or Q;

X₁₁₃ is Y, D or N;

X₁₁₄ is Y or F;

X₁₁₅ is S or N;

X₁₁₆ is Y or F;

X₁₁₇ is F or Y;

X₁₁₈ is R, Y, F or H;

X₁₁₉ is L or M;

X₁₁₉ is W or L;

X₁₂₀ is S or R;

X₁₂₁ is T or S;

X₁₂₂ is F or Y;

X₁₂₃ is S or T;

X₁₂₄ is S or R;

X₁₂₅ is T or D;

X₁₂₆ is V or M;

X₁₂₇ is S or Y;

X₁₂₈ is R or S;

X₁₂₉ is S or F; and

X₁₃₀ is F or Y.
The antigen binding protein of claim 1, comprising one or more of:
(a) a light chain CDR1 sequence comprising one or more of:
(i) a light chain CDR1 that differs by seven, six, five, four, three, two or one amino acid additions, substitutions, deletions and combinations thereof from a CDR1 sequence of one or more of CDRL1-1 to CDRL1-81 of Table 3B (SEQ ID NOs 814-893, respectively, in order of appearance);

(ii) RASX₆₂SX₆₃X₆₄X₆₅X₆₆X₆₇X₆₈A (SEQ ID NO: 1591);

(iii) RX₆₉SQX₇₀IX₇₁X₇₂YLN (SEQ ID NO: 1602);

(iv) GGNX₇₃IGSX₇₄X₇₅VX₇₆ (SEQ ID NO: 1612);

(v) RASQX₇₇IRNDLX₇₈ (SEQ ID NO: 1616);

(vi) RSSQSLX₇₉X₈₀X₈₁DX₈₂GX₈₃TYLD (SEQ ID NO: 1622);

(vii) SGX₈₄X₈₅LGDKYX₈₆X₈₇ (SEQ ID NO: 1629);

viii) QASQX₈₈IX₈₉X₉₀X₉₁LN (SEQ ID NO: 1635); (ix)RASQX₉₂IX₉₃ X₉₄WLX₉₅ (SEQ ID NO: 1640);

(x) SGSSSNIGX₉₆NYVX₉₇ (SEQ ID NO: 1645);

(xi) RASX₉₈DISNYLA (SEQ ID NO: 1648);

(xii) RASQX₉₉VX₁₀₀SSYX₁₀₁V (SEQ ID NO: 1652);

(xiii) RSSQSLX₁₀₂HSNGX₁₀₃NYLD (SEQ ID NO: 1656);

(xiv) RASQTX₁₀₄RNX₁₀₅YLA (SEQ ID NO: 1659);

(xv) RSSX₁₀₆X₁₀₇LVYSDGNTYLN (SEQ ID NO: 1662); and

(xvi) SGDAX₁₀₈PKKYAX₁₀₉ (SEQ ID NO: 1665);

(b) a light chain CDR2 sequence comprising one or more of:
(i) a light chain CDR2 that differs by three, two or one amino acid additions, substitutions, deletions and combinations thereof from a CDR2 sequence of one or more of of CDRL2-1 to CDRL2-53 of Table 3B (SEQ ID NOs 894-946, respectively, in order of appearance);

(ii)X₃₉ASSLX₄₀X₄₁ (SEQ ID NO: 1517);

(iii) GX₄₂S X₄₃RX₄₄T (SEQ ID NO: 1525);

(iv) GAFSRAX₄₅ (SEQ ID NO: 1528);

(v) X₄₆DX₄₇KRPS (SEQ ID NO: 1533);

(vi) TLSX₄₈RAS (SEQ ID NO: 1536);

(vii) AASNLQX₄₉ (SEQ ID NO: 1539);

(viii) GX₅₀SNRAX₅₁ (SEQ ID NO: 1543);

(ix) X₅₂ASX₅₃LQS (SEQ ID NO: 1548);

