Method of treating or ameliorating metabolic disorders using binding proteins for gastric inhibitory peptide receptor (GIPR) in combination with GLP-1 agonists

Abstract

To provide methods of treating metabolic diseases and disorders using an antigen binding protein specific for the GIPR polypeptide.SOLUTION: In various embodiments, a metabolic disease or disorder is type 2 diabetes, obesity, dyslipidemia, elevated glucose levels, elevated insulin levels and diabetic nephropathy. In certain embodiments, the antigen binding protein is administered in combination with a GLP-1 receptor agonist.SELECTED DRAWING: None

Description

[Technical field]

[0001] The present disclosure relates to the treatment or amelioration of metabolic disorders such as type 2 diabetes, elevated glucose levels, elevated insulin levels, obesity, non-alcoholic fatty liver disease, or cardiovascular disease using antigen binding proteins specific for the gastric inhibitory peptide receptor (GIPR). [Background technology]

[0002] Glucose-dependent insulinotropic polypeptide (GIP) is a single peptide of 42 amino acids secreted from K cells in the small intestine (duodenum and jejunum). Human GIP is generated by processing proGIP, which is a precursor of 153 amino acids encoded by a gene located on chromosome 17q (Inagaki et al., Mol Endocrinol 1989;3:1014-1021, Fehmann et al. Endocr Rev.1995;16:390-410). GIP was previously called gastric inhibitory polypeptide.

[0003] GIP secretion is induced by food intake. GIP exerts many physiological effects on tissues, including promoting fat storage in adipocytes and pancreatic islet β-cell function and glucose-dependent insulin secretion. GIP and glucagon-like polypeptide-1 (GLP-1) are known as insulin secretion stimulating factors ("incretins"). Intact GIP is rapidly degraded by DPPIV to an inactive form. In type 2 diabetes patients, the insulin secretion stimulating effect of GIP is lost, but the incretin effect of GLP-1 remains intact (Nauck et al. J. Clinc. Invest. 1993;91:301-307).

[0004] The GIP receptor (GIPR) is a member of the secretin-glucagon family of G protein-coupled receptors (GPCRs) with an extracellular N-terminus, seven transmembrane domains, and an intracellular C-terminus. The N-terminal extracellular domain of this family of receptors is usually glycosylated and forms the recognition and binding domain of the receptor. GIPR is highly expressed in many tissues, including the pancreas, intestine, adipose tissue, heart, pituitary gland, adrenal cortex, and brain (Usdin et al., Endocrinology. 1993, 133:2861-2870). Human GIPR contains 466 amino acids and is encoded by a gene located on chromosome 19q13.3 (Gremlich et al., Diabetes. 1995; 44:1202-8; Volz et al., FEBS Lett. 1995, 373:23-29). Studies suggest that in humans, rats, and mice, GIP receptor variants of different lengths are generated as a result of alternative splicing of the mRNA.

[0005] GIPR knockout mice (Gipr - / - ) are resistant to high-fat diet-induced weight gain and have improved insulin sensitivity and lipid profile (Yamada et al., Diabetes. 2006, 55:S86; Miyawaki et al. Nature Med. 2002, 8:738-742). Furthermore, a novel small molecule GIPR antagonist, SKL-14959, prevents obesity and insulin resistance (Diabetologia 2008, 51:S373, 44th EASD Annual meeting poster).

[0006] Glucagon-like peptide-1 ("GLP-1") is a 31 amino acid peptide derived from the proglucagon gene. GLP-1 is secreted by intestinal L-cells and is released in response to food intake to induce insulin secretion from pancreatic β-cells (Diabetes 2004,53:S3,205-214). In addition to its incretin action, GLP-1 also reduces glucagon secretion, delays gastric emptying, and reduces caloric intake (Diabetes Care,2003,26(10):2929-2940). GLP-1 exerts its action by activating the GLP-1 receptor, which is a class B G protein-coupled receptor (Endocrinology.1993,133(4):1907-10). GLP-1's function is limited by its rapid degradation by the DPP-IV enzyme, resulting in a half-life of approximately 2 minutes. Long-acting GLP-1 receptor agonists, such as exenatide, liraglutide, and dulaglutide, have been recently developed and are currently in clinical use to improve glycemic control in patients with type 2 diabetes. In addition, GLP-1 receptor agonists also promote weight loss and reduction of blood pressure and plasma cholesterol levels in patients (Bioorg.Med.Chem.Lett 2013,23:4011-4018).

[0007] Collectively, these links to obesity and insulin resistance imply that inhibiting GIPR, either as monotherapy or in combination with GLP-1, may be a useful approach for therapeutic intervention. Summary of the Invention [Means for solving the problem]

[0008] In one aspect, the disclosure provides a method of treating a subject having a metabolic disorder, the method comprising administering to the subject a therapeutically effective amount of an antigen binding protein that specifically binds to a protein having an amino acid sequence that has at least 90% amino acid sequence identity to the amino acid sequence of GIPR. In one aspect, the present invention is directed to a method of treating a subject having a metabolic disorder, the method comprising administering to the subject a therapeutically effective amount of a GLP-1 receptor agonist and a therapeutically effective amount of a GIPR antagonist that specifically binds to a protein having an amino acid sequence that has at least 90% amino acid sequence identity to the amino acid sequence of GIPR. In one embodiment, the metabolic disorder is a disorder of glucose metabolism. In another embodiment, the glucose metabolic disorder comprises hyperglycemia, and administration of the antigen binding protein reduces plasma glucose. In another embodiment, the glucose metabolic disorder comprises hyperinsulinemia, and administration of the antigen binding protein reduces plasma insulin. In another embodiment, the glucose metabolic disorder comprises impaired glucose tolerance, and administration of the antigen binding protein improves glucose tolerance. In another embodiment, the glucose metabolism disorder comprises insulin resistance, and administration of the antigen binding protein reduces insulin resistance. In another embodiment, the glucose metabolism disorder comprises diabetes. In another embodiment, the subject is obese. In another embodiment, administration of the antigen binding protein reduces body weight in the obese subject. In another embodiment, administration of the antigen binding protein reduces weight gain in the obese subject. In another embodiment, administration of the antigen binding protein reduces body fat mass in the obese subject. In another embodiment, the glucose metabolism disorder comprises insulin resistance, and administration of the antigen binding protein reduces insulin resistance in the obese subject. In another embodiment, administration of the antigen binding protein reduces hepatic steatosis in an obese subject with advanced hepatic steatosis. In another embodiment, administration of the antigen binding protein reduces liver fat content in an obese subject with increased liver fat content.

[0009] In one aspect, the present invention is directed to a method of treatment, the method comprising administration to a subject having symptoms of a metabolic disorder of at least one GLP-1 receptor agonist in a therapeutically effective amount in combination with administration of at least one GIPR antagonist that provides a sustained beneficial effect when administered to the subject.

[0010] In one embodiment, the administration of at least one GLP-1 receptor agonist in combination with the administration of at least one GIPR antagonist provides a sustained beneficial effect on at least one symptom of a metabolic disorder.

[0011] In one embodiment, a therapeutically effective amount of a GLP-1 receptor agonist and a therapeutically effective amount of a GIPR antagonist are combined prior to administration to a subject.

[0012] In one embodiment, a therapeutically effective amount of a GLP-1 receptor agonist and a therapeutically effective amount of a GIPR antagonist are administered sequentially to a subject.

[0013] In one embodiment, the therapeutically effective amount of the GLP-1 receptor agonist and the therapeutically effective amount of the GIPR antagonist are synergistically effective amounts.

[0014] In one embodiment, the molar ratio of the GLP-1 receptor agonist to the GIPR antagonist is about 1:1 to 1:110, about 1:1 to 1:100, about 1:1 to 1:75, about 1:1 to 1:50, about 1:1 to 1:25, about 1:1 to 1:10, about 1:1 to 1:5, and about 1: 1. In one embodiment, the molar ratio of the GIPR antagonist to the GLP-1 receptor agonist is about 1:1 to 1:110, about 1:1 to 1:100, about 1:1 to 1:75, about 1:1 to 1:50, about 1:1 to 1:25, about 1:1 to 1:10, and about 1:1 to 1:5.

[0015] In one embodiment, the GLP-1 receptor agonist and the GIPR antagonist are used in combination in a therapeutically effective molar ratio of about 1:1.5 to 1:150, preferably 1:2 to 1:50.

[0016] In one embodiment, the GLP-1 receptor agonist and the GIPR antagonist are present in dosages that are at least about 1.1-1.4 times, 1.5 times, 2 times, 3 times, 4 times, 5 times, 6 times, 7 times, 8 times, 9 times, or 10 times less than the dosages that would be required to treat the condition and / or disease with each compound alone.

[0017] In one embodiment, the GLP-1 receptor agonist is GLP-1(7-37) or a GLP-1(7-37) analogue.

[0018] In one embodiment, the GLP-1 receptor agonist is selected from the group consisting of exenatide, liraglutide, lixisenatide, albiglutide, dulaglutide, semaglutide, and taspoglutide.

[0019] In one embodiment, the GLP-1 receptor agonist is GLP-1(7-37) (SEQ ID NO: 1244), GLP-1(7-36)-NH2 (SEQ ID NO: 1245), liraglutide, albiglutide, taspoglutide, dulaglutide, semaglutide, LY2428757, desamino-His 7 ,Arg 26 ,Lys 34 (N ε -(γ-Glu(N-α-hexadecanoyl)))-GLP-1(7-37) (core peptide disclosed as SEQ ID NO: 1282), desamino-His 7 ,Arg 26 ,Lys 34 (N ε -octanoyl)-GLP-1(7-37) (SEQ ID NO: 1283), Arg 26,34 ,Lys 38 (N ε -(ω-carboxypentadecanoyl)-GLP-1(7-38) (SEQ ID NO: 1284), Arg 26,34 ,Lys36 (N ε -(γ-Glu(N-α-hexadecanoyl)))-GLP-1(7-36) (core peptide disclosed as SEQ ID NO: 1285), Aib 8,35 ,Arg 26,34 ,Phe 31 -GLP-1(7-36)) (SEQ ID NO: 1246), HXaa8EGTFTSDVSSYLEXaa 22 Xaa 23 AAKEFIXaa 30 WLXaa 33 Xaa 34 G Xaa 36 Xaa 37 (Xaa3 is A, V, or G; 22 is G, K, or E; Xaa 23 is Q or K, and Xaa 30 is A or E, and Xaa 33 is V or K, and Xaa 34 is K, N, or R; Xaa 36 is R or G, and Xaa 37 is G, H, P, or absent) (SEQ ID NO: 1247), Arg 34 -GLP-1(7-37) (SEQ ID NO: 1248), Glu 30 -GLP-1(7-37) (SEQ ID NO: 1249), Lys 22 -GLP-1(7-37) (SEQ ID NO: 1250), Gly 8,36 ,Glu 22 -GLP-1(7-37) (SEQ ID NO: 1251), Val 8 ,Glu 22 ,Gly 36 -GLP-1(7-37) (SEQ ID NO: 1252), Gly 8,36 ,Glu 22 ,Lys 33 ,Asn 34 -GLP-1(7-37) (SEQ ID NO: 1253), Val 8 ,Glu 22 ,Lys 33 ,Asn 34 ,Gly 36 -GLP-1(7-37) (SEQ ID NO: 1254), Gly 8,36 ,Glu 22,Pro 37 -GLP-1(7-37) (SEQ ID NO: 1255), Val 8 ,Glu 22 ,Gly 36 Pro 37 -GLP-1(7-37) (SEQ ID NO: 1256), Gly 8,36 ,Glu 22 ,Lys 33 ,Asn 34 ,Pro 37 -GLP-1(7-37) (SEQ ID NO: 1257), Val 8 ,Glu 22 ,Lys 33 ,Asn 34 ,Gly 36 ,Pro 37 -GLP-1(7-37) (SEQ ID NO: 1258), Gly 8,36 ,Glu 22 -GLP-1(7-36) (SEQ ID NO: 1259), Val 8 ,Glu 22 ,Gly 36 -GLP-1(7-36) (SEQ ID NO: 1260), Val 8 ,Glu 22 ,Asn 34 ,Gly 36 -GLP-1(7-36) (SEQ ID NO: 1261), and Gly 8,36 ,Glu 22 ,Asn 34 - GLP-1(7-36) (SEQ ID NO: 1262).

[0020] In another embodiment, the subject is a mammal. In another embodiment, the subject is a human. In another embodiment, the GIPR is a human GIPR. In another embodiment, the administration is by parenteral injection. In another embodiment, the administration is by subcutaneous injection.

[0021] In another aspect, the present disclosure provides an antigen binding protein that specifically binds to human GIPR polypeptide and inhibits the activation of GIPR by GIP ligand.In one embodiment, the antigen binding protein inhibits the binding of GIP ligand to GIPR.In another embodiment, the antigen binding protein is a human antigen binding protein.In another embodiment, the antigen binding protein is a human antibody.In another embodiment, the antigen binding protein is a monoclonal antibody.

[0022] In another aspect, the present disclosure provides a pharmaceutical composition comprising at least one antigen binding protein according to any one of the preceding embodiments.

[0023] In another aspect, the disclosure provides a nucleic acid molecule encoding an antigen binding protein according to any one of the preceding embodiments.

[0024] In another aspect, the present disclosure provides a vector comprising a nucleic acid molecule encoding an antigen binding protein according to any one of the preceding embodiments.

[0025] In another aspect, the disclosure provides a host cell comprising a nucleic acid molecule encoding an antigen binding protein according to any one of the preceding embodiments, or a host cell comprising a vector comprising a nucleic acid molecule encoding an antigen binding protein according to any one of the preceding embodiments. In another aspect, the disclosure provides an antigen binding protein expressed by the vector that specifically binds to a human GIPR polypeptide.

[0026] In another aspect, the disclosure provides a method of preparing an antigen binding protein according to any one of the preceding embodiments, the method comprising expressing the antigen binding protein in a host cell that secretes the antigen binding protein, followed by purification of the antigen binding protein from the cell culture medium. In another aspect, the disclosure provides an antigen binding protein that specifically binds to a human GIPR polypeptide, purified from the host cell.

[0027] In another aspect, the disclosure provides an antigen binding protein of any one of the preceding embodiments, or a pharmaceutical composition of any one of the preceding embodiments, for use in therapy. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0028] The present disclosure provides a method for treating metabolic disorders such as disorders of glucose metabolism (e.g., type 2 diabetes, elevated glucose levels, elevated insulin levels, dyslipidemia, metabolic syndrome (syndrome X or insulin resistance syndrome), diabetes, metabolic acidosis, type 1 diabetes, obesity, and conditions exacerbated by obesity) by blocking or interfering with the biological activity of GIP. In one embodiment, an isolated human GIPR binding protein is administered in a therapeutically effective amount to a subject in need thereof. Methods of administration and delivery are also provided.

[0029] Recombinant methods for producing polypeptides and nucleic acids used herein, including the Examples, are generally those set forth in Sambrook et al., Molecular Cloning: A Laboratory Manual (Cold Spring Harbor Laboratory Press, 1989), or Current Protocols in Molecular Biology (Ausubel et al., eds., Green Publishers Inc. and Wiley and Sons 1994), both of which are incorporated herein by reference for all purposes.

[0030] The section headings used herein are for organizational purposes only and are not to be construed as limiting the subject matter described.

[0031] As used herein, unless otherwise defined, scientific and technical terms used in connection with this application shall have the meanings commonly understood by those of ordinary skill in the art. Further, unless the context otherwise requires, singular terms shall include the plural and plural terms shall include the singular.

[0032] Generally, the nomenclature used in connection with, and techniques for, cell and tissue culture, molecular biology, immunology, microbiology, genetics, and protein and nucleic acid chemistry, and hybridization described herein are well known and commonly used in the art. Unless otherwise indicated, the methods and techniques of the present application are generally performed according to conventional methods well known in the art, and such methods and techniques are described in various general and more specific references cited and discussed throughout this specification. See, for example, Sambrook et al., Molecular Cloning: A Laboratory Manual, 3rd ed., Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY (2001); Ausubel et al., Current Protocols in Molecular Biology, Greene Publishing Associates (1992); and Harlow and Lane Antibodies: A Laboratory Manual Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY (1990). These documents are incorporated herein by reference. Enzymatic reactions and purification techniques are performed according to manufacturer's instructions, 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 well known and commonly used in the art. Standard techniques may be used for the purposes of chemical syntheses, chemical analyses, pharmaceutical preparation, formulation, and delivery, and treatment of patients.

[0033] It is to be understood that this invention is not limited to the particular methodology, protocols, and reagents, etc. described herein and as such may vary. The terminology used herein is for the purpose of describing particular embodiments only and is not intended to limit the scope of the disclosure, which is defined only by the claims.

[0034] Except as otherwise noted in the examples, all numbers indicating quantities of ingredients or reaction conditions used herein should be understood to be modified in all instances by the term "about." The term "about" when used in connection with percentages can mean ±1%.

[0035] As used herein, "a" and "an" follow their convention and mean "one or more" unless otherwise noted.

[0036] As used herein, the terms "amino acid" and "residue" are used interchangeably and when used in the context of a peptide or polypeptide refer to both naturally occurring and synthetic amino acids, as well as amino acid analogs, amino acid mimetics, and non-naturally occurring amino acids that are chemically similar to the naturally occurring amino acids.

[0037] "Naturally occurring amino acids" are those amino acids encoded by the genetic code, as well as those amino acids encoded by the genetic code that are synthesized and subsequently modified (e.g., hydroxyproline, γ-carboxyglutamate, and O-phosphoserine). Amino acid analogs are compounds that have the same basic chemical structure as a naturally occurring amino acid, i.e., an alpha carbon bonded to a hydrogen, a carboxyl group, an amino group, and an R group, e.g., homoserine, norleucine, methionine sulfoxide, methylmethionine sulfonium. Such analogs may have modified R groups (e.g., norleucine) or modified peptide backbones, but will retain the same basic chemical structure as a naturally occurring amino acid.

[0038] "Amino acid mimetics" are chemical compounds that have a structure that is different from the general chemical structure of an amino acid, but that functions in a manner similar to a naturally occurring amino acid. Examples include methacryloyl or acryloyl derivatives of amides, β-amino acids, γ-amino acids, δ-amino acids (such as piperidine-4-carboxylic acid), and the like.

[0039] A "non-naturally occurring amino acid" is a compound that has the same basic chemical structure as a naturally occurring amino acid, but is not incorporated into a growing polypeptide chain by the translation complex. "Non-naturally occurring amino acids" also include, but are not limited to, amino acids that arise by modification (e.g., post-translational modification) of naturally encoded amino acids (including, but not limited to, the 20 common amino acids), but that are not themselves naturally incorporated into a growing polypeptide chain by the translation complex. An exemplary list of non-naturally occurring amino acids that can be inserted into or used in place of wild-type residues in a polypeptide sequence includes, but is not limited to, β-amino acids, homoamino acids, cyclic amino acids, and amino acids with side chain derivatization. Examples include 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 (NMe R), 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-guanidinophenylalanine (Guf), glycyrrhizin (abbreviated as "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-methylvaline (NMeVal), N-α-methylleucine (NMeLeu), Nα-methylnorleucine (NMeNle), cyclopentylglycine (Cpg), cyclohexylglycine (Chg), acetylarginine (acetylarg), α,β-diaminopropionic acid (Dpr), α,γ-diaminobutanoic acid (Dab), diaminopropionic acid (Dap), cyclohexylalanine (C ha), 4-methyl-phenylalanine (MePhe), β,β-diphenyl-alanine (BiPhA), aminobutanoic acid (Abu), 4-phenyl-phenylalanine (or biphenylalanine, 4Bip), α-amino-isobutanoic acid (Aib), beta-alanine, beta-aminopropionic acid, piperidine acid, aminocaproic acid, aminoheptanoic acid, aminopimelic acid, desmosine, diaminopimelic acid, N-ethylglycine, N-ethylasparagine, hydroxylysine, allo-hydroxylysine, isodesmosine, allo-isoleucine, N Included are 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, as well as derivatized forms of any of those specifically mentioned, which may be in the L- or D-form, with the abbreviations listed in parentheses.

[0040] The term "isolated nucleic acid molecule" refers to a single- or double-stranded polymer of deoxyribonucleotide or ribonucleotide bases, read from the 5' to the 3' end (e.g., a GIPR nucleic acid sequence provided herein) or an analog thereof, which is free of at least about 50 percent of the polypeptides, peptides, lipids, carbohydrates, polynucleotides, or other materials that are naturally found with the nucleic acid when the total nucleic acid is isolated from a cellular source. Preferably, an isolated nucleic acid molecule is substantially free of any other contaminating nucleic acid molecules or other molecules that are found in the nucleic acid's natural environment and that would interfere with its use in the production of a polypeptide, or its therapeutic, diagnostic, prophylactic, or research uses.

[0041] The term "isolated polypeptide" refers to a polypeptide (e.g., a GIPR polypeptide sequence provided herein, or an antigen binding protein of the invention) that has been removed from at least about 50 percent of the polypeptides, peptides, lipids, carbohydrates, polynucleotides, or other materials that are naturally found with the polypeptide when it is isolated from a cellular source. Preferably, an isolated polypeptide is substantially free of any other contaminating polypeptides or other contaminants that are found in its natural environment and that would interfere with its therapeutic, diagnostic, prophylactic, or research use.

[0042] The term "encoding" refers to a polynucleotide sequence that codes for one or more amino acids. The term does not require a start or stop codon.

[0043] The terms "identical" and "percent identity" in the context of two or more nucleic acid or polypeptide sequences refer to two or more sequences or subsequences that are identical. "Percent identity" refers to the percent of residues that are identical 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. In such calculations, gaps in the alignment, if any, can be addressed by a specific mathematical model or computer program (i.e., an "algorithm"). Methods that can be used to calculate the identity of 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) SIAM J. Applied Math. 48:1073.

[0044] In calculating percent identity, the sequences being compared are aligned in a manner that maximizes the match between the sequences. A 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 two polypeptides or polynucleotides for which percent sequence identity is to be determined. The sequences are aligned so that their respective amino acid or nucleotide matches are optimal (the "match span" is determined by the algorithm). A gap opening penalty (calculated as 3× average diagonal element, where the “average diagonal element” is the average of the diagonal elements of the comparison matrix used. A “diagonal element” is the score or number assigned to each perfect amino acid match by a particular comparison matrix) and gap extension penalty (usually 1 / 10th the gap opening penalty), as well as a comparison matrix such as PAM250 or BLOSUM62, are used in conjunction with the algorithm. In certain embodiments, a standard comparison matrix (for the PAM250 comparison matrix, see Dayhoff et al., (1978) Atlas of Protein Sequence and Structure 5:345-352; for the BLOSUM62 comparison matrix, see Henikoff et al., (1992) Proc. Natl. Acad. Sci. USA 89:10915-10919) are also used by the algorithm.

[0045] Recommended parameters for determining percent identity of a polypeptide or nucleotide sequence using the GAP program include the following: Algorithm: Needleman et al., 1970, J. Mol. Biol. 48:443-453 Comparison matrix: BLOSUM62 by Henikoff et al., 1992, supra Gap penalty: 12 (exception: no penalty for end gaps) Gap length penalty: 4 Similarity tolerance: 0

[0046] When using a certain alignment scheme for aligning two amino acid sequences, only a short region may match in the two sequences.This short aligned region may have very high sequence identity even if there is no significant relationship between the two full-length sequences.Therefore, if desired, the alignment method selected (for example, GAP program) can be adjusted to obtain an alignment spanning at least 50 consecutive amino acids in target polypeptide.

[0047] The terms "GIPR polypeptide" and "GIPR protein" are used interchangeably and refer to naturally occurring wild-type polypeptides expressed in mammals, such as humans or mice, including naturally occurring alleles (e.g., naturally occurring allelic forms of human GIPR proteins). For purposes of the present disclosure, the term "GIPR polypeptide" can be used interchangeably to refer to any full-length GIPR polypeptide, such as SEQ ID NO: 1201 (consisting of 466 amino acid residues and encoded by the nucleotide sequence of SEQ ID NO: 1202), or SEQ ID NO: 1203 (consisting of 430 amino acid residues and encoded by the nucleic acid sequence of SEQ ID NO: 1204), or SEQ ID NO: 1205 (consisting of 493 amino acid residues and encoded by the nucleic acid sequence of SEQ ID NO: 1206), or SEQ ID NO: 1207 (consisting of 460 amino acid residues and encoded by the nucleic acid sequence of SEQ ID NO: 1208), or SEQ ID NO: 1209 (consisting of 230 amino acid residues and encoded by the nucleic acid sequence of SEQ ID NO: 1210).

[0048] The term "GIPR polypeptide" also encompasses GIPR polypeptides in which a naturally occurring GIPR polypeptide sequence (e.g., SEQ ID NO: 1201, SEQ ID NO: 1203, or SEQ ID NO: 1205) has been modified. Such modifications include one or more amino acid substitutions, including but not limited to, substitutions with non-naturally occurring amino acids, non-naturally occurring amino acid analogs, and amino acid mimetics.

[0049] In various embodiments, the GIPR polypeptide comprises an amino acid sequence that is at least about 85 percent identical to a naturally occurring GIPR polypeptide (e.g., SEQ ID NO: 1201, SEQ ID NO: 1203, or SEQ ID NO: 1205). In other embodiments, the GIPR polypeptide comprises an amino acid sequence that is at least about 90 percent identical, or about 95 percent, about 96 percent, about 97 percent, about 98 percent, or about 99 percent identical to a naturally occurring GIPR polypeptide amino acid sequence (e.g., SEQ ID NO: 1201, SEQ ID NO: 1203, or SEQ ID NO: 1205). Such a GIPR polypeptide preferably, but not necessarily, has at least one activity of a wild-type GIPR polypeptide, such as the ability to bind to GIP. The present invention also encompasses nucleic acid molecules that encode such GIPR polypeptide sequences.

[0050] The term "GIPR activity assay" (also referred to as "GIPR functional assay") refers to an assay that can be used to measure the activity of GIP or GIP-binding protein in a cellular context. In one embodiment, the "activity" (or "functional") assay can be a cAMP assay in GIPR-expressing cells (GIP can induce cAMP signaling), and the activity of GIP / GIPR-binding protein can be measured in the presence / absence of GIP ligand, in which case IC50 / EC50 and degree of inhibition / activation can be obtained (Biochemical and Biophysical Research Communications (2002) 290:1420-1426). In another embodiment, the "activity" (or "function") assay can be an insulin secretion assay in pancreatic beta cells (where GIP can induce glucose-dependent insulin secretion) and the activity of the GIP / GIPR binding protein can be measured in the presence / absence of a GIP ligand, in which case the IC50 / EC50 and degree of inhibition / activation can be obtained (Biochemical and Biophysical Research Communications (2002) 290:1420-1426).

[0051] The term "GIPR binding assay" refers to an assay that can be used to measure the binding of GIP to GIPR. In one embodiment, the "GIPR binding assay" can be an assay that uses FMAT or FACS to measure the binding of fluorescently labeled GIP to GIPR-expressing cells, and the activity of GIP / GIPR binding protein can be measured by displacing the binding of fluorescently labeled GIP to GIPR-expressing cells. In another embodiment, the "GIPR binding assay" can be an assay that measures the binding of radiolabeled GIP to GIPR-expressing cells, and the activity of GIP / GIPR binding protein can be measured by displacing the binding of radiolabeled GIP to GIPR-expressing cells (Biochimica et Biophysica Acta (2001) 1547: 143-155).

[0052] The terms "GIP", "gastric inhibitory polypeptide", "glucose-dependent insulinotropic peptide", and "GIP ligand" are used interchangeably and refer to naturally occurring wild-type polypeptides expressed in mammals, such as humans or mice, including naturally occurring alleles (e.g., naturally occurring allelic forms of the human GIP protein). For purposes of this disclosure, the term "GIP" can be used interchangeably to refer to any mature GIP polypeptide.

[0053] The sequence of the 42 amino acids of mature human GIP is: YAEGTFISDY SIAMDKIHQQ DFVNWLLAQK GKKNDWKHNI TQ (SEQ ID NO: 1211) and [ka] It is encoded by the DNA sequence:

[0054] The sequence of the 42 amino acids of mature mouse GIP is: YAEGTFISDY SIAMDKIRQQ DFVNWLLAQR GKKSDWKHNI TQ (sequence number 1213) and [ka] It is encoded by the DNA sequence:

[0055] The sequence of the 42 amino acids of mature rat GIP is: YAEGTFISDY SIAMDKIRQQ DFVNWLLAQK GKKNDWKHNL TQ (SEQ ID NO: 1215) and [ka] It is encoded by the DNA sequence:

[0056] As used herein, "antigen binding protein" refers to any protein that specifically binds to a particular target antigen, such as a GIPR polypeptide (e.g., a human GIPR polypeptide such as those provided in SEQ ID NO: 1201, SEQ ID NO: 1203, or SEQ ID NO: 1205). The term encompasses intact antibodies comprising at least two full-length heavy chains and two full-length light chains, as well as derivatives, mutants, fragments, and variants thereof. Examples of antibody fragments include Fab fragments, Fab' fragments, F(ab')2 fragments, and Fv fragments. Antigen binding proteins also include domain antibodies, such as nanobodies and scFvs, which are further described below.

[0057] In general, a GIPR antigen-binding protein is said to "specifically bind" to its target antigen, GIPR, when the antigen-binding protein exhibits essentially background binding to non-GIPR molecules. However, an antigen-binding protein that specifically binds to GIPR may cross-react with GIPR polypeptides from different species. Typically, a GIPR antigen-binding protein has a dissociation constant (KD) of ≦10 as measured via surface plasma resonance techniques (e.g., BIACore, GE-Healthcare Uppsala, Sweden) or equilibrium exclusion techniques (KinExA, Sapidyne, Boise, Idaho). -7 M. A GIPR antigen binding protein specifically binds to human GIPR when it has a KD of ≦5×10 as measured using the methods described. -9 M specifically binds human GIPR with "high affinity" and has a KD of ≦5×10 as measured using the described method -10 When M, it specifically binds to human GIPR with "very high affinity."

[0058] "Antigen binding region" refers to a protein or a portion of a protein that specifically binds to a particular antigen. For example, that portion of an antigen binding protein that contains amino acid residues that interact with an antigen and confer its specificity and affinity for the antigen to the antigen is referred to as the "antigen binding region". An antigen binding region typically comprises one or more "complementary binding regions" ("CDRs") of an immunoglobulin, single-chain immunoglobulin, or camelid antibody. Certain antigen binding regions also comprise one or more "framework" regions. "CDRs" are amino acid sequences that contribute to the specificity and affinity of antigen binding. "Framework" regions can facilitate binding between the antigen binding region and the antigen by helping to maintain the proper conformation of the CDRs.

[0059] A "recombinant protein," including a recombinant GIPR antigen binding protein, is a protein prepared using recombinant techniques, i.e., through the expression of a recombinant nucleic acid as described herein. Methods and techniques for producing recombinant proteins are well known in the art.

[0060] The term "antibody" refers to an intact immunoglobulin of any isotype, or a fragment thereof that can compete with the intact antibody for specific binding to a target antigen, including, for example, chimeric antibodies, humanized antibodies, fully human antibodies, and bispecific antibodies. Thus, an "antibody" is a type of antigen-binding protein. An intact antibody will generally contain at least two full-length heavy chains and two full-length light chains. An antibody may be derived from only a single source, or may be "chimeric," i.e., different portions of the antibody may be derived from two different antibodies, as further described below. Antigen-binding proteins, antibodies, or binding fragments may be produced in hybridomas, by recombinant DNA techniques, or by enzymatic or chemical cleavage of intact antibodies.

[0061] The term "light chain" as used with respect to an antibody or fragment thereof includes a full-length light chain and fragments thereof having sufficient variable region sequence to confer binding specificity. A full-length light chain contains a variable region domain (VL) and a constant region domain (CL). The variable region domain of the light chain is located at the amino-terminus of the polypeptide. Light chains include kappa chains and lambda chains.

[0062] The term "heavy chain" as used with respect to an antibody or fragment thereof includes a full-length heavy chain and fragments thereof having sufficient variable region sequence to confer binding specificity. A full-length heavy chain contains a variable region domain (VH) and three constant region domains (CH1, CH2, and CH3). The VH domain is located at the amino-terminus of the polypeptide, the CH domain is located at the carboxyl-terminus, and CH3 is located closest to the carboxy-terminus of the polypeptide. The heavy chain can be of any isotype, including IgG (including IgG1, IgG2, IgG3, and IgG4 subtypes), IgA (including IgA1 and IgA2 subtypes), IgM, and IgE.

[0063] As used herein, the term "immunologically functional fragment" (or simply "fragment") of an antibody or immunoglobulin chain (heavy or light chain) is an antigen-binding protein that lacks at least some of the amino acids present in the full-length chain, but that contains a portion of an antibody (regardless of how that portion is obtained or synthesized) that has the ability to specifically bind to an antigen. Such a fragment is biologically active in that it specifically binds to a target antigen and can compete with other antigen-binding proteins, including intact antibodies, for specific binding to a given epitope.

[0064] Such biologically active fragments may be produced by recombinant DNA techniques, or by enzymatic or chemical cleavage of antigen-binding proteins, including intact antibodies. Immunologically functional immunoglobulin fragments include, but are not limited to, Fab fragments, Fab' fragments, and F(ab')2 fragments.

[0065] In another embodiment are Fvs, domain antibodies, and scFvs, which may be derived from the antibodies of the invention.

[0066] It is further contemplated that a functional portion of the antigen binding proteins disclosed herein, such as, for example, one or more CDRs, can be covalently linked to a second protein or small molecule to create a therapeutic agent that is directed to a specific target in the body, has bifunctional therapeutic properties, or has extended serum half-life.

[0067] A "Fab fragment" is composed of one light chain and the CH1 and variable regions of one heavy chain. The heavy chain of a Fab molecule cannot form disulfide bonds with another heavy chain molecule.

[0068] The "Fc" region comprises two heavy chain fragments comprising the CH2 and CH3 domains of an antibody. The two heavy chain fragments are held together by two or more disulfide bonds and hydrophobic interactions of the CH3 domain.

[0069] In certain embodiments, one or more amino acid modifications may be introduced into the Fc region of an antibody provided herein, thereby generating an Fc region variant. The Fc region variant may comprise a human Fc region sequence (e.g., a human IgG1 Fc region, a human IgG2 Fc region, a human IgG3 Fc region, or a human IgG4 Fc region) that contains an amino acid modification (e.g., a substitution) at one or more amino acid positions.

[0070] In certain embodiments, the present invention contemplates antibody variants that retain some, but not all, effector functions, making them desirable candidates for applications where in vivo antibody half-life is important, but certain effector functions (e.g., complement and ADCC) are unnecessary or deleterious. In vitro and / or in vivo cytotoxicity assays may be performed to confirm reduced / depleted CDC and / or ADCC activity. For example, Fc receptor (FcR) binding assays may be performed to confirm that the antibody lacks Fc.gamma.R binding (and thus likely lacks ADCC activity) but retains FcRn binding ability. The primary cells for mediating ADCC, NK cells, express only Fc(RIII, whereas monocytes express Fc(RI, Fc(RII and Fc(RIII). FcR expression on hematopoietic cells is summarized in Table 3 on page 464 of Ravetch and Kinet, Annu. Rev. Immunol. 9:457-492 (1991). Non-limiting examples of in vitro assays to assess ADCC activity of a molecule of interest include those described in U.S. Pat. No. 5,500,362 (see, e.g., Hellstrom, I. et al. Proc. Nat'l Acad. Sci. USA 83:7059-7063 (1986)), and Hellstrom, I. et al., Proc. Nat'l Acad. Sci. USA 82:1499-1502 (1985); 5,821,337 (Bruggemann, M. et al., Proc. Nat'l Acad. Sci. USA 82:1499-1502 (1985); al., J. Exp. Med. 166:1351-1361 (1987). Alternatively, non-radioactive assay methods may be used (see, e.g., ACTI.TM. Non-Radioactive Cytotoxicity Assay for Flow Cytometry (Cell Technology, Inc. Mountain View, Calif.); and CytoTox 96.RTM. Non-Radioactive Cytotoxicity Assay (Promega, Madison, Wis.). Useful effector cells for such assays include peripheral blood mononuclear cells (PBMC) and natural killer (NK) cells.Alternatively, or in addition, ADCC activity of the molecule of interest may be assessed in vivo, e.g., in an animal model such as that disclosed in Clynes et al. Proc. Nat'l Acad. Sci. USA 95:652-656 (1998). Similarly, C1q binding assays may be performed to confirm that the antibody is unable to bind C1p and thus lacks CDC activity. See, e.g., C1q and C3c binding ELISAs in WO 2006 / 029879 and WO 2005 / 100402. To assess complement activation, CDC assays can be performed (see, e.g., Gazzano-Santoro et al., J. Immunol. Methods 202:163 (1996); Cragg, MS et al., Blood 101:1045-1052 (2003); and Cragg, MS and MJ Glennie, Blood 103:2738-2743 (2004)). FcRn binding and in vivo clearance / half-life determinations can also be performed using methods known in the art (see, e.g., Petkova, SB et al., Int'l. Immunol. 18(12):1759-1769 (2006)).

[0071] In some embodiments, one or more amino acid modifications may be introduced into the Fc portion of the antibodies provided herein to increase IgG binding to the neonatal Fc receptor. In certain embodiments, the antibody contains the following three mutations according to EU numbering: M252Y, S254T, and T256E ("YTE mutations") (U.S. Pat. No. 8,697,650; Dall'Acqua et al., Journal of Biological Chemistry 281(33):23514-23524 (2006). In certain embodiments, the YTE mutation does not affect the ability of the antibody to bind to its cognate antigen. In certain embodiments, the YTE mutation increases the serum half-life of the antibody as compared to the native (i.e., non-YTE mutant) antibody. In some embodiments, the YTE mutation increases the serum half-life of the antibody by 3-fold as compared to the native (i.e., non-YTE mutant) antibody. In some embodiments, the YTE mutation increases the serum half-life of the antibody by 3-fold as compared to the native (i.e., non-YTE mutant) antibody. In some embodiments, the YTE mutation increases the serum half-life of the antibody by 2-fold compared to the native (i.e., non-YTE mutant) antibody. In some embodiments, the YTE mutation increases the serum half-life of the antibody by at least 5-fold compared to the native (i.e., non-YTE mutant) antibody. In some embodiments, the YTE mutation increases the serum half-life of the antibody by at least 10-fold compared to the native (i.e., non-YTE mutant) antibody. See, e.g., U.S. Pat. No. 8,697,650; see also Dall'Acqua et al., Journal of Biological Chemistry 281(33):23514-23524 (2006).

[0072] In certain embodiments, the YTE variants provide a means of modulating the antibody-dependent cell-mediated cytotoxicity (ADCC) activity of the antibody. In certain embodiments, the YTEO variants provide a means of modulating the ADCC activity of humanized IgG antibodies against human antigens. See, e.g., U.S. Pat. No. 8,697,650; see also Dall'Acqua et al., Journal of Biological Chemistry 281(33):23514-23524 (2006). In certain embodiments, the YTE variants allow for simultaneous modulation of serum half-life, tissue distribution, and antibody activity (e.g., ADCC activity of IgG antibodies). See, e.g., U.S. Pat. No. 8,697,650; see also Dall'Acqua et al., Journal of Biological Chemistry 281(33):23514-23524 (2006).

[0073] Antibodies with reduced effector function include those with substitutions at one or more of Fc region residues 238, 265, 269, 270, 297, 327, and 329 according to EU numbering (U.S. Pat. No. 6,737,056). Such Fc variants include Fc variants with substitutions at two or more of amino acid positions 265, 269, 270, 297, and 327 according to EU numbering (e.g., so-called "DANA" Fc variants with substitutions at residues 265 and 297 according to EU numbering to alanine (i.e., D265A and N297A according to EU numbering)) (U.S. Pat. No. 7,332,581). In certain embodiments, the Fc variant comprises the following two amino acid substitutions: D265A and N297A. In certain embodiments, the Fc variant consists of the following two amino acid substitutions: D265A and N297A.

[0074] In certain embodiments, the proline at position 329 (EU numbering) of the wild-type human Fc region (P329) is substituted with glycine, or arginine, or an amino acid residue large enough to disrupt the proline sandwich in the Fc / Fc gamma receptor interface formed between P329 of Fc and tryptophan residues W87 and W110 of FcgRIII (Sondermann et al.: Nature 406, 267-273 (20 Jul. 2000)). In a further embodiment, the at least one additional amino acid substitution in the Fc variant is S228P, E233P, L234A, L235A, L235E, N297A, N297D, or P331S, and in yet another embodiment, said at least one additional amino acid substitution is L234A and L235A in a human IgG1 Fc region, or S228P and L235E in a human IgG4 Fc region, all according to EU numbering (U.S. Pat. No. 8,969,526, all incorporated by reference).

