Method of treating or ameliorating metabolic disorders using GLP-1 receptor agonists conjugated to antagonists for gastric inhibitory peptide receptor (GIPR)

Abstract

To provide a composition comprising an antigen binding protein that specifically binds to human GIPR for use in a method of treating metabolic diseases and disorders.SOLUTION: In one aspect, a composition comprises: a) an antibody or functional fragment thereof that specifically binds to human GIPR, where the antibody or functional fragment thereof comprises a cysteine or amino acid substitution of non-canonical amino acid at one or more conjugation sites; and b) a GLP-1 receptor agonist, where the GLP-1 receptor agonist is conjugated to the antibody or functional fragment thereof through a side chain of the cysteine residue or non-canonical amino acid residue substituted at the one or more conjugation sites.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, cardiovascular disease or diabetic nephropathy using a GLP-1 receptor agonist conjugated to an antigen binding protein 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 of pro-GIP, a 153-amino acid precursor 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 numerous physiological effects on tissues, including promoting fat storage in adipocytes and stimulating pancreatic islet β-cell function and glucose-dependent insulin secretion. GIP and glucagon-like polypeptide-1 (GLP-1) are known as insulinotropic factors ("incretins"). Intact GIP is rapidly degraded by DPPIV to an inactive form. In type 2 diabetic patients, the insulinotropic effect of GIP is lost, whereas 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, forming 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 profiles (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] Taken together, these associations with obesity and insulin resistance suggest that inhibiting GIPR may be a useful approach for therapeutic intervention.

[0007] 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, upon food ingestion, induces 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 effects by activating the GLP-1 receptor, a class B G protein-coupled receptor (Endocrinology 1993, 133(4):1907-10). The function of GLP-1 is limited by rapid degradation by the DPP-IV enzyme, resulting in a half-life of approximately 2 minutes. Long-acting GLP-1 receptor agonists (GLP-1RAs), such as exenatide, liraglutide, and dulaglutide, have recently been developed and are currently being used clinically to improve glycemic control in patients with type 2 diabetes. Furthermore, GLP-1RAs also promote weight loss and reductions in blood pressure and plasma cholesterol levels in patients (Bioorg. Med. Chem. Lett 2013, 23:4011-4018). [Prior art documents] [Non-patent literature]

[0008] [Non-Patent Document 1] Inagaki et al.,Mol Endocrinol 1989;3:1014-1021 [Non-patent document 2] Fehmann et al. Endocr Rev.1995;16:390-410 [Non-patent document 3] Nauck et al.J.Clinc.Invest.1993;91:301-307 [Non-patent document 4] Usdin et al.,Endocrinology.1993,133:2861-2870 [Non-Patent Document 5] Gremlich et al.,Diabetes.1995;44:1202-8 [Non-patent document 6] Volz et al.,FEBS Lett.1995,373:23-29 [Non-Patent Document 7] Yamada et al.,Diabetes.2006,55:S86 [Non-patent document 8] Miyawaki et al.Nature Med.2002,8:738-742 [Non-Patent Document 9] Diabetologia 2008,51:S373,44th EASD Annual meeting poster [Non-Patent Document 10] Diabetes 2004,53:S3,205-214 [Non-Patent Document 11] Diabetes Care,2003,26(10):2929-2940 [Non-Patent Document 12] Endocrinology.1993,133(4):1907-10 [Non-Patent Document 13] Bioorg.Med.Chem.Lett 2013,23:4011-4018 Summary of the Invention [Means for solving the problem]

[0009] 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 a composition comprising an antigen binding protein that specifically binds to human GIPR, wherein the antigen binding protein is conjugated to a GLP-1 receptor agonist. In one embodiment, the metabolic disorder is a disorder of glucose metabolism. In another embodiment, the glucose metabolism disorder comprises hyperglycemia, and administration of the antigen binding protein reduces plasma glucose. In another embodiment, the glucose metabolism disorder comprises hyperinsulinemia, and administration of the antigen binding protein reduces plasma insulin. In another embodiment, the glucose metabolism 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 composition reduces the body weight of the obese subject. In another embodiment, administration of the composition reduces weight gain in the obese subject. In another embodiment, administration of the composition reduces body fat mass in the obese subject. In another embodiment, the impaired glucose metabolism comprises insulin resistance, and administration of the composition reduces insulin resistance in the obese subject. In another embodiment, administration of the composition reduces hepatic steatosis in an obese subject with advanced hepatic steatosis. In another embodiment, administration of the composition reduces liver fat content in an obese subject with increased liver fat content.

[0010] In one embodiment, the composition comprises an antibody or functional fragment thereof that specifically binds to a protein having an amino acid sequence with at least 90% amino acid sequence identity to the amino acid sequence of human GIPR, wherein the antibody or functional fragment thereof comprises an amino acid substitution of a cysteine ​​or a non-standard amino acid at one or more conjugation sites selected from the group consisting of D70 of the antibody light chain for reference sequence SEQ ID NO: 455, E276 of the antibody heavy chain for reference sequence SEQ ID NO: 612, and T363 of the antibody heavy chain for reference sequence SEQ ID NO: 612, and a GLP-1 receptor agonist conjugated to the antibody or functional fragment through the side chain of the substituted cysteine ​​residue or non-standard amino acid residue at the one or more conjugation sites.

[0011] In one embodiment, the human GIPR has a sequence comprising a sequence selected from the group consisting of SEQ ID NO: 3141, SEQ ID NO: 3143, and SEQ ID NO: 3145. In one embodiment, the antibody or functional fragment thereof is a monoclonal antibody, a recombinant antibody, a human antibody, a humanized antibody, a chimeric antibody, a multispecific antibody, or an antibody fragment thereof. In one embodiment, the antibody fragment is a Fab fragment, a Fab' fragment, or a F(ab')2 fragment. In one embodiment, the antibody or functional fragment thereof is of the IgG1, IgG2, IgG3, or IgG4 type. In one embodiment, the antibody or functional fragment thereof inhibits binding of GIP to the extracellular portion of human GIPR.