(x) DNX₅₃KRPS (SEQ ID NO: 1551);

(xi) DX₅₄SNLET (SEQ ID NO: 1554);

(xii) LX₅₅SNRAS (SEQ ID NO: 1557);

(xiii) QX₅₆NX₅₇RPS (SEQ ID NO: 1561);

(xiv) RDRNRPS (SEQ ID NO: 1562);

(xv) X₅₈DSNRPS (SEQ ID NO: 1565);

(xvi) DDSDRPS (SEQ ID NO: 1566);

(xvii) AX₅₉SSLQS (SEQ ID NO: 1569);

(xviii) TX₆₀SSLQS (SEQ ID NO: 1572); and

(xix) KX₆₁SNWDS (SEQ ID NO: 1575);

(c) a heavy chain CDR1 sequence comprising one or more of:
(i) a heavy chain CDR1 that differs by three, two or one amino acid additions, substitutions, deletions and combinations thereof from a CDR1 sequence of one or more of CDRH1-1 to CDRH1-53 of Table 3A (SEQ ID NOs 603-655, respectively, in order of appearance);

(ii) SGX₁₇₀X₁₇₁TWX₁₇₂ (SEQ ID NO: 1775);

(iii)X₁₇₃YYWX₁₇₄ (SEQ ID NO: 1781);

(iv)X₁₇₅X₁₇₆GMS (SEQ ID NO: 1786);

(v) SYX₁₇₇MX₁₇₈ (SEQ ID NO: 1790);

(vi)X₁₇₉YYX₁₈₀H (SEQ ID NO: 1796);

(vii) SYGX₁₈₁H (SEQ ID NO: 1799);

(viii) NYX₁₈₂MX₁₈₃ (SEQ ID NO: 1803);

(ix)X₁₈₄YWIG (SEQ ID NO: 1806);

(x) GYX₁₈₅MH (SEQ ID NO: 1809);

(xi)SX₁₈₆DIX₁₈₇ (SEQ ID NO: 1813);

(xii) X₁₈₈YAMS (SEQ ID NO: 1816);

(xiii) NAWMS (SEQ ID NO: 1817);

(xiv) SSSYYWG (SEQ ID NO: 1818);

(xv) X₁₈₉YYWN (SEQ ID NO: 1821);

(xvi) SNSAX₁₉₀WN (SEQ ID NO: 1824); and

(xvii) X₁₉₁YDMH (SEQ ID NO: 1827);

(d) a heavy chain CDR2 sequence comprising one or more of
(i) a heavy chain CDR2 that differs by nine, eight, seven, six, five, four, three, two or one amino acid additions, substitutions, deletions and combinations thereof from a CDR2 sequence of one or more of CDRH2-1 to CDRH2-77 of Table 3A (SEQ ID NOs 656-732, respectively, in order of appearance);

(ii) X₁₃₁X₁₃₂X₁₃₃X₁₃₄X₁₃₅GX₁₃₆X₁₃₇X₁₃₈X₁₃₉NPSLKS (SEQ ID NO: 1716);

(iii)X₁₄₀IX₁₄₁X₁₄₂DGX₁₄₃NX₁₄₄X₁₄₅X₁₄₆ADSVKG (SEQ ID NO: 1732);

(iv)WX₁₄₇NPX₁₄₈SGX₁₄₉TX₁₅₀YAQKFX₁₅₁G (SEQ ID NO: 1741);

(v) EINHSX₁₅₂X₁₅₃TNYNPSLKS (SEQ ID NO: 1744);

(vi) IIYPGDSX₁₅₄TRYSPSFQG (SEQ ID NO: 1747);

(vii) SISSSSX₁₅₅YX₁₅₆YYX₁₅₇DSX₁₅₈KG (SEQ ID NO: 1751);

(viii) RIX₁₅₉ X₁₆₀KTDGGTTX₁₆₁Y AAPVKG (SEQ ID NO: 1755);