[0075] In certain embodiments, the polypeptide comprises an Fc variant of a wild-type human IgG Fc region, the polypeptide having P329 of the human IgG Fc region substituted with glycine, the Fc variant comprising at least two additional amino acid substitutions at L234A and L235A of the human IgG1 Fc region or S228P and L235E of the human IgG4 Fc region, the residues being numbered according to EU numbering (U.S. Pat. No. 8,969,526, incorporated by reference in its entirety). In certain embodiments, the polypeptide comprising the P329G substitution, the L234A substitution, and the L235A substitution (EU numbering) exhibits reduced affinity for human Fc.gamma.RIIIA and human Fc.gamma.RIIA for downregulation of ADCC to at least 20% of that induced by a polypeptide comprising a wild-type human IgG Fc region, and / or for downregulation of ADCP (U.S. Pat. No. 8,969,526, incorporated by reference in its entirety).

[0076] In a specific embodiment, a polypeptide comprising an Fc variant of a wild-type human Fc polypeptide comprises the following triple mutation: an amino acid substitution at position Pro329, an L234A mutation, and an L235A mutation according to EU numbering (P329 / LALA) (U.S. Patent No. 8,969,526, incorporated by reference in its entirety). In a specific embodiment, the polypeptide comprises the following amino acid substitutions: P329G, L234A, and L235A according to EU numbering.

[0077] Certain antibody variants with improved or diminished binding to FcRs have been described (see, e.g., U.S. Pat. No. 6,737,056; WO 2004 / 056312; and Shields et al., J. Biol. Chem. 9(2):6591-6604 (2001)).

[0078] In certain embodiments, the antibody variant comprises an Fc region with one or more amino acid substitutions that improve ADCC (e.g., substitutions at positions 298, 333, and / or 334 (EU numbering) of the Fc region).

[0079] In some embodiments, modifications are made in the Fc region that result in altered (i.e., improved or decreased) C1q binding and / or complement dependent cytotoxicity (CDC), e.g., as described in U.S. Pat. No. 6,194,551, WO 99 / 51642, and Idusogie et al. J Immunol. 164:4178-4184 (2000).

[0080] Antibodies with extended half-lives and improved binding to the neonatal Fc receptor (FcRn), which is responsible for the transfer of maternal IgG to the fetus (Guyer et al., J. Immunol. 117:587 (1976) and Kim et al., J. Immunol. 24:249 (1994)), have been described in U.S. Patent Application Publication No. 2005 / 0014934A1 (Hinton et al.).

[0081] The antibody comprises an Fc region having one or more substitutions that improve binding of the Fc region to FcRn. Such Fc variants include those having a substitution at one or more of the following residues in the Fc region: 238, 256, 265, 272, 286, 303, 305, 307, 311, 312, 317, 340, 356, 360, 362, 376, 378, 380, 382, ​​413, 424, or 434 (e.g., a substitution at Fc region residue 434 according to EU numbering (U.S. Pat. No. 7,371,826)). See also Duncan & Winter, Nature 322:738-40 (1988); U.S. Pat. No. 5,648,260; U.S. Pat. No. 5,624,821; and WO 94 / 29351 for other examples of Fc region variants.

[0082] A "Fab' fragment" comprises one light chain and a portion of one heavy chain containing the VH domain, the CH1 domain, and also the region between the CH1 and CH2 domains, such that an F(ab')2 molecule can be formed by the formation of interchain disulfide bonds between the two heavy chains of the two Fab' fragments.

[0083] An "F(ab')2 fragment" comprises two light chains and two heavy chains that contain a portion of the constant region between the CH1 and CH2 domains, such that an interchain disulfide bond is formed between the two heavy chains. Thus, an F(ab')2 fragment is composed of two Fab' fragments held together by disulfide bonds between the two heavy chains.

[0084] The "Fv region" comprises the variable regions from both the heavy and light chains, but lacks the constant regions.

[0085] A "single chain antibody" or "scFv" is an Fv molecule in which the heavy and light chain variable regions are linked by a flexible linker to form a single polypeptide chain that forms the antigen-binding region. scFvs are discussed in detail in International Patent Application Publication No. WO 88 / 01649, and U.S. Patent Nos. 4,946,778 and 5,260,203, the disclosures of which are incorporated by reference.

[0086] A "domain antibody" or "single-chain immunoglobulin" is an immunologically functional immunoglobulin fragment that contains only the variable region of a heavy chain or only the variable region of a light chain. Examples of domain antibodies include Nanobodies®. In some cases, two or more VH regions are covalently linked via a peptide linker to create a bivalent domain antibody. The two VH regions of a bivalent domain antibody may target the same or different antigens.

[0087] A "bivalent antigen-binding protein" or a "bivalent antibody" comprises two antigen-binding regions. In some cases, the two binding regions have the same antigen specificity. Bivalent antigen-binding proteins and bivalent antibodies may be bispecific, see below.

[0088] A "multispecific antigen binding protein" or "multispecific antibody" is one that targets multiple antigens or epitopes.

[0089] A "bispecific", "dual-specific" or "bifunctional" antigen-binding protein or antibody is a hybrid antigen-binding protein or antibody, respectively, that has two different antigen-binding sites. Bispecific antigen-binding proteins and antibodies are a type of multispecific antigen-binding protein or antibody, which may be generated by a variety of methods, including but not limited to, fusion of hybridomas or linking of Fab' fragments. See, for example, 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 may be present on the same or different protein targets.

[0090] The term "compete" when used in the context of an antigen binding protein (e.g., an antibody), means that competition between antigen binding proteins is determined by an assay in which the antigen binding protein (e.g., an antibody or an immunologically functional fragment thereof) blocks or inhibits specific binding of a reference antigen binding protein to a common antigen (e.g., GIPR or a fragment thereof) under test. Various types of competitive binding assays can be used, such as solid-phase direct or indirect radioimmunoassays (RIAs), solid-phase direct or indirect enzyme immunoassays (EIAs), sandwich competition assays (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 label assays, solid-phase direct label sandwich assays (see, e.g., Harlow and Lane, 1988, Antibodies, A Laboratory Manual, Cold Spring Harbor Press), solid-phase direct label RIA using I-125 labels (see, e.g., Morel et al., 1988, Antibodies, A Laboratory Manual, Cold Spring Harbor Press), and the like. 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 label RIA (Moldenhauer et al., 1990, Scand. J. Immunol. 32:77-82) can be used. Typically, such assays use purified antigen bound to a solid surface or cells bearing either such antigen, an unlabeled test antigen binding protein, and 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. Typically, the test antigen binding protein is present in excess. Additional details regarding methods for determining competitive binding are provided in the Examples herein.Typically, when a competing antigen binding protein is present in excess, the competing antigen binding protein will inhibit specific binding of the reference antigen binding protein to a common antigen by at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, or at least 75%. In some cases, binding is inhibited by at least 80%, at least 85%, at least 90%, at least 95%, or at least 97% or more.

[0091] The term "antigen" refers to a molecule or portion of a molecule that is capable of being bound by a selective binding agent, such as an antigen binding protein (including, for example, an antibody), and that can be used in an animal to generate antibodies capable of binding to that antigen. An antigen can have one or more epitopes that are capable of interacting with different antigen binding proteins (e.g., antibodies).

[0092] The term "epitope" is a portion of a molecule that is bound by an antigen-binding protein (e.g., an antibody). The term includes any determinant that has the ability to specifically bind to an antigen-binding protein, such as an antibody. Epitopes can be contiguous or discontinuous (discontinuous) (e.g., in a polypeptide, amino acid residues that are not contiguous with each other in the polypeptide sequence but have connections in the molecule are subject to binding by an antigen-binding protein). A conformational epitope is an epitope that is present in the conformation of the active protein but not in the denatured protein. In certain embodiments, an epitope can be mimetic in that it contains a three-dimensional structure similar to the epitope used to generate the antigen-binding protein, but does not contain, or contains only some of, the amino acid residues found in the epitope used to generate the antigen-binding protein. Epitopes are most often present in proteins, but in some cases can be present in other types of molecules, such as nucleic acids. Epitopic determinants may include chemically active surface groupings of molecules, such as amino acids, sugar side chains, phosphate groups, or sulfonyl groups, and may have specific three-dimensional structural characteristics and / or specific charge characteristics. In general, an antigen-binding protein specific for a particular target antigen will preferentially recognize epitopes present on the target antigen in a complex mixture of proteins and / or macromolecules.

[0093] As used herein, "substantially pure" means that the molecule of the stated species is the predominant species present, i.e., it is more abundant on a molar basis than any other individual species in the same mixture. In certain embodiments, a substantially pure molecule is a composition in which the target species constitutes at least 50% (on a molar basis) of all macromolecular species present. In other embodiments, a substantially pure composition will constitute at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% of all macromolecular species present in the composition. In other embodiments, the target species is purified to essential homogeneity such that contaminating species cannot be detected in the composition by conventional detection methods, and thus the composition consists of a single detectable macromolecular species.

[0094] The term "treatment" refers to any indication of success in treating or ameliorating an injury, condition, or disease state, including any objective or subjective parameter, such as alleviation, mitigation, or reduction of symptoms, or improvement in the patient's tolerance of the injury, condition, or disease state, slowing the rate of deterioration or decline, less debilitating end points of deterioration, or improvement in the patient's physical or mental well-being. The treatment or amelioration of symptoms may be based on objective or subjective parameters, including the results of a physical examination, a neuropsychiatric examination, and / or a psychiatric evaluation. For example, certain methods presented herein successfully treat cardiovascular disease, such as atherosclerosis, by reducing the incidence of cardiovascular disease, inducing remission of cardiovascular disease, and / or ameliorating symptoms associated with cardiovascular disease.

[0095] An "effective amount" is generally an amount sufficient to reduce the severity and / or frequency of a symptom, eliminate a symptom and / or its underlying cause, prevent the onset of a symptom and / or its underlying cause, and / or ameliorate or treat damage caused by or associated with a disease condition (e.g., diabetes, obesity, dyslipidemia, elevated glucose levels, elevated insulin levels, or diabetic nephropathy). In some embodiments, an effective amount is a therapeutically effective amount or a prophylactically effective amount. A "therapeutically effective amount" is an amount sufficient to treat a disease condition (e.g., atherosclerosis) or symptom, particularly a condition or symptom associated with a disease condition, or to prevent, prevent, delay, or reverse the progression of a disease condition or any other undesirable symptoms associated with a disease, however caused. A "prophylactically effective amount" is an amount of a pharmaceutical composition that, when administered to a subject, will have the intended prophylactic effect, such as preventing or delaying the onset (or recurrence) of a disease condition, or reducing the likelihood of the onset (or recurrence) of a disease condition or associated symptoms. The full therapeutic or prophylactic effect does not necessarily result from administration of a single dose, and may occur only after administration of a series of doses. Thus, a therapeutically or prophylactically effective amount may be administered in one or more administrations.

[0096] As used herein, the terms "therapeutically effective dose" and "therapeutically effective amount" refer to an amount of GIPR binding protein that elicits a biological or medicinal response in a tissue system, animal, or human being sought by a researcher, physician, or other clinician, including reduction or amelioration of symptoms of the disease or disorder being treated, i.e., an amount of GIPR binding protein that supports an observable level of one or more desired biological or medicinal responses, such as reduced blood glucose, insulin, triglyceride, or cholesterol levels, weight loss, or improved glucose tolerance, energy expenditure, or insulin sensitivity.

[0097] The term "polynucleotide" or "nucleic acid" includes both single-stranded and double-stranded nucleotide polymers. The nucleotides that make up a polynucleotide can be ribonucleotides or deoxyribonucleotides, or modified forms of either type of nucleotide. Modifications include base modifications such as bromouridine and inosine derivatives, ribose modifications such as 2',3'-dideoxyribose, and modifications of internucleotide linkages such as phosphorothioates, phosphorodithioates, phosphoroselenoates, phosphorodiselenoates, phosphoroanilothioates, phosphoroaniladates, and phosphoroamidates.

[0098] The term "oligonucleotide" refers to a polynucleotide comprising 200 or fewer nucleotides. In some embodiments, the oligonucleotide is 10-60 bases in length. In other embodiments, the oligonucleotide is 12, 13, 14, 15, 16, 17, 18, 19, or 20-40 nucleotides in length. The oligonucleotide can be single-stranded or double-stranded, for example, for use in constructing mutant genes. The oligonucleotide can be a sense oligonucleotide or an antisense oligonucleotide. The oligonucleotide can include a label, including a radiolabel, a fluorescent label, a hapten, or an antigenic label for detection assays. The oligonucleotide may be used, for example, as a PCR primer, a cloning primer, or a hybridization probe.

[0099] An "isolated nucleic acid molecule" refers to DNA or RNA of genomic, mRNA, cDNA, or synthetic origin, or any combination thereof, where the isolated polynucleotide is unaccompanied by all or a portion of polynucleotides found in nature, or linked to polynucleotides with which it is not naturally linked. For purposes of this disclosure, a "nucleic acid molecule comprising" a particular nucleotide sequence should be understood not to encompass an intact chromosome. An isolated nucleic acid molecule that "comprises" a particular nucleic acid sequence may, in addition to the particular sequence, include sequences encoding up to 10 or even up to 20 other proteins or portions thereof, or may include operably linked regulatory sequences that control expression of the coding region of the described nucleic acid sequence, and / or may include vector sequences.

[0100] Unless otherwise stated, the left end of any single-stranded polynucleotide sequence discussed herein is the 5' end, and the left direction of a double-stranded polynucleotide sequence is referred to as the 5' direction. The direction in which nascent RNA transcripts are added from 5' to 3' is referred to as the transcription direction. The sequence region present on the DNA strand with the same sequence as the RNA transcript and located 5' to the 5' end of the RNA transcript is referred to as the "upstream sequence". The sequence region present on the DNA strand with the same sequence as the RNA transcript and located 3' to the 3' end of the RNA transcript is referred to as the "downstream sequence".

[0101] 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 may depend on the host organism. In certain embodiments, control sequences for prokaryotes may include a promoter, a ribosomal binding site, and a transcription termination sequence. For example, control sequences for eukaryotes may include a promoter containing one or more recognition sites for transcription factors, a transcription enhancer sequence, and a transcription termination sequence. "Control sequences" may include leader sequences and / or fusion partner sequences.

[0102] The term "vector" refers to any molecule or entity (eg, nucleic acid, plasmid, bacteriophage, or virus) used to introduce protein-coding information into a host cell.

[0103] 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 cooperation with the host cell) the expression of one or more heterologous coding regions operably linked thereto. Expression constructs may include, but are not limited to, sequences that affect or control transcription, translation, and, if introns are present, affect RNA splicing of the coding regions operably linked thereto.

[0104] 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 specific functions under appropriate conditions. For example, a control sequence "operably linked" to a protein coding sequence in a vector is ligated thereto such that expression of the protein coding sequence is achieved under conditions compatible with the transcriptional activity of the control sequences.

[0105] The term "host cell" refers to a cell that has been transformed with a nucleic acid sequence and thereby expresses a gene of interest. The term includes the progeny of a parent cell, whether or not the progeny is identical in morphology or genetic make-up to the original parent cell, so long as the gene of interest is present.

[0106] The terms "polypeptide" or "protein" are used interchangeably herein to refer to a polymer of amino acid residues. The terms apply to amino acid polymers in which one or more amino acid residues are analogs or mimetics of the corresponding naturally occurring amino acids, as well as to naturally occurring amino acid polymers. The terms can also include amino acid polymers that have been modified, for example, by the addition of carbohydrate residues to form glycoproteins, or by phosphorylation. Polypeptides and proteins can be produced by naturally occurring and non-naturally occurring cells, or by genetically engineered or recombinant cells, and include molecules having the amino acid sequence of a naturally occurring protein, or molecules having one or more amino acid deletions, additions, and / or substitutions of the naturally occurring sequence. The terms "polypeptide" and "protein" specifically encompass GIPR antigen binding proteins, antibodies, or sequences having one or more amino acid deletions, additions, and / or substitutions of antigen binding proteins. The term "polypeptide fragment" refers to a polypeptide having an amino-terminal deletion, a carboxyl-terminal deletion, and / or an internal deletion compared to the full-length protein. Such fragments may also contain modified amino acids compared to the full-length protein. In certain embodiments, the fragments are about 5-500 amino acids in length. For example, the fragments may be at least 5, at least 6, at least 8, at least 10, at least 14, at least 20, at least 50, at least 70, at least 100, at least 110, at least 150, at least 200, at least 250, at least 300, at least 350, at least 400, or at least 450 amino acids in length. Useful polypeptide fragments include immunologically functional fragments of antibodies that contain the binding domain.

[0107] The term "isolated protein" means that the subject protein is (1) free from at least some of the other proteins with which it is normally expected to be found; (2) essentially free from other proteins from the same source, e.g., the same species; (3) expressed by cells from a different species; (4) free from at least about 50 percent of the polynucleotides, lipids, carbohydrates, or other materials that naturally accompany it; (5) operably associated (by covalent or non-covalent interactions) with polypeptides that are not naturally associated with it; or (6) not naturally occurring. 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. Such isolated proteins can be encoded by genomic DNA, cDNA, mRNA, or other RNA of synthetic origin, or any combination thereof. Preferably, an isolated protein is substantially free of proteins or polypeptides or other contaminants that are found in its natural environment and that would interfere with its therapeutic, diagnostic, prophylactic, research, or other use.

[0108] A "variant" of a polypeptide (e.g., an antigen-binding protein such as an antibody) includes an amino acid sequence in which one or more amino acid residues have been inserted, deleted, and / or substituted in the amino acid sequence compared to another polypeptide sequence. Variants include fusion proteins.

[0109] A "derivative" of a polypeptide is a polypeptide (e.g., an antigen binding protein such as an antibody) that has been chemically modified in some way that differs from a mutant by insertion, deletion, or substitution, for example, through conjugation to another chemical moiety.

[0110] The term "natural origin" as used throughout this specification in connection with biological material such as polypeptides, nucleic acids, host cells, and the like, refers to material that is found in nature.

[0111] As used herein, a "subject" or "patient" can be any mammal. In an exemplary embodiment, the subject or patient is a human.

[0112] As disclosed herein, the GIPR polypeptide described by this disclosure can be engineered and / or produced using standard molecular biology methodology. In various examples, the nucleic acid sequence encoding GIPR can include all or part of SEQ ID NO:1, SEQ ID NO:3, or SEQ ID NO:5, and can be isolated and / or amplified from genomic DNA or cDNA using appropriate oligonucleotide primers. Primers can be designed based on the nucleic acid and amino acid sequences provided herein according to standard (RT)-PCR amplification techniques. The amplified GIPR nucleic acid can then be cloned into an appropriate vector and characterized by DNA sequence analysis.

[0113] Oligonucleotides for use as probes in isolating or amplifying all or a portion of the GIPR sequences provided herein can be designed and generated using standard synthetic techniques, such as, for example, an automated DNA synthesizer, or can be isolated from longer DNA sequences.

[0114] The sequence of the 466 amino acids of human GIPR is (Volz et al., FEBS Lett. 373:23-29 (1995), NCBI Reference Sequence: NP_0001555): [ka] and is encoded by the following DNA sequence (NCBI Reference Sequence: NM_000164): [ka] [ka]

[0115] The 430 amino acid isoform of human GIPR (isoform X1) was predicted by automated computational analysis and has the sequence (NCBI reference sequence XP_005258790): [ka] and is encoded by the DNA sequence: [ka]

[0116] A 493 amino acid isoform of human GIPR is generated by alternative splicing and has the sequence (Gremlich et al., Diabetes 44:1202-8 (1995), UniProtKB sequence identifier: P48546-2): [ka] and is encoded by the DNA sequence: [ka]

[0117] The sequence of the 460 amino acids of mouse GIPR is (NCBI Reference Sequence: NP_001074284, uniprotKB / Swiss-Prot Q0P543-1) (see Vassilatis et al., PNAS USA 2003, 100:4903-4908). [ka] and is encoded by the following DNA sequence (NCBI Reference Sequence: NM_001080815): [ka]

[0118] A 230 amino acid isoform of mouse GIPR is generated by alternative splicing and has the sequence (Gerhard et al., Genome Res, 14:2121-2127 (2004); NCBI Reference Sequence: AAI20674): [ka] and is encoded by the DNA sequence: [ka]

[0119] The term "GIPR polypeptide" as used herein encompasses naturally occurring GIPR polypeptide sequences, such as, for example, the human amino acid sequences of SEQ ID NO: 1201, SEQ ID NO: 1203, or SEQ ID NO: 1205. However, the term "GIPR polypeptide" also encompasses polypeptides that include an amino acid sequence that differs from the amino acid sequence of a naturally occurring GIPR polypeptide sequence, such as, for example, SEQ ID NO: 1201, SEQ ID NO: 1203, or SEQ ID NO: 1205, by one or more amino acids, such that the sequence is at least 85% identical to SEQ ID NO: 1201, SEQ ID NO: 1203, or SEQ ID NO: 1205. GIPR polypeptides can be generated by using naturally occurring or non-naturally occurring amino acids and introducing one or more amino acid substitutions (conservative or non-conservative) for specific positions in a GIPR polypeptide.

[0120] In a "conservative amino acid substitution," a naturally occurring amino acid residue (i.e., a residue found at a given position in a wild-type GIPR polypeptide sequence) may be substituted with a non-naturally occurring residue (i.e., a residue not found at a given position in a wild-type GIPR polypeptide sequence) such that there is little or no effect on the polarity or charge of the amino acid residue at that position. Conservative amino acid substitutions also encompass non-naturally occurring amino acid residues that are typically incorporated by chemical peptide synthesis rather than by synthesis in a biological system. These include peptidomimetics and other forms in which amino acid moieties are reversed or inverted.

[0121] Naturally occurring residues can be grouped into classes based on the following common side chain properties: (1) Hydrophobic: Norleucine, Met, Ala, Val, Leu, Ile, (2) Neutral hydrophilic: Cys, Ser, Thr, (3) Acidic: Asp, Glu, (4) Basic: Asn, Gln, His, Lys, Arg, (5) Residues that affect chain orientation: Gly, Pro, and (6) Aromatic: Trp, Tyr, Phe.

[0122] Additional taxa of amino acids can also be constructed using the principles described, for example, in Creighton (1984) PROTEINS: STRUCTURE AND MOLECULAR PROPERTIES (2d Ed. 1993), W.H. Freeman and Company. In some cases, it may be useful to further characterize the substitution based on two or more of such properties (e.g., substitution with a "small polar" residue such as a Thr residue may be a highly conservative substitution in the appropriate circumstances).

[0123] Conservative substitutions may involve the exchange of a member of one such class for another member of the same class. Non-conservative substitutions may involve the exchange of a member of one such class for a member of another class.

[0124] Synthetic, rare, or modified amino acid residues known to have similar physiochemical properties to those in the above classes can be used as "conservative" replacements for certain amino acid residues in the sequence. For example, a D-Arg residue can serve as a replacement for a typical L-Arg residue. There may be cases where a particular replacement can be described in terms of more than one of the above classes (e.g., replacement with a small, hydrophobic residue means replacement of one amino acid with a residue(s) found in both of the above classes, or with another synthetic, rare, or modified residue known in the art to have similar physiochemical properties to such a residue that meets both definitions).

[0125] Nucleic acid sequences encoding GIPR polypeptides provided herein include those that are degenerate to SEQ ID NO:1201, SEQ ID NO:1203, or SEQ ID NO:1205, and those that encode polypeptide variants of SEQ ID NO:1201, SEQ ID NO:1203, or SEQ ID NO:1205 from other aspects of the disclosure.

[0126] To express the GIPR nucleic acid sequences provided herein, a suitable coding sequence, such as, for example, SEQ ID NO: 1201, SEQ ID NO: 1203, or SEQ ID NO: 1205, can be cloned into a suitable vector according to standard cloning and expression techniques, and after introduction into a suitable host, the sequence can be expressed to produce the encoded polypeptide, as known in the art (e.g., as described in Sambrook, J., Fritsh, EF, and Maniatis, T. Molecular Cloning: A Laboratory Manual 2nd, ed., Cold Spring Harbor Laboratory, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, 1989). The present invention also relates to such vectors comprising the nucleic acid sequences according to the present invention.

[0127] "Vector" refers to a delivery vehicle that (a) facilitates expression of a nucleic acid sequence encoding a polypeptide, (b) facilitates production of a polypeptide therefrom, (c) facilitates transduction / transformation of a target cell therewith, (d) facilitates replication of a nucleic acid sequence, (e) facilitates stability of the nucleic acid, (f) facilitates detection of the nucleic acid and / or transformed / transduced cells, and / or (g) otherwise confers advantageous biological and / or physiochemical functions to the nucleic acid encoding the polypeptide. The vector can be any suitable vector, including chromosomal vectors, non-chromosomal vectors, and synthetic nucleic acid vectors (nucleic acid sequences comprising an appropriate set of expression control elements). Examples of such vectors include derivatives of SV40, bacterial plasmids, phage DNA, baculovirus, yeast plasmids, vectors derived from combinations of plasmids and phage DNA, and viral nucleic acid (RNA or DNA) vectors.

[0128] Recombinant expression vectors can be designed to express GIPR proteins in prokaryotic cells (e.g., E. coli) or eukaryotic cells (e.g., insect cells, yeast cells, or mammalian cells using baculovirus expression vectors). In one embodiment, the host cell is a mammalian non-human host cell. Representative host cells include hosts typically used for cloning and expression, such as Escherichia coli strains TOP10F', TOP10, DH10B, DH5a, HB101, W3110, BL21(DE3), and BL21(DE3)pLysS, BLUESCRIPT (Stratagene), mammalian cell lines CHO, CHO-K1, HEK293, 293-EBNApIN vectors (Van Heeke & Schuster, J. Biol. Chem. 264:5503-5509 (1989)), pET vectors (Novagen, Madison, MA). Alternatively, the recombinant expression vector can be transcribed and translated in vitro, for example, using a T7 promoter regulatory sequence and T7 polymerase and an in vitro translation system. Preferably, the vector contains a promoter upstream of the cloning site that contains the nucleic acid sequence encoding the polypeptide. Examples of promoters that can be switched on and off include the lac promoter, the T7 promoter, the trc promoter, the tac promoter, and the trp promoter.

[0129] Thus, vectors are provided herein that contain a nucleic acid sequence encoding GIPR and promote the expression of recombinant GIPR. In various embodiments, the vectors contain operably linked nucleotide sequences that regulate the expression of GIPR. The vectors may contain or be associated with any suitable promoters, enhancers, and other expression promoting elements. Examples of such elements include a strong expression promoter (e.g., human CMV IE promoter / enhancer, RSV promoter, SV40 promoter, SL3-3 promoter, MMTV promoter, or HIV LTR promoter, EF1 alpha promoter, CAG promoter), an effective poly(A) termination sequence, an origin of replication for the plasmid product in E. coli, an antibiotic resistance gene as a selectable marker, and / or a convenient cloning site (e.g., polylinker). The vectors may also contain an inducible promoter as opposed to a constitutive promoter such as CMV IE. In one aspect, a nucleic acid is provided that contains a GIPR polypeptide coding sequence operably linked to a tissue-specific promoter that promotes the expression of the sequence in metabolically relevant tissues such as liver tissue or pancreatic tissue.

[0130] In another aspect of the disclosure, host cells are provided that contain the GIPR nucleic acids and vectors disclosed herein. In various embodiments, the vector or nucleic acid is integrated into the host cell genome, while in other embodiments, the vector or nucleic acid is present extrachromosomally.

[0131] Recombinant cells, such as yeast cells, bacterial cells (e.g., E. coli), and mammalian cells (e.g., immortalized mammalian cells) containing such nucleic acids, vectors, or a combination of either or both, are provided. In various embodiments, cells are provided that contain a non-integrated nucleic acid, such as a plasmid, cosmid, phagemid, or linear expression element, that includes a sequence encoding a GIPR polypeptide for expression.

[0132] A vector containing a nucleic acid sequence encoding a GIPR polypeptide provided herein can be introduced into a host cell by transformation or transduction. Methods for transforming cells with expression vectors are well known.

[0133] The nucleic acid encoding the GIPR can be placed and / or delivered to a host cell or host animal via a viral vector. Any suitable viral vector with this capability can be used. The viral vector can include any number of viral polynucleotides, alone or in combination with one or more viral proteins that facilitate the delivery, replication, and / or expression of the nucleic acid of the present invention in a desired host cell. The viral vector can be a polynucleotide that includes all or part of the viral genome, a viral protein / nucleic acid complex, a virus-like particle (VLP), or an intact viral particle that includes a viral nucleic acid and a nucleic acid encoding a GIPR polypeptide. The viral vector that is a viral particle can include a wild-type viral particle or a modified viral particle. The viral vector can be a vector that requires the presence of another vector or wild-type virus for replication and / or expression, such as an adenoviral vector amplicon (e.g., the viral vector can be a helper-dependent virus). Typically, such viral vectors consist of wild-type viral particles or viral particles whose protein and / or nucleic acid content has been modified to increase transgene capacity or aid in gene transfer and / or expression of nucleic acid (examples of such vectors include herpesvirus / AAV amplicons). Typically, viral vectors are similar to and / or derived from viruses that normally infect humans. Suitable viral vector particles in this regard include, for example, adenovirus vector particles (including any virus of the Adenoviridae family or any virus derived from the Adenoviridae family), adeno-associated virus vector particles (AAV vector particles) or other parvoviruses and parvovirus vector particles, papillomavirus vector particles, flavivirus vectors, alphavirus vectors, herpesvirus vectors, poxvirus vectors, retrovirus vectors (including lentivirus vectors).

[0134] GIPR polypeptides expressed as described herein can be isolated using standard protein purification methods. GIPR polypeptides can be isolated from cells that naturally express them, or from cells that have been engineered to express GIPR, such as cells that do not naturally express GIPR.

[0135] Protein purification methods that can be used to isolate GIPR polypeptides, as well as related materials and reagents, are known in the art. Additional purification methods that can be useful for isolating GIPR polypeptides can be found in references such as Bootcov MR, 1997, Proc. Natl. Acad. Sci. USA 94:11514-9, Fairlie WD, 2000, Gene 254:67-76.

[0136] Provided herein is an antagonist antigen binding protein that binds to GIPR, including human GIPR (hGIPR).In one embodiment, human GIPR has a sequence such as that shown in SEQ ID NO: 1201.In another embodiment, human GIPR has a sequence such as that shown in SEQ ID NO: 1203.In another embodiment, human GIPR has a sequence such as that shown in SEQ ID NO: 1205.

[0137] The antigen binding proteins provided are polypeptides incorporating and / or linked to one or more complementarity determining regions (CDRs) as described herein. In some antigen binding proteins, the CDRs are incorporated into "framework" regions that orient the CDR(s) so that the appropriate antigen binding properties of the CDR(s) are achieved. Certain antigen binding proteins described herein are antibodies or are derived from antibodies. In other antigen binding proteins, the CDR sequences are incorporated into different types of protein scaffolds. The various structures are further described below.

[0138] The antigen binding proteins disclosed herein have a variety of utilities. The antigen binding proteins are useful, for example, in specific binding assays, affinity purification of GIPR, and screening assays to identify other antagonists of GIPR activity. Other uses of the antigen binding proteins include, for example, diagnosis of diseases or conditions associated with GIPR, and screening assays to determine the presence or absence of GIPR. Given that the antigen binding proteins provided are antagonists, the GIPR antigen binding proteins are valuable in therapeutic methods useful for reducing weight gain, improving glucose tolerance, reducing insulin levels, and reducing cholesterol and triglyceride levels, even while food intake is maintained or increased, % body fat mass is increased, and % lean mass is increased. Thus, the antigen binding proteins have utility, for example, in the treatment and prevention of diabetes, such as type 2 diabetes, obesity, dyslipidemia, elevated glucose levels, or elevated insulin levels.

[0139] A variety of selective binding agents are provided that are useful for regulating the activity of GIPR.These agents include, for example, antigen-binding proteins (e.g., scFv, domain antibodies, and polypeptides with antigen-binding regions) that contain antigen-binding domains and specifically bind to GIPR polypeptides, particularly human GIPR.Some of these agents are, for example, useful for enhancing the activity of GIPR, and can activate one or more activities associated with GIPR.

[0140] In general, the antigen binding proteins provided typically comprise one or more (e.g., one, two, three, four, five, or six) CDRs described herein. In some cases, the antigen binding protein comprises (a) a polypeptide structure and (b) one or more CDRs inserted and / or linked to the polypeptide structure. The polypeptide structure may take a variety of different forms. For example, the polypeptide structure may be or may comprise a framework of a naturally occurring antibody or a fragment or variant thereof, or may be entirely synthetic in nature. Examples of various polypeptide structures are further described below.

[0141] In certain embodiments, the polypeptide structure of the antigen-binding protein is an antibody or is derived from an antibody. Thus, certain examples of antigen-binding proteins provided include, but are not limited to, monoclonal antibodies, bispecific antibodies, minibodies, domain antibodies (such as Nanobodies®), synthetic antibodies (sometimes referred to herein as "antibody mimetics"), chimeric antibodies, humanized antibodies, human antibodies, antibody fusions, and portions or fragments thereof. In some cases, the antigen-binding protein is an immunological fragment of a complete antibody (e.g., Fab, Fab', F(ab')2). In other cases, the antigen-binding protein is an scFv that uses CDRs derived from an antibody of the invention.

[0142] The antigen binding proteins provided herein specifically bind to human GIPR. In certain embodiments, the antigen binding proteins specifically bind to human GIPR comprising or consisting of the amino acid sequence of SEQ ID NO: 1201. In certain embodiments, the antigen binding proteins specifically bind to human GIPR comprising or consisting of the amino acid sequence of SEQ ID NO: 1203. In certain embodiments, the antigen binding proteins specifically bind to human GIPR comprising or consisting of the amino acid sequence of SEQ ID NO: 1205.

[0143] The antigen binding proteins provided are antagonists and typically have one, two, three, four, five, six, seven, or all eight of the following properties: (a) Ability to block or reduce binding of GIP to GIPR, the levels of which can be measured by methods such as binding studies with radioactively or fluorescently labeled ligands or methods described herein (e.g., cAMP assays or other functional assays). Reductions can be at least 10%, at least 25%, at least 50%, at least 100%, or more, compared to pre-treatment levels of SEQ ID NO:1201, SEQ ID NO:1203, or SEQ ID NO:1205 under comparable conditions. (b) the ability to reduce blood glucose; (c) the ability to improve glucose tolerance; (d) the ability to improve insulin sensitivity; (e) the ability to lose weight or reduce weight gain; (f) the ability to reduce body fat mass or reduce inflammation in adipose tissue; (g) the ability to reduce fasting insulin levels; (h) the ability to reduce circulating cholesterol levels; (i) the ability to reduce circulating triglyceride levels; (j) the ability to reduce hepatic steatosis or reduce triglyceride levels in the liver; (k) the ability to reduce AST, ALT, and / or ALP levels.

[0144] In one embodiment, the GIPR antigen binding protein has one or more of the following activities: (a) binds to human GIPR, resulting in a KD of ≦200 nM, ≦150 nM, ≦100 nM, ≦50 nM, ≦10 nM, ≦5 nM, ≦2 nM, or ≦1 nM, as measured, for example, via surface plasma resonance or equilibrium exclusion binding techniques. (b) a half-life in human serum of at least 3 days; Some of the antigen binding proteins provided have a binding rate (ka) for GIPR of at least 10, e.g., as measured as described below. 4 / Mx seconds, at least 10 5 / Mx seconds or at least 10 6 / Mx seconds. Certain antigen-binding proteins provided have slow dissociation or off-rates. Some antigen-binding proteins have dissociation rates of, for example, 1x10 -2 seconds -1 , or 1x10 -3 seconds -1 , or 1x10 -4 seconds -1 , or 1x10 -5 seconds -1 In certain embodiments, the antigen binding protein has a KD (equilibrium binding affinity) of less than 25 pM, less than 50 pM, less than 100 pM, less than 500 pM, less than 1 nM, less than 5 nM, less than 10 nM, less than 25 nM, or less than 50 nM.

[0145] The assay can also measure the binding of an antigen-binding protein to its target as EC50 (the concentration of antigen-binding protein that gives half of the maximum response when bound to the target).The EC50 for the anti-GIPR antigen-binding protein of the present invention can be determined by incubating various concentrations of the antigen-binding protein with cells expressing GIPR.The anti-GIPR antigen-binding protein of the present invention can have an EC50 of less than 200nM, 150nM, 125nM, 100nM, 90nM, 80nM, 70nM, 60nM, 50nM, 40nM, or 30nM.

[0146] IC50 (half maximal inhibitory concentration: a measure of the effectiveness of an antigen-binding protein in inhibiting a specific biological or biochemical function) can also be used to measure the activity of anti-GIPR antigen-binding proteins. IC50 can be measured using a functional assay. For example, such an assay can be used to quantitatively determine cAMP in HEK293T cells expressing human GIPR or cynomolgus GIPR. Binding of GIP causes a conformational change in GIPR, stimulating G protein to activate adenylate cyclase, which results in the production of cAMP from ATP. Binding of an antibody to GIPR prevents binding of GIP to GIPR, resulting in a decrease in cAMP. This can be measured by a cAMP assay. Anti-GIPR antigen binding proteins of the invention may have an IC50 of less than 200nM, 150nM, 125nM, 100nM, 90nM, 80nM, 70nM, 60nM, 50nM, 40nM, 30nM, 29nM, 28nM, 27nM, 26nM, 25nM, 24nM, 23nM, 22nM, 21mM, 20nM, 19nM, 18nM, 17nM, 16nM, 15nM, 14nM, 13nM, 12nM, 11mM, 10nM, 9nM, 8nM, 7nM, 6nM, 5nM, 4nM, 3nM, 2nM, or 1nM.

[0147] In another aspect, antigen binding proteins are provided that have a half-life in vitro or in vivo (e.g., when administered to a human subject) of at least 1 day. In one embodiment, the antigen binding protein has a half-life of at least 3 days. In various other embodiments, the antigen binding protein has a half-life of 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 30, 40, 50, or 60 days or more. In another embodiment, the antigen binding protein is derivatized or modified to increase its half-life compared to underivatized or unmodified antibodies. In another embodiment, point mutations are included in the antigen binding protein to increase serum half-life. Further details regarding such variants and derivatized forms are provided below.

[0148] Some of the antigen-binding proteins provided have a structure typically associated with naturally occurring antibodies. Such antibody structural units typically include one or more tetramers, each of which is composed of two identical couplets of polypeptide chains, although some mammalian species also produce antibodies with 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 of which consists of approximately 90-110 amino acids and exhibits a characteristic folding pattern. These domains are the basic units that make up an antibody polypeptide. The amino-terminal portion of each chain typically includes a variable domain that is responsible for antigen recognition. The carboxy-terminal portion is more evolutionarily conserved compared to the other end of the chain and is referred to as the "constant region" or "C region". Human light chains are generally classified as kappa and lambda light chains, each of which contains one variable and one constant domain. Heavy chains are typically classified as mu, delta, gamma, alpha, or epsilon chains, which 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, IgA and IgD isotypes contain four heavy chains and four light chains, IgG and IgE isotypes contain two heavy chains and two light chains, and IgM isotypes contain five heavy chains and five light chains. The C region of a heavy chain typically contains one or more domains that may be responsible for effector function. The number of constant region domains in a heavy chain will depend on the isotype. IgG heavy chains, for example, each contain three C region domains known as CH1, CH2, and CH3.The antibodies provided may have any of these isotypes and subtypes. In certain embodiments, the GIPR antibody is of the IgG1, IgG2, or IgG4 subtype. The terms "GIPR antibody" and "anti-GIPR antibody" are used interchangeably throughout this application and the figures. Both terms refer to an antibody that binds to GIPR.

[0149] In full-length 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 or more amino acids. See, e.g., Fundamental Immunology, 2nd ed., Ch. 7 (Paul, W., ed.) 1989, New York: Raven Press, incorporated herein by reference in its entirety for all purposes. The variable regions of each light / heavy chain pair typically form the antigen-binding site.

[0150] For the antibodies provided herein, the variable regions of immunoglobulin chains generally exhibit the same overall structure, including relatively conserved framework regions (FR) linked by three hypervariable regions (more often referred to as "complementarity determining regions" or CDRs). The CDRs from the two chains of each heavy / light chain pair described above are typically aligned by the framework regions to form a structure that specifically binds to a particular epitope present in the GIPR. These elements are typically present in the following order from N-terminus to C-terminus in both naturally occurring light and heavy chain variable regions: FR1, CDR1, FR2, CDR2, FR3, CDR3, and FR4. A numbering system has been devised to assign numbers to the amino acids that occupy each position in these domains. This numbering system is defined in Kabat Sequences of Proteins of Immunological Interest (1987 and 1991, NIH, Bethesda, Md.), or Chothia & Lesk, 1987, J. Mol. Biol. 196:901-917; Chothia et al., 1989, Nature 342:878-883.

[0151] Sequence information for particular antibodies prepared and identified as described in the Examples below is summarized in Table 1. Thus, in one embodiment, the antigen binding protein is an antibody having the CDR sequences, variable domain sequences, and light and heavy chain sequences identified in one of the rows of Table 1.