[0012] In one embodiment, the GLP-1 receptor agonist is GLP-1(7-37) or a GLP-1(7-37) analog. In one embodiment, the GLP-1 receptor agonist is selected from the group consisting of exenatide, liraglutide, lixisenatide, albiglutide, dulaglutide, semaglutide, and taspoglutide. In one embodiment, the GLP-1 receptor agonist is selected from the group consisting of GLP-1(7-37) (SEQ ID NO: 3184); GLP-1(7-36)-NH2 (SEQ ID NO: 3185); liraglutide, albiglutide, taspoglutide, dulaglutide, semaglutide; LY2428757; exendin-4 (SEQ ID NO: 3163); exendin-3 (SEQ ID NO: 3164); Leu 14 - exendin-4 (SEQ ID NO: 3165); Leu 14 ,Phe 25 - exendin-4 (SEQ ID NO: 3166); Leu 14 ,Ala 19 ,Phe 25 - Exendin-4 (SEQ ID NO: 3167); Exendin-4(1-30) (SEQ ID NO: 3168); Leu 14 - exendin-4(1-30) (SEQ ID NO: 3169); Leu 14 ,Phe 25 - Exendin-4(1-30) (SEQ ID NO: 3170); Leu 14 ,Ala 19 ,Phe 25 - Exendin-4(1-30) (SEQ ID NO: 3171); Exendin-4(1-28) (SEQ ID NO: 3172); Leu 14 - exendin-4(1-28) (SEQ ID NO: 3173); Leu 14 ,Phe 25 - exendin-4(1-28) (SEQ ID NO: 3174); Leu 14 ,Ala 19 ,Phe 25 - exendin-4(1-28) (SEQ ID NO: 3175); Leu 14 ,Lys 17,20 ,Ala 19 ,Glu 21 ,Phe 25 ,Gln 28- exendin-4 (SEQ ID NO: 3176); Leu 14 ,Lys 17,20 ,Ala 19 ,Glu 21 ,Gln 28 - Exendin-4 (SEQ ID NO: 3177); OctylGly 14 ,Gln 28 - exendin-4 (SEQ ID NO: 3178); Leu 14 ,Gln 28 ,Octyl Gly 34 - Exendin-4 (SEQ ID NO: 3179); Phe 4 ,Leu 14 ,Gln 28 ,Lys 33 ,Glu 34 ,Ile 35,36 ,Ser 37 - Exendin-4(1-37) (SEQ ID NO: 3180); Phe 4 ,Leu 14 ,Lys 17,20 ,Ala 19 ,Glu 21 ,Gln 28 - Exendin-4 (SEQ ID NO: 3181); Val 11 ,Ile 13 ,Leu 14 ,Ala 16 ,Lys 21 ,Phe 25 -Exendin-4 (SEQ ID NO: 3182); Exendin-4-Lys 40 (SEQ ID NO: 3183); GLP-1(7-37) (SEQ ID NO: 3184); GLP-1(7-36)-NH2 (SEQ ID NO: 3185); Aib 8,35 ,Arg 26,34 ,Phe 31 -GLP-1(7-36)) (SEQ ID NO: 3186); HXaa8EGTFTSDVSSYLEXaa 22 Xaa 23 AAKEFIXaa 30 WLXaa 33 Xaa 34 GXaa 36 Xaa 37 (Wherein Xaa8 is A, V or G, Xaa 22 is G, K or E, and Xaa23 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, and Xaa 36 is R or G, and Xaa 37 is G, H, P or absent) (SEQ ID NO: 3187); Arg 34 -GLP-1(7-37) (SEQ ID NO: 3188); Glu 30 -GLP-1(7-37) (SEQ ID NO: 3189); Lys 22 -GLP-1(7-37) (SEQ ID NO: 3190); Gly 8,36 ,Glu 22 -GLP-1(7-37) (SEQ ID NO: 3191); Val 8 ,Glu 22 ,Gly 36 -GLP-1(7-37) (SEQ ID NO: 3192); Gly 8,36 ,Glu 22 ,Lys 33 ,Asn 34 -GLP-1(7-37) (SEQ ID NO: 3193); Val 8 ,Glu 22 ,Lys 33 ,Asn 34 ,Gly 36 -GLP-1(7-37) (SEQ ID NO: 3194); Gly 8,36 ,Glu 22 ,Pro 37 -GLP-1(7-37) (SEQ ID NO: 3195); Val 8 ,Glu 22 ,Gly 36 ,Pro 37 -GLP-1(7-37) (SEQ ID NO: 3196); Gly 8,36 ,Glu 22 ,Lys 33 ,Asn 34 ,Pro 37 -GLP-1(7-37) (SEQ ID NO: 3197); Val 8 ,Glu 22 ,Lys 33 ,Asn 34 ,Gly 36 ,Pro 37-GLP-1(7-37) (SEQ ID NO: 3198); Gly 8,36 ,Glu 22 -GLP-1(7-36) (SEQ ID NO: 3199); Val 8 ,Glu 22 ,Gly 36 -GLP-1(7-36) (SEQ ID NO: 3200); Val 8 ,Glu 22 ,Asn 34 ,Gly 36 -GLP-1(7-36) (SEQ ID NO: 3201); Gly 8,36 ,Glu 22 ,Asn 34 -GLP-1(7-36) (SEQ ID NO: 3202); GLP-1 analog (SEQ ID NO: 3206); GLP-1 analog (SEQ ID NO: 3207); [N e -(17-carboxyheptadecanoic acid)Lys 20 ]Exendin-4-NH2 (SEQ ID NO: 3208); [N e -(17-carboxyheptadecanoyl)Lys 32 ] Exendin-4-NH2 (SEQ ID NO: 3209); [Desamino-His 1 ,N e -(17-carboxyheptadecanoyl)Lys 20 ] exendin-4-NH2 (SEQ ID NO: 3210); [Arg 12,27 ,NLe 14 ,N e -(17-carboxy-heptadecanoyl)Lys 32 ]Exendin-4-NH2 (SEQ ID NO: 3211); [N e -(19-carboxy-nonadecanoylamino)Lys 20 ]-exendin-4-NH2 (SEQ ID NO: 3212); [N e -(15-carboxypentadecanoylamino)Lys 20 ]-exendin-4-NH2 (SEQ ID NO: 3213); [N e -(13-carboxytridecanoylamino)Lys 20 ]Exendin-4-NH2 (SEQ ID NO: 3214); [N e -(11-carboxy-undecanoyl-amino)Lys 20] Exendin-4-NH2 (SEQ ID NO: 3215); Exendin-4-Lys 40 (e-MPA)-NH2 (SEQ ID NO: 3216); Exendin-4-Lys 40 (e-AEEA-AEEA-MPA)-NH2 (SEQ ID NO: 3217); Exendin-4-Lys 40 (e-AEEA-MPA)-NH2 (SEQ ID NO: 3218); Exendin-4-Lys 40 (e-MPA)-albumin (SEQ ID NO: 3219); exendin-4-Lys 40 (e-AEEA-AEEA-MPA)-albumin (SEQ ID NO: 3220); Exendin-4-Lys 40 (e-AEEA-MPA)-albumin (SEQ ID NO: 3221); 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: 3222) (SEQ ID NO: 3222); desamino-His 7 ,Arg 26 ,Lys 34 (N ε -octanoyl)-GLP-1(7-37) (SEQ ID NO: 3223); Arg 26,34 ,Lys 38 (N ε -(ω-carboxypentadecanoyl)-GLP-1(7-38) (SEQ ID NO: 3224); Arg 26,34 ,Lys 36 (N ε -(γ-Glu(N-α-hexadecanoyl)))-GLP-1(7-36) (core peptide disclosed as SEQ ID NO: 3225) (SEQ ID NO: 3225); [Aib 8 ;Lys 37 ]GLP-1_(7-37) (SEQ ID NO: 3226); [Aib 8 ,Lys 26 ]GLP-1_(7-37) (SEQ ID NO: 3227); [Aib 8,22 ;Lys 36 ]GLP-1(7-36)-amide (SEQ ID NO: 3228); [Aib 8,22 ;BLeu32 ;Lys 36 ]GLP-1(7-36)-amide (SEQ ID NO: 3229); [Aib 8,22 ;Lys 37 ]GLP-1(7-37)-amide (SEQ ID NO: 3230); [Aib 8,22 ;BLeu 32 ;Lys 37 ]GLP-1(7-37)-amide (SEQ ID NO: 3231); [Aib 8,22 ;aMeLeu 32 ;Lys 37 ]GLP-1(7-37)-amide (SEQ ID NO: 3232); [Aib 8,22 ;AMEF 12 ;Lys 37 ]GLP-1(7-37)-amide (SEQ ID NO: 3233); [Aib 8,22 ;BLeu 16 ;Lys 37 ]GLP-1(7-37)-amide (SEQ ID NO: 3234); [Aib 8,22 ;Gly 36 ;Lys 37 ]GLP-1(7-37)-amide (SEQ ID NO: 3235); [Aib 8,22 ;Lys 33,37 ;Asn 34 ;Gly 36 ]GLP-1(7-37)-amide (SEQ ID NO: 3236); [Aib 8,22 ;Lys 33 ;Asn 34 ;Gly 36 ;Aeea 37 ]GLP-1(7-37)-Aeea-Lys-amide (SEQ ID NO: 3237); [Aib 8,22 ;Gly 36 ]GLP-1(7-37) (SEQ ID NO: 3238); cyclo[E 23 -K 27 ][Aib 8 ;Gly 36 ]GLP-1(7-37) (SEQ ID NO: 3239); cyclo[E 22 -K 26 ][Aib 8 ;Gly 36 ;Lys 37]GLP-1(7-37)-amide (SEQ ID NO: 3240); [Aib 8,22 ]-GLP-1(7-22)-Ex4(17-39) (SEQ ID NO: 3241); [Gly 8,36 ;Glu 22 ]GLP-1(7-37) (SEQ ID NO: 3242); [Aib 8 ;Glu 22 ;Gly 36 ]GLP-1(7-37)-amide (SEQ ID NO: 3243); [Aib 8 ;Tyr 16 ;Glu 22 ;Gly 36 ]GLP-1_(7-37) (SEQ ID NO: 3244); [Aib 8 ;Lys 18,33 ;Glu 22,23,30 ;Val 25 ;Arg 26 ;Leu 27 ;Asn 34 ;Gly 36 ]GLP-1(7-37) (SEQ ID NO: 3245); [Aib 8 ;Lys 18,33 ;Glu 22,23,30 ;Leu 27 ;Asn 34 ;Gly 36 ]GLP-1(7-37) (SEQ ID NO: 3246); [Aib 8 ;Lys 18 ;Glu 22,23,30 ;Leu 27 ;Gly 36 ]GLP-1(7-37) (SEQ ID NO: 3247); [Aib 8,22 ;Ile 9 ;Gly 36 ]GLP-1_(7-36) (SEQ ID NO: 3248); and [Aib 8,22 ;Glu 15 ;Gly 36 ]GLP-1_(7-36) (SEQ ID NO: 3249).