(ix) GISGSSAGTYYADSVGK (SEQ ID NO: 1756);

(x) VISX₁₆₂SGGX₁₆₃TYYADSVKG (SEQ ID NO: 1759);

(xi) RTYYRSKWYNDYAVSVKS (SEQ ID NO: 1760);

(xii) RIYX₁₆₄SGSTNYNPSLX₁₆₅X₁₆₆ (SEQ ID NO: 1763); and

(xiii) WMNPYSGSTGX₁₆₇AQX₁₆₈FQX₁₆₉ (SEQ ID NO: 1766);
wherein
X₃₉ is A or S;

X₄₀ is Q or K;

X₄₁ is S or F;

X₄₂ is A or T;

X₄₃ is S, T, A, R or N;

X₄₄ is A or D;

X₄₅ is S or T;

X₄₆ is Q, R or E;

X₄₇ is T or S;

X₄₈ is Y or F;

X₄₉ is R or S;

X₅₀ is A or S;

X₅₁ is I or T;

X₅₂ is D or G;

X₅₃ is S, T or N;

X₅₃ is N or D;

X₅₄ is A or V;

X₅₅ is G or D;

X₅₆ is D or N;

X₅₇ is K or E;

X₅₈ is S or C;

X₅₉ is S or V;

X₆₀ is A or T;

X₆₁ is V or G;

X₆₂ is P or Q;

X₆₃ is V, I or F;

X₆₄ is S, R or absent;

X₆₅ is S, R or N;

X₆₆ is D, S, N or M;

X₆₇ is I, Y, H, Q, N or S;

X₆₈ is L or V;

X₆₉ is A or T;

X₇₀ is I, S, T or N;

X₇₁ is R, S or N;

X₇₂ is R, S, N, or I;

X₇₃ is N, or D;

X₇₄ is Y, I or K;

X₇₅ is N, S, T or A;

X₇₆ is H or Q;

X₇₇ is D or G;

X₇₈ is G or A;

X₇₉ is L or F;

X₈₀ is N or D;

X₈₁ is S or N;

X₈₂ is A or D;

X₈₃ is T, D or N;

X₈₄ is N or D;

X₈₅ is K, E or N;

X₈₆ is V or A;

X₈₇ is C or F;

X₈₈ isG or D;

X₈₉ is S, K N or T;

X₉₀ is N, K or I;

X₉₁ is Y or F;

X₉₂ is D or G;

X₉₃ isD or S;

X₉₄ is R or S;

X₉₅ is V or A;

X₉₆ is N, I or D;

X₉₇ is A or S;

X₉₈ is Q or H.;

X₉₉ is R or S;

X₁₀₀ is P or A;

X₁₀₁ is I or L;

X₁₀₂ is L or Q;

X₁₀₃ is Y or F.

X₁₀₄ is V or I;

X₁₀₅ is N or S;

X₁₀₆ is Q or P;

X₁₀₇ is R or S;

X₁₀₈ is L or V;

X₁₀₉ is Y or N;

X₁₃₁ is N, F, Y, S or M;

X₁₃₂ is I or L;

X₁₃₃ is Y or F;

X₁₃₄ is Y, H or D;

X₁₃₅ is S or T;

X₁₃₆ is T, G, S or T;

X₁₃₇ is T or A;

X₁₃₈ is Y, N or H;

X₁₃₉ is F or Y;

X₁₄₀ is L, G, I or V;

X₁₄₁ is W or S;

X₁₄₂ is Y, D or N;

X₁₄₃ is S or D;

X₁₄₄ is K or N;

X₁₄₅ is Y, N, D, or H;

X₁₄₆ is Y or H;

X₁₄₇ is I or M;

X₁₄₈ is P, N, S or D;

X₁₄₉ is A, G or D;

X₁₅₀ isN, K, or D;

X₁₅₁ is R, H or Q;

X₁₅₂ is E or G;

X₁₅₃ is N or T;

X₁₅₄ is D or E;

X₁₅₅ is T or S;

X₁₅₆ is I or E;

X₁₅₇ is A or V;

X₁₅₈ is V or L.