[0152] The variable light chain sequences, variable heavy chain sequences, light chain sequences, heavy chain sequences, CDRL1 sequences, CDRL2 sequences, CDRL3 sequences, CDRH1 sequences, CDRH2 sequences, and CDRH3 sequences of the antibodies and fragments thereof of the invention have been assigned SEQ ID NOs, which are shown in Table 1. The polynucleotides encoding the variable light chain sequences, variable heavy chain sequences, light chain sequences, heavy chain sequences, CDRL1 sequences, CDRL2 sequences, CDRL3 sequences, CDRH1 sequences, CDRH2 sequences, and CDRH3 sequences of the antibodies and fragments thereof of the invention have also been assigned SEQ ID NOs, which are shown in Table 2. The antigen binding proteins of the invention can be identified by SEQ ID NOs, but also by construct names (e.g., 2C2.005) or identifier numbers (e.g., iPS:336175). The antigen binding proteins identified in Tables 1-5 below can be grouped into families based on construct names. For example, the "4B1 family" includes construct 4B1, construct 4B1.010, construct 4B1.011, construct 4B1.012, construct 4B1.013, construct 4B1.014, construct 4B1.015, and construct 4B1.016.

[0153] The various light and heavy chain variable regions provided herein are shown in Table 3. Each of these variable regions can be added to a heavy or light chain constant region to form the heavy and light chains, respectively, of a complete antibody. Furthermore, each of the heavy and light chain sequences thus generated can be combined to form the structure of a complete antibody.

[0154] [Table 1]

[0155] [Table 2]

[0156] [Table 3]

[0157]

Table 4

[0158]

Table 5

[0159]

Table 6

[0160]

Table 7

[0161]

Table 8

[0162]

Table 9

[0163]

Table 10

[0164]

Table 11

[0165]

Table 12

[0166]

Table 13

[0167]

Table 14

[0168]

Table 15

[0169]

Table 16

[0170]

Table 17

[0171]

Table 18

[0172]

Table 19

[0173]

Table 20

[0174]

Table 21

[0175]

Table 22

[0176]

Table 23

[0177]

Table 24

[0178]

Table 25

[0179]

Table 26

[0180]

Table 27

[0181]

Table 28

[0182]

Table 29

[0183]

Table 30

[0184]

Table 31

[0185]

Table 32

[0186]

Table 33

[0187]

Table 34

[0188]

Table 35

[0189]

Table 36

[0190]

Table 37

[0191]

Table 38

[0192]

Table 39

[0193]

Table 40

[0194]

Table 41

[0195]

Table 42

[0196]

Table 43

[0197]

Table 44

[0198]

Table 45

[0199]

Table 46

[0200]

Table 47

[0201]

Table 48

[0202]

Table 49

[0203]

Table 50

[0204]

Table 51

[0205]

Table 52

[0206]

Table 53

[0207]

Table 54

[0208]

Table 55

[0209]

Table 56

[0210]

Table 57

[0211]

Table 58

[0212]

Table 59

[0213]

Table 60

[0214]

Table 61

[0215]

Table 62

[0216]

Table 63

[0217]

Table 64

[0218]

Table 65

[0219]

Table 66

[0220]

Table 67

[0221]

Table 68

[0222]

Table 69

[0223]

Table 70

[0224]

Table 71

[0225]

Table 72

[0226]

Table 73

[0227]

Table 74

[0228]

Table 75

[0229]

Table 76

[0230]

Table 77

[0231]

Table 78

[0232]

Table 79

[0233]

Table 80

[0234]

Table 81

[0235]

Table 82

[0236]

Table 83

[0237]

Table 84

[0238]

Table 85

[0239]

Table 86

[0240]

Table 87

[0241]

Table 88

[0242]

Table 89

[0243]

Table 90

[0244]

Table 91

[0245]

Table 92

[0246]

Table 93

[0247]

Table 94

[0248]

Table 95

[0249]

Table 96

[0250]

Table 97

[0251]

Table 98

[0252]

Table 99

[0253]

Table 100

[0254]

Table 101

[0255]

Table 102

[0256]

Table 103

[0257]

Table 104

[0258]

Table 105

[0259]

Table 106

[0260]

Table 107

[0261]

Table 108

[0262]

Table 109

[0263]

Table 110

[0264]

Table 111

[0265]

Table 112

[0266]

Table 113

[0267]

Table 114

[0268]

Table 115

[0269]

Table 116

[0270]

Table 117

[0271]

Table 118

[0272]

Table 119

[0273]

Table 120

[0274]

Table 121

[0275]

Table 122

[0276]

Table 123

[0277]

Table 124

[0278]

Table 125

[0279]

Table 126

[0280]

Table 127

[0281]

Table 128

[0282]

Table 129

[0283]

Table 130

[0284]

Table 131

[0285]

Table 132

[0286]

Table 133

[0287]

Table 134

[0288]

Table 135

[0289]

Table 136

[0290]

Table 137

[0291]

Table 138

[0292]

Table 139

[0293]

Table 140

[0294]

Table 141

[0295]

Table 142

[0296]

Table 143

[0297]

Table 144

[0298]

Table 145

[0299]

Table 146

[0300]

Table 147

[0301]

Table 148

[0302]

Table 149

[0303]

Table 150

[0304]

Table 151

[0305] In one embodiment, the antibody or fragment thereof is selected from the group consisting of SEQ ID NOs: 723, 727, 731, 735, 739, 743, 747, 751, 755, 759, 763, 767, 771, 775, 779, 783, 787, 791, 795, 799, 803, 807, 811, 815, 819, 823, 827, 831, 835, 839, 843, 847, 851, 855, 859, 86 In one embodiment, the invention comprises a light chain variable region comprising a sequence selected from the group consisting of: 3, 867, 871, 875, 879, 883, 887, 891, 895, 899, 903, 907, 911, 915, 919, 923, 927, 931, 935, 939, 943, 947, 951, 955, 959, 1286, 1296, 1306, 1316, 1326, 1336, 1346, and 1356. In one embodiment, the antibody or fragment thereof is selected from the group consisting of SEQ ID NOs: 724, 728, 732, 736, 740, 744, 748, 752, 756, 760, 764, 768, 772, 776, 780, 784, 788, 792, 796, 800, 804, 808, 812, 816, 820, 824, 828, 832, 836, 840, 844, 848, 852, 856, 860, 86 4, 868, 872, 876, 880, 884, 888, 892, 896, 900, 904, 908, 912, 916, 920, 924, 928, 932, 936, 940, 944, 948, 952, 956, 960, 1287, 1297, 1307, 1317, 1327, 1337, 1347, and 1357.In one embodiment, the antibody or fragment thereof is selected from the group consisting of SEQ ID NOs: 723, 727, 731, 735, 739, 743, 747, 751, 755, 759, 763, 767, 771, 775, 779, 783, 787, 791, 795, 799, 803, 807, 811, 815, 819, 823, 827, 831, 835, 839, 843, 847, 851, 855, 859, 860, 861, 862, 863, 864, 865, 866, 867, 871, 871, 875, 879, 883, 887, 881, 882, 883, 884, 885, 886, 887, 888, 889, 890, 900, 901, 902, 903, 904, 905, 906, 907, 908, 909, 910, 911, 912, 913, 914, 915, 916, 917, 918, 919, 920, 921, 922, 923, 924, 925, 926, 927, 928, 929, 930, 931, 932, 933, 934, 935, 936, 937, 938 59, 863, 867, 871, 875, 879, 883, 887, 891, 895, 899, 903, 907, 911, 915, 919, 923, 927, 931, 935, 939, 943, 947, 951, 955, 959, 1286, 1296, 1306, 1316, 1326, 1336, 1346, and 1356. Light chain variable regions and SEQ ID NOs: 724, 728, 732, 736, 740, 744, 748, 752, 756, 760, 764, 768, 772, 776, 780, 784, 788, 792, 796, 800, 804, 808, 812, 816, 820, 824, 828, 832, 836, 840, 844, 848, 852, 856, 860, 864, 868, 870, 872, 874, 876, 878, 879, 880, 882, 884, 886, 888, 889, 890, 900, 902, 904, 906, 908, 910, 912, 916, 920, 924, 928, 932, 936, 940, 944, 948, 952, 956, 956, 960, 964, 968, 970, 972, 976, 980, 984, 988, 990, 992, 996, 1000, 1002, 1004, 1006, 1008, 1006, 1008, 1009, 1010, 1011, 1012, 1013, 1014, 1015, 1016, 1017 and a heavy chain variable region comprising a sequence selected from the group consisting of: 72, 876, 880, 884, 888, 892, 896, 900, 904, 908, 912, 916, 920, 924, 928, 932, 936, 940, 944, 948, 952, 956, 960, 1287, 1297, 1307, 1317, 1327, 1337, 1347, and 1357.

[0306] In one embodiment, the antibody or fragment thereof comprises a light chain variable region comprising SEQ ID NO:723 and a heavy chain variable region comprising SEQ ID NO:724, a light chain variable region comprising SEQ ID NO:727 and a heavy chain variable region comprising SEQ ID NO:728, a light chain variable region comprising SEQ ID NO:731 and a heavy chain variable region comprising SEQ ID NO:732, a light chain variable region comprising SEQ ID NO:735 and a heavy chain variable region comprising SEQ ID NO:736, a light chain variable region comprising SEQ ID NO:739 and a heavy chain variable region comprising SEQ ID NO:740, a light chain variable region comprising SEQ ID NO:743 and a heavy chain variable region comprising SEQ ID NO:744, a light chain variable region comprising SEQ ID NO:747 and a heavy chain variable region comprising SEQ ID NO:748, A heavy chain variable region comprising SEQ ID NO: 748, a light chain variable region comprising SEQ ID NO: 751 and a heavy chain variable region comprising SEQ ID NO: 752, a light chain variable region comprising SEQ ID NO: 755 and a heavy chain variable region comprising SEQ ID NO: 756, a light chain variable region comprising SEQ ID NO: 759 and a heavy chain variable region comprising SEQ ID NO: 760, a light chain variable region comprising SEQ ID NO: 763 and a heavy chain variable region comprising SEQ ID NO: 764, a light chain variable region comprising SEQ ID NO: 767 and a heavy chain variable region comprising SEQ ID NO: 768, a light chain variable region comprising SEQ ID NO: 771 and a heavy chain variable region comprising SEQ ID NO: 772, a light chain variable region comprising SEQ ID NO: 775 and a heavy chain variable region comprising SEQ ID NO: 776 a light chain variable region comprising SEQ ID NO: 779 and a heavy chain variable region comprising SEQ ID NO: 780; a light chain variable region comprising SEQ ID NO: 783 and a heavy chain variable region comprising SEQ ID NO: 784; a light chain variable region comprising SEQ ID NO: 787 and a heavy chain variable region comprising SEQ ID NO: 788; a light chain variable region comprising SEQ ID NO: 791 and a heavy chain variable region comprising SEQ ID NO: 792; a light chain variable region comprising SEQ ID NO: 795 and a heavy chain variable region comprising SEQ ID NO: 796; a light chain variable region comprising SEQ ID NO: 799 and a heavy chain variable region comprising SEQ ID NO: 800; a light chain variable region comprising SEQ ID NO: 803 and a heavy chain variable region comprising SEQ ID NO: 804 a light chain variable region comprising SEQ ID NO:807 and a heavy chain variable region comprising SEQ ID NO:808; a light chain variable region comprising SEQ ID NO:811 and a heavy chain variable region comprising SEQ ID NO:812; a light chain variable region comprising SEQ ID NO:815 and a heavy chain variable region comprising SEQ ID NO:816; a light chain variable region comprising SEQ ID NO:819 and a heavy chain variable region comprising SEQ ID NO:820; a light chain variable region comprising SEQ ID NO:823 and a heavy chain variable region comprising SEQ ID NO:824; a light chain variable region comprising SEQ ID NO:827 and a heavy chain variable region comprising SEQ ID NO:828; a light chain variable region comprising SEQ ID NO:831 and a heavy chain variable region comprising SEQ ID NO:832;a light chain variable region comprising SEQ ID NO:835 and a heavy chain variable region comprising SEQ ID NO:836; a light chain variable region comprising SEQ ID NO:839 and a heavy chain variable region comprising SEQ ID NO:840; a light chain variable region comprising SEQ ID NO:843 and a heavy chain variable region comprising SEQ ID NO:844; a light chain variable region comprising SEQ ID NO:847 and a heavy chain variable region comprising SEQ ID NO:848; a light chain variable region comprising SEQ ID NO:851 and a heavy chain variable region comprising SEQ ID NO:852; a light chain variable region comprising SEQ ID NO:855 and a heavy chain variable region comprising SEQ ID NO:856; a light chain variable region comprising SEQ ID NO:859 and a heavy chain variable region comprising SEQ ID NO:860; a light chain variable region comprising SEQ ID NO:863 and a heavy chain variable region comprising SEQ ID NO:864; a light chain variable region comprising SEQ ID NO:867 and a heavy chain variable region comprising SEQ ID NO:868; a light chain variable region comprising SEQ ID NO:871 and a heavy chain variable region comprising SEQ ID NO:872; a light chain variable region comprising SEQ ID NO:875 and a heavy chain variable region comprising SEQ ID NO:876; a light chain variable region comprising SEQ ID NO:879 and a heavy chain variable region comprising SEQ ID NO:880; a light chain variable region comprising SEQ ID NO:883 and a heavy chain variable region comprising SEQ ID NO:884; a light chain variable region comprising SEQ ID NO:887 and a heavy chain variable region comprising SEQ ID NO:888; a light chain variable region comprising SEQ ID NO:891 and a heavy chain variable region comprising SEQ ID NO:892; a light chain variable region comprising SEQ ID NO:895 and a heavy chain variable region comprising SEQ ID NO:896; a light chain variable region comprising SEQ ID NO:899 and a heavy chain variable region comprising SEQ ID NO:900; a light chain variable region comprising SEQ ID NO:903 and a heavy chain variable region comprising SEQ ID NO:904; a light chain variable region comprising SEQ ID NO:907 and a heavy chain variable region comprising SEQ ID NO:908; a light chain variable region comprising SEQ ID NO:911 and a heavy chain variable region comprising SEQ ID NO:912; a light chain variable region comprising SEQ ID NO:915 and a heavy chain variable region comprising SEQ ID NO:916; a light chain variable region comprising SEQ ID NO:919 and a heavy chain variable region comprising SEQ ID NO:920; a light chain variable region comprising SEQ ID NO:923 and a heavy chain variable region comprising SEQ ID NO:924; a light chain variable region comprising SEQ ID NO:927 and a heavy chain variable region comprising SEQ ID NO:928; a light chain variable region comprising SEQ ID NO:931 and a heavy chain variable region comprising SEQ ID NO:932; a light chain variable region comprising SEQ ID NO:935 and a heavy chain variable region comprising SEQ ID NO:936; a light chain variable region comprising SEQ ID NO:939 and a heavy chain variable region comprising SEQ ID NO:940; a light chain variable region comprising SEQ ID NO:943 and a heavy chain variable region comprising SEQ ID NO:944;A light chain variable region comprising SEQ ID NO:947 and a heavy chain variable region comprising SEQ ID NO:948, a light chain variable region comprising SEQ ID NO:951 and a heavy chain variable region comprising SEQ ID NO:952, a light chain variable region comprising SEQ ID NO:955 and a heavy chain variable region comprising SEQ ID NO:956, a light chain variable region comprising SEQ ID NO:959 and a heavy chain variable region comprising SEQ ID NO:960, a light chain variable region comprising SEQ ID NO:1286 and a heavy chain variable region comprising SEQ ID NO:1287, a light chain variable region comprising SEQ ID NO:1296 and a heavy chain variable region comprising SEQ ID NO:1297, a light chain variable region comprising SEQ ID NO:1306 and a heavy chain variable region comprising SEQ ID NO: The combination of light chain variable regions and heavy chain variable regions selected from the group consisting of a heavy chain variable region comprising SEQ ID NO: 1307, a light chain variable region comprising SEQ ID NO: 1316 and a heavy chain variable region comprising SEQ ID NO: 1317, a light chain variable region comprising SEQ ID NO: 1326 and a heavy chain variable region comprising SEQ ID NO: 1327, a light chain variable region comprising SEQ ID NO: 1336 and a heavy chain variable region comprising SEQ ID NO: 1337, a light chain variable region comprising SEQ ID NO: 1346 and a heavy chain variable region comprising SEQ ID NO: 1347, and a light chain variable region comprising SEQ ID NO: 1356 and a heavy chain variable region comprising SEQ ID NO: 1357.

[0307] In one embodiment, the antibody or fragment thereof is selected from the group consisting of SEQ ID NOs: 721, 725, 729, 733, 737, 741, 745, 749, 753, 757, 761, 765, 769, 773, 777, 781, 785, 789, 793, 797, 801, 805, 809, 813, 817, 821, 825, 829, 833, 837, 841, 845, 849, 853, 857, 861, 865, 866, 867, 868, 869, 870, 871, 872, 873, 874, 875, 876, 877, 878, 879, 880, 881, 882, 883, 884, 885, 886, 887, 888, 889, 890, 900, 901, 902, 903, 904, 905, 906, 907, 908, 909, 910, 911, 912, 913, 914, 915, 916, 917, 918, 919, 920, 921, 922, 923, 924, 925, 926, 927, 928, 929, 930, 931, 932, 933, 934, 935 69, 873, 877, 881, 885, 889, 893, 897, 901, 905, 909, 913, 917, 921, 925, 929, 933, 937, 941, 945, 949, 953, 955, 1377, 1379, 1381, 1383, 1385, 1387, 1389, and 1391. In one embodiment, the antibody or fragment thereof is selected from the group consisting of SEQ ID NOs: 722, 726, 730, 734, 738, 742, 746, 750, 754, 758, 762, 766, 770, 774, 778, 782, 786, 790, 794, 798, 802, 806, 810, 814, 818, 822, 826, 830, 834, 838, 842, 846, 850, 854, 858, 862, 866, 870, 874, 878, 882, 886, 890, 894, 898, 902, 906, 910, 914, 918, 922, 926, 930, 934, 938, 942, 946, 950, 954, 958, 1378, 1380, 1382, 1384, 1386, 1388, 1390, and 1392.In one embodiment, the antibody or fragment thereof is selected from the group consisting of SEQ ID NOs: 721, 725, 729, 733, 737, 741, 745, 749, 753, 757, 761, 765, 769, 773, 777, 781, 785, 789, 793, 797, 801, 805, 809, 813, 817, 821, 825, 829, 833, 837, 841, 845, 849, From the group consisting of 853, 857, 861, 865, 869, 873, 877, 881, 885, 889, 893, 897, 901, 905, 909, 913, 917, 921, 925, 929, 933, 937, 941, 945, 949, 953, 955, 1377, 1379, 1381, 1383, 1385, 1387, 1389, and 1391 Light chain variable regions encoded by selected polynucleotide sequences and SEQ ID NOs: 722, 726, 730, 734, 738, 742, 746, 750, 754, 758, 762, 766, 770, 774, 778, 782, 786, 790, 794, 798, 802, 806, 810, 814, 818, 822, 826, 830, 834, 838 , 842, 846, 850, 854, 858, 862, 866, 870, 874, 878, 882, 886, 890, 894, 898, 902, 906, 910, 914, 918, 922, 926, 930, 934, 938, 942, 946, 950, 954, 958, 1378, 1380, 1382, 1384, 1386, 1388, 1390, . and a heavy chain variable region encoded by a polynucleotide sequence selected from the group consisting of SEQ ID NO: 722, SEQ ID NO: 725, and SEQ ID NO: 726, SEQ ID NO: 729, and SEQ ID NO: 730, SEQ ID NO: 733, and SEQ ID NO: 734, SEQ ID NO: 737, and SEQ ID NO: 738, SEQ ID NO: 741, and SEQ ID NO: 742, a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 745 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 746; a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 749 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 750; a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 753 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 754; a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 757 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 758; a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 761 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 762; a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 765 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 766;a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO:769 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO:770; a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO:773 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO:774; a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO:777 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO:778; a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO:781 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO:782; a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO:785 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO:786; a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO:789 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO:790; a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO:793 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO:794, a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO:797 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO:798, a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO:801 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO:802, a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO:805 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO:806, a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO:809 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO:810, a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO:813 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO:814, a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO:817 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO:818,A light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO:821 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO:822; a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO:825 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO:826; a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO:829 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO:830; a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO:833 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO:834; a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO:837 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO:838; a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO:841 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO:842; a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO:845 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO:846 a heavy chain variable region encoded by a nucleotide sequence, a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO:849 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO:850, a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO:853 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO:854, a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO:857 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO:858, a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO:861 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO:862, a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO:865 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO:866, a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO:869 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO:870, a polynucleotide sequence comprising SEQ ID NO:873,a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO:873 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO:874; a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO:877 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO:878; a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO:881 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO:882; a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO:885 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO:886; a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO:889 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO:890; a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO:893 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO:894; a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO:897 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO:898 a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO:901 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO:902; a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO:905 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO:906; a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO:909 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO:910; a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO:913 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO:914; a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO:917 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO:918; a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO:921 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO:922;A light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO:925 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO:926, a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO:929 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO:930, a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO:933 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO:934, a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO:937 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO:938, a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO:941 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO:942, a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO:945 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO:946, a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO:949 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO:949. a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO:950; a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO:953 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO:954; a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO:957 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO:958; a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO:1377 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO:1378; a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO:1379 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO:1380; a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO:1381 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO:1382; a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO:1383 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO:1384;The combination of a light chain variable region and a heavy chain variable region selected from the group consisting of a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1385 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1386, a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1387 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1388, a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1389 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1390, a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1391 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1392.

[0308] Some antigen binding proteins comprise a variable light domain and a variable heavy domain set forth in one of the rows directed to one of the antibodies set forth in Table 3. In some cases, the antigen binding protein comprises two identical variable light domains and two identical variable heavy domains derived from one of the antibodies set forth in Table 3. Some of the antigen binding proteins provided comprise a variable light domain and a variable heavy domain as set forth in one of the rows directed to one of the antibodies set forth in Table 3, with the exception that one or both of the domains differs from the sequence specified in that table by only 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15 amino acid residues, each such sequence difference being independently either a single amino acid deletion, insertion, or substitution, and such deletions, insertions, and / or substitutions result in no more than 1, no more than 2, no more than 3, no more than 4, no more than 5, no more than 6, no more than 7, no more than 8, no more than 9, no more than 10, no more than 11, no more than 12, no more than 13, no more than 14, or no more than 15 amino acids being altered compared to the variable domain sequence specified in Table 3. In one embodiment the antigen binding protein comprises a variable region sequence from Table 3, but lacking the N-terminal methionine. Other antigen binding proteins also comprise a variable light domain and a variable heavy domain as set forth in one of the rows directed to one of the antibodies set forth in Table 3, except that one or both of the domains differ from the sequences specified in that table in that the heavy chain variable domain and / or the light chain variable domain comprises or consists of an amino acid sequence that has at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to the amino acid sequence of the heavy chain variable domain sequence or light chain variable domain sequence identified in Table 3.

[0309] In another embodiment, the antigen binding protein consists solely of a variable light domain or variable heavy domain from an antibody listed in Table 3. In yet another embodiment, the antigen binding protein comprises two or more identical variable heavy domains or two or more identical variable light domains from those listed in Table 3. Such domain antibodies may be fused or linked together via a linker, as described in more detail below. Domain antibodies may also be fused or linked to one or more molecules (e.g., PEG or albumin) to extend their half-life.

[0310] In certain embodiments, the antigen binding protein is desirably an antibody with reduced viscosity, which may be generated by altering the sequences of framework regions and / or Fc domains known to be associated with high viscosity.

[0311] Such low viscosity antigen-binding proteins include antibodies, the VH1|1-18 germline subfamily sequence contains one or more substitutions selected from 82R, 94S, and 95R; the VH3|3-33 germline subfamily sequence contains one or more of the substitutions 1E, 17G, and 85A; the VK3|L16 germline subfamily sequence comprises one or more substitutions selected from 4L, 13L, 76D, 95R, 97E, and 98P; the VK3|L6 germline subfamily sequence comprises one or more substitutions selected from 76D and 95R; the Fc domain sequence comprises one or more substitutions selected from 253A, 440K, and 439E; Antibodies were listed, in which the C-terminus of the Fc domain comprises a sequence selected from KP, KKP, KKKP, and E.

[0312] All of the above preferred viscosity-reducing amino acid substitutions in the variable regions are designated by the Aho numbering system. All viscosity-reducing residues in conserved regions such as Fc are designated by the EU numbering system.

[0313] Other antigen binding proteins provided are variants of antibodies formed by the combination of heavy and light chains shown in Table 3, and include a light chain and / or a heavy chain that each have at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity to the amino acid sequences of such chains. In some cases, such antibodies include at least one heavy chain and one light chain, while in other cases, the variant form includes two identical light chains and two identical heavy chains.

[0314] The various combinations of heavy chain variable regions may be combined with any of the various combinations of light chain variable regions.

[0315] In additional embodiments, the isolated antigen binding protein provided herein is a human antibody comprising a sequence as shown in Table 3, and is of the IgG1, IgG2, IgG3, or IgG4 type.

[0316] The antigen binding proteins disclosed herein are polypeptides into which one or more CDRs have been grafted, inserted, and / or linked. An antigen binding protein may have one, two, three, four, five, or six CDRs. Thus, an antigen binding protein may 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 CDRH3 and CDRL3. Particular light chain and heavy chain CDRs are identified in Table 4A and Table 4B, respectively.

[0317] The complementarity determining regions (CDRs) and framework regions (FRs) of a given antibody may be identified using the system described by Kabat et al. in Sequences of Proteins of Immunological Interest, 5th Ed., US Dept. of Health and Human Services, PHS, NIH, NIH Publication no. 91-3242, 1991. Certain antibodies disclosed herein contain one or more amino acid sequences that are identical to or have substantial sequence identity with one or more of the amino acid sequences of the CDRs shown in Tables 4A and 4B. For such CDRs, the system described by Kabat et al., supra, is used.

[0318] The structure and properties of the CDRs contained in naturally occurring antibodies are described above. Briefly, in traditional antibodies, the CDRs are integrated into frameworks in the heavy and light chain variable regions where they constitute the regions responsible for binding and recognizing the antigen. The variable regions comprise at least three heavy or light chain CDRs (see Kabat et al., 1991, Sequences of Proteins of Immunological Interest, Public Health Service NIH, Bethesda, MD, supra; see also Chothia and Lesk, 1987, J. Mol. Biol. 196:901-917; Chothia et al., 1989, Nature 342:877-883) within framework regions (designated framework regions 1-4 (FR1, FR2, FR3, and FR4) by Kabat et al., 1991, supra; see also Chothia and Lesk, 1987, J. Mol. Biol. 196:901-917; Chothia et al., 1989, Nature 342:877-883). However, the CDRs provided herein may be used to define the antigen-binding domain of a traditional antibody structure, as well as be incorporated into a variety of other polypeptide structures as described herein.

[0319] In one embodiment, the antibody or fragment thereof comprises CDRL1, CDRL2, CDRL3, CDRH1, CDRH2, and CDRH3. In one embodiment, the antibody or fragment thereof comprises SEQ ID NO: 4, 10, 16, 22, 28, 34, 40, 46, 52, 58, 64, 70, 76, 82, 88, 94, 100, 106, 112, 118, 124, 130, 136, 142, 148, 154, 160, 166, 172, 178, 184, 190, 196, 202, 208, 214, 220, 230, 240, 250, 260, 270, 280, 290, 300, 310, 320, 330, 340, 350, 360, 370, 380, 390, 410, 420, 430, 440, 450, 460, 470, 480, 500, 510, 520, 530, 540, 550, 560, 570, 580, 590, 600, 610, 620, 630, 640, 650, 660, 670, 680, 700, 710, 720, 730, 740, 750, 760, 770, 780, 800, 810, 820, 830, 840, 85 , 226, 232, 238, 244, 250, 256, 262, 268, 274, 280, 286, 292, 298, 304, 310, 316, 322, 328, 334, 340, 346, 352, 358, 1290, 1300, 1310, 1320, 1330, 1340, and 1350. In one embodiment, the antibody or fragment thereof is selected from the group consisting of SEQ ID NOs: 5, 11, 17, 23, 29, 35, 41, 47, 53, 59, 65, 71, 77, 83, 89, 95, 101, 107, 113, 119, 125, 131, 137, 143, 149, 155, 161, 167, 173, 179, 185, 191, 197, 203, 209, 215, 221 , 227, 233, 239, 245, 251, 257, 263, 269, 275, 281, 287, 293, 299, 305, 311, 317, 323, 329, 335, 341, 347, 353, 359, 1291, 1301, 1311, 1321, 1331, 1341, and 1351. In one embodiment, the antibody or fragment thereof is selected from the group consisting of SEQ ID NOs: 6, 12, 18, 24, 30, 36, 42, 48, 54, 60, 66, 72, 78, 84, 90, 96, 102, 108, 114, 120, 126, 132, 138, 144, 150, 156, 162, 168, 174, 180, 186, 192, 198, 204, 210, 216, 222 , 228, 234, 240, 246, 252, 258, 264, 270, 276, 282, 288, 294, 300, 306, 312, 318, 324, 330, 336, 342, 348, 354, 360, 1292, 1302, 1312, 1322, 1332, 1342, and 1352.The antibodies or fragments thereof may be selected from the group consisting of SEQ ID NOs: 364, 370, 376, 382, ​​388, 394, 400, 406, 412, 418, 424, 430, 436, 442, 448, 454, 460, 466, 472, 478, 484, 490, 496, 502, 508, 514, 520, 526, 532, 538, 544, 550, 556, 562, 568, 574, and a CDRH1 comprising a sequence selected from the group consisting of 580, 586, 592, 598, 604, 610, 616, 622, 628, 634, 640, 646, 652, 658, 664, 670, 676, 682, 688, 694, 700, 706, 712, 718, 1293, 1303, 1313, 1323, 1333, 1343, and 1353. In one embodiment, the antibody or fragment thereof is selected from the group consisting of SEQ ID NOs: 365, 371, 377, 383, 389, 395, 401, 407, 413, 419, 425, 431, 437, 443, 449, 455, 461, 467, 473, 479, 485, 491, 497, 503, 509, 515, 521, 527, 533, 539, 545, 551, 557, 563, 569 , 575, 581, 587, 593, 599, 605, 611, 617, 623, 629, 635, 641, 647, 653, 659, 665, 671, 677, 683, 689, 695, 701, 707, 713, 719, 1294, 1304, 1314, 1324, 1334, 1344, and 1354. In one embodiment, the antibody or fragment thereof is selected from the group consisting of SEQ ID NOs: 366, 372, 378, 384, 390, 396, 402, 408, 414, 420, 426, 432, 438, 444, 450, 456, 462, 468, 474, 480, 486, 492, 498, 504, 510, 516, 522, 528, 534, 540, 546, 552, 558, 564, 570, 572, 576, 578, 580, 582, 584, 586, 590, 592, 596, 600, 602, 604, 606, 608, 610, 614, 616, 620, 626, 626, 632, 638, 644, 650, 656, 650, 662, 668, 674, 674, 680, 686, 692, 698, 704, 710, 722, 730, 740, 752, 760, 770, 780, 790, 802, 810, 820, 830, 840, 852, 860, 870, 880, 890, 902, 910, 920, 930, 940, 952, 960, 970 , 576, 582, 588, 594, 600, 606, 612, 618, 624, 630, 636, 642, 648, 654, 660, 666, 672, 678, 684, 690, 696, 702, 708, 714, 720, 1295, 1305, 1315, 1325, 1335, 1345, and 1355. In one embodiment, the antibody or fragment thereof comprises a CDRH3 comprising a sequence selected from the group consisting of CDRL1, CDRL2,and CDRH3, wherein each of CDRL1, CDRL2, CDRL3, CDRH1, CDRH2, and CDRH3 is set forth in SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:364, SEQ ID NO:365, and SEQ ID NO:366; SEQ ID NO:10, SEQ ID NO:11, SEQ ID NO:12, SEQ ID NO:370, SEQ ID NO:371, and SEQ ID NO:372; SEQ ID NO:16, SEQ ID NO:17, SEQ ID NO:18, SEQ ID NO:376, SEQ ID NO:377, and SEQ ID NO:378; SEQ ID NO:22, SEQ ID NO:23, SEQ ID NO:24, SEQ ID NO:382, SEQ ID NO:383, and and SEQ ID NO:384; SEQ ID NO:28, SEQ ID NO:29, SEQ ID NO:30, SEQ ID NO:388, SEQ ID NO:389, and SEQ ID NO:390; SEQ ID NO:34, SEQ ID NO:35, SEQ ID NO:36, SEQ ID NO:394, SEQ ID NO:395, and SEQ ID NO:396; SEQ ID NO:40, SEQ ID NO:41, SEQ ID NO:42, SEQ ID NO:400, SEQ ID NO:401, and SEQ ID NO:402; SEQ ID NO:46, SEQ ID NO:47, SEQ ID NO:48, SEQ ID NO:406, SEQ ID NO:407, and SEQ ID NO:408; SEQ ID NO:52, SEQ ID NO:53, SEQ ID NO:54, SEQ ID NO:412, SEQ ID NO:413, and SEQ ID NO:414; SEQ ID NO:5 8, SEQ ID NO:59, SEQ ID NO:60, SEQ ID NO:418, SEQ ID NO:419, and SEQ ID NO:420; SEQ ID NO:64, SEQ ID NO:65, SEQ ID NO:66, SEQ ID NO:424, SEQ ID NO:425, and SEQ ID NO:426; SEQ ID NO:70, SEQ ID NO:71, SEQ ID NO:72, SEQ ID NO:430, SEQ ID NO:431, and SEQ ID NO:432; SEQ ID NO:76, SEQ ID NO:77, SEQ ID NO:78, SEQ ID NO:436, SEQ ID NO:437, and SEQ ID NO:438; SEQ ID NO:82, SEQ ID NO:83, SEQ ID NO:84, SEQ ID NO:442, SEQ ID NO:443, and SEQ ID NO:444; SEQ ID NO:88, SEQ ID NO:89, SEQ ID NO:9 0, SEQ ID NO:448, SEQ ID NO:449, and SEQ ID NO:450; SEQ ID NO:94, SEQ ID NO:95, SEQ ID NO:96, SEQ ID NO:454, SEQ ID NO:455, and SEQ ID NO:456; SEQ ID NO:100, SEQ ID NO:101, SEQ ID NO:102, SEQ ID NO:460, SEQ ID NO:461, and SEQ ID NO:462; SEQ ID NO:106, SEQ ID NO:107, SEQ ID NO:108, SEQ ID NO:466, SEQ ID NO:467, and SEQ ID NO:468; SEQ ID NO:112, SEQ ID NO:113, SEQ ID NO:114, SEQ ID NO:472, SEQ ID NO:473, and SEQ ID NO:474; SEQ ID NO:118, SEQ ID NO:119, SEQ ID NO:120,SEQ ID NO:478, SEQ ID NO:479, and SEQ ID NO:480; SEQ ID NO:124, SEQ ID NO:125, SEQ ID NO:126, SEQ ID NO:484, SEQ ID NO:485, and SEQ ID NO:486; SEQ ID NO:130, SEQ ID NO:131, SEQ ID NO:132, SEQ ID NO:490, SEQ ID NO:491, and SEQ ID NO:492; SEQ ID NO:136, SEQ ID NO:137, SEQ ID NO:138, SEQ ID NO:496, SEQ ID NO:497, and SEQ ID NO:498; SEQ ID NO:142, SEQ ID NO:143, SEQ ID NO:144, SEQ ID NO:502, SEQ ID NO:503, and SEQ ID NO:504; SEQ ID NO:148, SEQ ID NO:149, SEQ ID NO:150, SEQ ID NO:508, SEQ ID NO:509, and SEQ ID NO:510; SEQ ID NO:154, SEQ ID NO:155, SEQ ID NO:156, SEQ ID NO:514, SEQ ID NO:515, and SEQ ID NO:516; SEQ ID NO:160, SEQ ID NO:161, SEQ ID NO:162, SEQ ID NO:520, SEQ ID NO:521, and SEQ ID NO:522; SEQ ID NO:166, SEQ ID NO:167, SEQ ID NO:168, SEQ ID NO:526, SEQ ID NO:527, and SEQ ID NO:528; SEQ ID NO:172, SEQ ID NO:173, SEQ ID NO:174, SEQ ID NO:532, SEQ ID NO:533, and SEQ ID NO:534; SEQ ID NO:178, SEQ ID NO:179, SEQ ID NO:180, SEQ ID NO:538, SEQ ID NO:539, and SEQ ID NO:540; SEQ ID NO:184, SEQ ID NO:185, SEQ ID NO:186, SEQ ID NO:544, SEQ ID NO:545, and SEQ ID NO:546; SEQ ID NO:190, SEQ ID NO:191, SEQ ID NO:192, SEQ ID NO:550, SEQ ID NO:551, and SEQ ID NO:552; SEQ ID NO:196, SEQ ID NO:197, SEQ ID NO:198, SEQ ID NO:556, SEQ ID NO:557, and SEQ ID NO:558; SEQ ID NO:202, SEQ ID NO:203, SEQ ID NO:204, SEQ ID NO:562, SEQ ID NO:563, and SEQ ID NO:564; SEQ ID NO:208, SEQ ID NO:209, SEQ ID NO:210, SEQ ID NO:568, SEQ ID NO:569, and SEQ ID NO:570; SEQ ID NO:214, SEQ ID NO:215, SEQ ID NO:216, SEQ ID NO:574, SEQ ID NO:575, and SEQ ID NO:576; SEQ ID NO:220, SEQ ID NO:221, SEQ ID NO:222, SEQ ID NO:580, SEQ ID NO:581, and SEQ ID NO:582; SEQ ID NO:226, SEQ ID NO:227, SEQ ID NO:228, SEQ ID NO:586, SEQ ID NO:587, and SEQ ID NO:588; SEQ ID NO:232, SEQ ID NO:233, SEQ ID NO:234, SEQ ID NO:592, SEQ ID NO:593, and SEQ ID NO:594; SEQ ID NO:238, SEQ ID NO:239, SEQ ID NO:240,SEQ ID NO:598, SEQ ID NO:599, and SEQ ID NO:600; SEQ ID NO:244, SEQ ID NO:245, SEQ ID NO:246, SEQ ID NO:604, SEQ ID NO:605, and SEQ ID NO:606; SEQ ID NO:250, SEQ ID NO:251, SEQ ID NO:252, SEQ ID NO:610, SEQ ID NO:611, and SEQ ID NO:612; SEQ ID NO:256, SEQ ID NO:257, SEQ ID NO:258, SEQ ID NO:616, SEQ ID NO:617, and SEQ ID NO:618; SEQ ID NO:262, SEQ ID NO:263, SEQ ID NO:264, SEQ ID NO:622, SEQ ID NO:623, and SEQ ID NO: No. 624; SEQ ID NO: 268, SEQ ID NO: 269, SEQ ID NO: 270, SEQ ID NO: 628, SEQ ID NO: 629, and SEQ ID NO: 630; SEQ ID NO: 274, SEQ ID NO: 275, SEQ ID NO: 276, SEQ ID NO: 634, SEQ ID NO: 635, and SEQ ID NO: 636; SEQ ID NO: 280, SEQ ID NO: 281, SEQ ID NO: 282, SEQ ID NO: 640, SEQ ID NO: 641, and SEQ ID NO: 642; SEQ ID NO: 286, SEQ ID NO: 287, SEQ ID NO: 288, SEQ ID NO: 646, SEQ ID NO: 647, and SEQ ID NO: 648; SEQ ID NO: 292, SEQ ID NO: 293, SEQ ID NO:294, SEQ ID NO:652, SEQ ID NO:653, and SEQ ID NO:654; SEQ ID NO:298, SEQ ID NO:299, SEQ ID NO:300, SEQ ID NO:658, SEQ ID NO:659, and SEQ ID NO:660; SEQ ID NO:304, SEQ ID NO:305, SEQ ID NO:306, SEQ ID NO:664, SEQ ID NO:665, and SEQ ID NO:666; SEQ ID NO:310, SEQ ID NO:311, SEQ ID NO:312, SEQ ID NO:670, SEQ ID NO:671, and SEQ ID NO:672; SEQ ID NO:316, SEQ ID NO:317, SEQ ID NO:318, SEQ ID NO:676, SEQ ID NO:6 77, and SEQ ID NO:678; SEQ ID NO:322, SEQ ID NO:323, SEQ ID NO:324, SEQ ID NO:682, SEQ ID NO:683, and SEQ ID NO:684; SEQ ID NO:328, SEQ ID NO:329, SEQ ID NO:330, SEQ ID NO:688, SEQ ID NO:689, and SEQ ID NO:690; SEQ ID NO:334, SEQ ID NO:335, SEQ ID NO:336, SEQ ID NO:694, SEQ ID NO:695, and SEQ ID NO:696; SEQ ID NO:340, SEQ ID NO:341, SEQ ID NO:342, SEQ ID NO:700, SEQ ID NO:701, and SEQ ID NO:702; SEQ ID NO:346, SEQ ID NO:347, SEQ ID NO:348, SEQ ID NO:706, SEQ ID NO:707, and SEQ ID NO:708; SEQ ID NO:352, SEQ ID NO:353, SEQ ID NO:354, SEQ ID NO:712, SEQ ID NO:713, and SEQ ID NO:714; SEQ ID NO:358, SEQ ID NO:359, SEQ ID NO:360, SEQ ID NO:718, SEQ ID NO:719, and SEQ ID NO:720; SEQ ID NO:1290, SEQ ID NO:1291, SEQ ID NO:1292, SEQ ID NO:1293, SEQ ID NO:1294, and SEQ ID NO:1295; SEQ ID NO:1300, SEQ ID NO:1301, SEQ ID NO:1302, SEQ ID NO:1303, SEQ ID NO:1304, and SEQ ID NO:1305; SEQ ID NO:1310, SEQ ID NO:1311, SEQ ID NO:1312, SEQ ID NO:1313, SEQ ID NO:1314, and SEQ ID NO: SEQ ID NO:1315; SEQ ID NO:1320, SEQ ID NO:1321, SEQ ID NO:1322, SEQ ID NO:1323, SEQ ID NO:1324, and SEQ ID NO:1325; SEQ ID NO:1330, SEQ ID NO:1331, SEQ ID NO:1332, SEQ ID NO:1333, SEQ ID NO:1334, and SEQ ID NO:1335; SEQ ID NO:1340, SEQ ID NO:1341, SEQ ID NO:1342, SEQ ID NO:1343, SEQ ID NO:1344, and SEQ ID NO:1345; SEQ ID NO:1350, SEQ ID NO:1351, SEQ ID NO:1352, SEQ ID NO:1353, SEQ ID NO:1354, and SEQ ID NO:1355; and SEQ ID NO:1360, SEQ ID NO:1361, SEQ ID NO:1362, SEQ ID NO:1363, SEQ ID NO:1364, and SEQ ID NO:1365.