[0013] In one embodiment, the GLP-1(7-37) or GLP-1(7-37) analog is conjugated to the antibody or fragment thereof at K26, K36, K37, K39 or a residue corresponding to the C-terminal amine group of the analog.

[0014] In one embodiment, the GLP-1 receptor agonist is conjugated to the antibody or fragment thereof via a peptide linker comprising a sequence selected from the group consisting of (Gly3Ser)2, (Gly4Ser)2, (Gly3Ser)3, (Gly4Ser)3, (Gly3Ser)4, (Gly4Ser)4, (Gly3Ser)5, (Gly4Ser)5, (Gly3Ser)6 and (Gly4Ser)6. [Brief explanation of the drawings]

[0015] [Figure 1] 1 is a reaction scheme for the production of anti-GIPR / GLP-1 peptide conjugates. [Figure 2A] GLP-1 receptor agonist activity in CHOK1-GLP-1R H20 recombinant cells and GIPR antagonist activity in 293HEK-GIPR #10 recombinant cells. [Figure 2B] GLP-1 receptor agonist activity in CHOK1-GLP-1R H20 recombinant cells and GIPR antagonist activity in 293HEK-GIPR #10 recombinant cells. [Figure 2C] GLP-1 receptor agonist activity in CHOK1-GLP-1R H20 recombinant cells and GIPR antagonist activity in 293HEK-GIPR #10 recombinant cells. [Figure 2D] GLP-1 receptor agonist activity in CHOK1-GLP-1R H20 recombinant cells and GIPR antagonist activity in 293HEK-GIPR #10 recombinant cells. [Figure 2E] GLP-1 receptor agonist activity in CHOK1-GLP-1R H20 recombinant cells and GIPR antagonist activity in 293HEK-GIPR #10 recombinant cells. [Figure 2F] GLP-1 receptor agonist activity in CHOK1-GLP-1R H20 recombinant cells and GIPR antagonist activity in 293HEK-GIPR #10 recombinant cells. [Figure 2G]GLP-1 receptor agonist activity in CHOK1-GLP-1R H20 recombinant cells and GIPR antagonist activity in 293HEK-GIPR #10 recombinant cells. [Figure 2H] GLP-1 receptor agonist activity in CHOK1-GLP-1R H20 recombinant cells and GIPR antagonist activity in 293HEK-GIPR #10 recombinant cells. [Figure 2I] GLP-1 receptor agonist activity in CHOK1-GLP-1R H20 recombinant cells and GIPR antagonist activity in 293HEK-GIPR #10 recombinant cells. [Figure 2J] GLP-1 receptor agonist activity in CHOK1-GLP-1R H20 recombinant cells and GIPR antagonist activity in 293HEK-GIPR #10 recombinant cells. [Figure 2K] GLP-1 receptor agonist activity in CHOK1-GLP-1R H20 recombinant cells and GIPR antagonist activity in 293HEK-GIPR #10 recombinant cells. [Figure 2L] GLP-1 receptor agonist activity in CHOK1-GLP-1R H20 recombinant cells and GIPR antagonist activity in 293HEK-GIPR #10 recombinant cells. [Figure 2M] GLP-1 receptor agonist activity in CHOK1-GLP-1R H20 recombinant cells and GIPR antagonist activity in 293HEK-GIPR #10 recombinant cells. [Figure 2N] GLP-1 receptor agonist activity in CHOK1-GLP-1R H20 recombinant cells and GIPR antagonist activity in 293HEK-GIPR #10 recombinant cells. [Figure 2O] GLP-1 receptor agonist activity in CHOK1-GLP-1R H20 recombinant cells and GIPR antagonist activity in 293HEK-GIPR #10 recombinant cells. [Figure 2P]GLP-1 receptor agonist activity in CHOK1-GLP-1R H20 recombinant cells and GIPR antagonist activity in 293HEK-GIPR #10 recombinant cells. [Figure 2Q] GLP-1 receptor agonist activity in CHOK1-GLP-1R H20 recombinant cells and GIPR antagonist activity in 293HEK-GIPR #10 recombinant cells. [Figure 2R] GLP-1 receptor agonist activity in CHOK1-GLP-1R H20 recombinant cells and GIPR antagonist activity in 293HEK-GIPR #10 recombinant cells. [Figure 2S] GLP-1 receptor agonist activity in CHOK1-GLP-1R H20 recombinant cells and GIPR antagonist activity in 293HEK-GIPR #10 recombinant cells. [Figure 2T] GLP-1 receptor agonist activity in CHOK1-GLP-1R H20 recombinant cells and GIPR antagonist activity in 293HEK-GIPR #10 recombinant cells. [Figure 3A] The GLP-1 agonist activity of the 2G10 conjugate is more potent in cells expressing both GLP-1R and GIPR (CHOK1 hGLP-1R / hGIPR M1) compared to cells expressing only GLP-1R (CHOK1 hGLP-1R H20). [Figure 3B] The GLP-1 agonist activity of the 2G10 conjugate is more potent in cells expressing both GLP-1R and GIPR (CHOK1 hGLP-1R / hGIPR M1) compared to cells expressing only GLP-1R (CHOK1 hGLP-1R H20). [Figure 3C] The GLP-1 agonist activity of the 2G10 conjugate is more potent in cells expressing both GLP-1R and GIPR (CHOK1 hGLP-1R / hGIPR M1) compared to cells expressing only GLP-1R (CHOK1 hGLP-1R H20). [Figure 4A]In vivo activity in db / db mice - effects on glucose and body weight. [Figure 4B] In vivo activity in db / db mice - effects on glucose and body weight. [Figure 4C] In vivo activity in db / db mice - effects on glucose and body weight. [Figure 4D] In vivo activity in db / db mice - effects on glucose and body weight. [Figure 4E] In vivo activity in db / db mice - effects on glucose and body weight. [Figure 4F] In vivo activity in db / db mice - effects on glucose and body weight. [Figure 4G] In vivo activity in db / db mice - effects on glucose and body weight. [Figure 4H] In vivo activity in db / db mice - effects on glucose and body weight. [Figure 4I] In vivo activity in db / db mice - effects on glucose and body weight. [Figure 4J] In vivo activity in db / db mice - effects on glucose and body weight. [Figure 4K] In vivo activity in db / db mice - effects on glucose and body weight. [Figure 4L] In vivo activity in db / db mice - effects on glucose and body weight. [Figure 4M] In vivo activity in db / db mice - effects on glucose and body weight. [Figure 4N] In vivo activity in db / db mice - effects on glucose and body weight. [Figure 4O] In vivo activity in db / db mice - effects on glucose and body weight. [Figure 4P] In vivo activity in db / db mice - effects on glucose and body weight. [Figure 4Q] In vivo activity in db / db mice - effects on glucose and body weight. [Figure 4R] In vivo activity in db / db mice - effects on glucose and body weight. [Figure 5A] In vivo GLP-1 receptor agonist and GIPR antagonist activity of 5G12 and 2G10 chemical conjugates in C57B16 mice (glucose tolerance test). [Figure 5B] In vivo GLP-1 receptor agonist and GIPR antagonist activity of 5G12 and 2G10 chemical conjugates in C57B16 mice (glucose tolerance test). [Figure 5C] In vivo GLP-1 receptor agonist and GIPR antagonist activity of 5G12 and 2G10 chemical conjugates in C57B16 mice (glucose tolerance test). [Figure 5D] In vivo GLP-1 receptor agonist and GIPR antagonist activity of 5G12 and 2G10 chemical conjugates in C57B16 mice (glucose tolerance test). [Figure 6A] In vivo GLP-1 receptor agonist and GIPR antagonist activity (plasma glucose and body weight changes) of 5G12 and 2G10 chemical conjugates in DIO mice. [Figure 6B] In vivo GLP-1 receptor agonist and GIPR antagonist activity (plasma glucose and body weight changes) of 5G12 and 2G10 chemical conjugates in DIO mice. [Figure 7] GLP-1R-GIPR bispecific conjugate 1273 reduces body weight (% change from baseline) in male idiopathically obese cynomolgus monkeys. [Figure 8] GLP-1R-GIPR bispecific conjugate 1273 reduces total energy intake (% change from baseline) in male idiopathically obese cynomolgus monkeys. [Figure 9] GLP-1R-GIPR bispecific conjugate 1273 reduces fasting insulin (% change from baseline) in male idiopathic obese cynomolgus monkeys. [Figure 10] GLP-1R-GIPR bispecific conjugate 1273 reduces fasting triglycerides in male idiopathically obese cynomolgus monkeys (% change from baseline). [Figure 11] GLP-1R-GIPR bispecific conjugate 1273 reduces fasting total cholesterol (% change from baseline) in male idiopathically obese cynomolgus monkeys. [Figure 12] GLP-1R-GIPR bispecific conjugate 1264 reduces body weight (% change from baseline) in male idiopathically obese cynomolgus monkeys. [Figure 13] GLP-1R-GIPR bispecific conjugate 1264 reduces total energy intake (% change from baseline) in male idiopathically obese cynomolgus monkeys. [Figure 14] GLP-1R-GIPR bispecific conjugate 1264 reduces fasting insulin (% change from baseline) in male idiopathic obese cynomolgus monkeys. [Figure 15] GLP-1R-GIPR bispecific conjugate 1264 reduces fasting triglycerides in male idiopathically obese cynomolgus monkeys (% change from baseline). [Figure 16] GLP-1R-GIPR bispecific conjugate 1264 reduces fasting total cholesterol (% change from baseline) in male idiopathically obese cynomolgus monkeys. [Figure 17] GLP-1R-GIPR bispecific conjugates 1248 and 1257 reduce body weight (% change from baseline) in male idiopathically obese cynomolgus monkeys. [Figure 18]GLP-1R-GIPR bispecific conjugates 1248 and 1257 reduce total energy intake (% change from baseline) in male idiopathically obese cynomolgus monkeys. [Figure 19] GLP-1R-GIPR bispecific conjugates 1248 and 1257 reduce fasting insulin (% change from baseline) in male idiopathic obese cynomolgus monkeys. [Figure 20] GLP-1R-GIPR bispecific conjugates 1248 and 1257 reduce fasting triglycerides (% change from baseline) in male idiopathically obese cynomolgus monkeys. [Figure 21] GLP-1R-GIPR bispecific conjugates 1248 and 1257 reduce fasting cholesterol (% change from baseline) in male idiopathically obese cynomolgus monkeys. [Figure 22A] Binding affinity of anti-GIPR / GLP-1R bispecific conjugates. [Figure 22B] Binding affinity of anti-GIPR / GLP-1R bispecific conjugates. [Figure 23] The anti-GIPR / GLP-1R bispecific conjugate 1248 promotes potent GLP-1R and GIPR internalization in cells expressing both GLP-1R and GIPR. [Figure 24A] Time course and dose response of anti-GIPR / GLP-1R bispecific conjugates in a GLP-1R internalization assay. [Figure 24B] Time course and dose response of anti-GIPR / GLP-1R bispecific conjugates in a GLP-1R internalization assay. [Figure 25A] GLP-1R, GIPR and the bispecific conjugate co-localize upon internalization. [Figure 25B] GLP-1R, GIPR and the bispecific conjugate co-localize upon internalization. [Figure 25C]GLP-1R, GIPR and the bispecific conjugate co-localize upon internalization. [Figure 25D] GLP-1R, GIPR and the bispecific conjugate co-localize upon internalization. [Figure 26A] Inhibition of receptor internalization reduces 1248-induced cAMP production. [Figure 26B] Inhibition of receptor internalization reduces 1248-induced cAMP production. [Figure 27A] The bispecific conjugate binds to GLP-1R and GIPR in FACS analysis. [Figure 27B] The bispecific conjugate binds to GLP-1R and GIPR in FACS analysis. [Figure 27C] The bispecific conjugate binds to GLP-1R and GIPR in FACS analysis. [Figure 28] The anti-GIPR / GLP-1R bispecific conjugate stimulates calcium flux. [Figure 29A] The anti-GIPR / GLP-1R bispecific conjugate 1248 promotes β-arrestin recruitment through both the GLP-1R and GIPR pathways. [Figure 29B] The anti-GIPR / GLP-1R bispecific conjugate 1248 promotes β-arrestin recruitment through both the GLP-1R and GIPR pathways. [Figure 30] The anti-GIPR / GLP-1R bispecific conjugate 1248 stimulates insulin secretion in human pancreatic islets. DETAILED DESCRIPTION OF THE INVENTION

[0016] The present disclosure provides methods 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), glycosuria, metabolic acidosis, diabetic neuropathy, diabetic nephropathy, diabetic retinopathy, diabetic cardiomyopathy, type 1 diabetes, obesity, and conditions exacerbated by obesity) by blocking or interfering with the biological activity of GIP. In one embodiment, a therapeutically effective amount of an isolated human GIPR binding protein conjugated to a GLP-1 receptor agonist is administered to a subject in need thereof. Methods of administration and delivery are also provided.