X₁₅₉ is K or I;

X₁₆₀ is S or G;

X₁₆₁ is D or E;

X₁₆₂ is D or G;

X₁₆₃ is S or D;

X₁₆₄ is I or T;

X₁₆₅ is E or K;

X₁₆₆ is N or S;

X₁₆₇ is Y or L;

X₁₆₈ is N or R;

X₁₆₉ is G or D;

X₁₇₀ is V, G, N or D;

X₁₇₁ is Y or N;

X₁₇₂ is N, S or T;

X₁₇₃ is T, S or G;

X₁₇₄ is S or T;

X₁₇₅ is S, T or F;

X₁₇₆ is Y or F;

X₁₇₇ is A or S;

X₁₇₈ is S, N or M;

X₁₇₉ is Y or G;

X₁₈₀ is I, L, K or T;

X₁₈₁ is L or I;

X₁₈₂ is G or N;

X₁₈₃ is H, R or M;

X₁₈₄ is S or G;

X₁₈₅ is Y or F;

X₁₈₆ is Y or H;

X₁₈₇ is N or D;

X₁₈₈ is N or H;

X₁₈₉ is D or S;

X₁₉₀ is T or A;

X₁₉₁ is S or T;

(e) the light chain CDR1 of (a) and the light chain CDR2 of (b);

(f) the light chain CDR1 of (a) and the heavy chain CDR1 of (c);

(g) the light chain CDR1 of (a) and the heavy chain CDR2 of (d);

(h) the light chain CDR1 (a) and the heavy chain CDR1 of (c);

(i) the heavy chain CDR1 of (c) and the heavy chain CDR2 of (d);

(j) the light chain CDR2 of (b) and the heavy chain CDR2 of (d);

(k) the light chain CDR1 of (a), the light chain CDR2 of (b), and the heavy chain CDR1 of (c);

(l) the light chain CDR2 of (b), the heavy CDR1 of (c), and the heavy chain CDR2 of (d);

(m) the light chain CDR1 of (a), the heavy chain CDR1 of (c), and the heavy chain CDR2 of (d); and

(n) the light chain CDR1 of (a), the light chain CDR2 of (b), the heavy chain CDR2 of (c), and the heavy chain CDR2 of (d).
The antigen binding protein of claim 1, comprising one or more of:
(a) a light chain variable domain comprising one or more of;
(i) a light chain CDR1 sequence selected from CDRL1-1 to CDRL1-81 of Table 3B (SEQ ID NOs 814-893, respectively, in order of appearance);

(ii) a light chain CDR2 sequence selected from CDRL2-1 to CDRL2-53 of Table 3B (SEQ ID NOs 894-946, respectively, in order of appearance);

(iii) a light chain CDR3 sequence selected from CDRL3-1 to CDRL3-75 of Table 3B (SEQ ID NOs 947-1020, respectively, in order of appearance);

(b) a heavy chain variable domain comprising one or more of:
(i) a heavy chain CDR1 sequence selected from CDRH1-1 to CDRH1-53 of Table 3A (SEQ ID NOs 603-655, respectively, in order of appearance);

(ii) a heavy chain CDR2 sequence selected from CDRH2-1 to CDRH2-77 of Table 3A (SEQ ID NOs 656-732, respectively, in order of appearance);

(iii) a heavy chain CDR3 sequence selected from CDRH3-1 to CDRH3-81 of Table 3A (SEQ ID NOs 733-813, respectively, in order of appearance); and

(c) a combination comprising a light chain variable domain of (a) and a heavy chain variable domain of (b).
The antigen binding protein of claim 1, wherein the antigen binding protein comprises one or more of:
(a) a light chain variable domain sequence comprising one or more of:
(i) V_L1-V_L100 of Table 2A (SEQ ID NOs 217-315, respectively, in order of appearance);