[0320] In another embodiment, the antigen binding protein comprises one, two, three, four, five, or six variants of the CDRs set out in Tables 4A and 4B that have at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to the CDR sequences set out in Tables 4A and 4B, respectively. Some antigen binding proteins comprise one, two, three, four, five, or six CDRs set out in Tables 4A and 4B that, individually or collectively, differ from the CDRs set out in the tables by no more than one, no more than two, no more than three, no more than four, or no more than five amino acids.

[0321] In various other embodiments, the antigen binding protein is derived from such an antibody. For example, in one aspect, the antigen binding protein comprises one, two, three, four, five, or all six of the CDRs set forth in one of the rows directed to any of the specific antibodies set forth in Tables 4A and 4B. In another aspect, the antigen binding protein comprises one, two, three, four, five, or six mutated forms of the CDRs set forth in one of the rows directed to the antibodies of Tables 4A and 4B, wherein the CDRs have at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to the CDR sequences set forth in Tables 4A and 4B, respectively. Some antigen binding proteins comprise one, two, three, four, five or six CDRs set forth in one of the rows of Table 4A and Table 4B, which differ from the CDRs set forth in these tables by no more than one, no more than two, no more than three, no more than four or no more than five amino acids, respectively. In another embodiment, an antigen binding protein comprises all six CDRs set forth in the rows of Table 4A and Table 4B, where the total number of amino acid changes to the CDRs collectively is no more than one, no more than two, no more than three, no more than four or no more than five amino acids.

[0322] In one embodiment, the antibody or fragment thereof is selected from the group consisting of SEQ ID NOs: 963, 967, 971, 975, 979, 983, 987, 991, 995, 999, 1003, 1007, 1011, 1015, 1019, 1023, 1027, 1031, 1035, 1039, 1043, 1047, 1051, 1055, 1059, 1063, 1067, 1071, 1075, 1079, 1083, 1087, 1091, 1095, 1099, and a light chain comprising a sequence selected from the group consisting of: 1103, 1107, 1111, 1115, 1119, 1123, 1127, 1131, 1135, 1139, 1143, 1147, 1151, 1155, 1159, 1163, 1167, 1171, 1175, 1179, 1183, 1187, 1191, 1195, 1199, 1288, 1298, 1308, 1318, 1328, 1338, 1348, and 1358. In one embodiment, the antibody or fragment thereof is selected from the group consisting of SEQ ID NOs: 964, 968, 972, 976, 980, 984, 988, 992, 996, 1000, 1004, 1008, 1012, 1016, 1020, 1024, 1028, 1032, 1036, 1040, 1044, 1048, 1052, 1056, 1060, 1064, 1068, 1072, 1076, 1080, 1084, 1088, 1092, 1096, 1100 , 1104, 1108, 1112, 1116, 1120, 1124, 1128, 1132, 1136, 1140, 1144, 1148, 1152, 1156, 1160, 1164, 1168, 1172, 1176, 1180, 1184, 1188, 1192, 1196, 1200, 1289, 1299, 1309, 1319, 1329, 1339, 1349, and 1359. In one embodiment, the antibody or fragment thereof is selected from the group consisting of SEQ ID NOs: 963, 967, 971, 975, 979, 983, 987, 991, 995, 999, 1003, 1007, 1011, 1015, 1019, 1023, 1027, 1031, 1035, 1039, 1043, 1047, 1051, 1055, 1059, 1063, 1067, 1071, 1075, 1079, 1083, 1087, 1091, 1095, 1099, 1103, 1107, 1111, 1115, 1119, 1123, 1127, 1131, 1135, 1139, 1143,1147, 1151, 1155, 1159, 1163, 1167, 1171, 1175, 1179, 1183, 1187, 1191, 1195, 1199, 1288, 1298, 1308, 1318, 1328, 1338, 1348, and 1358; and a light chain comprising a sequence selected from the group consisting of SEQ ID NOs: 964, 968, 972, 976, 980, 984, 988, 992, 996, 1000, 1004, 1008, 1012, 1016, 1020, 1024, 1028, 1032, 1036, 1040, 1044, 1048, 1052, 1056. , 1060, 1064, 1068, 1072, 1076, 1080, 1084, 1088, 1092, 1096, 1100, 1104, 1108, 1112, 1116, 1120, 1124, 1128, 1132, 1136, 1140, 1144, 1148, 1152, 1156, 1160, 1164, 1168, 1172, 1176, 1180, 1184, 1188, 1192, 1196, 1200, 1289, 1299, 1309, 1319, 1329, 1339, 1349, and 1359. In one embodiment, the antibody or fragment thereof comprises a light chain comprising SEQ ID NO:963 and a heavy chain comprising SEQ ID NO:964, a light chain comprising SEQ ID NO:967 and a heavy chain comprising SEQ ID NO:968, a light chain comprising SEQ ID NO:971 and a heavy chain comprising SEQ ID NO:972, a light chain comprising SEQ ID NO:975 and a heavy chain comprising SEQ ID NO:976, a light chain comprising SEQ ID NO:979 and a heavy chain comprising SEQ ID NO:980, a light chain comprising SEQ ID NO:983 and a heavy chain comprising SEQ ID NO:984, a light chain comprising SEQ ID NO:987 and a heavy chain comprising SEQ ID NO:988, a light chain comprising SEQ ID NO:991 and a heavy chain comprising SEQ ID NO:992, a light chain comprising SEQ ID NO:995 and a heavy chain comprising SEQ ID NO:996, a light chain comprising SEQ ID NO:997 and a heavy chain comprising SEQ ID NO:998, a light chain comprising SEQ ID NO:999 and a heavy chain comprising SEQ ID NO:999, a light chain comprising SEQ ID NO:998 and a heavy chain comprising SEQ ID NO:999, a light chain comprising SEQ ID NO:999 and a heavy chain comprising SEQ ID NO:990, a light chain comprising SEQ ID NO:999 and a heavy chain comprising SEQ ID NO:991, a light chain comprising SEQ ID NO:999 and a heavy chain comprising SEQ ID NO:991, a light chain comprising SEQ ID NO:999 and a heavy chain comprising SEQ ID NO:992, a light chain comprising SEQ ID NO:999 and a heavy chain comprising SEQ ID NO:993, a light chain comprising SEQ ID NO:999 and a heavy chain comprising SEQ ID NO:994, a light chain comprising SEQ ID NO:999 and a heavy chain comprising SEQ ID NO:995, a light chain comprising SEQ ID a light chain comprising SEQ ID NO: 999 and a heavy chain comprising SEQ ID NO: 1000; a light chain comprising SEQ ID NO: 1003 and a heavy chain comprising SEQ ID NO: 1004; a light chain comprising SEQ ID NO: 1007 and a heavy chain comprising SEQ ID NO: 1008; a light chain comprising SEQ ID NO: 1011 and a heavy chain comprising SEQ ID NO: 1012; a light chain comprising SEQ ID NO: 1015 and a heavy chain comprising SEQ ID NO: 1016; a light chain comprising SEQ ID NO: 1019 and a heavy chain comprising SEQ ID NO: 1020; a light chain comprising SEQ ID NO: 1023 and a heavy chain comprising SEQ ID NO: 1024; a light chain comprising SEQ ID NO: 1027 and a heavy chain comprising SEQ ID NO: 1028;A light chain comprising SEQ ID NO:1031 and a heavy chain comprising SEQ ID NO:1032, a light chain comprising SEQ ID NO:1035 and a heavy chain comprising SEQ ID NO:1036, a light chain comprising SEQ ID NO:1039 and a heavy chain comprising SEQ ID NO:1040, a light chain comprising SEQ ID NO:1043 and a heavy chain comprising SEQ ID NO:1044, a light chain comprising SEQ ID NO:1047 and a heavy chain comprising SEQ ID NO:1048, a light chain comprising SEQ ID NO:1051 and a heavy chain comprising SEQ ID NO:1052, a light chain comprising SEQ ID NO:1055 and a heavy chain comprising SEQ ID NO:1056, a light chain comprising SEQ ID NO:1059 and a heavy chain comprising SEQ ID NO:1060, a light chain comprising SEQ ID NO:1063 and and a heavy chain comprising SEQ ID NO:1064, a light chain comprising SEQ ID NO:1067 and a heavy chain comprising SEQ ID NO:1068, a light chain comprising SEQ ID NO:1071 and a heavy chain comprising SEQ ID NO:1072, a light chain comprising SEQ ID NO:1075 and a heavy chain comprising SEQ ID NO:1076, a light chain comprising SEQ ID NO:1079 and a heavy chain comprising SEQ ID NO:1080, a light chain comprising SEQ ID NO:1083 and a heavy chain comprising SEQ ID NO:1084, a light chain comprising SEQ ID NO:1087 and a heavy chain comprising SEQ ID NO:1088, a light chain comprising SEQ ID NO:1091 and a heavy chain comprising SEQ ID NO:1092, a light chain comprising SEQ ID NO:1095 and a heavy chain comprising SEQ ID NO:1096, a light chain comprising SEQ ID NO:1099 and a heavy chain comprising SEQ ID NO:1100, a light chain comprising SEQ ID NO:1103 and a heavy chain comprising SEQ ID NO:1104, a light chain comprising SEQ ID NO:1107 and a heavy chain comprising SEQ ID NO:1108, a light chain comprising SEQ ID NO:1111 and a heavy chain comprising SEQ ID NO:1112, a light chain comprising SEQ ID NO:1115 and a heavy chain comprising SEQ ID NO:1116, a light chain comprising SEQ ID NO:1119 and a heavy chain comprising SEQ ID NO:1120, a light chain comprising SEQ ID NO:1123 and a heavy chain comprising SEQ ID NO:1124, a light chain comprising SEQ ID NO:1127 and a heavy chain comprising SEQ ID NO:1128, a light chain comprising SEQ ID NO:1131, and a heavy chain comprising SEQ ID NO:1132, a light chain comprising SEQ ID NO:1135 and a heavy chain comprising SEQ ID NO:1136, a light chain comprising SEQ ID NO:1139 and a heavy chain comprising SEQ ID NO:1140, a light chain comprising SEQ ID NO:1143 and a heavy chain comprising SEQ ID NO:1144, a light chain comprising SEQ ID NO:1147 and a heavy chain comprising SEQ ID NO:1148, a light chain comprising SEQ ID NO:1151 and a heavy chain comprising SEQ ID NO:1152, a light chain comprising SEQ ID NO:1155 and a heavy chain comprising SEQ ID NO:1156, a light chain comprising SEQ ID NO:1159 and a heavy chain comprising SEQ ID NO:1160, a light chain comprising SEQ ID NO:1163 and a heavy chain comprising SEQ ID NO:1164,a light chain comprising SEQ ID NO:1167 and a heavy chain comprising SEQ ID NO:1168; a light chain comprising SEQ ID NO:1171 and a heavy chain comprising SEQ ID NO:1172; a light chain comprising SEQ ID NO:1175 and a heavy chain comprising SEQ ID NO:1176; a light chain comprising SEQ ID NO:1179 and a heavy chain comprising SEQ ID NO:1180; a light chain comprising SEQ ID NO:1183 and a heavy chain comprising SEQ ID NO:1184; a light chain comprising SEQ ID NO:1187 and a heavy chain comprising SEQ ID NO:1188; a light chain comprising SEQ ID NO:1191 and a heavy chain comprising SEQ ID NO:1192; a light chain comprising SEQ ID NO:1195 and a heavy chain comprising SEQ ID NO:1196; a light chain comprising SEQ ID NO:1199 and a heavy chain comprising SEQ ID NO:1200; The combination of light and heavy chains is selected from the group consisting of a light chain comprising SEQ ID NO: 1288 and a heavy chain comprising SEQ ID NO: 1289, a light chain comprising SEQ ID NO: 1298 and a heavy chain comprising SEQ ID NO: 1299, a light chain comprising SEQ ID NO: 1308 and a heavy chain comprising SEQ ID NO: 1309, a light chain comprising SEQ ID NO: 1318 and a heavy chain comprising SEQ ID NO: 1319, a light chain comprising SEQ ID NO: 1328 and a heavy chain comprising SEQ ID NO: 1329, a light chain comprising SEQ ID NO: 1338 and a heavy chain comprising SEQ ID NO: 1339, a light chain comprising SEQ ID NO: 1348 and a heavy chain comprising SEQ ID NO: 1349, and a light chain comprising SEQ ID NO: 1358 and a heavy chain comprising SEQ ID NO: 1359.

[0323] In one embodiment, the antibody or fragment thereof is selected from the group consisting of SEQ ID NOs: 961, 965, 969, 973, 977, 981, 985, 989, 993, 997, 1001, 1005, 1009, 1013, 1017, 1021, 1025, 1029, 1033, 1037, 1041, 1045, 1049, 1053, 1057, 1061, 1065, 1069, 1073, 1077, 1081, 1085, 1089, 1093, 1097, 1101, 1102, 1104, 1106, 1108, 1109, 1110, 1111, 1112, 1113, 1114, 1115, 1116, 1117, 1118, 1119, 1120, 1121, 1122, 1123, 1124, 1125, 1126, 1127, 1128, 1130, 1131, 1132, 1133, 1134, 1135, 1136, 1137, 1138, 1139, 1140, 1141, 1142, 1143, 1144, 1145, 1146, 1147, 1148, 1150, 1151, 1152, 1153, 05, 1109, 1113, 1117, 1121, 1125, 1129, 1133, 1137, 1141, 1145, 1149, 1153, 1157, 1161, 1165, 1169, 1173, 1177, 1181, 1185, 1189, 1193, 1197, 1361, 1363, 1365, 1367, 1369, 1371, 1373, and 1375. In one embodiment, the antibody or fragment thereof is selected from the group consisting of SEQ ID NOs: 962, 966, 970, 974, 978, 982, 986, 990, 994, 998, 1002, 1006, 1010, 1014, 1018, 1022, 1026, 1030, 1034, 1038, 1042, 1046, 1050, 1054, 1058, 1062, 1066, 1070, 1074, 1078, 1082, 1086, 1090, 1094, 1098, 1102, 1104, 1106, 1108, 1106, 1108, 1108, 1104, 1106, 1108 ... 06, 1110, 1114, 1118, 1122, 1126, 1130, 1134, 1138, 1142, 1146, 1150, 1154, 1158, 1162, 1166, 1170, 1174, 1178, 1182, 1186, 1190, 1194, 1198, 1362, 1364, 1366, 1368, 1370, 1372, 1374, and 1376. In one embodiment, the antibody or fragment thereof is selected from the group consisting of SEQ ID NOs: 961, 965, 969, 973, 977, 981, 985, 989, 993, 997, 1001, 1005, 1009, 1013, 1017, 1021, 1025, 1029, 1033, 1037, 1041, 1045, 1049, 1053, 1057, 1061, 1065, 1069, 1073, 1077, 1081, 1085, 1089, 1093, 1097, 1101, 1105, 1109, 1113, 1117,1121, 1125, 1129, 1133, 1137, 1141, 1145, 1149, 1153, 1157, 1161, 1165, 1169, 1173, 1177, 1181, 1185, 1189, 1193, 1197, 1361, 1363, 1365, 1367, 1369, 1371, 1373, and 1375; and a light chain encoded by a polynucleotide sequence selected from the group consisting of SEQ ID NOs: 962, 966, 970, 974, 978, 982, 986, 990, 994, 998, 1002, 1006, 1010, 1014, 1018, 1022, 1026, 1030, 1034, 1036, 1038, 1040, 1042, 1044, 1046, 1048, 1049, 1050, 1051, 1052, 1053, 1054, 1055, 1056, 1057, 1060, 1061, 1062, 1063, 1064, 1065, 1066, 1067, 1068, 1070, 1072, 1074, 1076, 1078, 1080, 1082, 1084, 1086, 1088, 1089, 38, 1042, 1046, 1050, 1054, 1058, 1062, 1066, 1070, 1074, 1078, 1082, 1086, 1090, 1094, 1098, 1102, 1106, 1110, 1114, 1118, 1122, 1126, 1130, 1134, 1138, 1142, 1 and a heavy chain comprising a sequence selected from the group consisting of: 146, 1150, 1154, 1158, 1162, 1166, 1170, 1174, 1178, 1182, 1186, 1190, 1194, 1198, 1362, 1364, 1366, 1368, 1370, 1372, 1374, and 1376. In one embodiment, the antibody or fragment thereof comprises a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:961 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:962; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:965 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:966; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:969 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:970; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:973 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:974; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:977 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:978; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:981 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:982;a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:985 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:986; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:989 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:990; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:993 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:994; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:997 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:998; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:1001 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:1002; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:1005 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:1006; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:1009 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:1010; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:1013 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:1014; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:1017 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:1018; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:1021 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:1022; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:1025 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:1026; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:1029 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:1030; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:1033 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:1034; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:1037 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:1038;a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:1041 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:1042; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:1045 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:1046; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:1049 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:1050; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:1053 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:1054; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:1057 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:1058; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:1061 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:1062; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:1065 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:1066; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:1069 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:1070; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:1073 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:1074; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:1077 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:1078; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:1081 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:1082; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:1085 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:1086; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:1089 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:1090; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:1093 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:1094;a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:1097 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:1098; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:1101 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:1102; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:1105 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:1106; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:1109 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:1110; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:1113 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:1114; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:1117 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:1118; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:1121 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:1122; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:1125 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:1126; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:1129 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:1130; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:1133 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:1134; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:1137 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:1138; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:1141 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:1142; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:1145 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:1146; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:1149 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:1150;A light chain encoded by a polynucleotide sequence comprising SEQ ID NO:1153 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:1154; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:1157 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:1158; SEQ ID NO:1161; and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:1162; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:1165 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:1166; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:1169 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:1170; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:1173 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:1174; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:1177 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:1178; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:1181 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:1182; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:1185 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:1186; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:1189 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:1190; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:1193 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:1194; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:1197 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:1198; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:1361 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:1362; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:1363 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:1364; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:1365 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:1366; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:1367 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:1368;The combination of light and heavy chain variable regions is selected from the group consisting of a light chain encoded by a polynucleotide sequence comprising SEQ ID NO: 1369 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO: 1370, a light chain encoded by a polynucleotide sequence comprising SEQ ID NO: 1371 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO: 1372, a light chain encoded by a polynucleotide sequence comprising SEQ ID NO: 1373 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO: 1374, and a light chain encoded by a polynucleotide sequence comprising SEQ ID NO: 1375 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO: 1376.

[0324] In some embodiments, the invention includes antibodies that bind to GIPR, where the antibodies bind to GIPR and reduce the likelihood that GIPR will bind to GIP.

[0325] In another aspect, the antigen binding protein comprises a full length light chain and a full length heavy chain as set forth in one of the rows directed to one of the antibodies set forth in Table 5. Some of the antigen binding proteins provided comprise a full length light chain and a full length heavy chain as set forth in one of the rows directed to one of the antibodies set forth in Table 5, with the exception that one or both of the chains differs from the sequence specified in that table by only 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15 amino acid residues, each such sequence difference being independently either a single amino acid deletion, insertion, or substitution, and such deletions, insertions, and / or substitutions result in no more than 1, no more than 2, no more than 3, no more than 4, no more than 5, no more than 6, no more than 7, no more than 8, no more than 9, no more than 10, no more than 11, no more than 12, no more than 13, no more than 14, or no more than 15 amino acids being altered compared to the full length sequence specified in Table 5. In one embodiment, the antigen binding protein comprises a full length light chain and / or a full length heavy chain from Table 5, but lacking the N-terminal methionine. In one embodiment, the antigen binding protein comprises a full length light chain and / or a full length heavy chain from Table 5, but lacking the C-terminal lysine. Other antigen binding proteins also comprise a full length light chain and a full length heavy chain as set forth in one of the rows directed to one of the antibodies set forth in Table 5, except that one or both of the chains differ from the sequences identified in that table in that the light and / or heavy chain comprises or consists of an amino acid sequence that has at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to the amino acid sequence of the light or heavy chain sequence identified in Table 5.

[0326] In another embodiment, the antigen binding protein consists solely of a light chain polypeptide or a heavy chain polypeptide as shown in Table 5.

[0327] In yet another aspect, the antigen binding protein comprising the CDRs, variable domains, and / or full length sequences set forth in Table 3, Table 4A, Table 4B, and Table 5 is a monoclonal antibody, a chimeric antibody, a humanized antibody, a human antibody, a multispecific antibody, or an antibody fragment thereof. In another embodiment, the antibody fragment of the isolated antigen binding protein provided herein is a Fab fragment, a Fab' fragment, a F(ab')2 fragment, an Fv fragment, a diabody, or an scFv based on an antibody having a sequence set forth in Table 5.

[0328] In yet another embodiment, an isolated antigen binding protein provided in Table 5 can be coupled to a labeling group and allowed to compete for binding to GIPR with an antigen binding protein that is one of the isolated antigen binding proteins provided herein.

[0329] In another embodiment, antigen binding proteins are provided that compete with one of the exemplary antibodies or functional fragments above for specific binding to human GIPR (e.g., SEQ ID NO: 1201). Such antigen binding proteins may bind to the same epitope as one of the antigen binding proteins described herein, or may bind overlapping epitopes. Antigen binding proteins and fragments that compete with the exemplary antigen binding proteins are expected to exhibit similar functional properties. Exemplary antigen binding proteins and fragments include those described above, including those having heavy and light chains, variable region domains, and CDRs contained in Table 3, Table 4A, Table 4B, and Table 5. Thus, as a specific example, the antigen binding proteins provided include: An antibody that competes with an antibody having all six CDRs described in Table 4A and Table 4B against any of the antibodies described in Table 4A and Table 4B; competes with an antibody having the VH and VL as set forth in Table 3; or Competing with an antibody having two light chains and two heavy chains specified against any of the antibodies in Table 5; Includes:

[0330] The antigen binding protein provided includes a monoclonal antibody that binds to GIPR. The monoclonal antibody may be produced using any technique known in the art, for example, by immortalizing the spleen cells collected from the transgenic animal after the completion of the immunization schedule. The spleen cells may be immortalized using any technique known in the art, for example, by fusing the spleen cells with myeloma cells to produce hybridomas. The myeloma cells used in the fusion procedure to produce hybridomas are preferably non-antibody producing, have high fusion efficiency, and have enzyme deficiencies that make them unable to grow in certain selective media that support the growth of only the desired fused cells (hybridomas). Examples of cell lines suitable for use in mouse fusions include Sp-20, P3-X63 / Ag8, P3-X63-Ag8.653, NS1 / 1.Ag41, Sp210-Ag14, FO, NSO / U, MPC-11, MPC11-X45-GTG1.7, and S194 / 5XXO Bul, and examples of cell lines used in rat fusions include R210.RCY3, Y3-Ag1.2.3, IR983F, and 4B210. Other cell lines useful for cell fusions are U-266, GM1500-GRG2, LICR-LON-HMy2, and UC729-6.

[0331] In some cases, a hybridoma cell line is generated by immunizing an animal (e.g., a transgenic animal with human immunoglobulin sequences) with a GIPR immunogen, collecting spleen cells from the immunized animal, generating hybridoma cells by fusing the collected spleen cells to a myeloma cell line, establishing a hybridoma cell line from the hybridoma cells, and identifying a hybridoma cell line that produces an antibody that binds to a GIPR polypeptide. Such hybridoma cell lines, and the anti-GIPR monoclonal antibodies they produce, are aspects of the present application.

[0332] Monoclonal antibodies secreted by the hybridoma cell line can be purified using any technique known in the art. The hybridomas or mAbs may be further screened to identify mAbs with particular properties, such as the ability to increase GIPR activity.

[0333] Chimeric and humanized antibodies based on the aforementioned sequences are also provided. Monoclonal antibodies for use as therapeutic agents may be modified in various ways before use. One example is a chimeric antibody, which is an antibody consisting of protein segments from different antibodies that are covalently linked to generate a functional immunoglobulin light chain or immunoglobulin heavy chain or an immunologically functional portion thereof. Generally, a portion of such heavy and / or light chain is identical or homologous to a corresponding sequence in an antibody from a particular species or belonging to a particular antibody class or subclass, while the remainder of the chain(s) is identical or homologous to a corresponding sequence in an antibody from another species or belonging to another antibody class or subclass. For methods relating to chimeric antibodies, see, for example, U.S. Pat. No. 4,816,567, and Morrison et al., 1985, Proc. Natl. Acad. Sci. USA 81:6851-6855, which are incorporated herein by reference. CDR grafting is described, for example, in U.S. Patent Nos. 6,180,370, 5,693,762, 5,693,761, 5,585,089, and 5,530,101.

[0334] In general, the goal in preparing chimeric antibodies is to create chimeras in which the number of amino acids derived from the intended patient species is maximized. One example is a "CDR-grafted" antibody, which contains one or more complementarity determining regions (CDRs) derived from a particular species or belonging to a particular antibody class or subclass, while the remainder of the antibody chain(s) is identical or homologous to the corresponding sequence in an antibody derived from another species or belonging to another antibody class or subclass. For use in humans, variable regions from rodent antibodies or CDRs selected from rodent antibodies are often grafted into a human antibody, thereby replacing the naturally occurring variable regions or CDRs of the human antibody.

[0335] One useful type of chimeric antibody is a "humanized" antibody. Generally, humanized antibodies are generated from monoclonal antibodies that are first produced in non-human animals. Certain amino acid residues in the monoclonal antibody are modified to be homologous to corresponding residues in a human antibody of the corresponding isotype, and these modified amino acid residues are typically from non-antigen-recognizing portions of the antibody. Humanization can be performed using a variety of methods, for example, by replacing the corresponding regions of a human antibody with at least a portion of a rodent variable region (see, for example, U.S. Patent Nos. 5,585,089 and 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).

[0336] In one embodiment, the CDRs of the light and heavy chain variable regions of the antibodies provided herein are grafted onto framework regions (FRs) from an antibody from the same or a different species. For example, the heavy and light chain variable regions V H 1. V H 2. V H 3. V H 4. V H 5. VH 6. V H 7. V H 8. V H 9. V H 10. V H 11. V H 12 and / or V L 1 and V L The CDRs of the two can be grafted into a consensus human FR. To create a consensus human FR, a consensus amino acid sequence can be identified by aligning FRs from several human heavy or light chain amino acid sequences. In other embodiments, the FRs of the heavy or light chains disclosed herein are exchanged with FRs from different heavy or light chains. In one aspect, rare amino acids in the FRs of the heavy and light chains of the GIPR antibody are not exchanged, and the remaining FR amino acids are exchanged. A "rare amino acid" is a particular amino acid that is present in a position where it is not normally found in a FR. Alternatively, the variable region grafted from one heavy or light chain can be used with a constant region different from the constant region of that particular heavy or light chain disclosed herein. In other embodiments, the grafted variable region is part of a single chain Fv antibody.

[0337] In certain embodiments, constant regions derived from a species other than human can be used together with human variable region(s) to generate hybrid antibodies.

[0338] Fully human GIPR antibodies are also provided. Methods are available for preparing fully human antibodies specific to a given antigen without exposing the human to the antigen ("fully human antibodies"). One particular means provided for carrying out the generation of fully human antibodies is the "humanization" of the mouse humoral immune system. The introduction of human immunoglobulin (Ig) loci into mice with inactivated endogenous Ig genes is one means for producing fully human monoclonal antibodies (mAbs) in mice, animals that can be immunized with any desired antigen. The use of fully human antibodies can minimize immunogenic and allergic responses that may result from the administration of mouse mAbs or mouse-derived mAbs as therapeutic agents to humans.

[0339] Fully human antibodies can be produced by immunizing a transgenic animal (usually a mouse) that does not produce endogenous immunoglobulins and has the capacity to produce a repertoire of human antibodies. Antigens for this purpose typically have six or more consecutive amino acids and are optionally conjugated to a carrier such as a hapten. See, for example, 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, a transgenic animal is made by disabling the endogenous mouse immunoglobulin loci that code for mouse heavy and light immunoglobulin chains therein and inserting human genomic DNA containing loci encoding human heavy and light chain proteins into a large fragment of the mouse genome. The partially modified animals do not have a full complement of human immunoglobulin loci and can then be crossed to produce animals with all of the desired immune system modifications. Upon administration of an immunogen, these transgenic animals produce antibodies immunospecific for the immunogen, but which have human rather than murine amino acid sequences, including the variable regions. See, e.g., WO 96 / 33735 and WO 94 / 02602, for additional details of such methods. Additional methods relating to transgenic mice for the preparation of human antibodies are described in U.S. Pat. Nos. 5,545,807, 6,713,610, 6,673,986, 6,162,963, 5,545,807, 6,300,129, 6,255,458, 5,877,397, 5,874,299, and 5,545,806, PCT Publication Nos. WO 91 / 10741, WO 90 / 04036, and EP 546073 B1 and EP 546073 A1.

[0340] The above transgenic mice, referred to herein as "HuMab" mice, contain a human immunoglobulin gene minilocus encoding unrearranged human heavy chain (mu and gamma) and kappa light chain immunoglobulin sequences, together with targeted mutations that inactivate the endogenous mu and kappa chain loci (Lonberg et al., 1994, Nature 368:856-859). Thus, these mice have reduced expression of murine IgM or [kappa] and its expression in response to immunization, and the introduced human heavy and light chain transgenes undergo class switching and somatic mutation to generate high affinity human IgG [kappa] monoclonal antibodies (Lonberg et al., supra; Lonberg and Huszar, 1995, Intern. Rev. Immunol. 13:65-93; Harding and Lonberg, 1995, Ann. NY Acad. Sci. 764:536-546). HuMab mouse preparation was performed by Taylor et al., 1992, Nucleic Acids Research 20:6287-6295, Chen et al., 1993, International Immunology 5:647-656, Tuaillon et al., 1994, J. Immunol. 152:2912-2920, Lonberg et al., 1994, Nature 368:856-859, Lonberg,1994, Handbook of Exp.Pharmacology 113:49-101, Taylor et al.,1994, International Immunology 6:579-591, Lonberg and Huszar,1995, Intern.Rev.Immunol.13:65-93, Harding and Lonberg, 1995, Ann.NY Acad. Sci. 764:536-546; Fishwild et al., 1996, Nature Biotechnology 14:845-851, which references are incorporated herein by reference in their entireties for all purposes.See also U.S. Patent Nos. 5,545,806, 5,569,825, 5,625,126, 5,633,425, 5,789,650, 5,877,397, 5,661,016, 5,814,318, 5,874,299, and 5,770,429, as well as U.S. Patent No. 5,545,807, International Publication Nos. WO 93 / 1227, WO 92 / 22646, and WO 92 / 03918, the disclosures of all of which are incorporated herein by reference in their entireties and for all purposes. Such techniques for the production of human antibodies in transgenic mice are also disclosed in WO98 / 24893, and Mendez et al., 1997, Nature Genetics 15:146-156, which are incorporated herein by reference. For example, transgenic mouse strains HCo7 and HCo12 can be used to generate human monoclonal antibodies against GIPR. Further details regarding the production of human antibodies using transgenic mice are provided below.

[0341] Using hybridoma technology, antigen-specific human mAbs with desired specificity can be produced and selected from transgenic mice such as those described above. Such antibodies may be cloned and expressed using appropriate vectors and host cells. Alternatively, antibodies can be harvested from cultured hybridoma cells.

[0342] Fully human antibodies can also be obtained from phage display libraries (disclosed 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, which is incorporated herein by reference.

[0343] The GIPR binding protein may also be a variant, mimetic, derivative, or oligomer based on the structure of the GIPR antigen binding protein having the above CDRs, variable regions, and / or full length chains.

[0344] In one embodiment, for example, the antigen binding protein is a mutated form of the antigen binding protein disclosed above. For example, some of the antigen binding proteins have one or more conservative amino acid substitutions in the heavy or light chain, the variable region, or one or more of the CDRs.

[0345] Naturally occurring amino acids can be divided into classes based on the following 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 affect chain orientation: Gly, Pro, and 6) Aromatics: Trp, Tyr, Phe.

[0346] Conservative amino acid substitutions may involve the exchange of one member of such a class with another member of the same class. Conservative amino acid substitutions may also include non-naturally occurring amino acid residues, which are typically incorporated by chemical peptide synthesis rather than by synthesis in biological systems. These include peptidomimetics and other forms in which amino acid moieties are reversed or inverted.

[0347] Non-conservative substitutions may involve the exchange of a member of one of the above classes for a member from another class. Such substituted residues may be introduced into regions of the antibody that are homologous to human antibodies, or into the non-homologous regions of the molecule.

[0348] According to certain embodiments, when making such changes, the hydropathic index of amino acids may be taken into account: the hydropathic profile of a protein is calculated by assigning a numerical value (the "hydropathy index") to each amino acid and then iteratively averaging these values ​​along the peptide chain. Each amino acid is assigned a hydropathic index based on its hydrophobicity and charge characteristics: 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).

[0349] The importance of hydropathic profile in conferring interactive biological function to 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 with similar hydropathic indexes or hydropathic scores and still retain similar biological activity. In certain embodiments, when making changes based on hydropathic index, substitutions of amino acids whose hydropathic index is within ±2 are included. In some aspects, within ±1 are included, and in other aspects, within ±0.5 are included.

[0350] It is also understood in the art that substitutions of like amino acids can be made efficiently on the basis of hydrophilicity, particularly when the intended biologically functional protein or peptide thereby created is for use in an immunological embodiment, as is the case here. In certain embodiments, the greatest local average hydrophilicity of a protein is governed by the hydrophilicity of its adjacent amino acids, and correlates with its immunogenicity and antigen binding or immunogenicity, i.e., a biological property of the protein.

[0351] The following amino acid residues have been assigned hydrophilicity values: 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 certain embodiments, when making changes based on similar hydrophilicity values, substitutions of amino acids whose hydrophilicity values ​​are within ±2 are included. In other embodiments, within ±1 is included, and in still other embodiments, within ±0.5 is included. In some cases, epitopes may be identified from the primary amino acid sequence based on hydrophilicity. Such regions are also referred to as "epitope core regions."

[0352] Table 6 shows exemplary conservative amino acid substitutions.

[0353] [Table 152]

[0354] Those skilled in the art will be able to determine suitable variants of the polypeptides presented herein using well-known techniques. Those skilled in the art will be able to identify suitable regions of the molecule that can be altered without impairing activity by targeting regions that are not believed to be important for activity. Those skilled in the art will also be able to identify residues and portions of the molecule that are conserved between similar polypeptides. In additional embodiments, even regions that may not be important for biological activity or structure may be subject to conservative amino acid substitutions without impairing biological activity or detrimentally affecting the polypeptide structure.

[0355] Additionally, one skilled in the art can evaluate structure-function studies that identify residues important for activity or structure in similar polypeptides. Consideration of such comparisons allows prediction of the importance in a protein of amino acid residues that correspond to amino acid residues important for activity or structure in similar proteins. One skilled in the art can select chemically similar amino acid substitutions for those amino acid residues predicted to be important.

[0356] The three-dimensional structure of similar polypeptides and the amino acid sequence related to that structure can also be analyzed by those skilled in the art. Taking such information into account, the alignment of the amino acid residues of an antibody with respect to its three-dimensional structure can be predicted by those skilled in the art. Since amino acid residues predicted to be present on the surface of a protein may be involved in important interactions with other molecules, those skilled in the art can select such residues without making radical changes. Furthermore, those skilled in the art can generate test mutants containing single amino acid substitutions at each desired amino acid residue position. These mutants can then be screened using an assay for GIPR activity, thus providing information on which amino acids can be changed and which should not be changed. In other words, based on the information gathered from such routine experiments, those skilled in the art can easily determine amino acid positions where additional substitutions should be avoided, either alone or in combination with other mutations.

[0357] Many scientific publications have been devoted to predicting 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. In addition, computer programs are now available to aid in the prediction of secondary structure. One method of predicting secondary structure is based on homology modeling. For example, two polypeptides or proteins with greater than 30% sequence identity or greater than 40% similarity may have similar structural topologies. The recent expansion of the protein structural database (PDB) has improved the predictability of secondary structure, including the potential number of folds contained in a polypeptide or protein structure. See Holm et al., 1999, Nucl. Acid. Res. 27:244-247. It has been suggested that the number of folds present in a given polypeptide or protein is limited, and that the accuracy of structure prediction will improve dramatically once a definitive number of structures are solved (Brenner et al., 1997, Curr. Op. Struct. Biol. 7:369-376).

[0358] Additional methods for 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).

[0359] In some embodiments, amino acid substitutions are made that result in (1) reduced susceptibility to proteolysis, (2) reduced susceptibility to oxidation, (3) altered binding affinity to form protein complexes, (4) altered binding affinity to a ligand or antigen, and / or (4) imparting or altering other physicochemical or functional properties to such polypeptides. For example, single or multiple amino acid substitutions (in certain embodiments, conservative amino acid substitutions) may be made in the naturally occurring sequence. Substitutions may be made in portions located outside the domain(s) that form intermolecular contacts in antibodies. In such embodiments, conservative amino acid substitutions that do not substantially alter the structural characteristics of the parent sequence (e.g., replacement of one or more amino acids that do not disrupt the secondary structure that characterizes the parent or native antigen-binding protein) may be used. Examples of art-recognized polypeptide secondary and tertiary structures are described in Proteins, Structures and Molecular Principles (Creighton, Ed.), 1984, WH New York: Freeman and Company, Introduction to Protein Structure (Branden and Tooze, eds.), 1991, New York: Garland Publishing, and Thornton et al., 1991, Nature 354:105, each of which is incorporated herein by reference.

[0360] Preferred antibody variants further include cysteine ​​variants, in which one or more cysteine ​​residues in the parent or native amino acid sequence are deleted or replaced with another amino acid (e.g., serine). Cysteine ​​variants are particularly useful when the antibody must be refolded into a biologically active conformation. The number of cysteine ​​residues in cysteine ​​variants may be reduced compared to native antibodies, and typically may even be a number that minimizes interactions resulting from unpaired cysteines.

[0361] The disclosed heavy and light chains, variable region domains, and CDRs can be used to prepare polypeptides that contain an antigen-binding region that can specifically bind to GIPR. For example, one or more CDRs can be incorporated into a molecule (e.g., a polypeptide) covalently or non-covalently to prepare an immunoadhesin. The CDR(s) can be incorporated into the immunoadhesin as part of a larger polypeptide chain, the CDR(s) can be covalently linked to another polypeptide chain in the immunoadhesin, or the CDR(s) can be incorporated into the immunoadhesin non-covalently. The CDR(s) allow the immunoadhesin to specifically bind to a particular antigen of interest (e.g., a GIPR polypeptide or an epitope thereof).

[0362] Mimetics based on the variable region domains and CDRs described herein (e.g., "peptide mimetics" or "peptidomimetics") are also provided. Such 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 all purposes. Peptide mimetics that are structurally similar to therapeutically useful peptides may be used to achieve similar therapeutic or prophylactic effects. Such compounds are often developed with the aid of computerized molecular modeling. In general, a peptidomimetic is a protein that is structurally similar to an antibody that exhibits a desired biological activity (such as the ability to specifically bind to GIPR herein), but has one or more peptide bonds optionally replaced by bonds selected from -CH2NH-, -CH2S-, -CH2-CH2-, -CH-CH- (cis and trans), -COCH2-, -CH(OH)CH2-, and -CH2SO- by methods well known in the art. In certain embodiments, one or more amino acids of a consensus sequence may be systematically substituted with a D-amino acid of the same type (e.g., D-lysine in place of L-lysine) to generate a protein with improved stability. Furthermore, controlled peptides containing the consensus sequence or substantially identical variants of the consensus sequence may be generated by methods known in the art (Rizo and Gierasch, 1992, Ann. Rev. Biochem. 61:387, which is incorporated herein by reference), for example, by adding internal cysteine ​​residues capable of forming intramolecular disulfide bridges that cyclize the peptide.