[0017] Recombinant methods for 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.

[0018] The section headings used herein are for organizational purposes only and are not to be construed as limiting the subject matter described.

[0019] As used herein, unless otherwise defined, scientific and technical terms used in connection with this application shall have the meanings commonly understood by those skilled in the art. Further, unless the context otherwise requires, singular terms shall include the plural and plural terms shall include the singular.

[0020] Generally, the nomenclatures and techniques used in connection with 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 specified, the methods and techniques of the present application are generally performed according to conventional methods well known in the art, as described in various general and more specific references cited and discussed throughout the 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 references are incorporated herein by reference. Enzymatic reactions and purification techniques are performed according to manufacturer's specifications, as commonly accomplished in the art, or as described herein. The terminology used in connection with, and the laboratory procedures and techniques of, analytical chemistry, synthetic organic chemistry, and medicinal and pharmaceutical chemistry described herein are well-known and commonly used in the art. Standard techniques may be used for chemical syntheses, chemical analyses, pharmaceutical preparation, formulation, and delivery, and treatment of patients.

[0021] 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 solely by the claims.

[0022] 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." When used in connection with percentages, the term "about" can mean ±1%.

[0023] Unless otherwise specified, as used herein, "a" and "an" are used conventionally to mean "one or more."

[0024] 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.

[0025] "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 later 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, such as homoserine, norleucine, methionine sulfoxide, and methylmethionine sulfonium. Such analogs may have modified R groups (e.g., norleucine) or modified peptide backbones, but retain the same basic chemical structure as a naturally occurring amino acid.

[0026] "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-imino acids, γ-imino acids, δ-imino acids (such as piperidine-4-carboxylic acid), and the like.

[0027] A "non-naturally occurring amino acid" is a compound that has the same basic chemical structure as a naturally occurring amino acid, but that 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 a naturally encoded amino acid (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 side chain derivatized amino acids. 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, piperidinic 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 given in parentheses.

[0028] 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, from which at least about 50 percent of the polypeptides, peptides, lipids, carbohydrates, polynucleotides, or other materials naturally found with the nucleic acid when the total nucleic acid is isolated from a cellular source are removed. 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 polypeptide production, or its therapeutic, diagnostic, prophylactic, or research uses.

[0029] The term "isolated polypeptide" refers to a polypeptide (e.g., a GIPR polypeptide sequence provided herein, or an antigen binding protein of the invention) from which 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 have been removed. 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.

[0030] A composition of the invention is a "conjugate" or "conjugated" molecule, whether comprising a GLP-1 receptor agonist of the invention covalently linked, attached or bound to another anti-GIPR antigen binding protein of the invention, either directly or indirectly through a linker moiety, or conjugated by chemical means (e.g., post-translationally or post-synthetically).

[0031] The term "encoding" refers to a polynucleotide sequence that encodes one or more amino acids. The term does not require a start or stop codon.

[0032] 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., "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.

[0033] In calculating percent identity, the sequences being compared are aligned in a way 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 to optimize their respective amino acid or nucleotide matches (the "match span" is determined by the algorithm). A gap opening penalty (calculated as 3 x average diagonal element, where "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 (typically 1 / 10 of 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, standard comparison matrices (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.

[0034] Recommended parameters for determining percent identity of polypeptide or nucleotide sequences using the GAP program include the following: Algorithm: Needleman et al., 1970, J. Mol. Biol. 48:443-453 Comparison matrix: Henikoff et al., 1992, supra Gap penalty: 12 (except for end gaps, which have no penalty) Gap length penalty: 4 Similarity threshold: 0

[0035] When using a specific alignment scheme for aligning two amino acid sequences, only a short region may be identical 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 the alignment spanning at least 50 consecutive amino acids in the target polypeptide.

[0036] The terms "GIPR polypeptide" and "GIPR protein" are used interchangeably and refer to a naturally occurring wild-type polypeptide expressed in a mammal, such as a human or mouse, including naturally occurring alleles (e.g., naturally occurring allelic forms of a human GIPR protein). 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: 3141 (consisting of 466 amino acid residues and encoded by the nucleotide sequence of SEQ ID NO: 3142), or SEQ ID NO: 3143 (consisting of 430 amino acid residues and encoded by the nucleic acid sequence of SEQ ID NO: 3144), or SEQ ID NO: 3145 (consisting of 493 amino acid residues and encoded by the nucleic acid sequence of SEQ ID NO: 3146), or SEQ ID NO: 3147 (consisting of 460 amino acid residues and encoded by the nucleic acid sequence of SEQ ID NO: 3148), or SEQ ID NO: 3149 (consisting of 230 amino acid residues and encoded by the nucleic acid sequence of SEQ ID NO: 3150).

[0037] The term "GIPR polypeptide" also encompasses GIPR polypeptides in which a naturally occurring GIPR polypeptide sequence (e.g., SEQ ID NO: 3141, SEQ ID NO: 3143, or SEQ ID NO: 3145) 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.

[0038] In various embodiments, the GIPR polypeptide comprises an amino acid sequence that is at least about 90 percent identical to a naturally occurring GIPR polypeptide (e.g., SEQ ID NO: 3141, SEQ ID NO: 3143, or SEQ ID NO: 3145). In other embodiments, the GIPR polypeptide comprises an amino acid sequence that is 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: 3141, SEQ ID NO: 3143, or SEQ ID NO: 3145). Such a GIPR polypeptide preferably, but not necessarily, possesses 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 encoding such GIPR polypeptide sequences.

[0039] 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 proteins in a cellular context. In one embodiment, the "activity" (or "functional") assay can be a cAMP assay in GIPR-expressing cells (where GIP can induce cAMP signals), and the activity of GIP / GIPR-binding proteins can be measured in the presence / absence of a 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).

[0040] 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 using FMAT or FACS to measure the binding of fluorescently labeled GIP to GIPR-expressing cells, and the activity of the GIP / GIPR-binding protein can be measured by the displacement of the binding of fluorescently labeled GIP to GIPR-expressing cells. In another embodiment, the "GIPR binding assay" can be an assay to measure the binding of radiolabeled GIP to GIPR-expressing cells, and the activity of the GIP / GIPR-binding protein can be measured by the displacement of the binding of radiolabeled GIP to GIPR-expressing cells (Biochimica et Biophysica Acta (2001) 1547:143-155).

[0041] 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.