(ii) amino acids having a sequence at least 80% identical to a light chain variable domain sequence comprising one or more of V_L1-V_L100 of Table 2A (SEQ ID NOs 217-315, respectively, in order of appearance);

(iii) a sequence of amino acids encoded by a polynucleotide sequence that is at least 80% identical to a polynucleotide sequence encoding the light chain variable domain sequence comprising one or more of V_L1-Y_L100 of Table 2A(SEQ ID NOs 217-315, respectively, in order of appearance);

(b) a heavy chain variable domain sequence comprising one or more of:
(i) V_H1-V_H94 of Table 2B (SEQ ID NOs 316-409, respectively, in order of appearance);

(ii) a sequence of amino acids that is at least 80% identical to a heavy chain variable domain sequence comprising one or more of V_H1-V_H94 of Table 2B 9SEQ ID NOs 316-409, respectively, in order of appearance);

(iii) a sequence of amino acids encoded by a polynucleotide sequence that is at least 80% identical to a polynucleotide sequence encoding the heavy chain variable domain sequence of V_H1-V_H94 of Table 2B (SEQ ID NOs 316-409, respectively, in order of appearance); and

(c) a combination comprising a light chain variable domain of (a) and a heavy chain variable domain of (b),
such as an antigen binding protein in which the light chain variable domain and the heavy chain variable domain comprise one or more of: V_L1 and V_H1; V_L2 and V_H1; V_L3 and V_H2 or V_H3; V_L4 and V_H4; V_L5 and V_H5; V_L6 and V_H6; V_L7 and V_H6; V_L8 and V_H7 or V_H8; V_L9 and V_H9; V_L10 and V_H9; V_L11 and V_H 10; V_L12 and V_H11; V_L13 and V_H12; V_L13 and V_H14; V_L14 and V_H13; V_L15 and V_H14; V_L16 and V_H15; V_L17 and V_H16; V_L18 and V_H17; V_L19 and V_H18; V_L20 and V_H19; V_L21 and V_H20; V_L22 and V_H21; V_L23 and V_H22; V_L24 and V_H23; V_L25 and V_H24; V_L26 and V_H25; V_L27 and V_H26; V_L28 and V_H27; V_L29 and V_H28; V_L30 and V_H29; V_L31 and V_H30; V_L32 and V_H31; V_L33 and V_H32; V_L34 and V_H33; V_L35 and V_H34; V_L36 and V_H35; V_L37 and V_H36; V_L38 and V_H37; V_L39 and V_H38; V_L40 and V_H39; V_L41 and V_H40; V_L42 and V_H41; V_L43 and V_H42; V_L44 and V_H43; V_L45 and V_H44; V_L46 and V_H45; V_L47 and V_H46; V_L48 and V_H47; V_L49 and V_H48; V_L50 and V_H49; V_L51 and V_H50; V_L 52 and V_H51; V_L53 and V_H52; V_L54 and V_H53; V_L55 and V_H54; V_L56 and V_H54; V_L57 and V_H54; V_L58 and V_H55; V_L59 and V_H56; V_L60 and V_H57; V_L61 and V_H58; V_L62 and V_H59; V_L63 and V_H60; V_L64 and V_H1; V_L65 and V_H62; V_L66 and V_H63; V_L67 and V_H64; V_L68 and V_H65; V_L69 and V_H66; V_L70 and V_H67; V_L71 and V_H68; V_L72 and V_H69; V_L73 and V_H70; V_L74 and V_H70; V_L75 and V_H70; V_L76 and V_H71; V_L77 and V_H72; V_L78 and V_H73; V_L79 and V_H74; V_L80 and V_H75; V_L81 and V_H76; V_L82 and V_H77; V_L83 and V_H78; V_L84 and V_H79; V_L85 and V_H80; V_L86 and V_H81; V_L87 and V_H82; V_L88 and V_H86; V_L89 and V_H83; V_L90 and V_H84; V_L91 and V_H85; V_L92 and V_H87; V_L93 and V_H88; V_L94 and V_H88; V_L95 and V_H89; V_L96 and V_H90; V_L97 and V_H91; V_L98 and V_H92; V_L99 and V_H93; and V_L100 and V_H94.
The antigen binding protein of claim 6, further comprising:
(a) the kappa light chain constant sequence of SEQ ID NO: 12