[0363] Derivatives of the antigen binding proteins described herein are also provided. Derivatized antigen binding proteins may include any molecule or substance that confers a desired property to an antibody or fragment, such as increased half-life in a particular application. Derivatized antigen binding proteins may include, for example, a detectable (or labeling) moiety (e.g., a radioactive, colorimetric, antigenic, or enzymatic molecule, a detectable bead (such as a magnetic or electron-dense (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 medicamentously active moiety), or a molecule that improves the stability of the antigen binding protein for a particular application (e.g., administration to a subject, such as a human subject, or other in vivo or in vitro use). Examples of molecules that can be used to derivatize antigen binding proteins 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 the single chain polypeptides described herein that are pegylated. In one embodiment, the antigen binding protein is conjugated or linked to transthyretin (TTR) or a TTR variant. The TTR or TTR variant can be chemically modified, for example, with a chemical selected from the group consisting of dextran, poly(n-vinylpyrrolidone), polyethylene glycol, propropylene glycol homopolymer, polypropylene oxide / ethylene oxide copolymer, polyoxyethylated polyol, and polyvinyl alcohol.

[0364] Other derivatives include covalent or aggregate complexes of the GIPR antigen binding protein with other proteins or polypeptides, such as by expression of recombinant fusion proteins that include heterologous polypeptides fused to the N-terminus or C-terminus of the GIPR antigen binding protein. For example, the conjugated peptide can be a heterologous signal (or leader) polypeptide, such as the yeast alpha-factor leader, or a peptide such as an epitope tag. Fusion proteins containing the GIPR antigen binding protein can include an added peptide (e.g., poly-His) to facilitate purification or identification of the GIPR antigen binding protein. The GIPR antigen binding protein can also be linked to the FLAG peptide, as described in Hopp et al., 1988, Bio / Technology 6:1204 and U.S. Pat. No. 5,011,912. The FLAG peptide is highly antigenic and provides an epitope that is reversibly bound by a specific monoclonal antibody (mAb), allowing for rapid assays and facilitating purification of the expressed recombinant protein. Reagents useful for preparing fusion proteins in which a FLAG peptide is fused to a given polypeptide are commercially available (Sigma, St. Louis, Mo.).

[0365] In some embodiments, the antigen binding protein comprises one or more labels. The term "labeling group" or "label" refers to any detectable label. Examples of suitable labeling groups include, but are not limited to, radioisotopes or radionuclides (e.g., 3 H, 14 C. 15 N, 35 S, 90 Y, 99 Tc, 111 In, 125 I, 131I), fluorescent groups (e.g., FITC, rhodamine, lanthanide phosphors), enzymatic groups (e.g., horseradish peroxidase, β-galactosidase, luciferase, alkaline phosphatase), chemiluminescent groups, biotin 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. A variety of methods for labeling proteins are known in the art and may be used as appropriate.

[0366] The term "effector group" refers to any group that is coupled to an antigen binding protein and acts as a cytotoxic agent. Examples of suitable effector groups are radioisotopes or radionuclides (e.g., 3 H, 14 C. 15 N, 35 S, 90 Y, 99 Tc, 111 In, 125 I, 131 I). Other suitable groups include toxins, therapeutic groups, and chemotherapeutic groups. Examples of suitable groups include calicheamicin, auristatin, geldanamycin, and maytansine. In some embodiments, the effector group is coupled to the antigen binding protein via spacer arms of various lengths to reduce potential steric hindrance.

[0367] Generally, labels are divided into different 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 secondary reporters (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 of labeling proteins are known in the art.

[0368] Particular labels include optical dyes, including but not limited to chromophores, phosphors, and fluorophores, the latter being often specific. Fluorophores can be either "small molecule" fluorophores or proteinaceous fluorophores.

[0369] By "fluorescent label" is intended any molecule that may be detected via its inherent fluorescent properties. Suitable fluorescent labels include, but are not limited to, fluorescein, rhodamine, tetramethylrhodamine, eosin, erythrosine, coumarin, methyl-coumarin, pyrene, malachite green, stilbene, Lucifer Yellow, Cascade BlueJ, Texas Red, IAEDANS, EDANS, BODIPY FL, LC Red640, Cy5, Cy5.5, LC Red705, Oregon green, Alexa-Fluor dyes (Alexa Fluor 350, Alexa Fluor 430, Alexa Fluor488, Alexa Fluor546, Alexa Fluor568, Alexa Fluor594, Alexa Fluor633, Alexa Fluor660, Alexa Fluor680), Cascade Blue, Cascade Yellow, and R-Phycoerythrin (PE) (Molecular Probes, Eugene, OR), FITC, rhodamine, and Texas Red. 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, which is expressly incorporated herein by reference.

[0370] Suitable proteinaceous fluorescent labels include, but are not limited to, green fluorescent proteins, including GFPs of Renilla species, Ptilosarcus species, or Aequorea species (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 24:462-471; Heim et al., 1996, Curr. Biol. 6:178-182), enhanced yellow fluorescent protein (EYFP, Clontech Labs., Inc.), luciferase (Ichiki et al., 1993, J. Immunol. 150:5408-5417), β-galactosidase (Nolan et al., 1997, J. Immunol. 150:5408-5417), and β-galactosidase (Nolan et al., 1998, J. Immunol. 150:5408-5417). al., 1988, Proc. Natl. Acad. Sci. USA 85:2603-2607), and Renilla (WO 92 / 15673, WO 95 / 07463, WO 98 / 14605, WO 98 / 26277, WO 99 / 49019, U.S. Patent Nos. 5,292,658, 5,418,155, 5,683,888, 5,741,668, 5,777,079, 5,804,387, 5,874,304, 5,876,995, 5,925,558).

[0371] Nucleic acids encoding the antigen binding proteins or portions thereof described herein are also provided, including nucleic acids encoding one or both chains of an antibody, or fragments, derivatives, muteins, or variants thereof, polynucleotides encoding only the heavy chain variable region or CDRs, polynucleotides sufficient for use as hybridization probes, PCR primers, or sequencing primers for identifying, analyzing, mutagenesis, or amplifying polynucleotides encoding the polypeptides, antisense nucleic acids for inhibiting expression of polynucleotides, and complementary sequences thereof. The nucleic acids may be of any length. The nucleic acid may be, for example, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 75, 100, 125, 150, 175, 200, 250, 300, 350, 400, 450, 500, 750, 1,000, 1,500, 3,000, 5,000, or more nucleotides in length, and / or may include one or more additional sequences, such as, for example, regulatory sequences, and / or may be part of a longer nucleic acid, such as, for example, a vector. The nucleic acid may be single-stranded or double-stranded and may include RNA and / or DNA nucleotides, and artificial variants thereof (e.g., peptide nucleic acids). Any of the variable regions provided herein may be added to these constant regions to form complete heavy and light chain sequences. However, it should be understood that these constant region sequences are provided only as specific examples. In some embodiments, the variable region sequences are linked to other constant region sequences known in the art.

[0372] Nucleic acids encoding a particular antigen-binding protein or a portion thereof (e.g., full-length antibody, heavy or light chain, variable domain, or CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, or CDRL3) may be isolated from B cells of mice immunized with GIPR or an immunogenic fragment thereof. The nucleic acids may be isolated by conventional procedures such as polymerase chain reaction (PCR). Phage display is another example of a known technique by which derivatives of antibodies and other antigen-binding proteins may be prepared. In one technique, polypeptides that are components of the antigen-binding protein of interest are expressed in any suitable recombinant expression system, and the expressed polypeptides are allowed to associate to form the antigen-binding protein.

[0373] In one embodiment, the nucleic acid hybridizes to other nucleic acids under specific hybridization conditions is further provided. Nucleic acid hybridization methods are well known in the art. For example, see Current Protocols in Molecular Biology, John Wiley & Sons, NY (1989), 6.3.1-6.3.6. Moderately stringent hybridization conditions as defined herein use a pre-wash solution that contains 5x sodium chloride / sodium citrate (SSC), 0.5% SDS, 1.0 mM EDTA (pH 8.0), a hybridization buffer that contains about 50% formamide, 6x SSC, and a hybridization temperature of about 55°C (or other similar hybridization solutions, such as those that contain about 50% formamide, are used at a hybridization temperature of 42°C), and washing conditions that are performed at 60°C in 0.5x SSC, 0.1% SDS. Stringent hybridization conditions involve hybridization in 6xSSC at 45° C., followed by one or more washes in 0.1xSSC, 0.2% SDS at 68° C. Moreover, one of skill in the art can manipulate the hybridization and / or wash conditions to increase or decrease the stringency of hybridization, such that nucleic acids that contain nucleotide sequences that are at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, or at least 99% identical to each other will typically remain hybridized to each other.

[0374] Basic parameters influencing the selection of hybridization conditions, and guidance for devising appropriate conditions, are provided, for example, by Sambrook, Fritsch, and Maniatis (supra, 2001, Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, and Current Protocols in Molecular Biology, 1995, Ausubel et al., eds., John Wiley & Sons, Inc., sections 2.10 and 6.3-6.4) and can be readily determined by one of skill in the art based, for example, on the length and / or base composition of the nucleic acid.

[0375] Changes can be introduced by mutagenesis into a nucleic acid, which results in an alteration in the amino acid sequence of a polypeptide (e.g., an antibody or antibody derivative) that the nucleic acid encodes. Mutations can be introduced using any technique known in the art. In one embodiment, one or more specific amino acid residues are altered, e.g., using a site-directed mutagenesis protocol. In another embodiment, one or more randomly selected residues are altered, e.g., using a random mutagenesis protocol. Regardless of how it is performed, the mutant polypeptides can be expressed and screened for desired properties.

[0376] Mutations can be introduced into a nucleic acid without significantly altering the biological activity of the polypeptide it encodes. For example, nucleotide substitutions leading to amino acid substitutions at non-essential amino acid residue positions can be performed. Alternatively, one or more mutations can be introduced into a nucleic acid that selectively alter the biological activity of the polypeptide it encodes. For example, the mutations can quantitatively or qualitatively alter the biological activity. Examples of quantitative alterations include increasing, reducing, or eliminating an activity. Examples of qualitative alterations include altering the antigen specificity of an antibody. In one embodiment, a nucleic acid encoding any of the antigen binding proteins described herein can be mutated to alter the amino acid sequence using molecular biology techniques well established in the art.

[0377] In another aspect, a nucleic acid molecule suitable for use as a primer or hybridization probe for the detection of a nucleic acid sequence is provided. The nucleic acid molecule may comprise only a portion of a nucleic acid sequence encoding a full-length polypeptide, e.g., a fragment that can be used as a probe or primer, or a fragment that encodes an active portion of the polypeptide.

[0378] Probes based on the sequence of a nucleic acid can be used to detect the nucleic acid or similar nucleic acids, such as multiple transcripts that code for one polypeptide. Probes can include labeling groups, such as radioisotopes, fluorescent compounds, enzymes, or enzyme cofactors. Such probes can be used to identify cells that express the polypeptide.

[0379] In another aspect, a vector is provided that comprises a nucleic acid encoding a polypeptide or a portion thereof (e.g., a fragment comprising one or more CDRs or one or more variable region domains). Examples of vectors include, but are not limited to, plasmids, viral vectors, non-episomal mammalian vectors, and expression vectors (e.g., recombinant expression vectors). A recombinant expression vector may comprise a nucleic acid in a form suitable for expression of the nucleic acid in a host cell. A recombinant expression vector comprises one or more regulatory sequences selected based on the host cell to be used for expression, and such regulatory sequences are operably linked to the nucleic acid sequence to be expressed. Regulatory sequences include those that direct constitutive expression of a nucleotide sequence in many types of host cells (e.g., SV40 early gene enhancer, Rous sarcoma virus promoter, and cytomegalovirus promoter), those that direct expression of a nucleotide sequence only in certain host cells (e.g., tissue-specific regulatory sequences, see Voss et al., 1986, Trends Biochem. Sci. 11:287; Maniatis et al., 1987, Science 236:1237, which is incorporated herein by reference in its entirety), as well as those that direct inducible expression of nucleotide sequences in response to a particular treatment or condition (e.g., the metallothionin promoter in mammalian cells, and tetracycline (tet)- and / or streptomycin-responsive promoters in both prokaryotic and eukaryotic systems, see ibid. One of ordinary skill in the art will appreciate that the design of the expression vector can depend on such factors as the choice of the host cell to be transformed, the level of expression of protein desired, and the like. The expression vector, upon introduction into a host cell, can produce a protein or peptide, including a fusion protein or peptide, encoded by the nucleic acid described herein.

[0380] In another aspect, a host cell into which a recombinant expression vector has been introduced is provided. The host cell can be any prokaryotic cell (e.g., E. coli) or eukaryotic cell (e.g., yeast, insect, or mammalian cell (e.g., CHO cell)). Vector DNA can be introduced into prokaryotic or eukaryotic cells via standard transformation or gene transfer techniques. It is known that stable gene transfer of mammalian cells depends on the expression vector and gene transfer technique used, and only a small fraction of cells may integrate the foreign DNA into their genome. To identify and select such integrants, a gene encoding a selectable marker (e.g., for antibiotic resistance) is generally introduced into the host cell along with the gene of interest. Preferred selectable markers include those that confer resistance to drugs such as G418, hygromycin, and methotrexate. Cells stably transfected with the introduced nucleic acid can be identified by drug selection, among other methods (e.g., cells into which the selectable marker gene has been integrated will survive, while other cells die).

[0381] Also provided herein are expression systems and constructs in the form of plasmids, expression vectors, transcription cassettes or expression cassettes which contain at least one of the above-described polynucleotides, as well as host cells containing such expression systems or constructs.

[0382] The antigen binding proteins provided herein may be prepared by any of a number of conventional techniques.For example, GIPR antigen binding proteins may be produced by recombinant expression systems using any technique known in the art.See, for example, Monoclonal Antibodies, Hybridomas: A New Dimension in Biological Analyses, Kennet et al. (eds.) Plenum Press, New York (1980), and Antibodies: A Laboratory Manual, Harlow and Lane (eds.), Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY (1988).

[0383] The antigen binding protein can be expressed in a hybridoma cell line (e.g., specifically, an antibody may be expressed in a hybridoma) or in a cell line other than a hybridoma. An expression construct encoding the antibody can be used to transform a mammalian host cell, an insect host cell, or a microbial host cell. Transformation can be carried out using any known method for introducing a polynucleotide into a host cell, including, for example, packaging the polynucleotide into a virus or bacteriophage and transducing the host cell with the construct by gene transfer procedures known in the art, exemplified by U.S. Patent Nos. 4,399,216, 4,912,040, 4,740,461, and 4,959,455. The optimal transformation procedure used will depend on which type of host cell is being transformed. Methods for introducing heterologous polynucleotides into mammalian cells are well known in the art and include, but are not limited to, dextran-mediated gene transfer, calcium phosphate precipitation, polybrene-mediated gene transfer, protoplast fusion, electroporation, encapsulation of the polynucleotide(s) into liposomes, mixing of nucleic acids with positively charged lipids, and direct microinjection of DNA into the nucleus.

[0384] The recombinant expression construct typically includes a nucleic acid molecule encoding a polypeptide that includes one or more of the following: one or more of the CDRs provided herein, a light chain constant region, a light chain variable region, a heavy chain constant region (e.g., a CDR), a light chain variable region, a light chain constant region, ... H 1. C H 2, and / or C H 3), and / or another scaffold portion of the GIPR antigen binding protein. Such nucleic acid sequences are inserted into a suitable expression vector using standard nucleic acid ligation techniques. In one embodiment, a heavy or light chain constant region is added to the C-terminus of the anti-GIPR specific heavy or light chain variable region and ligated into an expression vector. The vector is typically selected to be functional in the particular host cell to be used (i.e., the vector can be compatible with the host's cellular machinery and allow gene amplification and / or expression). In some embodiments, vectors are used that use protein-fragment complementation assays using protein reporters such as dihydrofolate reductase (see, e.g., U.S. Pat. No. 6,270,964, which is incorporated herein by reference). Suitable expression vectors can be purchased, for example, from Invitrogen Life Technologies or BD Biosciences (formerly "Clontech"). Other vectors useful for cloning and expressing antibodies and fragments include those described in Bianchi and McGrew, 2003, Biotech. Biotechnol. Bioeng. 84:439-44, which is incorporated herein by reference. Additional suitable expression vectors are discussed, for example, in Methods Enzymol., vol. 185 (DV Goeddel, ed.), 1990, New York: Academic Press.

[0385] Typically, expression vectors used in any of the host cells will include sequences for maintaining the plasmid and for cloning and expressing exogenous nucleotide sequences. Such sequences are collectively referred to as "flanking sequences" and 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 transcription termination sequence, a complete intron sequence including donor and acceptor splice sites, a sequence encoding a leader sequence for secretion of a polypeptide, a ribosome binding site, a polyadenylation sequence, a polylinker region for inserting a nucleic acid encoding a polypeptide to be expressed, and a selectable marker element. Each of these sequences is discussed below.

[0386] Optionally, the vector may contain a sequence encoding a "tag", i.e., an oligonucleotide molecule located at the 5' or 3' end of the sequence encoding the GIPR antigen-binding protein, encoding polyHis (such as hexaHis) or another "tag" for which there are commercially available antibodies, such as FLAG®, HA (influenza virus hemagglutinin), or myc. 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 GIPR antigen-binding protein from the host cell. Affinity purification can be achieved, for example, by column chromatography using antibodies against the tag as an affinity matrix. Optionally, the tag can then be removed from the purified GIPR antigen-binding protein by various means, such as using certain peptidases for cleavage.

[0387] The flanking sequences can be homologous (i.e., derived from the same species and / or strain as the host cell), heterologous (i.e., derived from a species other than the host cell's species or strain), hybrid (i.e., a combination of flanking sequences derived from multiple sources), synthetic, or naturally occurring. Thus, the source of the flanking sequences can be any prokaryotic or eukaryotic organism, any vertebrate or invertebrate organism, or any plant, provided that the flanking sequences are functional in and capable of being activated by the host's cellular machinery.

[0388] Flanking sequences useful in vectors may 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 restriction endonuclease digestion, and can be isolated from a suitable tissue source using a suitable restriction endonuclease. In some cases, all of the nucleotide sequences of the flanking sequences may be known. In this case, the flanking sequences may be synthesized using the methods described herein for nucleic acid synthesis or cloning.

[0389] Whether all or only a portion of the flanking sequences are known, the flanking sequences may be obtained using polymerase chain reaction (PCR) and / or by screening a genomic library with suitable probes, such as oligonucleotides and / or flanking sequence fragments from the same or another species. If the flanking sequences are unknown, fragments of DNA containing the flanking sequences may be isolated from a larger fragment of DNA that may, for example, contain coding sequences or even one or more additional genes. Isolation may be achieved by digestion with restriction endonucleases to generate the appropriate DNA fragments, followed by isolation using agarose gel purification, Qiagen® column chromatography (Chatsworth, Calif.), or other methods known to those of skill in the art. The selection of appropriate enzymes to achieve this purpose will be readily apparent to those of skill in the art.

[0390] Replication origins are typically part of commercially purchased prokaryotic expression vectors, and origins aid in the amplification of vectors in host cells. If the vector of choice does not contain an origin of replication site, one may be chemically synthesized based on a known sequence and ligated into the vector. For example, the origin of replication from plasmid pBR322 (New England Biolabs, Beverly, Mass.) is suitable for most Gram-negative bacteria, and various viral origins (e.g., SV40, polyoma, adenovirus, vesicular stomatitis virus (VSV), or papillomaviruses such as HPV or BPV) are useful for cloning vectors in mammalian cells. In general, origin of replication elements are not required for mammalian expression vectors (e.g., the SV40 origin is often used only because it also contains the viral early promoter).

[0391] Transcription termination sequences are typically located 3' to the end of a polypeptide-encoding region and serve to terminate transcription.Transcription termination sequences in prokaryotic cells are usually GC-rich fragments followed by poly-T sequences.While this sequence can be easily cloned from a library or even purchased commercially as part of a vector, it can also be easily synthesized using methods for nucleic acid synthesis, such as those described herein.

[0392] A selectable marker gene encodes a protein necessary for the survival and growth of a host cell grown in a selective medium. Typical selectable marker genes encode (a) a protein that confers resistance to antibiotics or other toxins, such as, for example, ampicillin, tetracycline, or kanamycin for prokaryotic host cells, (b) a protein that complements an auxotrophic deficiency of the cell, or (c) a protein that supplies a vital nutrient unavailable from a complex or defined medium. Particular selectable markers are the kanamycin resistance gene, the ampicillin resistance gene, and the tetracycline resistance gene. Conveniently, the neomycin resistance gene may be used for selection in both prokaryotic and eukaryotic host cells.

[0393] Other selectable genes may be used to amplify the genes to be expressed. Amplification is the process by which genes required for the production of a protein important for growth or cell survival are repeated in tandem in the chromosomes of successive generations of recombinant cells. Examples of selectable markers suitable for mammalian cells include the dihydrofolate reductase (DHFR) gene and the promoterless thymidine kinase gene. Mammalian cell transformants are placed under selection pressure where only the transformants are uniquely adapted to survive because of the presence of the selectable gene in the vector. Selection pressure is applied by culturing the transformed cells under conditions of successively increasing concentrations of the selection drug in the medium, thereby amplifying both the selectable gene and the DNA encoding another gene, such as an antigen-binding protein that binds to the GIPR polypeptide. As a result, the amount of polypeptide, such as an antigen-binding protein, synthesized from the amplified DNA increases.

[0394] A ribosome binding site is usually necessary for the initiation of translation 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.

[0395] In some cases, such as when glycosylation is desired in a eukaryotic host cell expression system, various pre- or pro-sequence manipulations may be performed to improve glycosylation or yield. For example, the peptidase cleavage site of a particular signal peptide may be modified or a pro-sequence may be added, which may also affect glycosylation. The final protein product may have one or more additional amino acids at position -1 (relative to the first amino acid of the mature protein) that may not have been completely removed upon expression and remain. For example, the final protein product may have one or two additional amino acid residues found in the peptidase cleavage site at the amino terminus. Alternatively, the use of several enzymatic cleavage sites may result in a slightly truncated form of the desired polypeptide if the enzyme cleaves at such a region in the mature polypeptide.

[0396] Expression and cloning typically involve a promoter that is recognized by the host organism and operably linked to the molecule encoding the GIPR antigen-binding protein. A promoter is a non-transcribed sequence located upstream (i.e., 5') of the start codon of a structural gene (generally within about 100-1000 bp) and controls transcription of the structural gene. Promoters are conventionally classified into one of two classes: inducible promoters and constitutive promoters. An inducible promoter causes an increase in the level of transcription from DNA under its control in response to some change in culture conditions, such as the presence or absence of a nutrient or a change in temperature. On the other hand, a constitutive promoter uniformly transcribes the gene to which it is operably linked, i.e., expression of the gene is little or not controlled at all. There are a great many well-known promoters that are recognized by various potential host cells. An appropriate promoter is operably linked to DNA encoding the heavy or light chain that constitutes the GIPR antigen-binding protein by removing the promoter from the source DNA by restriction enzyme digestion and inserting the desired promoter sequence into a vector.

[0397] Promoters suitable for use in yeast hosts are also well known in the art. Yeast enhancers are advantageously used with yeast promoters. Promoters suitable for use in mammalian host cells are well known, including, but not limited to, those derived from the genomes of viruses such as polyoma virus, fowlpox virus, adenovirus (such as adenovirus type 2), bovine papilloma virus, avian sarcoma virus, cytomegalovirus, retrovirus, hepatitis B virus, and simian virus 40 (SV40). Other suitable mammalian promoters include, for example, heat shock promoters and heterologous mammalian promoters such as actin promoter.

[0398] To increase the transcription of DNA encoding the light or heavy chain constituting the GIPR antigen-binding protein by higher eukaryotes, an enhancer sequence may be inserted into the vector. Enhancers are cis-acting DNA elements that act on a promoter to increase transcription, and are usually about 10 to 300 bp in length. Enhancers are found at both the 5' and 3' positions of a transcription unit, and are relatively orientation and position dependent. Several enhancer sequences are known that can be used from mammalian genes (e.g., globin, elastase, albumin, alpha-fetal protein, and insulin). However, typically enhancers derived from viruses are used. SV40 enhancers, cytomegalovirus early promoter enhancers, polyoma enhancers, and adenovirus enhancers are known in the art, and these enhancers are examples of enhancing elements for activating eukaryotic promoters. Enhancers may be located either 5' or 3' of the coding sequence in a vector, but are typically located at the 5' site of the promoter. To facilitate extracellular secretion of the antibody, a sequence encoding a suitable native or heterologous signal sequence (leader sequence or signal peptide) can be incorporated into the expression vector. The choice of signal peptide or leader depends on the type of host cell in which the antibody is to be produced, and the native signal sequence can be replaced with a heterologous signal sequence. Examples of signal peptides that are functional in mammalian host cells include the interleukin-7 (IL-7) signal sequence described in U.S. Pat. No. 4,965,195, the interleukin-2 receptor signal sequence described in Cosman et al., 1984, Nature 312:768, the interleukin-4 receptor signal peptide described in EP Patent No. 0367566, the interleukin-1 receptor type I signal peptide described in U.S. Pat. No. 4,968,607, and the interleukin-1 receptor type II signal peptide described in EP Patent No. 0460846.

[0399] In one embodiment, the leader sequence comprises SEQ ID NO: 1217 (MDMRVPAQLL GLLLLWLRGA RC), which is encoded by SEQ ID NO: 1218 (atggacatga gagtgcctgc acagctgctg ggcctgctgc tgctgtggct gagaggcgcc agatgc). In another embodiment, the leader sequence comprises SEQ ID NO: 1219 (MAWALLLLTL LTQGTGSWA), which is encoded by SEQ ID NO: 1220 (atggcctggg ctctgctgct cctcaccctc ctcactcagg gcacagggtc ctgggcc).

[0400] The expression vector provided may be constructed from a starting vector, such as a commercially available vector. Such vector may or may not contain all of the desired flanking sequences. If one or more of the flanking sequences described herein are not present in the vector initially, they may be obtained individually and ligated into the vector. The method used to obtain each flanking sequence is well known to those skilled in the art.

[0401] After constructing the vector and inserting the nucleic acid molecules encoding the light, heavy, or light and heavy chains that make up the GIPR antigen binding sequence into the appropriate sites of the vector, the completed vector may be inserted into a suitable host cell for amplification and / or polypeptide expression. Transformation of the selected host cell with the expression vector for the antigen binding protein may 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 depend in part on the type of host cell to be used. These and other suitable methods are well-known to those of skill in the art and are set forth, for example, in Sambrook et al., 2001, supra.

[0402] When cultured under appropriate conditions, the host cells will synthesize the antigen binding protein, which can be subsequently collected from the medium (if the host cells secrete it into the medium) or directly from the host cells that produce it (if it is not secreted). The selection of an appropriate host cell will depend on a variety of factors, such as the desired expression level, polypeptide modifications (such as glycosylation or phosphorylation) that are desirable or necessary for activity, and the accessibility of folding into a biologically active molecule.

[0403] 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, 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 many other cell lines. In certain embodiments, cell lines may be selected by determining which cell lines express high levels and constitutively produce antigen binding proteins with GIPR binding properties. In another embodiment, a cell line derived from the B cell lineage may be selected that does not produce its own antibody but has the ability to produce and secrete heterologous antibodies.

[0404] In one embodiment, the invention is directed to an antigen binding protein produced by a cell expressing one or more of the polynucleotides identified in Table 2, Table 3, Table 4, and Table 5.

[0405] In one embodiment, the GIPR binding protein is administered for chronic treatment, hi another embodiment, the binding protein is administered for acute treatment.

[0406] Pharmaceutical compositions comprising GIPR antigen binding proteins are also provided, and such pharmaceutical compositions may be utilized in any of the preventive and therapeutic methods disclosed herein. In one embodiment, a therapeutically effective amount of one or more antigen binding proteins, as well as pharma- ceutical acceptable diluents, carriers, solubilizers, emulsifiers, preservatives, and / or adjuvants are also provided. Acceptable formulation materials are those that are non-toxic to recipients at the dosages and concentrations employed.

[0407] In certain embodiments, pharmaceutical compositions may include formulation materials intended to modify, maintain, or preserve, for example, the pH, osmolarity, viscosity, clarity, color, isotonicity, odor, sterility, stability, dissolution or release rate, absorption, or permeability 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), antimicrobial agents, antioxidants (such as ascorbic acid, sodium sulfite, or sodium bisulfite), buffers (such as borate, bicarbonate, Tris-HCl, citrate, phosphate, or other organic acids), bulking agents (such as mannitol or glycine), chelating agents (such as ethylenediaminetetraacetic acid (EDTA)), complexing agents (such as caffeine, polyvinylpyrrolidone, beta-cyclodextrin, or hydroxypropyl-beta-cyclodextrin), bulking agents, monosaccharides, disaccharides, and other carbohydrates (such as glucose, mannose, or dextrin), proteins (such as serum albumin, gelatin, or immunoglobulins), colorants, flavoring agents, and diluents, emulsifiers, 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 pluronic, PEG, sorbitan esters, polysorbates (such as polysorbate 20), polysorbates, triton, tromethamine, lecithin, cholesterol, tyloxapal, etc.), stability enhancers (such as sucrose or sorbitol), osmotic pressure enhancers (such as alkali metal halides, preferably sodium chloride or potassium chloride, mannitol sorbitol), delivery vehicles, diluents, excipients, and / or pharmaceutical adjuvants.REMINGTON'S PHARMACEUTICAL SCIENCES, 18th Edition, (AR Genrmo, ed.), 1990, Mack Publishing Company provides additional details and options regarding suitable drugs that can be incorporated into pharmaceutical compositions.

[0408] In certain embodiments, optimal pharmaceutical compositions will be determined by those of skill in the art depending, for example, on the intended route of administration, delivery format, and desired dosage. See, for example, REMINGTON'S PHARMACEUTICAL SCIENCES, supra. In certain embodiments, such compositions may affect the physical state, stability, in vivo release rate, and in vivo excretion rate of the disclosed antigen binding proteins. In certain embodiments, the primary vehicle or carrier in the pharmaceutical composition may be either aqueous or non-aqueous in nature. For example, a suitable vehicle or carrier may be water for injection or saline. In certain embodiments, the GIPR antigen binding protein composition may be prepared for storage in the form of a lyophilized cake or aqueous solution by mixing the selected composition having the desired purity with optional formulation agents (REMINGTON'S PHARMACEUTICAL SCIENCES, supra). Furthermore, in certain embodiments, the GIPR antigen binding protein may be formulated as a lyophilizate using appropriate excipients such as sucrose.

[0409] The pharmaceutical composition may be selected for parenteral delivery. Alternatively, the composition may be selected for delivery via the digestive tract, such as by inhalation or orally. The preparation of such pharma- ceutically acceptable compositions is a technique known in the art.

[0410] The formulation components are preferably present in concentrations that are acceptable to the site of administration. In certain embodiments, buffers are used to maintain the composition at physiological pH or slightly lower, typically within a pH range of about 5 to about 8.

[0411] When parenteral administration is contemplated, the therapeutic composition may be provided in the form of a pyrogen-free parenterally acceptable aqueous solution containing the desired human GIPR antigen binding protein in a medicament acceptable vehicle. A particularly suitable vehicle for parenteral injection is sterile distilled water in which the GIPR antigen binding protein is formulated as a sterile, isotonic solution with appropriate preservation. In certain embodiments, the preparation may require formulating the desired molecule with a drug, such as an injectable microparticle, bioerodible particle, polymeric compound (such as polylactic acid or polyglycolic acid), bead, or liposome, which may provide controlled or sustained release of the product that may be delivered via depot injection. In certain embodiments, hyaluronic acid may be used to promote sustained circulation. In certain embodiments, an implantable drug delivery device may be used to introduce the desired antigen binding protein.

[0412] Certain pharmaceutical compositions are formulated for inhalation. In some embodiments, the GIPR antigen binding protein is formulated as a dry inhalable powder. In certain embodiments, the GIPR antigen binding protein inhalation solution may be formulated with a propellant for aerosol delivery. In certain embodiments, the solution may be nebulized. Pulmonary administration, and therefore formulation methods, are further described in International Patent Application No. PCT / US94 / 001875 (incorporated by reference), which describes pulmonary delivery of chemically modified proteins. Some formulations may be administered orally. GIPR antigen binding proteins administered in this manner may be formulated with or without carriers customarily used in the compounding of solid dosage forms such as tablets and capsules. In certain embodiments, capsules may be designed to release the active portion of the formulation at a time in the gastrointestinal tract that maximizes bioavailability and minimizes pre-systemic degradation. Additional agents may be included to facilitate absorption of the GIPR antigen binding protein. Diluents, flavorings, low melting point waxes, vegetable oils, lubricants, suspending agents, tablet disintegrating agents, and binders may also be employed.

[0413] Some pharmaceutical compositions contain one or more GIPR antigen binding proteins in a mixture in an effective amount with non-toxic excipients suitable for tablet manufacture. A solution in unit dose form may be prepared by dissolving tablets in sterile water or another suitable medium. Suitable excipients include, but are not limited to, inert diluents such as calcium carbonate, sodium carbonate or bicarbonate, lactose, or calcium phosphate, or binders such as starch, gelatin, or acacia, or lubricants such as magnesium stearate, stearic acid, or talc.

[0414] It will be clear to those skilled in the art that the pharmaceutical composition additionally includes formulations that include GIPR binding protein in sustained or controlled delivery formulations. Various other sustained or controlled delivery means formulation techniques, such as liposome carriers, bioerodible microparticles or porous beads, and depot injections, are also known to those skilled in the art. For example, see International Patent Application No. PCT / US93 / 00829 (incorporated by reference). Therein, the controlled release of porous polymeric microparticles for delivering pharmaceutical compositions is described. The sustained release preparation may include, for example, a semipermeable polymer matrix in the form of a shaped article, such as a film or a microcapsule. Sustained release matrices may include polyesters, hydrogels, polylactic acid (disclosed in U.S. Pat. No. 3,773,919 and European Patent 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-hydroxybutanoic acid (European Patent Publication No. EP 133,988). Sustained release compositions may also include liposomes, which can be prepared by any of several methods known in the art. See, for example, Eppstein et al., 1985, Proc. Natl. Acad. Sci. USA 82:3688-3692, European Patent Application Publication Nos. EP 036,676, EP 088,046, and EP 143,949, which are incorporated by reference.

[0415] The pharmaceutical composition used for in vivo administration is typically provided as a sterile preparation. Sterilization can be achieved by filtration through a sterile filtration membrane. When the composition is lyophilized, sterilization using this method can be carried out either before or after lyophilization and reconstitution. The composition for parenteral administration can be stored in lyophilized form or in solution. Parenteral compositions are generally filled into a container with a sterile access port, such as an intravenous solution bag or a vial with a stopper that can be pierced by a hypodermic injection needle.

[0416] In certain formulations, the concentration of the antigen binding protein is at least 10 mg / mL, at least 20 mg / mL, at least 30 mg / mL, at least 40 mg / mL, at least 50 mg / mL, at least 60 mg / mL, at least 70 mg / mL, at least 80 mg / mL, at least 90 mg / mL, at least 100 mg / mL, or at least 150 mg / mL. In one embodiment, the pharmaceutical composition comprises the antigen binding protein, a buffering agent, and a polysorbate. In another embodiment, the pharmaceutical composition comprises the antigen binding protein, a buffering agent, sucrose, and a polysorbate. An example pharmaceutical composition comprises 50-100 mg / mL of the antigen binding protein, 5-20 mM sodium acetate, 5-10% w / v sucrose, and 0.002-0.008% w / v polysorbate. Certain compositions, for example, contain 65-75 mg / mL of 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 ranges from 4.5 to 6. Other formulations have a pH of 5.0-5.5 (e.g., pH 5.0, 5.2, or 5.4).

[0417] Once the pharmaceutical composition is formulated, it may be stored in a sterile vial as a solution, suspension, gel, emulsion, solid, crystal, or dehydrated or lyophilized powder. Such formulations may be stored either in a ready-to-use form or in a form that is reconstituted prior to administration (e.g., lyophilized). Kits for obtaining single-dose administration units are also provided. Certain kits include a first container containing a dried protein and a second container containing an aqueous formulation. In certain embodiments, kits are provided that include single-chamber and multi-chamber pre-filled syringes (e.g., liquid syringes and lyosyringes). The therapeutically effective amount of the GIPR antigen binding protein-containing pharmaceutical composition to be used will depend, for example, on the nature and purpose of the treatment. Those skilled in the art will appreciate that dosage levels suitable for treatment will vary in part depending on the molecule to be delivered, the indication for which the GIPR antigen binding protein is to be used, the route of administration, and the size (weight, body surface, or organ size) and / or condition (age and general health) of the patient. In certain embodiments, the clinician may titer the dosage and modify the route of administration to obtain the optimal therapeutic effect.

[0418] The frequency of administration will depend on the pharmacokinetic parameters of the particular GIPR antigen binding protein in the formulation used. Typically, the clinician will administer the composition until a dosage is reached that achieves the desired effect. Thus, the composition may be administered as a single dose, or as two or more doses (which may or may not contain the same amount of the desired molecule) over a period of time, or as a continuous infusion via an implanted device or catheter. The appropriate dosage may be ascertained using appropriate dose-response data. In certain embodiments, the antigen binding protein may be administered to the patient for an extended period of time. In certain embodiments, the antigen binding protein is administered every 2 weeks, every month, every 2 months, every 3 months, every 4 months, every 5 months, or every 6 months.

[0419] The route of administration of the pharmaceutical composition follows known methods, for example, via injection orally, intravenously, intraperitoneally, intracerebrally (intracemdrally), intraventricularly, intramuscularly, intraocularly, intraarterially, intraportally, or intralesionally, by sustained release system or implanted device. In certain embodiments, the composition may be administered by bolus injection, continuously by infusion, or by implanted device.

[0420] The compositions may be administered locally via implantation of a membrane, sponge, or another suitable material to which the desired molecule is adsorbed or encapsulated. In certain embodiments, when an implantation device is used, the device may be implanted in any suitable tissue or organ, and delivery of the desired molecule may be via diffusion, time-release bolus, or continuous administration.

[0421] It may also be desirable to use the GIPR antigen binding protein pharmaceutical composition according to the disclosed ex vivo. In such cases, cells, tissues, or organs removed from a patient are exposed to the GIPR antigen binding protein pharmaceutical composition. The cells, tissues, and / or organs are then subsequently transplanted back into the patient.

[0422] A physician will be able to select appropriate therapeutic indications and target lipid levels according to the individual profile of a particular patient.One of the widely accepted standards for guiding the treatment of hyperlipidemia is the Third Report of the National Cholesterol Education Program (NCEP) Expert Panel on Detection, Evaluation, and Treatment of the High Blood Cholesterol in Adults (Adult Treatment Panel III) Final Report, National Institutes of Health, NIH Publication No. 02-5215 (2002), the printed publication of which is incorporated herein by reference in its entirety.

[0423] The efficacy of a particular dose can be assessed by reference to a biomarker or improvement in a particular physiological parameter. Examples of suitable biomarkers include the ratio of free cholesterol to plasma lipids, the ratio of free cholesterol to membrane proteins, the ratio of phosphatidylcholine to sphingomyelin, or HDL-C levels.

[0424] Also provided herein are compositions comprising GIPR antigen binding proteins and one or more additional therapeutic agents, as well as methods of administering such agents simultaneously or sequentially with GIPR antigen binding proteins for use in the preventive and therapeutic methods disclosed herein. The one or more additional agents can be co-formulated with GIPR antigen binding proteins or co-administered with GIPR antigen binding proteins. In general, the therapeutic methods, compositions, and compounds, along with the additional agents administered simultaneously, may be used in combination with other therapeutic methods in the treatment of various disease conditions.

[0425] In one aspect, the present invention is directed to a method of treating a subject having a metabolic disorder, the method comprising administering to the subject a therapeutically effective amount of a GLP-1 receptor agonist and a therapeutically effective amount of a GIPR antagonist that specifically binds to a protein having an amino acid sequence that has at least 90% amino acid sequence identity to the amino acid sequence of GIPR.

[0426] "GLP-1 receptor agonist" refers to a compound having GLP-1 receptor activity. Examples of such compounds include exendins, exendin analogs, exendin agonists, GLP-1(7-37), GLP-1(7-37) analogs, GLP-1(7-37) agonists, and the like. GLP-1 receptor agonist compounds may be optionally amidated. The terms "GLP-1 receptor agonist" and "GLP-1 receptor agonist compound" have the same meaning.

[0427] The term "exendin" includes naturally occurring (or synthetic versions of those naturally occurring) exendin peptides found in the salivary secretions of the Gila monster. Exendins of particular interest include exendin-3 and exendin-4. The exendins, exendin analogs, and exendin agonists for use in the methods described herein may optionally be amidated, and may be in the acid form, in the form of a pharma- ceutically acceptable salt, or in any other physiologically active form of the molecule.

[0428] In one embodiment, the molar ratio of the GLP-1 receptor agonist to the GIPR antagonist is about 1:1 to 1:110, about 1:1 to 1:100, about 1:1 to 1:75, about 1:1 to 1:50, about 1:1 to 1:25, about 1:1 to 1:10, about 1:1 to 1:5, and about 1: 1. In one embodiment, the molar ratio of the GIPR antagonist to the GLP-1 receptor agonist is about 1:1 to 1:110, about 1:1 to 1:100, about 1:1 to 1:75, about 1:1 to 1:50, about 1:1 to 1:25, about 1:1 to 1:10, and about 1:1 to 1:5.