[0042] The sequence of the 42 amino acids of mature human GIP is: YAEGTFISDY SIAMDKIHQQ DFVNWLLAQK GKKNDWKHNI TQ (SEQ ID NO: 3151) and the DNA sequence: [ka] is coded by

[0043] The sequence of the 42 amino acids of mature mouse GIP is: YAEGTFISDY SIAMDKIRQQ DFVNWLLAQR GKKSDWKHNI TQ (SEQ ID NO: 3153) and the DNA sequence: [ka] is coded by

[0044] The sequence of 42 amino acids of mature rat GIP is: YAEGTFISDY SIAMDKIRQQ DFVNWLLAQK GKKNDWKHNL TQ (SEQ ID NO: 3155) and the DNA sequence: [ka] is coded by

[0045] "GIPR antagonist" refers to a compound that reduces or inhibits GIP activation of GIPR. Such antagonists include chemically synthesized small molecules and antigen-binding proteins.

[0046] 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 that provided in SEQ ID NO: 3141, SEQ ID NO: 3143, or SEQ ID NO: 3145). 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.

[0047] Generally, 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 by surface plasma resonance (e.g., BIACore, GE-Healthcare Uppsala, Sweden) or equilibrium exclusion (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 described method. -9 M specifically binds human GIPR with "high affinity" when the KD is ≤ 5x10 as measured using the described method -10 When M, it specifically binds to human GIPR with "very high affinity."

[0048] "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 containing amino acid residues that interact with an antigen and confer its specificity and affinity for the antigen to the antigen-binding protein 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. A particular antigen-binding region also comprises one or more "framework" regions. "CDRs" are amino acid sequences that contribute to antigen-binding specificity and affinity. "Framework" regions can facilitate binding between the antigen-binding region and the antigen by helping to maintain the proper conformation of the CDRs.

[0049] 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.

[0050] 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 described further 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.

[0051] The term "light chain" as used in reference to an antibody or a fragment thereof includes a full-length light chain and a fragment 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.

[0052] The term "heavy chain" as used with respect to antibodies or fragments thereof includes full-length heavy chains 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 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.

[0053] 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 comprises a portion of an antibody (regardless of how that portion is obtained or synthesized) that retains 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.

[0054] 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.

[0055] In another embodiment are Fvs, domain antibodies and scFvs, which may be derived from the antibodies of the invention.

[0056] 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 attached 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.

[0057] A "Fab fragment" consists of one light chain and the CH1 and variable region of one heavy chain. The heavy chain of a Fab molecule cannot form disulfide bonds with another heavy chain molecule.

[0058] The "Fc" region comprises two heavy chain fragments containing the CH2 and CH3 domains of an antibody, held together by two or more disulfide bonds and hydrophobic interactions of the CH3 domains.

[0059] An "Fab' fragment" comprises one light chain and a portion of one heavy chain containing the VH domain, the CH1 domain, and the region between the CH1 and CH2 domains, such that an F(ab')2 molecule can be formed by interchain disulfide bond formation between the two heavy chains of two Fab' fragments.

[0060] An "F(ab')2 fragment" contains 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.

[0061] The "Fv region" comprises the variable regions from both the heavy and light chains, but lacks the constant regions.

[0062] 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. Pat. Nos. 4,946,778 and 5,260,203, the disclosures of which are incorporated by reference.

[0063] A "domain antibody" or "single-chain immunoglobulin" is an immunologically functional immunoglobulin fragment containing 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.

[0064] A "bivalent antigen-binding protein" or "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.

[0065] A "multispecific antigen-binding protein" or "multispecific antibody" is one that targets multiple antigens or epitopes.

[0066] A "bispecific," "dual-specific," or "bifunctional" antigen-binding protein or antibody is a hybrid antigen-binding protein or antibody, respectively, having two different antigen-binding sites. Bispecific antigen-binding proteins and antibodies are a type of multispecific antigen-binding protein or multispecific antibody and may be produced 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 reside on the same or different protein targets.

[0067] 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) under test blocks or inhibits specific binding of a reference antigen-binding protein to a common antigen (e.g., GIPR or a fragment thereof). 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., Stahl et al., 1983, Methods in Enzymology 9:242-253), solid-phase direct biotin-avidin EIAs (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 RIAs 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 labeling 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, it 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.

[0068] 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 (e.g., including 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).

[0069] The term "epitope" refers to the portion of a molecule that is bound by an antigen-binding protein (e.g., an antibody). The term includes any determinant capable of specifically binding 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 that have connections within the molecule are subject to binding by an antigen-binding protein). A conformational epitope is an epitope that is present in the conformation of an active protein but not in a denatured protein. In certain embodiments, an epitope can be mimetic in that it comprises a three-dimensional structure similar to the epitope used to generate the antigen-binding protein, but does not comprise, or comprises only some of, the amino acid residues found in the epitope used to generate the antigen-binding protein. Epitopes are most often found on proteins, but can occasionally be present on 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. Generally, 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.

[0070] As used herein, "substantially pure" means that the molecule of the specified 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.

[0071] The term "treatment" refers to any indication of success in treating or ameliorating an injury, condition, or pathology, including any objective or subjective parameter, such as relief, alleviation, or reduction of symptoms, or improved patient tolerance of the injury, condition, or pathology, slowing the rate of deterioration or decline, less debilitating endpoints of deterioration, or improvement in the patient's physical or mental well-being. Treatment or amelioration of symptoms may be based on objective or subjective parameters, including the results of a physical examination, neuropsychiatric testing, and / or psychiatric evaluation. For example, certain methods provided 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.

[0072] An "effective amount" is generally an amount sufficient to reduce the severity and / or frequency of symptoms, eliminate the symptoms and / or underlying causes, prevent the onset of the symptoms and / or their underlying causes, and / or ameliorate or treat damage caused by or associated with a disease state (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 state (e.g., atherosclerosis) or symptom, particularly a condition or symptom associated with a disease state, or to prevent, prevent, delay, or reverse the progression of a disease state or any other undesirable symptom 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 state or reducing the likelihood of the onset (or recurrence) of a disease state or associated symptom. The full therapeutic or prophylactic effect does not necessarily occur by 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 doses.

[0073] 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.

[0074] The term "polynucleotide" or "nucleic acid" includes both single-stranded and double-stranded nucleotide polymers. The nucleotides comprising 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 internucleotide bond modifications such as phosphorothioate, phosphorodithioate, phosphoroselenoate, phosphorodiselenoate, phosphoroanilothioate, phosphoroaniladate, and phosphoroamidate.

[0075] The term "oligonucleotide" refers to a polynucleotide containing 200 or fewer nucleotides. In some embodiments, oligonucleotides are 10-60 bases in length. In other embodiments, oligonucleotides are 12, 13, 14, 15, 16, 17, 18, 19, or 20-40 nucleotides in length. Oligonucleotides can be single-stranded or double-stranded, e.g., for use in constructing mutant genes. Oligonucleotides can be sense or antisense oligonucleotides. Oligonucleotides can contain a label, including a radiolabel, fluorescent label, hapten, or antigenic label for detection assays. Oligonucleotides can be used, e.g., as PCR primers, cloning primers, or hybridization probes.

[0076] An "isolated nucleic acid molecule" means 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 with which it is found in nature or is 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 to not encompass intact chromosomes. An isolated nucleic acid molecule "comprising" 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 recited nucleic acid sequence, and / or may include vector sequences.

[0077] Unless otherwise specified, the left-hand end of any single-stranded polynucleotide sequence discussed herein is the 5'-end, and the left-hand 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."