(b) the lambda light chain constant sequence of SEQ ID NO: 13

(c) the heavy chain constant sequence of SEQ ID NO: 11; or

(d)
(i) the kappa light chain constant sequence of SEQ ID NO: 12 or the lambda light chain constant sequence of SEQ ID NO: 13, and

(ii) the heavy chain constant sequence of SEQ ID NO: 11.
The antigen binding protein of claim 1, wherein the antigen binding protein is a human antibody, a humanized antibody, chimeric antibody, a monoclonal antibody, a polyclonal antibody, a recombinant antibody, an antigen-binding antibody fragment, a single chain antibody, a diabody, a triabody, a tetrabody, a Fab fragment, an F(fab')₂ fragment, a domain antibody, an IgD antibody, an IgE antibody, an IgM antibody, an IgG1 antibody, an IgG2 antibody, an IgG3 antibody, an IgG4 antibody, or an IgG4 antibody having at least one mutation in the hinge region.
The antigen binding protein of claim 1, that, when bound to to a complex comprising β-Klotho and at least one of (i) FGFR1c, (ii) FGFR2c and (iii) FGFR3c:
(a) binds to (i) β-Klotho; (ii) FGFR1c, FGFR2c, FGFR3c or FGFR4; or (iii) a complex comprising β-Klotho and one of FGFR1c, FGFR2c, FGFR3c, and FGFR4, with substantially the same Kd as a reference antibody;

(b) induces FGF21-like signaling of 10% or greater than the signaling induced by a wild-type FGF21 standard comprising the mature form of SEQ ID NO: 2 as measured in an ELK-luciferase reporter assay;

(c) exhibits an EC₅₀ of 10nM or less of FGF21-like signaling in an assay comprising one of:
(i) a FGFR1c/β-Klotho-mediated in vitro recombinant cell-based assay; and

(ii) an in vitro human adipocyte functional assay;

(d) exhibits an EC₅₀ of less than 10nM of agonistic activity on FGFR1c in the presence of β-Klotho in an in vitro recombinant FGFR1c receptor mediated reporter assay;

(e) exhibits an EC₅₀ of greater than 1µM of agonistic activity on FGFR1c in the absence of β-Klotho in an in vitro recombinant FGFR1c receptor mediated reporter assay;