[0429] In one embodiment, the GLP-1 receptor agonist and the GIPR antagonist are used in combination in a therapeutically effective molar ratio of about 1:1.5 to 1:150, preferably 1:2 to 1:50.

[0430] In one embodiment, the GLP-1 receptor agonist and the GIPR antagonist are present in dosages that are at least about 1.1-1.4 times, 1.5 times, 2 times, 3 times, 4 times, 5 times, 6 times, 7 times, 8 times, 9 times, or 10 times less than the dosages that would be required to treat the condition and / or disease with each compound alone.

[0431] In one embodiment, the GLP-1 receptor agonist is GLP-1(7-37) or a GLP-1(7-37) analogue.

[0432] In one embodiment, the GLP-1 receptor agonist is selected from the group consisting of exenatide, liraglutide, lixisenatide, albiglutide, dulaglutide, semaglutide, and taspoglutide.

[0433] In one aspect, the present invention is directed to a method of treatment, the method comprising administration to a subject having symptoms of a metabolic disorder of at least one GLP-1 receptor agonist in a therapeutically effective amount in combination with administration of at least one GIPR antagonist that provides a sustained beneficial effect when administered to the subject.

[0434] In one embodiment, the administration of at least one GLP-1 receptor agonist in combination with the administration of at least one GIPR antagonist provides a sustained beneficial effect on at least one symptom of a metabolic disorder.

[0435] In one embodiment, a therapeutically effective amount of a GLP-1 receptor agonist and a therapeutically effective amount of a GIPR antagonist are combined prior to administration to a subject.

[0436] In one embodiment, a therapeutically effective amount of a GLP-1 receptor agonist and a therapeutically effective amount of a GIPR antagonist are administered sequentially to a subject.

[0437] In one embodiment, the therapeutically effective amount of the GLP-1 receptor agonist and the therapeutically effective amount of the GIPR antagonist are synergistically effective amounts.

[0438] Exendin-4 (HGEGTFTSDLSKQMEEEAVRLFIEWLKNGGPSSGAPPPS-NH2 (SEQ ID NO: 1223)) is a peptide found in the saliva of the Gila monster (Heloderma suspectum), and exendin-3 (HSDGTFTSDLSKQMEEEAVRLFIEWLKNGGPSSGAPPPS-NH2 (SEQ ID NO: 1224)) is a peptide found in the saliva of the Mexican beaded lizard (Heloderma horridum). Exendins have some amino acid sequence similarity to some members of the glucagon-like peptide (GLP) family. For example, exendin-4 has about 53% sequence identity with glucagon-like peptide-1 (GLP-1) (7-37) (HAEGTFTSDVSSYLEGQAAKEFIAWLVKGRG (SEQ ID NO: 1244)). However, exendin-4 is not the Gila monster homolog of the mammalian proglucagon gene, which is transcribed from a different gene and from which GLP-1 is expressed. Furthermore, exendin-4 is not an analog of GLP-1(7-37), since sequence modification of the GLP-1 structure did not produce a synthetic exendin-4 peptide structure. Nielsen et al., Current Opinion in Investigational Drugs, 4(4):401-405(2003).

[0439] Synthetic exendin-4 is also known as exenatide and is commercially available as BYETTA® (Amylin Pharmaceuticals, Inc. and Eli Lilly and Company). A once-weekly formulation of exenatide is described in WO2005 / 102293, the disclosure of which is incorporated herein by reference.

[0440] "Exendin analog" refers to a peptide that induces the biological activity of an exendin reference peptide, preferably with a potency equal to or greater than the exendin reference peptide (e.g., exendin-4) or within 5 orders of magnitude (plus or minus) of potency compared to the exendin reference peptide, as assessed by measures known in the art, such as receptor binding and / or competition assays, such as those described, for example, by Hargrove et al., Regulatory Peptides, 141:113-119 (2007), the disclosure of which is incorporated herein by reference. Preferably, the exendin analog will bind with an affinity of less than 1 μM in such assays, more preferably with an affinity of less than 3 nM, less than 1 nM, or less than 0.1 nM. The term "exendin analog" may also be referred to as an "exendin agonist." In a preferred embodiment, the exendin analog is an exendin-4 analog.

[0441] Exendin analogs also include peptides described herein that have been chemically derivatized or modified, such as peptides with unnatural amino acid residues (e.g., taurine, β-amino acid residues, γ-amino acid residues, and D-amino acid residues), peptides with C-terminal functional group modifications such as amide, ester, and C-terminal ketone modifications, and peptides with N-terminal functional group modifications such as acylated amines, Schiff bases, or cyclizations, such as found in the amino acid pyroglutamic acid. Exendin analogs can also include other chemical moieties, such as peptidomimetics.

[0442] Examples of exendins and exendin analogs include exendin-4 (SEQ ID NO: 1223), exendin-3 (SEQ ID NO: 1224), Leu 14 -Exendin-4 (SEQ ID NO: 1225), Leu 14 ,Phe 25 -Exendin-4 (SEQ ID NO: 1226), Leu 14 ,Ala 19,Phe 25 -Exendin-4 (SEQ ID NO: 1227), Exendin-4(1-30) (SEQ ID NO: 1228), Leu 14 -Exendin-4(1-30) (SEQ ID NO: 1229), Leu 14 ,Phe 25 -Exendin-4(1-30) (SEQ ID NO: 1230), Leu 14 ,Ala 19 ,Phe 25 -Exendin-4(1-30) (SEQ ID NO: 1231), Exendin-4(1-28) (SEQ ID NO: 1232), Leu 14 -Exendin-4(1-28) (SEQ ID NO: 1233), Leu 14 ,Phe 25 -Exendin-4(1-28) (SEQ ID NO: 1234), Leu 14 ,Ala 19 ,Phe 25 -Exendin-4(1-28) (SEQ ID NO: 1235), Leu 14 ,Lys 17,20 ,Ala 19 ,Glu 21 ,Phe 25 ,Gln 28 -Exendin-4 (SEQ ID NO: 1236), Leu 14 ,Lys 17,20 ,Ala 19 ,Glu 21 ,Gln 28 -Exendin-4 (SEQ ID NO: 1237), OctylGly 14 ,Gln 28 -Exendin-4 (SEQ ID NO: 1238), Leu 14 ,Gln 28 ,OctylGly 34 -Exendin-4 (SEQ ID NO: 1239), Phe 4 ,Leu 14 ,Gln 28 ,Lys 33 ,Glu 34 ,Ile 35,36 ,Ser 37 -Exendin-4(1-37) (SEQ ID NO: 1240), Phe 4 ,Leu 14 ,Lys17,20 ,Ala 19 ,Glu 21 ,Gln 28 - Exendin-4 (SEQ ID NO: 1241), Val 11 ,Ile 13 ,Leu 14 ,Ala 16 ,Lys 21 ,Phe 25 -Exendin-4 (SEQ ID NO: 1242), Exendin-4-Lys 40 (SEQ ID NO: 1243), lixisenatide (Sanofi-Aventis / Zealand Pharma), CJC-1134 (ConjuChem, Inc.), [N e -(17-Carboxyheptadecanoic acid)Lys 20 ]Exendin-4-NH2 (SEQ ID NO: 1268), [N e -(17-carboxyhepta-decanoyl)Lys 32 ]Exendin-4-NH2 (SEQ ID NO: 1269), [Desamino-His 1 ,N e -(17-Carboxyheptadecanoyl)Lys 20 ]Exendin-4-NH2 (SEQ ID NO: 1270), [Arg 12,27 ,NLe 14 ,N e -(17-Carboxy-heptadecanoyl)Lys 32 ]Exendin-4-NH2 (SEQ ID NO: 1271), [N e -(19-carboxy-nonadecanoylamino)Lys 20 ]-Exendin-4-NH2 (SEQ ID NO: 1272), [N e -(15-carboxypentadecanoylamino)Lys 20 ]-Exendin-4-NH2 (SEQ ID NO: 1273), [N e -(13-carboxytridecanoylamino)Lys 20 ]Exendin-4-NH2 (SEQ ID NO: 1274), [N e -(11-Carboxy-undecanoyl-amino)Lys 20 ]Exendin-4-NH2 (SEQ ID NO: 1275), Exendin-4-Lys40 (e-MPA)-NH2 (SEQ ID NO: 1276), Exendin-4-Lys 40 (e-AEEA-AEEA-MPA)-NH2 (SEQ ID NO: 1277), Exendin-4-Lys 40 (e-AEEA-MPA)-NH2 (SEQ ID NO: 1278), Exendin-4-Lys 40 (e-MPA)-Albumin (SEQ ID NO: 1279), Exendin-4-Lys 40 (e-AEEA-AEEA-MPA)-Albumin (SEQ ID NO: 1280), Exendin-4-Lys 40 (e-AEEA-MPA)-albumin (SEQ ID NO: 1281), and the like. AEEA refers to [2-(2-amino)ethoxy)]acetic acid. EDA refers to ethylenediamine. MPA refers to maleimidopropionic acid. Exendin and exendin analogs may be optionally amidated.

[0443] In one embodiment, the GLP-1 receptor agonist compound is an exendin-4 analog having at least 80% sequence identity to exendin-4 (SEQ ID NO: 1223), an exendin-4 analog having at least 85% sequence identity to exendin-4 (SEQ ID NO: 1223), an exendin-4 analog having at least 90% sequence identity to exendin-4 (SEQ ID NO: 1223), or an exendin-4 analog having at least 95% sequence identity to exendin-4 (SEQ ID NO: 1223).

[0444] Other exendins and exendin analogs useful in the methods described herein include those described in WO98 / 05351, WO99 / 07404, WO99 / 25727, WO99 / 25728, WO99 / 40788, WO00 / 41546, WO00 / 41548, WO00 / 73331, WO01 / 51078, WO03 / 099314, U.S. Pat. No. 6,956,026, U.S. Pat. No. 6,506,724, U.S. Pat. No. 6,741,623, and U.S. Pat. Nos. 6,858,576, 6,872,700, 6,902,744, 7,157,555, 7,223,725, 7,220,721, U.S. Publication No. 2003 / 0036504, and U.S. Publication No. 2006 / 0094652, the disclosures of which are incorporated herein by reference in their entireties.

[0445] "GLP-1(7-37) analog" refers to a peptide that induces biological activity similar to that of GLP-1(7-37) as assessed by measures known in the art, such as receptor binding assays or in vivo blood glucose assays, such as those described, for example, by Hargrove et al., Regulatory Peptides, 141:113-119 (2007), the disclosure of which is incorporated herein by reference. In one embodiment, the term "GLP-1(7-37) analog" refers to a peptide having an amino acid sequence that has one, two, three, four, five, six, seven, or eight amino acid substitutions, insertions, deletions, or a combination of two or more thereof, compared to the amino acid sequence of GLP-1(7-37). In one embodiment, the GLP-1(7-37) analog is GLP-1(7-36)-NH2. GLP-1(7-37) analogs include the amidated form, the acid form, pharma- ceutically acceptable salt forms, and any other physiologically active form of the molecule.

[0446] Examples of GLP-1(7-37) and GLP-1(7-37) analogs include GLP-1(7-37) (SEQ ID NO: 1244), GLP-1(7-36)-NH2 (SEQ ID NO: 1245), liraglutide (VICTOZA® supplied by Novo Nordisk), albiglutide (SYNCRIA® supplied by GlaxoSmithKline), taspoglutide (Hoffman La-Roche), dulaglutide (also known as LY2189265 (Eli Lilly and Company)), LY2428757 (Eli Lilly and Company), desamino-His 7 ,Arg 26 ,Lys 34 (N ε -(γ-Glu(N-α-hexadecanoyl)))-GLP-1(7-37) (core peptide disclosed as SEQ ID NO: 1282), desamino-His 7 ,Arg 26 ,Lys 34 (N ε -octanoyl)-GLP-1(7-37) (SEQ ID NO: 1283), Arg 26,34 ,Lys 38 (N ε -(ω-carboxypentadecanoyl)-GLP-1(7-38) (SEQ ID NO: 1284), Arg 26,34 ,Lys 36 (N ε -(γ-Glu(N-α-hexadecanoyl)))-GLP-1(7-36) (core peptide disclosed as SEQ ID NO: 1285), Aib 8,35 ,Arg 26,34 ,Phe 31 -GLP-1(7-36)) (SEQ ID NO: 1246), HXaa8EGTFTSDVSSYLEXaa 22 Xaa 23 AAKEFIXaa 30 WLXaa 33 Xaa 34 G Xaa 36 Xaa 37 (Xaa3 is A, V, or G; 22 is G, K, or E; Xaa 23 is Q or K, and Xaa30 is A or E, and Xaa 33 is V or K, and Xaa 34 is K, N, or R; Xaa 36 is R or G, and Xaa 37 is G, H, P, or absent) (SEQ ID NO: 1247), Arg 34 -GLP-1(7-37) (SEQ ID NO: 1248), Glu 30 -GLP-1(7-37) (SEQ ID NO: 1249), Lys 22 -GLP-1(7-37) (SEQ ID NO: 1250), Gly 8,36 ,Glu 22 -GLP-1(7-37) (SEQ ID NO: 1251), Val 8 ,Glu 22 ,Gly 36 -GLP-1(7-37) (SEQ ID NO: 1252), Gly 8,36 ,Glu 22 ,Lys 33 ,Asn 34 -GLP-1(7-37) (SEQ ID NO: 1253), Val 8 ,Glu 22 ,Lys 33 ,Asn 34 ,Gly 36 -GLP-1(7-37) (SEQ ID NO: 1254), Gly 8,36 ,Glu 22 ,Pro 37 -GLP-1(7-37) (SEQ ID NO: 1255), Val 8 ,Glu 22 ,Gly 36 Pro 37 -GLP-1(7-37) (SEQ ID NO: 1256), Gly 8,36 ,Glu 22 ,Lys 33 ,Asn 34 ,Pro 37 -GLP-1(7-37) (SEQ ID NO: 1257), Val 8 ,Glu 22 ,Lys 33 ,Asn 34 ,Gly 36 ,Pro 37 -GLP-1(7-37) (SEQ ID NO: 1258), Gly 8,36,Glu 22 -GLP-1(7-36) (SEQ ID NO: 1259), Val 8 ,Glu 22 ,Gly 36 -GLP-1(7-36) (SEQ ID NO: 1260), Val 8 ,Glu 22 ,Asn 34 ,Gly 36 -GLP-1(7-36) (SEQ ID NO: 1261), Gly 8,36 ,Glu 22 ,Asn 34 -GLP-1(7-36) (SEQ ID NO: 1262). GLP-1(7-37) and GLP-1(7-37) analogs may each be optionally amidated.

[0447] In one embodiment, GLP-1(7-37) or a GLP-1(7-37) analog is covalently linked (directly or via a linking group) to the Fc portion of an immunoglobulin (e.g., IgG, IgE, IgG, and the like). For example, [ka] (Xaa 16 is P or E, and Xaa 17 is F, V, or A; Xaa 18 is L, E, or A; Xaa 80 is N or A, and Xaa 230 Any one of SEQ ID NOs: 25-40 may be covalently linked to the Fc portion of an immunoglobulin comprising the sequence (-GGGGS-) (SEQ ID NO: 1263). The linking group can be any chemical moiety (e.g., amino acids and / or chemical groups). In one embodiment, the linking group is (-GGGGS-) x (SEQ ID NO: 1264) (x is 1, 2, 3, 4, 5, or 6, preferably 2, 3, or 4, and more preferably 3). In one embodiment, [ka] The GLP-1(7-37) analog is covalently linked to the Fc portion of an immunoglobulin comprising the amino acid sequence:

[0448] In another embodiment, GLP-1(7-37) or a GLP-1(7-37) analog may be covalently linked (directly or via a linking group) to one or two polyethylene glycol molecules. For example, a GLP-1(7-37) analog may be 22 QAAKEFIAWLXaa 33 KGGPSSGAPPPC 45 C 46 -Z (Xaa8 is D-Ala, G, V, L, I, S, or T; 22 is G, E, D, or K; Xaa 33 is V or I, and Z is OH or NH2 (SEQ ID NO: 1266), optionally comprising: (i) one polyethylene glycol moiety covalently attached to C 45 or (ii) one polyethylene glycol moiety is covalently attached to C 46 or (iii) one polyethylene glycol moiety is added to C 45 and one polyethylene glycol moiety is C 46 In one embodiment, the GLP-1(7-37) analog is HVEGTFTSDVSSYLEEQAAKEFIAWLIKGGPSSGAPPPC 45 C 46 -NH2 (SEQ ID NO: 1267), optionally having (i) one polyethylene glycol moiety covalently attached to C4 or (ii) one polyethylene glycol moiety covalently attached to C 46 or (iii) one polyethylene glycol moiety is added to C 45 and one polyethylene glycol moiety is C 46 is added to.

[0449] In one embodiment, the GLP-1 receptor agonist compound is a peptide having at least 80% sequence identity to GLP-1(7-37) (SEQ ID NO: 1244), a peptide having at least 85% sequence identity to GLP-1(7-37) (SEQ ID NO: 1244), a peptide having at least 90% sequence identity to GLP-1(7-37) (SEQ ID NO: 1244), or a peptide having at least 95% sequence identity to GLP-1(7-37) (SEQ ID NO: 1244).

[0450] GLP-1 receptor agonist compounds may be prepared by processes well known in the art, such as peptide purification as described in Eng et al., J. Biol. Chem., 265:20259-62 (1990), standard solid-phase peptide synthesis techniques as described in Raufman et al., J. Biol. Chem., 267:21432-37 (1992), recombinant DNA techniques as described in Sambrook et al., Molecular Cloning: A Laboratory Manual, 2d Ed., Cold Spring Harbor (1989), and the like.

[0451] [Table 153]

[0452] [Table 154]

[0453] [Table 155]

[0454] [Table 156]

[0455] [Table 157]

[0456] AEEA refers to [2-(2-amino)ethoxy)]acetic acid.

[0457] EDA refers to ethylenediamine.

[0458] MPA refers to maleimidopropionic acid.

[0459] The present disclosure also provides pharmaceutical compositions comprising the GLP-1 receptor agonist compounds described herein and pharma- ceutical acceptable carriers.The GLP-1 receptor agonist compounds can be present in the pharmaceutical composition in a therapeutically effective amount, and can be present in an amount that provides the minimum plasma level of the GLP-1 receptor agonist compounds required for therapeutic efficacy.Such pharmaceutical compositions are known in the art and are described, for example, in U.S. Patent No. 7,521,423, U.S. Patent No. 7,456,254, WO2000 / 037098, WO2005 / 021022, WO2005 / 102293, WO2006 / 068910, WO2006 / 125763, WO2009 / 068910, U.S. Publication No. 2004 / 0106547, and the like, the disclosures of which are incorporated herein by reference.

[0460] Pharmaceutical compositions containing the GLP-1 receptor agonist compounds described herein may be provided for peripheral administration, such as parenteral (e.g., subcutaneous, intravenous, intramuscular), continuous infusion (e.g., intravenous drip, intravenous bolus, intravenous infusion), topical, nasal, or oral administration. Suitable pharmaceutical acceptable carriers and their formulations are described in standard formulation treatises, such as Remington's Pharmaceutical Sciences by Martin;and Wang et al., Journal of Parenteral Science and Technology, Technical Report No. 10, Supp. 42: 2S (1988). The GLP-1 receptor agonist compounds described herein can be provided in parenteral compositions for injection or infusion. The GLP-1 receptor agonist compounds can be suspended, for example, in water, inert oil, such as vegetable oil (e.g., sesame oil, peanut oil, olive oil, and the like), or other pharmaceutical acceptable carriers. In one embodiment, the compound is suspended in an aqueous carrier, such as an isotonic buffer solution having a pH of about 3.0 to 8.0 or about 3.0 to 5.0. The composition may be sterilized by conventional sterilization techniques or by filtration. The composition may contain pharma- ceutically acceptable auxiliary substances, such as pH buffers, as necessary to approximate physiological conditions.

[0461] Useful buffers include, for example, acetate buffers. A sustained release preparation in the form of a repository or "depot" may be used to deliver therapeutically effective amounts of the preparation to the bloodstream over a period of hours or days after subcutaneous injection, transdermal injection, or other delivery method. The desired isotonicity may be achieved using sodium chloride or other pharma- ceutically acceptable agents such as dextrose, boric acid, sodium tartrate, propylene glycol, polyols (such as mannitol and sorbitol), or other inorganic or organic solutes. In one embodiment, for intravenous infusion, the formulation may include (i) a GLP-1 receptor agonist compound, (2) sterile water, and, optionally, (3) sodium chloride, dextrose, or a combination thereof.

[0462] Carriers or excipients can also be used to facilitate administration of the GLP-1 receptor agonist compound. Examples of carriers and excipients include calcium carbonate, calcium phosphate, various sugars such as lactose, glucose, or sucrose, or various types of starch, cellulose derivatives, gelatin, vegetable oils, polyethylene glycols, and physiologically compatible solvents.

[0463] The GLP-1 receptor agonist compound can also be formulated as a pharma- ceutically acceptable salt (e.g., acid addition salt) and / or its complex. A pharma-ceutically acceptable salt is a salt that is non-toxic at the concentration at which it is administered. Pharmaceutically acceptable salts include acid addition salts such as those containing sulfate, hydrochloride, phosphate, sulfamate, acetate, citrate, lactate, tartrate, methanesulfonate, ethanesulfonate, benzenesulfonate, p-toluenesulfonate, cyclohexylsulfamate, and quinate. Pharmaceutically acceptable salts can be obtained from acids such as hydrochloric acid, sulfuric acid, phosphoric acid, sulfamic acid, acetic acid, citric acid, lactic acid, tartaric acid, malonic acid, methanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, cyclohexylsulfamic acid, and quinic acid. Such salts may be prepared, for example, by reacting the product in its free acid or base form with one or more equivalents of the appropriate base or acid in a solvent or medium in which the salt is insoluble or in a solvent such as water which is then removed by vacuum or lyophilization, or by exchanging an existing salt ion for another ion with a suitable ion exchange resin.

[0464] Examples of pharmaceutical formulations of GLP-1 receptor agonist compounds are described in U.S. Pat. No. 7,521,423, U.S. Pat. No. 7,456,254, U.S. Publication No. 2004 / 0106547, WO2006 / 068910, WO2006 / 125763, and the like, the disclosures of which are incorporated herein by reference.

[0465] The therapeutically effective amount of the GLP-1 receptor agonist compound described herein for use in the methods described herein will typically be about 0.01 μg to about 5 mg, about 0.1 μg to about 2.5 mg, about 1 μg to about 1 mg, about 1 μg to about 50 μg, or about 1 μg to about 25 μg. Alternatively, the therapeutically effective amount of the GLP-1 receptor agonist compound may be about 0.001 μg to about 100 μg based on a patient weight of 70 kg, or about 0.01 μg to about 50 μg based on a patient weight of 70 kg. The frequency of administration of such therapeutically effective doses may be once per day, twice per day, three times per day, once per week, twice per week, or once per month, depending on the formulation. The exact dose to be administered will be determined by the formulation, e.g., immediate release formulation or sustained release formulation. For transdermal, nasal, or oral dosage forms, the dosage may be increased by about 5-fold to about 10-fold.

[0466] In certain embodiments, the GLP-1 receptor agonist will be administered simultaneously with the GIPR antigen binding protein. In one embodiment, the GLP-1 receptor agonist will be administered after the GIPR antigen binding protein. In one embodiment, the GLP-1 receptor agonist will be administered before the GIPR antigen binding protein. In certain embodiments, the subject or patient will already be treated with a GLP-1 receptor agonist before receiving additional treatment with the GIPR antigen binding protein.

[0467] The GIPR antigen binding protein provided herein is useful for detecting GIPR in biological samples.For example, the GIPR antigen binding protein can be used in diagnostic assays, such as binding assays, for example, to detect and / or quantify expressed GIPR in serum.

[0468] The described antigen binding proteins can be used for diagnostic purposes to detect, diagnose, or monitor diseases and / or conditions associated with GIPR. The disclosed antigen binding proteins provide a means to detect the presence of GIPR in a sample using classical immunohistological methods known to those skilled in the art (e.g., Tijssen, 1993, Practice and Theory of Enzyme Immunoassays, Vol 15 (Eds RH Burdon and PH van Knippenberg, Elsevier, Amsterdam); Zola, 1987, Monoclonal Antibodies: A Manual of Techniques, pp. 147-158 (CRC Press, Inc.); Jalkanen et al., 1985, J. Cell. Biol. 101: 976-985; Jalkanen et al., 1987, J. Cell Biol. 105: 3087-3096). Detection of GIPR can be performed in vivo or in vitro.

[0469] The diagnostic applications provided herein include the use of antigen binding proteins to detect the expression of GIPR. Examples of methods useful in detecting the presence of GIPR include immunoassays such as enzyme-linked immunosorbent assay (ELISA) and radioimmunoassay (RIA).

[0470] For diagnostic applications, the antigen binding protein will typically be labeled with a detectable labeling group. Suitable labeling groups include, but are not limited to, radioisotopes or radionuclides (e.g., 3 H, 14 C. 15 N, 35 S, 90 Y, 99 Tc, 111 In, 125 I, 131I), fluorescent groups (e.g., FITC, rhodamine, lanthanide phosphors), enzymatic groups (e.g., horseradish peroxidase, β-galactosidase, luciferase, alkaline phosphatase), chemiluminescent groups, biotin 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. A variety of methods for labeling proteins are known in the art and may be used.

[0471] In some embodiments, GIPR antigen binding proteins are isolated and measured using techniques known in the art, see, e.g., Harlow and Lane, 1988, Antibodies: A Laboratory Manual, New York: Cold Spring Harbor (ed. 1991 and periodic supplements); John E. Coligan, ed., 1993, Current Protocols In Immunology New York: John Wiley & Sons.

[0472] In another aspect of the present disclosure, the detection of the presence of a test molecule that competes with the provided antigen binding protein for binding to GIPR is provided. In one example of such an assay, it is envisioned that the amount of free antigen binding protein is detected in a solution containing a certain amount of GIPR in the presence or absence of a test molecule. If the amount of free antigen binding protein (i.e., antigen binding protein that is not bound to GIPR) increases, it indicates that the test molecule has the ability to compete with the antigen binding protein for binding to GIPR. 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 the antigen binding protein.

[0473] GIPR binding protein can be used for the treatment, diagnosis or amelioration of metabolic conditions or disorders.In one embodiment, the metabolic disorder to be treated is diabetes, such as type 2 diabetes.In another embodiment, the metabolic condition or disorder is obesity.In other embodiments, the metabolic condition or disorder is dyslipidemia, elevated glucose level, elevated insulin level, or diabetic nephropathy. For example, metabolic conditions or disorders that can be treated or ameliorated using GIPR-binding peptides include conditions in human subjects having fasting blood glucose levels of 125 mg / dL or greater (e.g., 130 mg / dL, 135 mg / dL, 140 mg / dL, 145 mg / dL, 150 mg / dL, 155 mg / dL, 160 mg / dL, 165 mg / dL, 170 mg / dL, 175 mg / dL, 180 mg / dL, 185 mg / dL, 190 mg / dL, 195 mg / dL, 200 mg / dL, or greater than 200 mg / dL). Blood glucose levels can be determined in a fed or fasted state, or randomly. Metabolic conditions or disorders can also include conditions in which a subject is at increased risk of developing a metabolic condition. For human subjects, such conditions include fasting blood glucose levels of 100 mg / dL. Conditions that can be treated using pharmaceutical compositions that contain GIPR binding proteins can also be found in American Diabetes Association Standards of Medical Care in Diabetes Care-2011, American Diabetes Association, Diabetes Care Vol.34, No.Supplement 1, S11-S61, 2010, which is incorporated herein by reference.

[0474] In application, a therapeutically effective dose of GIPR binding protein can be administered to a patient in need thereof to treat metabolic disorders or conditions such as type 2 diabetes, elevated glucose levels, elevated insulin levels, dyslipidemia, obesity, or diabetic nephropathy. Administration can be performed as described herein, such as by intravenous (IV) injection, intraperitoneal (IP) injection, subcutaneous injection, intramuscular injection, or orally in the form of a tablet or liquid formulation. In some circumstances, a therapeutically effective or preferred dose of GIPR binding protein can be determined by a clinician. A therapeutically effective dose of GIPR binding protein will depend, among other things, on the administration schedule, the unit dose of the drug administered, whether the GIPR binding protein is administered in combination with other therapeutic agents, and the immune status and health of the recipient. As used herein, the term "therapeutically effective dose" refers to an amount of GIPR binding protein that elicits a biological or medicinal response in a tissue system, animal, or human being sought by a researcher, physician, or other clinician, including reduction or amelioration of symptoms of the disease or disorder being treated, i.e., an amount of GIPR binding protein that supports an observable level of one or more desired biological or medicinal responses, such as reduced blood glucose, insulin, triglyceride, or cholesterol levels, weight loss, or improved glucose tolerance, energy expenditure, or insulin sensitivity.

[0475] It should be noted that the therapeutically effective dose of GIPR binding protein can also vary according to desired outcome.Therefore, for example, in the situation where blood glucose level is indicated to be low, the dose of GIPR binding protein is accordingly higher than the dose where blood glucose level is desired to be relatively low.On the contrary, in the situation where blood glucose level is indicated to be high, the dose of GIPR binding protein is accordingly lower than the dose where blood glucose level is desired to be relatively high.

[0476] In various embodiments, the subject is a human with a blood glucose level of 100 mg / dL or greater and can be treated with a GIPR binding protein.

[0477] In one embodiment, the method of the present disclosure includes first measuring the baseline level of one or more metabolically related compounds, such as glucose, insulin, cholesterol, lipids, etc., in a subject. Then, a pharmaceutical composition comprising a GIPR binding protein is administered to the subject. After a desired period of time, the level of one or more metabolically related compounds (e.g., blood glucose, insulin, cholesterol, lipids) in the subject is measured again. The two levels can then be compared to determine the relative change in metabolically related compounds in the subject. Depending on the result of the comparison, another dose of a pharmaceutical composition comprising a GIPR binding protein can be administered to achieve the desired level of one or more metabolically related compounds.

[0478] It should be noted that the pharmaceutical composition comprising GIPR binding protein can be co-administered with another compound.The identity and characteristics of the compound co-administered with GIPR binding protein will depend on the nature of the disease state to be treated or ameliorated.A non-limiting list of examples of compounds that can be administered in combination with the pharmaceutical composition comprising GIPR binding protein includes rosiglitizone, pioglitizone, repaglinide, nateglitinide, metformin, exenatide, sitagliptin, pramlintide, glipizide, glimeprirideacarbose, and miglitol.

[0479] Kits for carrying out the disclosed methods are also provided. Such kits may include pharmaceutical compositions such as those described herein, including pharmaceutical compositions comprising nucleic acids encoding the peptides or proteins provided herein, vectors and cells comprising such nucleic acids, and compounds comprising such nucleic acids, which may be provided in a sterile container. Optionally, instructions on how to use the provided pharmaceutical compositions in treating metabolic disorders may be included or made available to patients or health care providers.

[0480] In one aspect, the kit comprises (a) a pharmaceutical composition comprising a therapeutically effective amount of a GIPR binding protein, and (b) one or more containers for the pharmaceutical composition. Such a kit may also include instructions for its use, which may be adapted to suit the metabolic disorder being treated. The instructions may describe the use and properties of the materials provided in the kit. In certain embodiments, the kit includes instructions for the patient to administer to treat metabolic disorders such as elevated glucose levels, elevated insulin levels, obesity, type 2 diabetes, dyslipidemia, or diabetic nephropathy.

[0481] The instructions may be printed on a substrate such as paper or plastic, and may be present in the kit as a package insert, on a label on a container of the kit or its components (e.g., associated with the packaging), etc. In other embodiments, the instructions are present as an electronic stored data file present on a suitable computer readable storage medium, for example, a CD-ROM, diskette, etc. In yet other embodiments, the instructions are not present in the kit, but rather a means is provided for obtaining the instructions from a remote source, such as using the Internet. An example of this embodiment is a kit that includes a web address where the instructions can be viewed and / or from which the instructions can be downloaded.

[0482] It will often be desirable for some or all of the kit components to be packaged in suitable packaging to maintain sterility. The kit components can be packaged in a kit storage element that provides an easy to handle single unit, e.g., a box or similar structure, which may or may not be an airtight container to further maintain the sterility of some or all of the kit components. EXAMPLES

[0483] Example 1 Materials and Methods - Yeast Display gBlock and degenerate codon primers (IDT DNA) were used for the construction of yeast-displayed molecules and libraries. Standard PCR and overlap assembly PCR were performed using Q5 HotStart polymerase (NEB). Homologous recombination of inserts and digested vectors previously purified with a PCR purification kit (Qiagen) was used for transformation into yeast strain BJ5464 (ATCC) by electroporation. Briefly, BJ5464 cells were harvested from YPD agar plates and grown overnight at 30°C in YPD medium. Cells were then expanded to a starting OD of 0.2 and grown for 6 hours at 30°C. After pelleting and resuspension at room temperature, 10 mM Tris, 100 mM LiOAc, 0.6 M sorbitol, 10 mM DTT was added, followed by incubation at RT for 30 minutes with gentle shaking at 220 rpm. Pellet the cells, wash with cold 1 M sorbitol, 1 mM CaCl2, and add 2 x 10 10Cells were resuspended at 10000 cells / mL. Electroporation of 0.5ug DNA in 5uL volume / cuget with 120uL cells using the following settings: 540V; 25uF; ∞ ohms. Cells were quickly rescued with 2mL YPD, 0.5M sorbitol, 0.5mM CaCl2. After 1 hour recovery at 30°C, cells were transferred to SCD glucose medium without leucine or uracil and passaged for 2 days at 30°C. Induction of displayed Fab molecules was performed by medium change to SCD galactose medium allowing activation of Gal10 promoter. Cells were induced for 48 hours at 20°C. To measure the amount of displayed molecules, Alexafluor 647-conjugated anti-huFab antibody was used to assess surface displayed molecules. Antigen binding was measured using biotin-conjugated GIPR ECD fragment 21-129aa together with streptavidin PE molecules. Fluorescence was measured using a BD Canto or sorted using an AriaII FACS. Yeast were sorted and then grown on SCD-leu-ura agar plates. After 2-3 days of growth at 30°C, clones were picked and grown in SCD-leu-ura medium. PCR was performed from yeast cultures using Phire Plant Direct PCR (ThermoFisher). PCR samples were then sequenced by Genewiz.

[0484] Molecular Biology / Golden Gate Assembly The PCR samples used for sequencing were also used as templates for molecular cloning. In the gated assembly (GGA) strategy, PCR was used to add compatible cloning ends. Briefly, GGA relies on type II restriction enzymes to cut and seamlessly join multiple DNA fragments. In this example, the multiple DNA fragments consist of a synthetic nucleic acid sequence (gBlock, Integrated DNA Technologies, Coralville, IA) encoding a signal, another gBlock encoding an anti-GIPR variable domain, an antibody constant domain fragment released from a part vector, and an expression vector backbone. Table 9 depicts these representative fragments.

[0485] [Table 158]

[0486] The GGA reaction consisted of 10 ng part 1, 10 ng PCR product, 10 ng part 2, 10 ng expression vector, 1 μl 10× Fast Digest Reaction Buffer+0.5 mM ATP (Thermo Fisher, Waltham, MA), 0.5 μl FastDigest Esp3I (Thermo Fisher, Waltham, MA), 1 μl T4 DNA Ligase (5 U / μl, Thermo Fisher, Waltham, MA), and water up to 10 μl. The reaction was carried out for 15 cycles consisting of a 2 min digestion step at 37°C and a 3 min ligation step at 16°C. The 15 cycles were followed by a final 5 min digestion step at 37°C and a 5 min enzyme inactivation step at 80°C. DNA amplification was performed using chemically competent Top10 cells (Invitrogen). Colonies were picked and grown overnight at 37° C. in 2XYT medium, then DNA was extracted and purified using the Turbo kit QiaRobot (Qiagen). DNA sequences were confirmed and then expressed in mammalian cells.

[0487] Mammalian Expression Monoclonal antibodies were stably expressed in suspension compatible with CHO-K1 cells together with the corresponding cDNA. Transfection was performed using Lipofectamine LTX (Thermo Fisher) according to the manufacturer's protocol. Briefly, a total of 2 μg of mammalian expression plasmid DNA was used at a 1:1 heavy / light chain ratio. For each, the plasmid DNA was added to 0.5 ml of OPTI-MEM (Thermo Fisher) and mixed. In a separate tube, 10 μl of Lipofectamine LTX was added to 0.5 ml of OPTI-MEM. These solutions were incubated for 5 minutes at room temperature. To form the transfection complex, the DNA and Lipofectamine LTX mixtures for each were combined and incubated for an additional 10 minutes at room temperature.

[0488] Log-phase CHO-K1 cells were pelleted by centrifugation (1200-1500 RPM for 5 min), washed once with 1x PBS (Thermo Fisher), and resuspended in OPTI-MEM to 1.5-2e6 viable cells / mL. For each transfection, 1 mL of the washed cells was added to the transfection complex in a 24-deep-well block. The plate was incubated at 36°C, 5% CO2 with shaking at 225 RPM for 6 min. To stop the transfection, 2 mL of growth medium was added to each flask and incubated for 48 h.

[0489] To initiate selection, cells were pelleted by centrifugation (1200-1500 RPM for 5 min) 48 hours after transfection and the medium was replaced with 4 mL of growth medium supplemented with antibiotics (puromycin 10 mg / L / hygromycin 600 mg / L). Selection medium was replaced 2-3 times per week, diluting cultures as needed to prevent overgrowth (<5-6e6 vc / mL) until cell viability and density were restored.

[0490] Production (100 mL) was carried out in shake flasks at 36° C. The product was inoculated at 2e6 vc / mL in production medium. Conditioned medium was harvested on day 7 by centrifugation followed by filtration (0.45 μm).

[0491] purification Molecules were purified from cell culture medium using an AKTA Purifier (GE Healthcare Life Sciences, Little Chalfont, Buckinghamshire, UK) tandem liquid chromatography system with 1 mL of MabSelect SuRe (MSS) HiTrap (GE Healthcare Life Sciences) as the first column and 5 mL of Desalting HiTrap (GE Healthcare Life Sciences) as the second column. The medium was loaded directly onto the MSS column, which was then washed with 8 column volumes (CV) of 25 mM Tris-HCl, 100 mM NaCl, pH 7.4, and eluted with 2 CV of 100 mM acetic acid. The MSS column eluate was automatically sent to a desalting column where the protein was eluted isocratically with 4 CV of 10 mM Na acetate, 150 mM NaCl, pH 5.2. Samples were sterile filtered through a 3.0 μm glass fiber / 0.2 μm Supor membrane (Pall Corporation, Port Washington, New York, USA).

[0492] Protein concentration determination The protein concentration of each purified molecule was determined by UV absorbance at 280 nm using a Multiskan GO microplate spectrophotometer (Thermo Fisher Scientific, Rockford, Illinois, USA).

[0493] Size exclusion chromatography analysis Samples were analyzed by size exclusion chromatography using an ACQUITY UPLC Protein BEH SEC column, 200 Å, 1.7 μm, 4.6 × 300 mm (Waters Corporation, Milford, Massachusetts, USA) run in 100 mM NaH2PO4, 50 mM NaCl, 7.5% ethanol, pH 6.9, with absorbance at 280 nm monitored at 0.4 mL / min on a Waters ACQUITY UPLC system (Waters Corporation).

[0494] LCMS method For LCMS analysis, 20 μg of material was denatured with 8 M guanidine HCl / TRIS pH 8.0 (Teknova, Hollister, CA) and reduced with 10 mM DTT (EMD Millipore, Darmstadt, Germany) for 20 min at 50 °C. Samples were acidified with trifluoroacetic acid and 3 μg was injected onto a Waters BEH reversed-phase C4 column using a Waters Acquity HPLC (Waters Corporation, Milford, MA). The column effluent was introduced into the electrospray source of a Xevo QTOF mass spectrometer (Waters Corporation, Milford, MA) and mass spectra were collected. Relevant spectra were deconvoluted using the MaxEnt algorithm within the Waters MassLynx software package. The resulting LC and HC mass spectra were compared to the theoretically calculated masses of each chain and indicated as pass / fail.

[0495] Tagg method: Thermal aggregation onset temperatures were determined by performing stepwise thermal denaturation and aggregation studies on an Avacta, Optim-1000. Data analysis and Tagg determinations were performed using Optim Analysis software (Igor version 6.31). Samples with concentrations greater than 1 mg / ml were normalized to 1.0 mg / ml with formulation buffer prior to Tagg analysis. A step thermal ramp was applied with a start temperature of 20° C. and a stop temperature of 90° C. The temperature step was 1° C. and the temperature hold time was 30 seconds. The exposure time was 500 ms, the center wavelength was 380 nm, and the slit was 250 μm.

[0496] cAMP method method This method is aimed at quantifying cAMP in HEK293T cells expressing human GIPR or cynomolgus GIPR. Binding of GIP induces a conformational change in GIPR, stimulating G protein to activate adenylate cyclase, which results in the production of cAMP from ATP. Binding of the antibody to GIPR prevents GIP binding to GIPR, which can be measured by a cAMP assay.