[0078] 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 particular 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.

[0079] The term "vector" refers to any molecule or entity (eg, nucleic acid, plasmid, bacteriophage, or virus) used to transfer protein-coding information into a host cell.

[0080] 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 (in cooperation with the host cell) direct and / or control 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.

[0081] 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 so that expression of the protein coding sequence is achieved under conditions compatible with the transcriptional activity of the control sequences.

[0082] 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.

[0083] The terms "polypeptide" and "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 a corresponding naturally occurring amino acid, 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 with 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 with one or more amino acid deletions, additions, and / or substitutions of antigen-binding proteins. The term "polypeptide fragment" refers to a polypeptide that has an amino-terminal deletion, a carboxyl-terminal deletion, and / or an internal deletion compared to the full-length protein. Such fragments can also contain modified amino acids compared to the full-length protein. In certain embodiments, fragments are between about 5 and 500 amino acids in length. For example, fragments can 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.

[0084] The term "isolated protein" means that the subject protein (1) is free from at least some other proteins with which it is normally expected to be found; (2) is essentially free from other proteins from the same source, e.g., the same species; (3) is expressed by cells from a different species; (4) has been removed from at least about 50 percent of the polynucleotides, lipids, carbohydrates, or other materials which naturally accompany it; (5) is operably associated (by covalent or noncovalent interactions) with polypeptides which are not naturally associated with it; or (6) is 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 as found in its natural environment that would interfere with its therapeutic, diagnostic, prophylactic, research, or other use.

[0085] 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 compared to another polypeptide sequence. Variants include fusion proteins.

[0086] 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.

[0087] The term "naturally occurring" 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.

[0088] As used herein, a "subject" or "patient" can be any mammal. In typical embodiments, the subject or patient is a human.

[0089] As disclosed herein, the GIPR polypeptides described by this disclosure can be engineered and / or produced using standard molecular biology methodologies. 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.

[0090] 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, e.g., an automated DNA synthesizer, or can be isolated from longer DNA sequences.

[0091] 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 the DNA sequence (NCBI reference sequence: NM_000164): [ka] [ka] is coded by

[0092] 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] having the DNA sequence: [ka] is coded by

[0093] 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] having the DNA sequence: [ka] is coded by

[0094] 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 the DNA sequence (NCBI reference sequence: NM_001080815): [ka] is coded by

[0095] 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] having the DNA sequence: [ka] is coded by

[0096] The term "GIPR polypeptide" as used herein encompasses naturally occurring GIPR polypeptide sequences, such as, for example, the human amino acid sequence of SEQ ID NO: 3141, SEQ ID NO: 3143, or SEQ ID NO: 3145. However, the term "GIPR polypeptide" also encompasses polypeptides comprising 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: 3141, SEQ ID NO: 3143, or SEQ ID NO: 3145, by one or more amino acids, such that the sequence is at least 90% identical to SEQ ID NO: 3141, SEQ ID NO: 3143, or SEQ ID NO: 3145. 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) into specific positions of a GIPR polypeptide.

[0097] 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.

[0098] 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 influence chain orientation: Gly, Pro, and (6) Aromatic: Trp, Tyr, Phe.

[0099] 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 substitutions based on two or more of such properties (e.g., substitutions with "small polar" residues such as Thr residues may be highly conservative substitutions in appropriate circumstances).

[0100] Conservative substitutions may involve exchanging a member of one such class for another member of the same class. Non-conservative substitutions may involve exchanging a member of one such class for a member of another class.

[0101] Synthetic, rare, or modified amino acid residues known to have similar physicochemical properties to those in the above classes can be used as "conservative" substitutions for particular amino acid residues in a sequence. For example, a D-Arg residue can serve as a substitute for a typical L-Arg residue. In some cases, a particular substitution can be described in terms of more than one of the above classes (e.g., substitution with a small, hydrophobic residue refers to substitution of one amino acid with another synthetic, rare, or modified residue known in the art to have similar physicochemical properties as a residue found in both of the above classes or as such a residue meeting both definitions).

[0102] Nucleic acid sequences encoding GIPR polypeptides provided herein include those that are degenerate to SEQ ID NO:3141, SEQ ID NO:3143, or SEQ ID NO:3145, and those that encode polypeptide variants of SEQ ID NO:3141, SEQ ID NO:3143, or SEQ ID NO:3145 from other embodiments of the disclosure.

[0103] To express the GIPR nucleic acid sequences provided herein, a suitable coding sequence, such as SEQ ID NO:3141, SEQ ID NO:3143, or SEQ ID NO:3145, 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; such techniques are known in the art (e.g., as described in Sambrook, J., Fritsh, E. F., 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 invention.

[0104] A "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 / transfected cells, and / or (g) otherwise confers a beneficial biological and / or physiochemical function 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.

[0105] 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. Exemplary 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 vector (Van Heeke & Schuster, J. Biol. Chem. 264:5503-5509 (1989)), pET vector (Novagen, Madison, WI). Wis. 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 containing the nucleic acid sequence encoding the polypeptide. Examples of switchable promoters include the lac promoter, T7 promoter, trc promoter, tac promoter, and trp promoter.

[0106] Thus, provided herein are vectors that contain a nucleic acid sequence encoding GIPR and promote the expression of recombinant GIPR. In various embodiments, the vector contains, operably linked to a nucleotide sequence that regulates the expression of GIPR. The vector may contain or be associated with any suitable promoter, enhancer, and other expression-promoting elements. Examples of such elements include a strong expression promoter (e.g., the human CMV IE promoter / enhancer, RSV promoter, SV40 promoter, SL3-3 promoter, MMTV promoter, or HIV LTR promoter, EF1 alpha promoter, or CAG promoter), an efficient 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., a polylinker). The vector 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-encoding sequence operably linked to a tissue-specific promoter that promotes expression of the sequence in metabolically relevant tissues, such as liver or pancreatic tissue.

[0107] 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.

[0108] 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 combinations 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.

[0109] A vector containing a nucleic acid sequence encoding a GIPR polypeptide provided herein can be introduced into a host cell by transformation or transfection. Methods for transforming cells with expression vectors are well known.

[0110] 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 contain 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 the desired host cell. The viral vector can be a polynucleotide comprising all or part of the viral genome, a viral protein / nucleic acid complex, a virus-like particle (VLP), or an intact viral particle comprising 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 to facilitate gene transfer and / or expression of nucleic acids (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).

[0111] 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.

[0112] 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 may 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.

[0113] Provided herein are antagonist antigen-binding proteins that bind to GIPR, including human GIPR (hGIPR).In one embodiment, human GIPR has a sequence such as that shown in SEQ ID NO: 3141.In another embodiment, human GIPR has a sequence such as that shown in SEQ ID NO: 3143.In another embodiment, human GIPR has a sequence such as that shown in SEQ ID NO: 3145.

[0114] In one aspect, the present invention is directed to a composition comprising an antibody or functional fragment thereof that specifically binds to human GIPR, the antibody or functional fragment thereof comprising an amino acid substitution of a cysteine ​​or non-standard amino acid at one or more conjugation sites, and a GLP-1 receptor agonist conjugated to the antibody or functional fragment thereof through the side chain of the substituted cysteine ​​residue or non-standard amino acid residue at one or more conjugation sites.