(f) competes for binding with a reference antibody to (i) β-Klotho; (ii) FGFR1c, FGFR2c, FGFR3c or FGFR4; or (iii) a complex comprising β-Klotho and one of FGFR1c, FGFR2c, FGFR3c, and FGFR4, wherein the reference antibody comprises a combination of light chain and heavy chain variable domain sequences selected from the group consisting of V_L1 and V_H1; V_L2 and V_H1; V_L3 and V_H2 or V_H3; V_L4 and V_H4; V_L5 and V_H5; V_L6 and V_H6; V_L7 and V_H6; V_L8 and V_H7 or V_H8; V_L9 and V_H9; V_L10 and V_H9; V_L11 and V_H 10; V_L12 and V_H11; V_L13 and V_H12; V_L13 and V_H14; V_L14 and V_H13; V_L15 and V_H14; V_L16 and V_H15; V_L17 and V_H16; V_L18 and V_H17; V_L19 and V_H18; V_L20 and V_H19; V_L21 and V_H20; V_L22 and V_H21; V_L23 and V_H22; V_L24 and V_H23; V_L25 and V_H24; V_L26 and V_H25; V_L27 and V_H26; V_L28 and V_H27; V_L29 and V_H28; V_L30 and V_H29; V_L31 and V_H30; V_L32 and V_H31; V_L33 and V_H32; V_L34 and V_H33; V_L35 and V_H34; V_L36 and V_H35; V_L37 and V_H36; V_L38 and V_H37; V_L39 and V_H38; V_L40 and V_H39; V_L41 and V_H40; V_L42 and V_H41; V_L43 and V_H42; V_L44 and V_H43; V_L45 and V_H44; V_L46 and V_H45; V_L47 and V_H46; V_L48 and V_H47; V_L49 and V_H48; V_L50 and V_H49; V_L51 and V_H50; V_L52 and V_H51; V_L53 and V_H52; V_L54 and V_H53; V_L55 and V_H54; V_L56 and V_H54; V_L57 and V_H54; V_L58 and V_H55; V_L59 and V_H56; V_L60 and V_H57; V_L61 and V_H58; V_L62 and V_H59; V_L63 and V_H60; V_L64 and V_H1; V_L65 and V_H62; V_L66 and V_H63; V_L67 and V_H64; V_L68 and V_H65; V_L69 and V_H66; V_L70 and V_H67; V_L71 and V_H68; V_L72 and V_H69; V_L73 and V_H70; V_L74 and V_H70; V_L75 and V_H70; V_L76 and V_H71; V_L77 and V_H72; V_L78 and V_H73; V_L79 and V_H74; V_L80 and V_H75; V_L81 and V_H76; V_L82 and V_H77; V_L83 and V_H78; V_L84 and V_H79; V_L85 and V_H80; V_L86 and V_H81; V_L87 and V_H82; V_L88 and V_H86; V_L89 and V_H83; V_L90 and V_H84; V_L91 and V_H85; V_L92 and V_H87; V_L93 and V_H88; V_L94 and V_H88; V_L95 and V_H89; V_L96 and V_H90; V_L97 and V_H91; V_L98 and V_H92; V_L99 and V_H93; and V_L100 and V_H94; and

(g) two or more of (a) - (f).
The antigen binding protein of claim 9, that, when bound to a complex comprising β-Klotho and at least one of (i) FGFR1c, (ii) FGFR2c and (iii) FGFR3c and:
(a) lowers blood glucose in an animal model;

(b) lowers serum lipid levels in an animal model;

(c) lowers insulin levels in an animal model; or

(d) two or more of (a) and (b) and (c).
The antigen binding protein of claim 1, wherein the antigen binding protein comprises one or more of:
(a) a heavy chain comprising one of H1-H94;

(b) a light chain comprising one of L1-L100; and

(c) a combination comprising a heavy chain of (a) and a light chain of (b).
A pharmaceutical composition comprising one or more antigen binding proteins of claims 1-11 in admixture with a pharmaceutically acceptable carrier thereof.
An isolated nucleic acid comprising a polynucleotide sequence encoding the light chain variable domain amino acid sequence, the heavy chain variable domain amino acid sequence, or both amino acid sequences, of an antigen binding protein of claims 1-13, e.g. wherein the encoded amino acid sequence comprises one or more of:
(a) V_L1-V_L100;

(b) V_H1-V_H94; and

(c) a combination comprising one or more sequences of (a) and one or more sequences of (b).
An isolated cell comprising the nucleic acid of claim 13 and/or an expression vector comprising the nucleic acid of claim 13.
A method of producing an antigen binding protein comprising incubating the host cell of claim 14 under conditions that allow it to express the antigen binding protein.
The isolated antigen binding protein according to any one of claims 1 to 11, or the pharmaceutical composition according to claim 12 for use in preventing or treating a condition which is treatable by lowering one or more of blood glucose, insulin or serum lipid levels such as type 2 diabetes, obesity, dyslipidemia, NASH, cardiovascular disease or metabolic syndrome.