[0497] This cAMP assay is an HTRF immunoassay designed to measure cAMP produced upon regulation of adenylyl cyclase activity by GPCR. The assay is based on the competition between natural cAMP produced by cells and cAMP labeled with the dye d2 for binding sites on a cAMP-specific monoclonal antibody labeled with Eu3+-cryptate. When the Eu3+-cryptate-conjugated antibody binds to the cAMP-d2 tracer, a light pulse at 337 nm excites the Eu3+-cryptate. The energy released by the excited Eu3+-cryptate is transferred by FRET to the d2 molecule on the cAMP tracer, which then emits light at 665 nm. The residual energy from the Eu3+-cryptate will produce light at 620 nm. In the absence of free cAMP, a maximum HTRF signal is achieved. Free cAMP produced by stimulated cells competes with the cAMP-d2 tracer for binding to the anti-cAMP Eu3+-cryptate, causing a decrease in the HTRF signal. The specific signal (i.e., energy transfer) is inversely proportional to the concentration of cAMP in the sample.

[0498] Experimental design Cells were maintained in DMEM 10% FBS, 1x sodium pyruvate, 1x penicillin-streptomycin-glutamine, 2-5 μg / mL puromycin. Prior to experiments, cells were washed with DBPS and dissociated with 0.5 mM EDTA in DPBS.

[0499] To establish the GIP response, a solution of GIP (H2O) was prepared, serially diluted 3-fold (Ham's F-12 with 0.1% BSA) and added to 30,000 recombinant cells in a 96-well black round-bottom plate. The cells were incubated in the presence of 0.5 mM IBMX for 30 min at 5% CO2 and 37°C. After incubation, 25 μL of d2-cAMP, 25 μL of Eu3+-cryptate-cAMP antibody were added, followed by incubation at room temperature for 1 h. The wavelengths at 665 nm and 620 nm were measured using an EnVision Multilabel Reader, and the ratio 665 / 620 was calculated.

[0500] To establish the test substance response, a solution of the antibody (10 mM sodium acetate, 9% sucrose, pH 5.2) was prepared, serially diluted 3-fold (Ham's F-12 with 0.1% BSA) and added to 30,000 recombinant cells in a 96-well black round-bottom plate and the cells were incubated for 30 minutes at 5% CO2 and 37°C. GIP was added to a final concentration of 50 pM. The cells were incubated in the presence of 0.5 mM IBMX for 30 minutes at 5% CO2 and 37°C. After incubation, 25 μL of d2-cAMP, 25 μL of Eu3+-cryptate-cAMP antibody were added, followed by incubation at room temperature for 1 hour. The wavelengths at 665 nm and 620 nm were measured using an EnVision Multilabel Reader, and the ratio 665 / 620 was calculated.

[0501] Graphs were generated by plotting the concentrations of GIP and antibody on the abscissa against the average of the two replicate values ​​for cAMP levels (665 / 620 ratio). The data points were then fitted with the log (agonist) vs. response-variable slope GraphPad Prism to obtain fitted values ​​for top, bottom, hill slope, and both EC50 and IC50 (Table 10).

[0502] Validation of GIPR binding to variants of 2G10 affinity matured hits using BIAcore instrument, reagents, and sensor chip surface preparation: Biacore T200, Biacore series S-CM5 sensor chip, amine coupling kit, surfactant P-20, 10 mM sodium acetate, pH 4.0, and 10 mM glycine pH 1.5 were from GE (Pittsburgh, PA). Phosphate buffered saline (PBS, 1×, no calcium chloride, no magnesium chloride) was from Thermo Fisher Scientific (Waltham, MA). Bovine serum albumin (BSA, fraction V, IgG-free) was from Sigma-Aldrich Corp. (St. Louis, MO). Goat anti-huFc antibody was from Jackson ImmunoResearch Inc. (West Grove, PA).

[0503] Preparation of the sensor chip surface Immobilization of goat anti-huFc antibody onto the S-CM5 sensor chip surface was performed according to the manufacturer's instructions. The running buffer for the Biacore instrument was 0.005% P20 / PBS 1x, pH 7.4, no calcium chloride, no magnesium chloride. Briefly, carboxyl groups on the sensor chip surface were activated by injecting 60 μL of a mixture containing 0.2 M N-ethyl-N'-(dimethylaminopropyl)carbodiimide (EDC) and 0.05 M N-hydroxysuccinimide (NHS). Goat anti-huFc antibody was diluted to 20 μg / ml in 10 mM sodium acetate, pH 4.0, and injected over the activated chip surface at 30 uL / min for 6 min. Excess reactive groups on the surface were deactivated by injecting 60 μL of 1 M ethanolamine. The final immobilization level was approximately 8000 resonance units (RU).

[0504] Binding assays were performed on the immobilized goat anti-huFc antibody surface with a Biacore T200. The experiment was performed at 25°C. The instrument running buffer was 0.005% P20 / PBS. Goat anti-huFc capture antibody was covalently attached to the sensor chip surface by standard amine coupling to flow cell channels 1-4. Anti-huGIPR antibody was diluted to approximately 10 nM in sample buffer (0.1 mg / ml BSA, 0.005% P20, PBS) and then captured on flow cell channels 2, 3, and 4. Flow cell channel 1 was as a blank reference surface with no anti-huGIPR antibody injected. The response range of the captured antibody was approximately 250 RU. 200 nM huGIPR ECD was then injected over the goat anti-huFc antibody captured anti-huGIPR antibody surface at a flow rate of 50 ul / min for 3 minutes. After 10 min of dissociation, each surface was regenerated with two 30 s injections of 10 mM glycine, pH 1.5. Sensorgrams were analyzed using Biacore T200 evaluation software (v.2.0). The ks (1 / sec) for each antibody are shown in Table 1...

Claims

1. 1. A method of treating a subject having a metabolic disorder, comprising administering to the subject a therapeutically effective amount of an antigen binding protein that specifically binds to a protein having an amino acid sequence that has at least 90% amino acid sequence identity to the amino acid sequence of GIPR.

2. The method of claim 1 , wherein the metabolic disorder is a disorder of glucose metabolism.

3. 3. The method of claim 2, wherein the glucose metabolism disorder comprises hyperglycemia and the administration reduces plasma glucose in the subject.

4. 3. The method of claim 2, wherein the glucose metabolism disorder comprises hyperinsulinemia and the administration reduces plasma insulin in the subject.

5. 3. The method of claim 2, wherein the glucose metabolism disorder comprises impaired glucose tolerance, and wherein the administration improves glucose tolerance in the subject.

6. 3. The method of claim 2, wherein the glucose metabolism disorder comprises insulin resistance and the administering reduces insulin resistance in the subject.

7. The method of claim 2 , wherein the glucose metabolism disorder comprises diabetes.

8. The method of claim 2 , wherein the subject is obese.

9. 10. The method of claim 8, wherein said administering reduces the body weight of said subject.

10. 10. The method of claim 8, wherein said administration reduces weight gain in said subject.

11. 10. The method of claim 8, wherein said administration reduces body fat mass in said subject.

12. 9. The method of claim 8, wherein the glucose metabolism disorder comprises insulin resistance and the administering reduces insulin resistance in the subject.

13. 10. The method of claim 8, wherein the subject has advanced hepatic steatosis and the administration reduces hepatic steatosis in the subject.

14. 10. The method of claim 8, wherein the subject has increased liver fat content and the administration reduces liver fat content in the subject.

15. The method of claim 1 , wherein the subject is a mammal.

16. The method of claim 1 , wherein the subject is a human.

17. 10. The method of claim 1, wherein the administration is by parenteral injection.

18. 10. The method of claim 1, wherein the administration is by subcutaneous injection.

19. An isolated antigen binding protein that specifically binds to a human gastric inhibitory peptide receptor (GIPR) polypeptide.

20. 20. The isolated antigen binding protein of claim 19, wherein the human GIPR has a sequence comprising a sequence selected from the group consisting of SEQ ID NO:1201, SEQ ID NO:1203, and SEQ ID NO:1205.

21. 21. The isolated antigen binding protein of claim 20, wherein the antigen binding protein is 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.

22. 22. The isolated antigen binding protein of claim 21, wherein the antibody fragment is a Fab fragment, a Fab' fragment, or a F(ab')2 fragment.

23. 22. The isolated antigen binding protein of claim 21 , wherein the antigen binding protein is a human antibody.

24. 22. The isolated antigen binding protein of claim 21 , wherein the antigen binding protein is a monoclonal antibody.

25. 22. The isolated antigen binding protein of claim 21 , wherein the antigen binding protein is of the IgG1, IgG2, IgG3, or IgG4 type.

26. 26. The isolated antigen binding protein of claim 25, wherein the antigen binding protein is of the IgG1 or IgG2 type.

27. 22. The isolated antigen binding protein of any of claims 21, wherein the antigen binding protein is coupled to a labeling group.

28. 22. The isolated antigen binding protein of claim 21, wherein the antigen binding protein inhibits binding of GIP to the extracellular portion of human GIPR.

29. [0023] The antigen binding protein is an antibody or fragment thereof, wherein the antibody comprises CDRL1, CDRL2, CDRL3, CDRH1, CDRH2, and CDRH3, and wherein CDRL1 is selected from the group consisting of SEQ ID NOs: 4, 10, 16, 22, 28, 34, 40, 46, 52, 58, 64, 70, 76, 82, 88, 94, 100, 106, 112, 118, 124, 130, 136, 142, 148, 154, 160, 166, 172, 178, 184, 190, 196, 202, 208, 214, 220, 226, 232, 238, 244, 250, 256, 262, 268, 270, 272, 274, 276, 278, 278, 279, 280, 282, 284, 286, 288, 289, 290, 300, 304, 306, 308, 310, 312, 314, 316, 318, 320, 326, 328, 330, 336, 332, 338, 340, 342, 346, 348, 350, 356, 358, 360, 362, 366, 370, 374, 376, 378, 378, 379, 380, 382 74, 280, 286, 292, 298, 304, 310, 316, 322, 328, 334, 340, 346, 352, 358, 1290, 1300, 1310, 1320, 1330, 1340, and 1350, and said CDRL2 comprises a sequence selected from the group consisting of SEQ ID NOs: 5, 11, 17, 23, 29, 35, 41, 47, 53, 59, 65, 71, 77, 83, 89, 95, 101, 107, 113, 119, 125, 131, 137, 143, 149, 155, 161, 167, 173, 179, 185, 191, 197, 203, 209, 210, 220, 230, 240, 250, 260, 270, 280, 286, 292, 298, 304, 310, 316, 322, 328, 334, 340, 346, 352, 358, 1290, 1300, 1310, 1320, 1330, 1340, and 1350, 15, 221, 227, 233, 239, 245, 251, 257, 263, 269, 275, 281, 287, 293, 299, 305, 311, 317, 323, 329, 335, 341, 347, 353, 359, 1291, 1301, 1311, 1321, 1331, 1341, and 1351, wherein said CDRL3 comprises a sequence selected from the group consisting of SEQ ID NOs: 6, 12, 18, 24, 30, 36, 42, 48, 54, 60, 66, 72, 78, 84, 90, 96, 102, 108, 114, 120, 126, 132, 138, 144, 150, 1 and wherein said CDRH1 comprises a sequence selected from the group consisting of SEQ ID NOs: 364, 370, 376, 382, ​​388, 394, 400, 406, 412, 418, 424,430, 436, 442, 448, 454, 460, 466, 472, 478, 484, 490, 496, 502, 508, 514, 520, 526, 532, 538, 544, 550, 556, 562, 568, 574, 580, 586, 592, 598, 604, 610, 616, 622, 628, 634, 640, 646, 652, 658, 664, 670, 676, 682, 688, 694, 700, 706, 712, 718, 1293, 1303, 1313, 1323, 1333, 1 and 1353, wherein said CDRH2 comprises a sequence selected from the group consisting of SEQ ID NOs: 365, 371, 377, 383, 389, 395, 401, 407, 413, 419, 425, 431, 437, 443, 449, 455, 461, 467, 473, 479, 485, 491, 497, 503, 509, 515, 521, 527, 533, 539, 545, 551, 557, 563, 569, 575, 581, 587, 593, 599, 605, 611, 617, 623, 629, 63 and said CDRH3 comprises a sequence selected from the group consisting of SEQ ID NOs: 366, 372, 378, 384, 390, 396, 402, 408, 414, 420, 426, 432, 438, 444, 450, 456, 462, 468, 474, 480, 486, 492, 498, 504 22. The isolated antigen binding protein of claim 21, comprising a sequence selected from the group consisting of: 510, 516, 522, 528, 534, 540, 546, 552, 558, 564, 570, 576, 582, 588, 594, 600, 606, 612, 618, 624, 630, 636, 642, 648, 654, 660, 666, 672, 678, 684, 690, 696, 702, 708, 714, 720, 1295, 1305, 1315, 1325, 1335, 1345, and 1355.

30. and wherein the antigen binding protein is an antibody or fragment thereof, and the antibody comprises CDRL1, CDRL2, CDRL3, CDRH1, CDRH2, and CDRH3, each of CDRL1, CDRL2, CDRL3, CDRH1, CDRH2, and CDRH3 being set forth in SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:364, SEQ ID NO:365, and SEQ ID NO:366; SEQ ID NO:10, SEQ ID NO:11, SEQ ID NO:12, SEQ ID NO:370, SEQ ID NO:371, and SEQ ID NO:372; SEQ ID NO:16, SEQ ID NO:17, SEQ ID NO:18, SEQ ID NO:376, SEQ ID NO:37 SEQ ID NO: 7, and SEQ ID NO: 378; SEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO: 24, SEQ ID NO: 382, ​​SEQ ID NO: 383, and SEQ ID NO: 384; SEQ ID NO: 28, SEQ ID NO: 29, SEQ ID NO: 30, SEQ ID NO: 388, SEQ ID NO: 389, and SEQ ID NO: 390; SEQ ID NO: 34, SEQ ID NO: 35, SEQ ID NO: 36, SEQ ID NO: 394, SEQ ID NO: 395, and SEQ ID NO: 396; SEQ ID NO: 40, SEQ ID NO: 41, SEQ ID NO: 42, SEQ ID NO: 400, SEQ ID NO: 401, and SEQ ID NO: 402; SEQ ID NO: 46, SEQ ID NO: 47, SEQ ID NO: 48, SEQ ID NO: 406, SEQ ID NO: 407, and SEQ ID NO: No. 408; SEQ ID NO:52, SEQ ID NO:53, SEQ ID NO:54, SEQ ID NO:412, SEQ ID NO:413, and SEQ ID NO:414; SEQ ID NO:58, SEQ ID NO:59, SEQ ID NO:60, SEQ ID NO:418, SEQ ID NO:419, and SEQ ID NO:420; SEQ ID NO:64, SEQ ID NO:65, SEQ ID NO:66, SEQ ID NO:424, SEQ ID NO:425, and SEQ ID NO:426; SEQ ID NO:70, SEQ ID NO:71, SEQ ID NO:72, SEQ ID NO:430, SEQ ID NO:431, and SEQ ID NO:432; SEQ ID NO:76, SEQ ID NO:77, SEQ ID NO:78, SEQ ID NO:436, SEQ ID NO:437, and SEQ ID NO:438; SEQ ID NO:82, SEQ ID NO:83, SEQ ID NO:84, SEQ ID NO:442, SEQ ID NO:443, and SEQ ID NO:444; SEQ ID NO:88, SEQ ID NO:89, SEQ ID NO:90, SEQ ID NO:448, SEQ ID NO:449, and SEQ ID NO:450; SEQ ID NO:94, SEQ ID NO:95, SEQ ID NO:96, SEQ ID NO:454, SEQ ID NO:455, and SEQ ID NO:456; SEQ ID NO:100, SEQ ID NO:101, SEQ ID NO:102, SEQ ID NO:460, SEQ ID NO:461, and SEQ ID NO:462; SEQ ID NO:106, SEQ ID NO:107, SEQ ID NO:108, SEQ ID NO:466, SEQ ID NO:467, and SEQ ID NO:468;SEQ ID NO:112, SEQ ID NO:113, SEQ ID NO:114, SEQ ID NO:472, SEQ ID NO:473, and SEQ ID NO:474; SEQ ID NO:118, SEQ ID NO:119, SEQ ID NO:120, SEQ ID NO:478, SEQ ID NO:479, and SEQ ID NO:480; SEQ ID NO:124, SEQ ID NO:125, SEQ ID NO:126, SEQ ID NO:484, SEQ ID NO:485, and SEQ ID NO:486; SEQ ID NO:130, SEQ ID NO:131, SEQ ID NO:132, SEQ ID NO:490, SEQ ID NO:491, and SEQ ID NO:492; SEQ ID NO:136, SEQ ID NO:137, SEQ ID NO:138, SEQ ID NO:496, SEQ ID NO:497, and SEQ ID NO:498; SEQ ID NO:142, SEQ ID NO:143, SEQ ID NO:144, SEQ ID NO:502, SEQ ID NO:503, and SEQ ID NO:504; SEQ ID NO:148, SEQ ID NO:149, SEQ ID NO:150, SEQ ID NO:508, SEQ ID NO:509, and SEQ ID NO:510; SEQ ID NO:154, SEQ ID NO:155, SEQ ID NO:156, SEQ ID NO:514, SEQ ID NO:515, and SEQ ID NO:516; SEQ ID NO:160, SEQ ID NO:161, SEQ ID NO:162, SEQ ID NO:520, SEQ ID NO:521, and SEQ ID NO:522; SEQ ID NO:166, SEQ ID NO:167, SEQ ID NO:168, SEQ ID NO:526, SEQ ID NO:527, and SEQ ID NO:528; SEQ ID NO:172, SEQ ID NO:173, SEQ ID NO:174, SEQ ID NO:532, SEQ ID NO:533, and SEQ ID NO:534; SEQ ID NO:178, SEQ ID NO:179, SEQ ID NO:180, SEQ ID NO:538, SEQ ID NO:539, and SEQ ID NO:540; SEQ ID NO:184, SEQ ID NO:185, SEQ ID NO:186, SEQ ID NO:544, SEQ ID NO:545, and SEQ ID NO:546; SEQ ID NO:190, SEQ ID NO:191, SEQ ID NO:192, SEQ ID NO:550, SEQ ID NO:551, and SEQ ID NO:552; SEQ ID NO:196, SEQ ID NO:197, SEQ ID NO:198, SEQ ID NO:556, SEQ ID NO:557, and SEQ ID NO:558; SEQ ID NO:202, SEQ ID NO:203, SEQ ID NO:204, SEQ ID NO:562, SEQ ID NO:563, and SEQ ID NO:564; SEQ ID NO:208, SEQ ID NO:209, SEQ ID NO:210, SEQ ID NO:568, SEQ ID NO:569, and SEQ ID NO:570; SEQ ID NO:214, SEQ ID NO:215, SEQ ID NO:216, SEQ ID NO:574, SEQ ID NO:575, and SEQ ID NO:576; SEQ ID NO:220, SEQ ID NO:221, SEQ ID NO:222, SEQ ID NO:580, SEQ ID NO:581, and SEQ ID NO:582; SEQ ID NO:226, SEQ ID NO:227, SEQ ID NO:228, SEQ ID NO:586, SEQ ID NO:587, and SEQ ID NO:588;SEQ ID NO:232, SEQ ID NO:233, SEQ ID NO:234, SEQ ID NO:592, SEQ ID NO:593, and SEQ ID NO:594; SEQ ID NO:238, SEQ ID NO:239, SEQ ID NO:240, SEQ ID NO:598, SEQ ID NO:599, and SEQ ID NO:600; SEQ ID NO:244, SEQ ID NO:245, SEQ ID NO:246, SEQ ID NO:604, SEQ ID NO:605, and SEQ ID NO:606; SEQ ID NO:250, SEQ ID NO:251, SEQ ID NO:252, SEQ ID NO:610, SEQ ID NO:611, and SEQ ID NO:612; SEQ ID NO:256, SEQ ID NO:257, SEQ ID NO:258, SEQ ID NO:616, SEQ ID NO:617, and SEQ ID NO:618; SEQ ID NO:262, SEQ ID NO:263, SEQ ID NO:264, SEQ ID NO:622, SEQ ID NO:623, and SEQ ID NO:624; SEQ ID NO:268, SEQ ID NO:269, SEQ ID NO:270, SEQ ID NO:628, SEQ ID NO:629, and SEQ ID NO:630; SEQ ID NO:274, SEQ ID NO:275, SEQ ID NO:276, SEQ ID NO:634, SEQ ID NO:635, and SEQ ID NO:636; SEQ ID NO:280, SEQ ID NO:281, SEQ ID NO:282, SEQ ID NO:640, SEQ ID NO:641, and SEQ ID NO:642; SEQ ID NO:286, SEQ ID NO:287, SEQ ID NO:288, SEQ ID NO:646, SEQ ID NO:647, and SEQ ID NO:648; SEQ ID NO:292, SEQ ID NO:293, SEQ ID NO:294, SEQ ID NO:652, SEQ ID NO:653, and SEQ ID NO:654; SEQ ID NO:298, SEQ ID NO:299, SEQ ID NO:300, SEQ ID NO:658, SEQ ID NO:659, and SEQ ID NO:660; SEQ ID NO:304, SEQ ID NO:305, SEQ ID NO:306, SEQ ID NO:664, SEQ ID NO:665, and SEQ ID NO:666; SEQ ID NO:310, SEQ ID NO:311, SEQ ID NO:312, SEQ ID NO:670, SEQ ID NO:671, and SEQ ID NO:672; SEQ ID NO:316, SEQ ID NO:317, SEQ ID NO:318, SEQ ID NO:676, SEQ ID NO:677, and SEQ ID NO:678; SEQ ID NO:322, SEQ ID NO:323, SEQ ID NO:324, SEQ ID NO:682, SEQ ID NO:683, and SEQ ID NO:684; SEQ ID NO:328, SEQ ID NO:329, SEQ ID NO:330, SEQ ID NO:688, SEQ ID NO:689, and SEQ ID NO:690; SEQ ID NO:334, SEQ ID NO:335, SEQ ID NO:336, SEQ ID NO:694, SEQ ID NO:695, and SEQ ID NO:696; SEQ ID NO:340, SEQ ID NO:341, SEQ ID NO:342, SEQ ID NO:700, SEQ ID NO:701, and SEQ ID NO:702; SEQ ID NO:346, SEQ ID NO:347, SEQ ID NO:348, SEQ ID NO:706, SEQ ID NO:707, and SEQ ID NO:708;SEQ ID NO:352, SEQ ID NO:353, SEQ ID NO:354, SEQ ID NO:712, SEQ ID NO:713, and SEQ ID NO:714; SEQ ID NO:358, SEQ ID NO:359, SEQ ID NO:360, SEQ ID NO:718, SEQ ID NO:719, and SEQ ID NO:720; SEQ ID NO:1290, SEQ ID NO:1291, SEQ ID NO:1292, SEQ ID NO:1293, SEQ ID NO:1294, and SEQ ID NO:1295; SEQ ID NO:1300, SEQ ID NO:1301, SEQ ID NO:1302, SEQ ID NO:1303, SEQ ID NO:1304, and SEQ ID NO:1305; SEQ ID NO:1310, SEQ ID NO:1311, SEQ ID NO:1312, SEQ ID NO:1313, SEQ ID NO:1314, and SEQ ID NO:1315; SEQ ID NO:1320, SEQ ID NO:1321, SEQ ID NO:1322, 22. The isolated antigen binding protein of claim 21, comprising a sequence selected from the group consisting of: SEQ ID NO:1323, SEQ ID NO:1324, and SEQ ID NO:1325; SEQ ID NO:1330, SEQ ID NO:1331, SEQ ID NO:1332, SEQ ID NO:1333, SEQ ID NO:1334, and SEQ ID NO:1335; SEQ ID NO:1340, SEQ ID NO:1341, SEQ ID NO:1342, SEQ ID NO:1343, SEQ ID NO:1344, and SEQ ID NO:1345; SEQ ID NO:1350, SEQ ID NO:1351, SEQ ID NO:1352, SEQ ID NO:1353, SEQ ID NO:1354, and SEQ ID NO:1355; and SEQ ID NO:1360, SEQ ID NO:1361, SEQ ID NO:1362, SEQ ID NO:1363, SEQ ID NO:1364, and SEQ ID NO:1365.

31. The antigen binding protein is an antibody or a fragment thereof, and the antibody or fragment thereof is selected from the group consisting of SEQ ID NOs: 723, 727, 731, 735, 739, 743, 747, 751, 755, 759, 763, 767, 771, 775, 779, 783, 787, 791, 795, 799, 803, 807, 811, 815, 819, 823, 827, 831, 835, 839, 843, 847, 85 a light chain comprising a sequence selected from the group consisting of: 1, 855, 859, 863, 867, 871, 875, 879, 883, 887, 891, 895, 899, 903, 907, 911, 915, 919, 923, 927, 931, 935, 939, 943, 947, 951, 955, 959, 1286, 1296, 1306, 1316, 1326, 1336, 1346, and 1356; Variable regions and SEQ ID NOs: 724, 728, 732, 736, 740, 744, 748, 752, 756, 760, 764, 768, 772, 776, 780, 784, 788, 792, 796, 800, 804, 808, 812, 816, 820, 824, 828, 832, 836, 840, 844, 848, 852, 856, 860, 864, 868, 872, 876, 880, 88 and a heavy chain variable region comprising a sequence selected from the group consisting of: 1, 4, 888, 892, 896, 900, 904, 908, 912, 916, 920, 924, 928, 932, 936, 940, 944, 948, 952, 956, 960, 1287, 1297, 1307, 1317, 1327, 1337, 1347, and 1357.

32. The antigen-binding protein is an antibody or a fragment thereof, and the antibody or fragment thereof is an antibody or a fragment thereof, the antibody or fragment thereof being selected from the group consisting of a light chain variable region comprising SEQ ID NO: 723 and a heavy chain variable region comprising SEQ ID NO: 724, a light chain variable region comprising SEQ ID NO: 727 and a heavy chain variable region comprising SEQ ID NO: 728, a light chain variable region comprising SEQ ID NO: 731 and a heavy chain variable region comprising SEQ ID NO: 732, a light chain variable region comprising SEQ ID NO: 735 and a heavy chain variable region comprising SEQ ID NO: 736, a light chain variable region comprising SEQ ID NO: 739 and a heavy chain variable region comprising SEQ ID NO: 740, a light chain variable region comprising SEQ ID NO: 743 and a heavy chain variable region comprising SEQ ID NO: 744 a light chain variable region comprising SEQ ID NO:747 and a heavy chain variable region comprising SEQ ID NO:748; a light chain variable region comprising SEQ ID NO:751 and a heavy chain variable region comprising SEQ ID NO:752; a light chain variable region comprising SEQ ID NO:755 and a heavy chain variable region comprising SEQ ID NO:756; a light chain variable region comprising SEQ ID NO:759 and a heavy chain variable region comprising SEQ ID NO:760; a light chain variable region comprising SEQ ID NO:763 and a heavy chain variable region comprising SEQ ID NO:764; a light chain variable region comprising SEQ ID NO:767 and a heavy chain variable region comprising SEQ ID NO:768; a light chain variable region comprising SEQ ID NO:771 and a heavy chain variable region comprising SEQ ID NO:772 , a light chain variable region comprising SEQ ID NO:775 and a heavy chain variable region comprising SEQ ID NO:776, a light chain variable region comprising SEQ ID NO:779 and a heavy chain variable region comprising SEQ ID NO:780, a light chain variable region comprising SEQ ID NO:783 and a heavy chain variable region comprising SEQ ID NO:784, a light chain variable region comprising SEQ ID NO:787 and a heavy chain variable region comprising SEQ ID NO:788, a light chain variable region comprising SEQ ID NO:791 and a heavy chain variable region comprising SEQ ID NO:792, a light chain variable region comprising SEQ ID NO:795 and a heavy chain variable region comprising SEQ ID NO:796, a light chain variable region comprising SEQ ID NO:799 and a heavy chain variable region comprising SEQ ID NO:800 , a light chain variable region comprising SEQ ID NO:803 and a heavy chain variable region comprising SEQ ID NO:804, a light chain variable region comprising SEQ ID NO:807 and a heavy chain variable region comprising SEQ ID NO:808, a light chain variable region comprising SEQ ID NO:811 and a heavy chain variable region comprising SEQ ID NO:812, a light chain variable region comprising SEQ ID NO:815 and a heavy chain variable region comprising SEQ ID NO:816, a light chain variable region comprising SEQ ID NO:819 and a heavy chain variable region comprising SEQ ID NO:820, a light chain variable region comprising SEQ ID NO:823 and a heavy chain variable region comprising SEQ ID NO:824, a light chain variable region comprising SEQ ID NO:827 and a heavy chain variable region comprising SEQ ID NO:828,a light chain variable region comprising SEQ ID NO:831 and a heavy chain variable region comprising SEQ ID NO:832; a light chain variable region comprising SEQ ID NO:835 and a heavy chain variable region comprising SEQ ID NO:836; a light chain variable region comprising SEQ ID NO:839 and a heavy chain variable region comprising SEQ ID NO:840; a light chain variable region comprising SEQ ID NO:843 and a heavy chain variable region comprising SEQ ID NO:844; a light chain variable region comprising SEQ ID NO:847 and a heavy chain variable region comprising SEQ ID NO:848; a light chain variable region comprising SEQ ID NO:851 and a heavy chain variable region comprising SEQ ID NO:852; a light chain variable region comprising SEQ ID NO:855 and a heavy chain variable region comprising SEQ ID NO:856; a light chain variable region comprising SEQ ID NO:859 and a heavy chain variable region comprising SEQ ID NO:860; a light chain variable region comprising SEQ ID NO:863 and a heavy chain variable region comprising SEQ ID NO:864; a light chain variable region comprising SEQ ID NO:867 and a heavy chain variable region comprising SEQ ID NO:868; a light chain variable region comprising SEQ ID NO:871 and a heavy chain variable region comprising SEQ ID NO:872; a light chain variable region comprising SEQ ID NO:875 and a heavy chain variable region comprising SEQ ID NO:876; a light chain variable region comprising SEQ ID NO:879 and a heavy chain variable region comprising SEQ ID NO:880; a light chain variable region comprising SEQ ID NO:883 and a heavy chain variable region comprising SEQ ID NO:884; a light chain variable region comprising SEQ ID NO:887 and a heavy chain variable region comprising SEQ ID NO:888; a light chain variable region comprising SEQ ID NO:891 and a heavy chain variable region comprising SEQ ID NO:892; a light chain variable region comprising SEQ ID NO:895 and a heavy chain variable region comprising SEQ ID NO:896; a light chain variable region comprising SEQ ID NO:899 and a heavy chain variable region comprising SEQ ID NO:900; a light chain variable region comprising SEQ ID NO:903 and a heavy chain variable region comprising SEQ ID NO:904; a light chain variable region comprising SEQ ID NO:907 and a heavy chain variable region comprising SEQ ID NO:908; a light chain variable region comprising SEQ ID NO:911 and a heavy chain variable region comprising SEQ ID NO:912; a light chain variable region comprising SEQ ID NO:915 and a heavy chain variable region comprising SEQ ID NO:916; a light chain variable region comprising SEQ ID NO:919 and a heavy chain variable region comprising SEQ ID NO:920; a light chain variable region comprising SEQ ID NO:923 and a heavy chain variable region comprising SEQ ID NO:924; a light chain variable region comprising SEQ ID NO:927 and a heavy chain variable region comprising SEQ ID NO:928; a light chain variable region comprising SEQ ID NO:931 and a heavy chain variable region comprising SEQ ID NO:932; a light chain variable region comprising SEQ ID NO:935 and a heavy chain variable region comprising SEQ ID NO:936; a light chain variable region comprising SEQ ID NO:939 and a heavy chain variable region comprising SEQ ID NO:940;a light chain variable region comprising SEQ ID NO:943 and a heavy chain variable region comprising SEQ ID NO:944, a light chain variable region comprising SEQ ID NO:947 and a heavy chain variable region comprising SEQ ID NO:948, a light chain variable region comprising SEQ ID NO:951 and a heavy chain variable region comprising SEQ ID NO:952, a light chain variable region comprising SEQ ID NO:955 and a heavy chain variable region comprising SEQ ID NO:956, a light chain variable region comprising SEQ ID NO:959 and a heavy chain variable region comprising SEQ ID NO:960, a light chain variable region comprising SEQ ID NO:1286 and a heavy chain variable region comprising SEQ ID NO:1287, a light chain variable region comprising SEQ ID NO:1296 and a heavy chain variable region comprising SEQ ID NO:1297, a light chain variable region comprising SEQ ID NO:1306, 22. The isolated antigen binding protein of claim 21, comprising a combination of light chain variable regions and heavy chain variable regions selected from the group consisting of: a light chain variable region comprising SEQ ID NO: 1316 and a heavy chain variable region comprising SEQ ID NO: 1317, a light chain variable region comprising SEQ ID NO: 1326 and a heavy chain variable region comprising SEQ ID NO: 1327, a light chain variable region comprising SEQ ID NO: 1336 and a heavy chain variable region comprising SEQ ID NO: 1337, a light chain variable region comprising SEQ ID NO: 1346 and a heavy chain variable region comprising SEQ ID NO: 1347, and a light chain variable region comprising SEQ ID NO: 1356 and a heavy chain variable region comprising SEQ ID NO: 1357.

33. wherein the antigen binding protein is an antibody, and the antibody is selected from the group consisting of SEQ ID NOs: 963, 967, 971, 975, 979, 983, 987, 991, 995, 999, 1003, 1007, 1011, 1015, 1019, 1023, 1027, 1031, 1035, 1039, 1043, 1047, 1051, 1055, 1059, 1063, 1067 , 1071, 1075, 1079, 1083, 1087, 1091, 1095, 1099, 1103, 1107, 111, and a light chain comprising a sequence selected from the group consisting of SEQ ID NOs: 964, 968, 972, 976, 980, 984, 988, 992, 996, 1000, 1004, 1008, 1012, 1016, 1020, 1024, 102 8, 1032, 1036, 1040, 1044, 1048, 1052, 1056, 1060, 1064, 1068, 1072, 1076, 1080, 1084, 1088, 1092, 1096, 1100, 1104, 1108, 1112, 1116, 1120, 1124, 1128, 1132, 1136, 1140, 1144, 1148, 1 and a heavy chain comprising a sequence selected from the group consisting of: 152, 1156, 1160, 1164, 1168, 1172, 1176, 1180, 1184, 1188, 1192, 1196, 1200, 1289, 1299, 1309, 1319, 1329, 1339, 1349, and 1359.

34. The antigen binding protein is an antibody, the antibody comprising a light chain comprising SEQ ID NO:963 and a heavy chain comprising SEQ ID NO:964, a light chain comprising SEQ ID NO:967 and a heavy chain comprising SEQ ID NO:968, a light chain comprising SEQ ID NO:971 and a heavy chain comprising SEQ ID NO:972, a light chain comprising SEQ ID NO:975 and a heavy chain comprising SEQ ID NO:976, a light chain comprising SEQ ID NO:979 and a heavy chain comprising SEQ ID NO:980, a light chain comprising SEQ ID NO:983 and a heavy chain comprising SEQ ID NO:984, a light chain comprising SEQ ID NO:987 and a heavy chain comprising SEQ ID NO:988, a light chain comprising SEQ ID NO:991 and a heavy chain comprising SEQ ID NO:992, a light chain comprising SEQ ID NO:995 and a heavy chain comprising SEQ ID NO:996, a light chain comprising SEQ ID NO:997 and a heavy chain comprising SEQ ID NO:998, a light chain comprising SEQ ID NO:998 and a heavy chain comprising SEQ ID NO:999, and a heavy chain comprising SEQ ID NO:996, a light chain comprising SEQ ID NO:999 and a heavy chain comprising SEQ ID NO:1000, a light chain comprising SEQ ID NO:1003 and a heavy chain comprising SEQ ID NO:1004, a light chain comprising SEQ ID NO:1007 and a heavy chain comprising SEQ ID NO:1008, a light chain comprising SEQ ID NO:1011 and a heavy chain comprising SEQ ID NO:1012, a light chain comprising SEQ ID NO:1015 and a heavy chain comprising SEQ ID NO:1016, a light chain comprising SEQ ID NO:1019 and a heavy chain comprising SEQ ID NO:1020, a light chain comprising SEQ ID NO:1023 and a heavy chain comprising SEQ ID NO:1024, a light chain comprising SEQ ID NO:1027 and a heavy chain comprising SEQ ID NO:1028 heavy chain, light chain comprising SEQ ID NO:1031 and heavy chain comprising SEQ ID NO:1032, light chain comprising SEQ ID NO:1035 and heavy chain comprising SEQ ID NO:1036, light chain comprising SEQ ID NO:1039 and heavy chain comprising SEQ ID NO:1040, light chain comprising SEQ ID NO:1043 and heavy chain comprising SEQ ID NO:1044, light chain comprising SEQ ID NO:1047 and heavy chain comprising SEQ ID NO:1048, light chain comprising SEQ ID NO:1051 and heavy chain comprising SEQ ID NO:1052, light chain comprising SEQ ID NO:1055 and heavy chain comprising SEQ ID NO:1056, light chain comprising SEQ ID NO:1059 and heavy chain comprising SEQ ID NO:1060, light chain comprising SEQ ID NO:1063 and a heavy chain comprising SEQ ID NO: 1064, a light chain comprising SEQ ID NO: 1067 and a heavy chain comprising SEQ ID NO: 1068, a light chain comprising SEQ ID NO: 1071 and a heavy chain comprising SEQ ID NO: 1072, a light chain comprising SEQ ID NO: 1075 and a heavy chain comprising SEQ ID NO: 1076, a light chain comprising SEQ ID NO: 1079 and a heavy chain comprising SEQ ID NO: 1080, a light chain comprising SEQ ID NO: 1083 and a heavy chain comprising SEQ ID NO: 1084, a light chain comprising SEQ ID NO: 1087 and a heavy chain comprising SEQ ID NO: 1088, a light chain comprising SEQ ID NO: 1091 and a heavy chain comprising SEQ ID NO: 1092, a light chain comprising SEQ ID NO: 1095 and a heavy chain comprising SEQ ID NO: 1096,a light chain comprising SEQ ID NO:1099 and a heavy chain comprising SEQ ID NO:1100; a light chain comprising SEQ ID NO:1103 and a heavy chain comprising SEQ ID NO:1104; a light chain comprising SEQ ID NO:1107 and a heavy chain comprising SEQ ID NO:1108; a light chain comprising SEQ ID NO:1111 and a heavy chain comprising SEQ ID NO:1112; a light chain comprising SEQ ID NO:1115 and a heavy chain comprising SEQ ID NO:1116; a light chain comprising SEQ ID NO:1119 and a heavy chain comprising SEQ ID NO:1120; a light chain comprising SEQ ID NO:1123 and a heavy chain comprising SEQ ID NO:1124; a light chain comprising SEQ ID NO:1127 and a heavy chain comprising SEQ ID NO:1128; a light chain comprising SEQ ID NO:1131 and a heavy chain comprising SEQ ID NO:1132, a light chain comprising SEQ ID NO:1135 and a heavy chain comprising SEQ ID NO:1136, a light chain comprising SEQ ID NO:1139 and a heavy chain comprising SEQ ID NO:1140, a light chain comprising SEQ ID NO:1143 and a heavy chain comprising SEQ ID NO:1144, a light chain comprising SEQ ID NO:1147 and a heavy chain comprising SEQ ID NO:1148, a light chain comprising SEQ ID NO:1151 and a heavy chain comprising SEQ ID NO:1152, a light chain comprising SEQ ID NO:1155 and a heavy chain comprising SEQ ID NO:1156, a light chain comprising SEQ ID NO:1159 and a heavy chain comprising SEQ ID NO:1160, a light chain comprising SEQ ID NO:1163 and a heavy chain comprising SEQ ID NO:1164, A light chain comprising SEQ ID NO:1167 and a heavy chain comprising SEQ ID NO:1168, a light chain comprising SEQ ID NO:1171 and a heavy chain comprising SEQ ID NO:1172, a light chain comprising SEQ ID NO:1175 and a heavy chain comprising SEQ ID NO:1176, a light chain comprising SEQ ID NO:1179 and a heavy chain comprising SEQ ID NO:1180, a light chain comprising SEQ ID NO:1183 and a heavy chain comprising SEQ ID NO:1184, a light chain comprising SEQ ID NO:1187 and a heavy chain comprising SEQ ID NO:1188, a light chain comprising SEQ ID NO:1191 and a heavy chain comprising SEQ ID NO:1192, a light chain comprising SEQ ID NO:1195 and a heavy chain comprising SEQ ID NO:1196, a light chain comprising SEQ ID NO:1199 and a heavy chain comprising SEQ ID NO:1197, a heavy chain comprising sequence number 1200, a light chain comprising SEQ ID NO: 1288 and a heavy chain comprising SEQ ID NO: 1289, a light chain comprising SEQ ID NO: 1298 and a heavy chain comprising SEQ ID NO: 1299, a light chain comprising SEQ ID NO: 1308 and a heavy chain comprising SEQ ID NO: 1309, a light chain comprising SEQ ID NO: 1318 and a heavy chain comprising SEQ ID NO: 1319, a light chain comprising SEQ ID NO: 1328 and a heavy chain comprising SEQ ID NO: 1329, a light chain comprising SEQ ID NO: 1338 and a heavy chain comprising SEQ ID NO: 1339, a light chain comprising SEQ ID NO: 1348 and a heavy chain comprising SEQ ID NO: 1349, and a light chain comprising SEQ ID NO: 1358 and a heavy chain comprising SEQ ID NO: 1359,22. The isolated antigen binding protein of claim 21, comprising a combination of light and heavy chains selected from the group consisting of:

35. A nucleic acid molecule encoding the antibody or fragment thereof according to any one of claims 29 to 34.

36. The nucleic acid molecule of claim 35 , wherein the nucleic acid molecule is operably linked to a control sequence.

37. A vector comprising the nucleic acid molecule of claim 36.

38. A host cell comprising the nucleic acid molecule of claim 37.

39. 39. An antibody or fragment thereof produced by the host cell of claim 38.

40. A method for preparing the antibody or fragment thereof according to any one of claims 29 to 34, comprising the step of preparing the antibody or fragment thereof from a host cell which secretes the antibody.

41. A pharmaceutical composition comprising at least one antibody or fragment thereof according to any one of claims 29 to 34 and a pharma- ceutically acceptable excipient.

42. An isolated antigen-binding protein that competes with an antibody or fragment thereof according to any one of claims 29 to 34 for binding to the extracellular portion of human GIPR.

43. A composition comprising a therapeutically effective amount of a GLP-1 receptor agonist and a therapeutically effective amount of a GIPR antagonist that specifically binds to a protein having an amino acid sequence that has at least 90% amino acid sequence identity to the amino acid sequence of GIPR.

44. 44. The composition of claim 43, wherein the molar ratio of the GLP-1 receptor agonist to the GIPR antagonist is about 1:1 to 1:110, about 1:1 to 1:100, about 1:1 to 1:75, about 1:1 to 1:50, about 1:1 to 1:25, about 1:1 to 1:10, about 1:1 to 1:5, and about 1:

1.

45. 44. The composition of claim 43, wherein the molar ratio of the GIPR antagonist to the GLP-1 receptor agonist is about 1:1 to 1:110, about 1:1 to 1:100, about 1:1 to 1:75, about 1:1 to 1:50, about 1:1 to 1:25, about 1:1 to 1:10, and about 1:1 to 1:

5.

46. 44. The composition of claim 43, wherein the GLP-1 receptor agonist is used in combination with the GIPR antagonist in a therapeutically effective molar ratio of about 1:1.5 to 1:150, preferably 1:2 to 1:

50.

47. 44. The composition of claim 43, wherein the GLP-1 receptor agonist and the GIPR antagonist are present in a dosage that is at least about 1.1 to 1.4 times, 1.5 times, 2 times, 3 times, 4 times, 5 times, 6 times, 7 times, 8 times, 9 times, or 10 times less than the dosage of each compound alone that would be required to treat the condition and / or disease.

48. 44. The composition of claim 43, wherein the GLP-1 receptor agonist is GLP-1(7-37) or a GLP-1(7-37) analogue.

49. 49. The composition of claim 48, wherein the GLP-1 receptor agonist is selected from the group consisting of exenatide, liraglutide, lixisenatide, albiglutide, dulaglutide, semaglutide, and taspoglutide.

50. The GLP-1 receptor agonist is selected from the group consisting of GLP-1(7-37) (SEQ ID NO: 1244), GLP-1(7-36)-NH ２ (SEQ ID NO: 1245), liraglutide, albiglutide, taspoglutide, dulaglutide, semaglutide, LY2428757, desamino-His ７ , Arg ２６ , Lys ３４ (N ε -(γ-Glu(N-α-hexadecanoyl)))-GLP-1(7-37) (core peptide disclosed as SEQ ID NO:1282), desamino-His ７ , Arg ２６ , Lys ３４ (N ε -octanoyl)-GLP-1(7-37) (SEQ ID NO: 1283), Arg ２６，３４ , Lys ３８ (N ε -(ω-carboxypentadecanoyl)-GLP-1(7-38) (SEQ ID NO: 1284), Arg ２６，３４ , Lys ３６ (N ε -(γ-Glu(N-α-hexadecanoyl)))-GLP-1(7-36) (core peptide disclosed as SEQ ID NO: 1285), Aib ８，３５ , Arg ２６，３４ , Phe ３１ -GLP-1(7-36)) (SEQ ID NO: 1246), HXaa ８ EGTFTSDVSSYLEXaa ２２ Xaa ２３ AAKEFIXaa ３０ WLXaa ３３ Xaa ３４ G Xaa ３６ Xaa ３７ (Xaa ８ is A, V, or G; ２２ is G, K, or E; ２３ is Q or K, and Xaa ３０ is A or E, and Xaa ３３ is V or K, and Xaa ３４ is K, N, or R; ３６ is R or G, and Xaa ３７ is G, H, P, or absent) (SEQ ID NO: 1247), Arg ３４ -GLP-1(7-37) (SEQ ID NO: 1248), Glu ３０ -GLP-1(7-37) (SEQ ID NO: 1249), Lys ２２ -GLP-1(7-37) (SEQ ID NO: 1250), Gly ８，３６ , Glu ２２ -GLP-1(7-37) (SEQ ID NO: 1251), Val ８ , Glu ２２ , Gly ３６ -GLP-1(7-37) (SEQ ID NO: 1252), Gly ８，３６ , Glu ２２ , Lys ３３ , Asn ３４ -GLP-1(7-37) (SEQ ID NO: 1253), Val ８ , Glu ２２ , Lys ３３ , Asn ３４ , Gly ３６ -GLP-1(7-37) (SEQ ID NO: 1254), Gly ８，３６ , Glu ２２ , Pro ３７ -GLP-1(7-37) (SEQ ID NO: 1255), Val ８ , Glu ２２ , Gly ３６ Pro ３７ -GLP-1(7-37) (SEQ ID NO: 1256), Gly ８，３６ , Glu ２２ , Lys ３３ , Asn ３４ , Pro ３７ -GLP-1(7-37) (SEQ ID NO: 1257), Val ８ , Glu ２２ , Lys ３３ , Asn ３４ , Gly ３６ , Pro ３７ -GLP-1(7-37) (SEQ ID NO: 1258), Gly ８，３６ , Glu ２２ -GLP-1(7-36) (SEQ ID NO: 1259), Val ８ , Glu ２２ , Gly ３６ -GLP-1(7-36) (SEQ ID NO: 1260), Val ８ , Glu ２２ , Asn ３４ , Gly ３６ -GLP-1(7-36) (SEQ ID NO: 1261), and Gly ８，３６ , Glu ２２ , Asn ３４ - GLP-1(7-36) (SEQ ID NO: 1262).

51. 44. The composition of claim 43, wherein the human GIPR has a sequence comprising a sequence selected from the group consisting of SEQ ID NO: 1201, SEQ ID NO: 1203, and SEQ ID NO: 1205.

52. The composition of claim 43, wherein the GIPR antagonist is an antigen binding protein.

53. 53. The composition of claim 52, wherein the antigen binding protein is 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.

54. 54. The composition of claim 53, wherein the antibody fragment is a Fab fragment, a Fab' fragment, or an F(ab')2 fragment.

55. 54. The composition of claim 53, wherein the antigen binding protein is a human antibody.

56. 54. The composition of claim 53, wherein the antigen binding protein is a monoclonal antibody.

57. 54. The composition of claim 53, wherein the antigen binding protein is of the IgG1, IgG2, IgG3, or IgG4 type.

58. 58. The composition of claim 57, wherein the antigen binding protein is of the IgG1 or IgG2 type.

59. 54. The composition of claim 53, wherein the antigen binding protein is coupled to a labeling group.

60. 54. The composition of claim 53, wherein the antigen binding protein inhibits binding of GIP to the extracellular portion of human GIPR.

61. [0023] The antigen binding protein is an antibody or fragment thereof, wherein the antibody comprises CDRL1, CDRL2, CDRL3, CDRH1, CDRH2, and CDRH3, and wherein CDRL1 is selected from the group consisting of SEQ ID NOs: 4, 10, 16, 22, 28, 34, 40, 46, 52, 58, 64, 70, 76, 82, 88, 94, 100, 106, 112, 118, 124, 130, 136, 142, 148, 154, 160, 166, 172, 178, 184, 190, 196, 202, 208, 214, 220, 226, 232, 238, 244, 250, 256, 262, 268, 270, 272, 274, 276, 278, 278, 279, 280, 282, 284, 286, 288, 289, 290, 300, 304, 306, 308, 310, 312, 314, 316, 318, 320, 326, 328, 330, 336, 332, 338, 340, 342, 346, 348, 350, 356, 358, 360, 362, 366, 370, 374, 376, 378, 378, 379, 380, 382 74, 280, 286, 292, 298, 304, 310, 316, 322, 328, 334, 340, 346, 352, 358, 1290, 1300, 1310, 1320, 1330, 1340, and 1350, and said CDRL2 comprises a sequence selected from the group consisting of SEQ ID NOs: 5, 11, 17, 23, 29, 35, 41, 47, 53, 59, 65, 71, 77, 83, 89, 95, 101, 107, 113, 119, 125, 131, 137, 143, 149, 155, 161, 167, 173, 179, 185, 191, 197, 203, 209, 210, 220, 230, 240, 250, 260, 270, 280, 286, 292, 298, 304, 310, 316, 322, 328, 334, 340, 346, 352, 358, 1290, 1300, 1310, 1320, 1330, 1340, and 1350, 15, 221, 227, 233, 239, 245, 251, 257, 263, 269, 275, 281, 287, 293, 299, 305, 311, 317, 323, 329, 335, 341, 347, 353, 359, 1291, 1301, 1311, 1321, 1331, 1341, and 1351, wherein said CDRL3 comprises a sequence selected from the group consisting of SEQ ID NOs: 6, 12, 18, 24, 30, 36, 42, 48, 54, 60, 66, 72, 78, 84, 90, 96, 102, 108, 114, 120, 126, 132, 138, 144, 150, 1 and wherein said CDRH1 comprises a sequence selected from the group consisting of SEQ ID NOs: 364, 370, 376, 382, ​​388, 394, 400, 406, 412, 418, 424,430, 436, 442, 448, 454, 460, 466, 472, 478, 484, 490, 496, 502, 508, 514, 520, 526, 532, 538, 544, 550, 556, 562, 568, 574, 580, 586, 592, 598, 604, 610, 616, 622, 628, 634, 640, 646, 652, 658, 664, 670, 676, 682, 688, 694, 700, 706, 712, 718, 1293, 1303, 1313, 1323, 133 3, 1343, and 1353, wherein said CDRH2 comprises a sequence selected from the group consisting of SEQ ID NOs: 365, 371, 377, 383, 389, 395, 401, 407, 413, 419, 425, 431, 437, 443, 449, 455, 461, 467, 473, 479, 485, 491, 497, 503, 509, 515, 521, 527, 533, 539, 545, 551, 557, 563, 569, 575, 581, 587, 593, 599, 605, 611, 617, 623, and said CDRH3 comprises a sequence selected from the group consisting of SEQ ID NOs: 366, 372, 378, 384, 390, 396, 402, 408, 414, 420, 426, 432, 438, 444, 450, 456, 462, 468, 474, 480, 486, 492, 493, 501, 502, 503, 504, 505, 506, 507, 510, 512, 513, 514, 515, 516, 517, 518, 519, 520, 521, 522, 523, 524, 525, 526, 527, 528, 529, 530, 531, 532, 533, 534, 535, 536, 537, 538, 539, 540, 541, 542, 543, 544, 545, 546, 547, 548, 549, 550, 556, 557, 558, 559, 560, 561, 562, 563, 564, 565, 566, 567, 568, 570, 571, 572, 573, 574, 575, 576, 577, 578, 579, 580, 581, 582, 583, 584, 54. The composition of claim 53, comprising a sequence selected from the group consisting of: 498, 504, 510, 516, 522, 528, 534, 540, 546, 552, 558, 564, 570, 576, 582, 588, 594, 600, 606, 612, 618, 624, 630, 636, 642, 648, 654, 660, 666, 672, 678, 684, 690, 696, 702, 708, 714, 720, 1295, 1305, 1315, 1325, 1335, 1345, and 1355.

62. and wherein the antigen binding protein is an antibody or fragment thereof, and the antibody comprises CDRL1, CDRL2, CDRL3, CDRH1, CDRH2, and CDRH3, each of CDRL1, CDRL2, CDRL3, CDRH1, CDRH2, and CDRH3 being set forth in SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:364, SEQ ID NO:365, and SEQ ID NO:366; SEQ ID NO:10, SEQ ID NO:11, SEQ ID NO:12, SEQ ID NO:370, SEQ ID NO:371, and SEQ ID NO:372; SEQ ID NO:16, SEQ ID NO:17, SEQ ID NO:18, SEQ ID NO:376, SEQ ID NO:37 SEQ ID NO: 7, and SEQ ID NO: 378; SEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO: 24, SEQ ID NO: 382, ​​SEQ ID NO: 383, and SEQ ID NO: 384; SEQ ID NO: 28, SEQ ID NO: 29, SEQ ID NO: 30, SEQ ID NO: 388, SEQ ID NO: 389, and SEQ ID NO: 390; SEQ ID NO: 34, SEQ ID NO: 35, SEQ ID NO: 36, SEQ ID NO: 394, SEQ ID NO: 395, and SEQ ID NO: 396; SEQ ID NO: 40, SEQ ID NO: 41, SEQ ID NO: 42, SEQ ID NO: 400, SEQ ID NO: 401, and SEQ ID NO: 402; SEQ ID NO: 46, SEQ ID NO: 47, SEQ ID NO: 48, SEQ ID NO: 406, SEQ ID NO: 407, and SEQ ID NO: No. 408; SEQ ID NO:52, SEQ ID NO:53, SEQ ID NO:54, SEQ ID NO:412, SEQ ID NO:413, and SEQ ID NO:414; SEQ ID NO:58, SEQ ID NO:59, SEQ ID NO:60, SEQ ID NO:418, SEQ ID NO:419, and SEQ ID NO:420; SEQ ID NO:64, SEQ ID NO:65, SEQ ID NO:66, SEQ ID NO:424, SEQ ID NO:425, and SEQ ID NO:426; SEQ ID NO:70, SEQ ID NO:71, SEQ ID NO:72, SEQ ID NO:430, SEQ ID NO:431, and SEQ ID NO:432; SEQ ID NO:76, SEQ ID NO:77, SEQ ID NO:78, SEQ ID NO:436, SEQ ID NO:437, and SEQ ID NO:438; SEQ ID NO:82, SEQ ID NO:83, SEQ ID NO:84, SEQ ID NO:442, SEQ ID NO:443, and SEQ ID NO:444; SEQ ID NO:88, SEQ ID NO:89, SEQ ID NO:90, SEQ ID NO:448, SEQ ID NO:449, and SEQ ID NO:450; SEQ ID NO:94, SEQ ID NO:95, SEQ ID NO:96, SEQ ID NO:454, SEQ ID NO:455, and SEQ ID NO:456; SEQ ID NO:100, SEQ ID NO:101, SEQ ID NO:102, SEQ ID NO:460, SEQ ID NO:461, and SEQ ID NO:462; SEQ ID NO:106, SEQ ID NO:107, SEQ ID NO:108, SEQ ID NO:466, SEQ ID NO:467, and SEQ ID NO:468;SEQ ID NO:112, SEQ ID NO:113, SEQ ID NO:114, SEQ ID NO:472, SEQ ID NO:473, and SEQ ID NO:474; SEQ ID NO:118, SEQ ID NO:119, SEQ ID NO:120, SEQ ID NO:478, SEQ ID NO:479, and SEQ ID NO:480; SEQ ID NO:124, SEQ ID NO:125, SEQ ID NO:126, SEQ ID NO:484, SEQ ID NO:485, and SEQ ID NO:486; SEQ ID NO:130, SEQ ID NO:131, SEQ ID NO:132, SEQ ID NO:490, SEQ ID NO:491, and SEQ ID NO:492; SEQ ID NO:136, SEQ ID NO:137, SEQ ID NO:138, SEQ ID NO:496, SEQ ID NO:497, and SEQ ID NO:498; SEQ ID NO:142, SEQ ID NO:143, SEQ ID NO:144, SEQ ID NO:502, SEQ ID NO:503, and SEQ ID NO:504; SEQ ID NO:148, SEQ ID NO:149, SEQ ID NO:150, SEQ ID NO:508, SEQ ID NO:509, and SEQ ID NO:510; SEQ ID NO:154, SEQ ID NO:155, SEQ ID NO:156, SEQ ID NO:514, SEQ ID NO:515, and SEQ ID NO:516; SEQ ID NO:160, SEQ ID NO:161, SEQ ID NO:162, SEQ ID NO:520, SEQ ID NO:521, and SEQ ID NO:522; SEQ ID NO:166, SEQ ID NO:167, SEQ ID NO:168, SEQ ID NO:526, SEQ ID NO:527, and SEQ ID NO:528; SEQ ID NO:172, SEQ ID NO:173, SEQ ID NO:174, SEQ ID NO:532, SEQ ID NO:533, and SEQ ID NO:534; SEQ ID NO:178, SEQ ID NO:179, SEQ ID NO:180, SEQ ID NO:538, SEQ ID NO:539, and SEQ ID NO:540; SEQ ID NO:184, SEQ ID NO:185, SEQ ID NO:186, SEQ ID NO:544, SEQ ID NO:545, and SEQ ID NO:546; SEQ ID NO:190, SEQ ID NO:191, SEQ ID NO:192, SEQ ID NO:550, SEQ ID NO:551, and SEQ ID NO:552; SEQ ID NO:196, SEQ ID NO:197, SEQ ID NO:198, SEQ ID NO:556, SEQ ID NO:557, and SEQ ID NO:558; SEQ ID NO:202, SEQ ID NO:203, SEQ ID NO:204, SEQ ID NO:562, SEQ ID NO:563, and SEQ ID NO:564; SEQ ID NO:208, SEQ ID NO:209, SEQ ID NO:210, SEQ ID NO:568, SEQ ID NO:569, and SEQ ID NO:570; SEQ ID NO:214, SEQ ID NO:215, SEQ ID NO:216, SEQ ID NO:574, SEQ ID NO:575, and SEQ ID NO:576; SEQ ID NO:220, SEQ ID NO:221, SEQ ID NO:222, SEQ ID NO:580, SEQ ID NO:581, and SEQ ID NO:582; SEQ ID NO:226, SEQ ID NO:227, SEQ ID NO:228, SEQ ID NO:586, SEQ ID NO:587, and SEQ ID NO:588;SEQ ID NO:232, SEQ ID NO:233, SEQ ID NO:234, SEQ ID NO:592, SEQ ID NO:593, and SEQ ID NO:594; SEQ ID NO:238, SEQ ID NO:239, SEQ ID NO:240, SEQ ID NO:598, SEQ ID NO:599, and SEQ ID NO:600; SEQ ID NO:244, SEQ ID NO:245, SEQ ID NO:246, SEQ ID NO:604, SEQ ID NO:605, and SEQ ID NO:606; SEQ ID NO:250, SEQ ID NO:251, SEQ ID NO:252, SEQ ID NO:610, SEQ ID NO:611, and SEQ ID NO:612; SEQ ID NO:256, SEQ ID NO:257, SEQ ID NO:258, SEQ ID NO:616, SEQ ID NO:617, and SEQ ID NO:618; SEQ ID NO:262, SEQ ID NO:263, SEQ ID NO:264, SEQ ID NO:622, SEQ ID NO:623, and SEQ ID NO:624; SEQ ID NO:268, SEQ ID NO:269, SEQ ID NO:270, SEQ ID NO:628, SEQ ID NO:629, and SEQ ID NO:630; SEQ ID NO:274, SEQ ID NO:275, SEQ ID NO:276, SEQ ID NO:634, SEQ ID NO:635, and SEQ ID NO:636; SEQ ID NO:280, SEQ ID NO:281, SEQ ID NO:282, SEQ ID NO:640, SEQ ID NO:641, and SEQ ID NO:642; SEQ ID NO:286, SEQ ID NO:287, SEQ ID NO:288, SEQ ID NO:646, SEQ ID NO:647, and SEQ ID NO:648; SEQ ID NO:292, SEQ ID NO:293, SEQ ID NO:294, SEQ ID NO:652, SEQ ID NO:653, and SEQ ID NO:654; SEQ ID NO:298, SEQ ID NO:299, SEQ ID NO:300, SEQ ID NO:658, SEQ ID NO:659, and SEQ ID NO:660; SEQ ID NO:304, SEQ ID NO:305, SEQ ID NO:306, SEQ ID NO:664, SEQ ID NO:665, and SEQ ID NO:666; SEQ ID NO:310, SEQ ID NO:311, SEQ ID NO:312, SEQ ID NO:670, SEQ ID NO:671, and SEQ ID NO:672; SEQ ID NO:316, SEQ ID NO:317, SEQ ID NO:318, SEQ ID NO:676, SEQ ID NO:677, and SEQ ID NO:678; SEQ ID NO:322, SEQ ID NO:323, SEQ ID NO:324, SEQ ID NO:682, SEQ ID NO:683, and SEQ ID NO:684; SEQ ID NO:328, SEQ ID NO:329, SEQ ID NO:330, SEQ ID NO:688, SEQ ID NO:689, and SEQ ID NO:690; SEQ ID NO:334, SEQ ID NO:335, SEQ ID NO:336, SEQ ID NO:694, SEQ ID NO:695, and SEQ ID NO:696; SEQ ID NO:340, SEQ ID NO:341, SEQ ID NO:342, SEQ ID NO:700, SEQ ID NO:701, and SEQ ID NO:702; SEQ ID NO:346, SEQ ID NO:347, SEQ ID NO:348, SEQ ID NO:706, SEQ ID NO:707, and SEQ ID NO:708;SEQ ID NO:352, SEQ ID NO:353, SEQ ID NO:354, SEQ ID NO:712, SEQ ID NO:713, and SEQ ID NO:714; SEQ ID NO:358, SEQ ID NO:359, SEQ ID NO:360, SEQ ID NO:718, SEQ ID NO:719, and SEQ ID NO:720; SEQ ID NO:1290, SEQ ID NO:1291, SEQ ID NO:1292, SEQ ID NO:1293, SEQ ID NO:1294, and SEQ ID NO:1295; SEQ ID NO:1300, SEQ ID NO:1301, SEQ ID NO:1302, SEQ ID NO:1303, SEQ ID NO:1304, and SEQ ID NO:1305; SEQ ID NO:1310, SEQ ID NO:1311, SEQ ID NO:1312, SEQ ID NO:1313, SEQ ID NO:1314, and SEQ ID NO:1315; SEQ ID NO:1320, SEQ ID NO:1321, SEQ ID NO:

54. The composition of claim 53, comprising a sequence selected from the group consisting of SEQ ID NO:1322, SEQ ID NO:1323, SEQ ID NO:1324, and SEQ ID NO:1325; SEQ ID NO:1330, SEQ ID NO:1331, SEQ ID NO:1332, SEQ ID NO:1333, SEQ ID NO:1334, and SEQ ID NO:1335; SEQ ID NO:1340, SEQ ID NO:1341, SEQ ID NO:1342, SEQ ID NO:1343, SEQ ID NO:1344, and SEQ ID NO:1345; SEQ ID NO:1350, SEQ ID NO:1351, SEQ ID NO:1352, SEQ ID NO:1353, SEQ ID NO:1354, and SEQ ID NO:1355; and SEQ ID NO:1360, SEQ ID NO:1361, SEQ ID NO:1362, SEQ ID NO:1363, SEQ ID NO:1364, and SEQ ID NO:1365.

63. The antigen binding protein is an antibody or a fragment thereof, and the antibody or fragment thereof is selected from the group consisting of SEQ ID NOs: 723, 727, 731, 735, 739, 743, 747, 751, 755, 759, 763, 767, 771, 775, 779, 783, 787, 791, 795, 799, 803, 807, 811, 815, 819, 823, 827, 831, 835, 839, 843, 847, A sequence selected from the group consisting of 851, 855, 859, 863, 867, 871, 875, 879, 883, 887, 891, 895, 899, 903, 907, 911, 915, 919, 923, 927, 931, 935, 939, 943, 947, 951, 955, 959, 1286, 1296, 1306, 1316, 1326, 1336, 1346, and 1356. and a light chain variable region comprising SEQ ID NOs: 724, 728, 732, 736, 740, 744, 748, 752, 756, 760, 764, 768, 772, 776, 780, 784, 788, 792, 796, 800, 804, 808, 812, 816, 820, 824, 828, 832, 836, 840, 844, 848, 852, 856, 860, 864, 868, 872, 87 and a heavy chain variable region comprising a sequence selected from the group consisting of: 6, 880, 884, 888, 892, 896, 900, 904, 908, 912, 916, 920, 924, 928, 932, 936, 940, 944, 948, 952, 956, 960, 1287, 1297, 1307, 1317, 1327, 1337, 1347, and 1357.

64. The antigen-binding protein is an antibody or a fragment thereof, and the antibody or fragment thereof is an antibody or a fragment thereof, the antibody or fragment thereof being selected from the group consisting of a light chain variable region comprising SEQ ID NO: 723 and a heavy chain variable region comprising SEQ ID NO: 724, a light chain variable region comprising SEQ ID NO: 727 and a heavy chain variable region comprising SEQ ID NO: 728, a light chain variable region comprising SEQ ID NO: 731 and a heavy chain variable region comprising SEQ ID NO: 732, a light chain variable region comprising SEQ ID NO: 735 and a heavy chain variable region comprising SEQ ID NO: 736, a light chain variable region comprising SEQ ID NO: 739 and a heavy chain variable region comprising SEQ ID NO: 740, a light chain variable region comprising SEQ ID NO: 743 and a heavy chain variable region comprising SEQ ID NO: 744 a light chain variable region comprising SEQ ID NO:747 and a heavy chain variable region comprising SEQ ID NO:748; a light chain variable region comprising SEQ ID NO:751 and a heavy chain variable region comprising SEQ ID NO:752; a light chain variable region comprising SEQ ID NO:755 and a heavy chain variable region comprising SEQ ID NO:756; a light chain variable region comprising SEQ ID NO:759 and a heavy chain variable region comprising SEQ ID NO:760; a light chain variable region comprising SEQ ID NO:763 and a heavy chain variable region comprising SEQ ID NO:764; a light chain variable region comprising SEQ ID NO:767 and a heavy chain variable region comprising SEQ ID NO:768; a light chain variable region comprising SEQ ID NO:771 and a heavy chain variable region comprising SEQ ID NO:772 , a light chain variable region comprising SEQ ID NO:775 and a heavy chain variable region comprising SEQ ID NO:776, a light chain variable region comprising SEQ ID NO:779 and a heavy chain variable region comprising SEQ ID NO:780, a light chain variable region comprising SEQ ID NO:783 and a heavy chain variable region comprising SEQ ID NO:784, a light chain variable region comprising SEQ ID NO:787 and a heavy chain variable region comprising SEQ ID NO:788, a light chain variable region comprising SEQ ID NO:791 and a heavy chain variable region comprising SEQ ID NO:792, a light chain variable region comprising SEQ ID NO:795 and a heavy chain variable region comprising SEQ ID NO:796, a light chain variable region comprising SEQ ID NO:799 and a heavy chain variable region comprising SEQ ID NO:800 , a light chain variable region comprising SEQ ID NO:803 and a heavy chain variable region comprising SEQ ID NO:804, a light chain variable region comprising SEQ ID NO:807 and a heavy chain variable region comprising SEQ ID NO:808, a light chain variable region comprising SEQ ID NO:811 and a heavy chain variable region comprising SEQ ID NO:812, a light chain variable region comprising SEQ ID NO:815 and a heavy chain variable region comprising SEQ ID NO:816, a light chain variable region comprising SEQ ID NO:819 and a heavy chain variable region comprising SEQ ID NO:820, a light chain variable region comprising SEQ ID NO:823 and a heavy chain variable region comprising SEQ ID NO:824, a light chain variable region comprising SEQ ID NO:827 and a heavy chain variable region comprising SEQ ID NO:828,a light chain variable region comprising SEQ ID NO:831 and a heavy chain variable region comprising SEQ ID NO:832; a light chain variable region comprising SEQ ID NO:835 and a heavy chain variable region comprising SEQ ID NO:836; a light chain variable region comprising SEQ ID NO:839 and a heavy chain variable region comprising SEQ ID NO:840; a light chain variable region comprising SEQ ID NO:843 and a heavy chain variable region comprising SEQ ID NO:844; a light chain variable region comprising SEQ ID NO:847 and a heavy chain variable region comprising SEQ ID NO:848; a light chain variable region comprising SEQ ID NO:851 and a heavy chain variable region comprising SEQ ID NO:852; a light chain variable region comprising SEQ ID NO:855 and a heavy chain variable region comprising SEQ ID NO:856; a light chain variable region comprising SEQ ID NO:859 and a heavy chain variable region comprising SEQ ID NO:860; a light chain variable region comprising SEQ ID NO:863 and a heavy chain variable region comprising SEQ ID NO:864; a light chain variable region comprising SEQ ID NO:867 and a heavy chain variable region comprising SEQ ID NO:868; a light chain variable region comprising SEQ ID NO:871 and a heavy chain variable region comprising SEQ ID NO:872; a light chain variable region comprising SEQ ID NO:875 and a heavy chain variable region comprising SEQ ID NO:876; a light chain variable region comprising SEQ ID NO:879 and a heavy chain variable region comprising SEQ ID NO:880; a light chain variable region comprising SEQ ID NO:883 and a heavy chain variable region comprising SEQ ID NO:884; a light chain variable region comprising SEQ ID NO:887 and a heavy chain variable region comprising SEQ ID NO:888; a light chain variable region comprising SEQ ID NO:891 and a heavy chain variable region comprising SEQ ID NO:892; a light chain variable region comprising SEQ ID NO:895 and a heavy chain variable region comprising SEQ ID NO:896; a light chain variable region comprising SEQ ID NO:899 and a heavy chain variable region comprising SEQ ID NO:900; a light chain variable region comprising SEQ ID NO:903 and a heavy chain variable region comprising SEQ ID NO:904; a light chain variable region comprising SEQ ID NO:907 and a heavy chain variable region comprising SEQ ID NO:908; a light chain variable region comprising SEQ ID NO:911 and a heavy chain variable region comprising SEQ ID NO:912; a light chain variable region comprising SEQ ID NO:915 and a heavy chain variable region comprising SEQ ID NO:916; a light chain variable region comprising SEQ ID NO:919 and a heavy chain variable region comprising SEQ ID NO:920; a light chain variable region comprising SEQ ID NO:923 and a heavy chain variable region comprising SEQ ID NO:924; a light chain variable region comprising SEQ ID NO:927 and a heavy chain variable region comprising SEQ ID NO:928; a light chain variable region comprising SEQ ID NO:931 and a heavy chain variable region comprising SEQ ID NO:932; a light chain variable region comprising SEQ ID NO:935 and a heavy chain variable region comprising SEQ ID NO:936; a light chain variable region comprising SEQ ID NO:939 and a heavy chain variable region comprising SEQ ID NO:940;a light chain variable region comprising SEQ ID NO:943 and a heavy chain variable region comprising SEQ ID NO:944, a light chain variable region comprising SEQ ID NO:947 and a heavy chain variable region comprising SEQ ID NO:948, a light chain variable region comprising SEQ ID NO:951 and a heavy chain variable region comprising SEQ ID NO:952, a light chain variable region comprising SEQ ID NO:955 and a heavy chain variable region comprising SEQ ID NO:956, a light chain variable region comprising SEQ ID NO:959 and a heavy chain variable region comprising SEQ ID NO:960, a light chain variable region comprising SEQ ID NO:1286 and a heavy chain variable region comprising SEQ ID NO:1287, a light chain variable region comprising SEQ ID NO:1296 and a heavy chain variable region comprising SEQ ID NO:1297, a light chain variable region comprising SEQ ID NO:1306 54. The composition of claim 53, comprising a combination of light chain variable regions and heavy chain variable regions selected from the group consisting of: a light chain variable region comprising SEQ ID NO: 1316 and a heavy chain variable region comprising SEQ ID NO: 1317, a light chain variable region comprising SEQ ID NO: 1326 and a heavy chain variable region comprising SEQ ID NO: 1327, a light chain variable region comprising SEQ ID NO: 1336 and a heavy chain variable region comprising SEQ ID NO: 1337, a light chain variable region comprising SEQ ID NO: 1346 and a heavy chain variable region comprising SEQ ID NO: 1347, and a light chain variable region comprising SEQ ID NO: 1356 and a heavy chain variable region comprising SEQ ID NO: 1357.

65. wherein the antigen binding protein is an antibody, and the antibody is selected from the group consisting of SEQ ID NOs: 963, 967, 971, 975, 979, 983, 987, 991, 995, 999, 1003, 1007, 1011, 1015, 1019, 1023, 1027, 1031, 1035, 1039, 1043, 1047, 1051, 1055, 1059, 1063, 10 67, 1071, 1075, 1079, 1083, 1087, 1091, 1095, 1099, 1103, 1107, 111, and a light chain comprising a sequence selected from the group consisting of SEQ ID NOs: 964, 968, 972, 976, 980, 984, 988, 992, 996, 1000, 1004, 1008, 1012, 1016, 1020, 102 4, 1028, 1032, 1036, 1040, 1044, 1048, 1052, 1056, 1060, 1064, 1068, 1072, 1076, 1080, 1084, 1088, 1092, 1096, 1100, 1104, 1108, 1112, 1116, 1120, 1124, 1128, 1132, 1136, 1140, 114 54. The composition of claim 53, comprising a heavy chain comprising a sequence selected from the group consisting of: 1148, 1152, 1156, 1160, 1164, 1168, 1172, 1176, 1180, 1184, 1188, 1192, 1196, 1200, 1289, 1299, 1309, 1319, 1329, 1339, 1349, and 1359.

66. The antigen binding protein is an antibody, the antibody comprising a light chain comprising SEQ ID NO:963 and a heavy chain comprising SEQ ID NO:964, a light chain comprising SEQ ID NO:967 and a heavy chain comprising SEQ ID NO:968, a light chain comprising SEQ ID NO:971 and a heavy chain comprising SEQ ID NO:972, a light chain comprising SEQ ID NO:975 and a heavy chain comprising SEQ ID NO:976, a light chain comprising SEQ ID NO:979 and a heavy chain comprising SEQ ID NO:980, a light chain comprising SEQ ID NO:983 and a heavy chain comprising SEQ ID NO:984, a light chain comprising SEQ ID NO:987 and a heavy chain comprising SEQ ID NO:988, a light chain comprising SEQ ID NO:991 and a heavy chain comprising SEQ ID NO:992, a light chain comprising SEQ ID NO:995 and a heavy chain comprising SEQ ID NO:996, a light chain comprising SEQ ID NO:997 and a heavy chain comprising SEQ ID NO:998, a light chain comprising SEQ ID NO:998 and a heavy chain comprising SEQ ID NO:999, and a heavy chain comprising SEQ ID NO:996, a light chain comprising SEQ ID NO:999 and a heavy chain comprising SEQ ID NO:1000, a light chain comprising SEQ ID NO:1003 and a heavy chain comprising SEQ ID NO:1004, a light chain comprising SEQ ID NO:1007 and a heavy chain comprising SEQ ID NO:1008, a light chain comprising SEQ ID NO:1011 and a heavy chain comprising SEQ ID NO:1012, a light chain comprising SEQ ID NO:1015 and a heavy chain comprising SEQ ID NO:1016, a light chain comprising SEQ ID NO:1019 and a heavy chain comprising SEQ ID NO:1020, a light chain comprising SEQ ID NO:1023 and a heavy chain comprising SEQ ID NO:1024, a light chain comprising SEQ ID NO:1027 and a heavy chain comprising SEQ ID NO:1028 heavy chain, light chain comprising SEQ ID NO:1031 and heavy chain comprising SEQ ID NO:1032, light chain comprising SEQ ID NO:1035 and heavy chain comprising SEQ ID NO:1036, light chain comprising SEQ ID NO:1039 and heavy chain comprising SEQ ID NO:1040, light chain comprising SEQ ID NO:1043 and heavy chain comprising SEQ ID NO:1044, light chain comprising SEQ ID NO:1047 and heavy chain comprising SEQ ID NO:1048, light chain comprising SEQ ID NO:1051 and heavy chain comprising SEQ ID NO:1052, light chain comprising SEQ ID NO:1055 and heavy chain comprising SEQ ID NO:1056, light chain comprising SEQ ID NO:1059 and heavy chain comprising SEQ ID NO:1060, light chain comprising SEQ ID NO:1063 and a heavy chain comprising SEQ ID NO: 1064, a light chain comprising SEQ ID NO: 1067 and a heavy chain comprising SEQ ID NO: 1068, a light chain comprising SEQ ID NO: 1071 and a heavy chain comprising SEQ ID NO: 1072, a light chain comprising SEQ ID NO: 1075 and a heavy chain comprising SEQ ID NO: 1076, a light chain comprising SEQ ID NO: 1079 and a heavy chain comprising SEQ ID NO: 1080, a light chain comprising SEQ ID NO: 1083 and a heavy chain comprising SEQ ID NO: 1084, a light chain comprising SEQ ID NO: 1087 and a heavy chain comprising SEQ ID NO: 1088, a light chain comprising SEQ ID NO: 1091 and a heavy chain comprising SEQ ID NO: 1092, a light chain comprising SEQ ID NO: 1095 and a heavy chain comprising SEQ ID NO: 1096,a light chain comprising SEQ ID NO:1099 and a heavy chain comprising SEQ ID NO:1100; a light chain comprising SEQ ID NO:1103 and a heavy chain comprising SEQ ID NO:1104; a light chain comprising SEQ ID NO:1107 and a heavy chain comprising SEQ ID NO:1108; a light chain comprising SEQ ID NO:1111 and a heavy chain comprising SEQ ID NO:1112; a light chain comprising SEQ ID NO:1115 and a heavy chain comprising SEQ ID NO:1116; a light chain comprising SEQ ID NO:1119 and a heavy chain comprising SEQ ID NO:1120; a light chain comprising SEQ ID NO:1123 and a heavy chain comprising SEQ ID NO:1124; a light chain comprising SEQ ID NO:1127 and a heavy chain comprising SEQ ID NO:1128; a light chain comprising SEQ ID NO:1131 and a heavy chain comprising SEQ ID NO:1132, a light chain comprising SEQ ID NO:1135 and a heavy chain comprising SEQ ID NO:1136, a light chain comprising SEQ ID NO:1139 and a heavy chain comprising SEQ ID NO:1140, a light chain comprising SEQ ID NO:1143 and a heavy chain comprising SEQ ID NO:1144, a light chain comprising SEQ ID NO:1147 and a heavy chain comprising SEQ ID NO:1148, a light chain comprising SEQ ID NO:1151 and a heavy chain comprising SEQ ID NO:1152, a light chain comprising SEQ ID NO:1155 and a heavy chain comprising SEQ ID NO:1156, a light chain comprising SEQ ID NO:1159 and a heavy chain comprising SEQ ID NO:1160, a light chain comprising SEQ ID NO:1163 and a heavy chain comprising SEQ ID NO:1164, A light chain comprising SEQ ID NO:1167 and a heavy chain comprising SEQ ID NO:1168, a light chain comprising SEQ ID NO:1171 and a heavy chain comprising SEQ ID NO:1172, a light chain comprising SEQ ID NO:1175 and a heavy chain comprising SEQ ID NO:1176, a light chain comprising SEQ ID NO:1179 and a heavy chain comprising SEQ ID NO:1180, a light chain comprising SEQ ID NO:1183 and a heavy chain comprising SEQ ID NO:1184, a light chain comprising SEQ ID NO:1187 and a heavy chain comprising SEQ ID NO:1188, a light chain comprising SEQ ID NO:1191 and a heavy chain comprising SEQ ID NO:1192, a light chain comprising SEQ ID NO:1195 and a heavy chain comprising SEQ ID NO:1196, a light chain comprising SEQ ID NO:1199 and a heavy chain comprising SEQ ID NO:1197, a heavy chain comprising sequence number 1200, a light chain comprising SEQ ID NO: 1288 and a heavy chain comprising SEQ ID NO: 1289, a light chain comprising SEQ ID NO: 1298 and a heavy chain comprising SEQ ID NO: 1299, a light chain comprising SEQ ID NO: 1308 and a heavy chain comprising SEQ ID NO: 1309, a light chain comprising SEQ ID NO: 1318 and a heavy chain comprising SEQ ID NO: 1319, a light chain comprising SEQ ID NO: 1328 and a heavy chain comprising SEQ ID NO: 1329, a light chain comprising SEQ ID NO: 1338 and a heavy chain comprising SEQ ID NO: 1339, a light chain comprising SEQ ID NO: 1348 and a heavy chain comprising SEQ ID NO: 1349, and a light chain comprising SEQ ID NO: 1358 and a heavy chain comprising SEQ ID NO: 1359,54. The composition of claim 53, comprising a combination of light and heavy chains selected from the group consisting of:

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