[0115] The antigen binding proteins provided are polypeptides incorporating and / or linked to one or more complementarity determining regions (CDRs) described herein. In some antigen binding proteins, the CDRs are incorporated into "framework" regions that orient the CDRs so that their proper antigen binding properties 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.

[0116] The antigen-binding proteins disclosed herein have a variety of utilities. They 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 antigen-binding proteins include, for example, diagnosing diseases or conditions associated with GIPR and screening assays to determine the presence or absence of GIPR. Given that the provided antigen-binding proteins are antagonists, 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 while maintaining or increasing food intake, increasing % body fat mass, and increasing % lean mass. Thus, antigen-binding proteins have utility in the treatment and prevention of, for example, diabetes, including type 2 diabetes, obesity, dyslipidemia, elevated glucose levels, or elevated insulin levels.

[0117] Various selective binding agents useful for regulating the activity of GIPR are provided. Such agents include, for example, antigen-binding proteins (e.g., scFv, domain antibodies, and polypeptides having antigen-binding regions) that contain an antigen-binding domain and specifically bind to GIPR polypeptides, particularly human GIPR. Some of these agents are useful, for example, for enhancing the activity of GIPR, and can activate one or more activities associated with GIPR.

[0118] 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 proteins comprise (a) a polypeptide structure and (b) one or more CDRs inserted into and / or linked to the polypeptide structure. The polypeptide structure can take a variety of different forms. For example, the polypeptide structure can be or comprise the framework of a naturally occurring antibody or a fragment or variant thereof, or can be entirely synthetic in nature. Examples of various polypeptide structures are described further below.

[0119] In certain embodiments, the polypeptide structure of the antigen-binding protein is or is derived from an antibody. Accordingly, examples of specific 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.

[0120] 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: 3141. In certain embodiments, the antigen binding proteins specifically bind to human GIPR comprising or consisting of the amino acid sequence of SEQ ID NO: 3143. In certain embodiments, the antigen binding proteins specifically bind to human GIPR comprising or consisting of the amino acid sequence of SEQ ID NO: 3145.

[0121] The antigen binding proteins that are 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, at levels measurable by methods such as, for example, binding studies with radioactively or fluorescently labeled ligands, or by methods described herein (e.g., cAMP assays or other functional assays). The reduction 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: 3141, SEQ ID NO: 3143, or SEQ ID NO: 3145 under equivalent 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 fatty liver or reduce triglyceride levels in the liver; (k) reducing AST, ALT and / or ALP levels;

[0122] 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 techniques. (b) a half-life in human serum of at least 3 days;

[0123] 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.

[0124] In another aspect, antigen binding proteins are provided that have an in vitro or in vivo (e.g., when administered to a human subject) half-life 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 an underivatized or unmodified antibody. 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.

[0125] Some of the antigen-binding proteins provided typically have a structure associated with naturally occurring antibodies. The structural unit of such antibodies typically comprises one or more tetramers, each 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 comprises one full-length "light" chain (in certain embodiments, approximately 25 kDa) and one full-length "heavy" chain (in certain embodiments, approximately 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 antibody polypeptides. The amino-terminal portion of each chain typically contains a variable domain responsible for antigen recognition. The carboxy-terminal portion is more evolutionarily conserved than 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 containing one variable and one constant domain. Heavy chains are typically classified as mu, delta, gamma, alpha, or epsilon, and these chains 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. For example, the heavy chain of an IgG contains three C region domains known as CH1, CH2, and CH3. The provided antibodies 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 figures. Both terms refer to an antibody that binds to GIPR.

[0126] 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.

[0127] For the antibodies provided herein, the variable regions of immunoglobulin chains generally exhibit the same overall structure, comprising relatively conserved framework regions (FRs) connected 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- 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 occupying 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.

[0128] 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.

[0129] 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 present invention have been assigned SEQ ID NOs, and these SEQ ID NOs are shown in Table 1. 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 present invention have also been assigned SEQ ID NOs, and these SEQ ID NOs are shown in Table 2. The antigen-binding proteins of the present invention can be identified by SEQ ID NOs, but can also be identified by construct name (e.g., 2C2.005) or identifier number (e.g., iPS:336175). The antigen-binding proteins identified in Tables 1 to 5 below can be grouped into families based on construct name. 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.

[0130] The various light chain variable regions and heavy chain variable regions provided herein are shown in Table 3. Each of these variable regions can be added to a heavy chain constant region or a light chain constant region to form the heavy and light chains, respectively, of a complete antibody. The heavy and light chain sequences thus generated can then be combined to form the structure of a complete antibody.

[0131] [Table 1]

[0132]

Table 2

[0133]

Table 3

[0134]

Table 4

[0135]

Table 5

[0136]

Table 6

[0137]

Table 7

[0138]

Table 8

[0139]

Table 9

[0140]

Table 10

[0141]

Table 11

[0142]

Table 12

[0143]

Table 13

[0144]

Table 14

[0145]

Table 15

[0146] Table 16

[0147] Table 17

[0148] Table 18

[0149] Table 19

[0150] Table 20

[0151] Table 21

[0152] Table 22

[0153] Table 23

[0154] Table 24

[0155] Table 25

[0156] Table 26

[0157] Table 27

[0158] Table 28

[0159] Table 29

[0160]

Table 30

[0161] Table 31

[0162] Table 32

[0163]

Table 33

[0164] Table 34

[0165] Table 35

[0166] Table 36

[0167] Table 37

[0168] Table 38

[0169] Table 39

[0170] Table 40

[0171] Table 41

[0172] Table 42

[0173] Table 43

[0174] Table 44

[0175] Table 45

[0176] Table 46

[0177] Table 47

[0178] Table 48

[0179] Table 49

[0180] Table 50

[0181] Table 51

[0182] Table 52

[0183] Table 53

[0184] Table 54

[0185] Table 55

[0186] Table 56

[0187] Table 57

[0188] Table 58

[0189] Table 59

[0190] Table 60

[0191] Table 61

[0192] Table 62

[0193] Table 63

[0194] Table 64

[0195] Table 65

[0196] Table 66

[0197] Table 67

[0198] Table 68

[0199] Table 69

[0200] Table 70

[0201] Table 71

[0202] Table 72

[0203] Table 73

[0204] Table 74

[0205] Table 75

[0206] Table 76

[0207] Table 77

[0208] Table 78

[0209] Table 79

[0210] Table 80

[0211] Table 81

[0212] Table 82

[0213] Table 83

[0214] Table 84

[0215] Table 85

[0216] Table 86

[0217] Table 87

[0218] Table 88

[0219] Table 89

[0220] Table 90

[0221]

Table 91

[0222] Table 92

[0223]

Table 93

[0224] Table 94

[0225]

Table 95

[0226] Table 96

[0227] Table 97

[0228] Table 98

[0229]

Table 99

[0230]

Table 100

[0231] Table 101

[0232] Table 102

[0233] Table 103

[0234] Table 104

[0235] Table 105

[0236] Table 106

[0237] Table 107

[0238] Table 108

[0239] Table 109

[0240] Table 110

[0241] Table 111

[0242] Table 112

[0243] Table 113

[0244] Table 114

[0245] Table 115

[0246] Table 116

[0247] Table 117

[0248] Table 118

[0249] Table 119

[0250] Table 120

[0251] Table 121

[0252] Table 122

[0253] Table 123

[0254] Table 124

[0255] Table 125

[0256] Table 126

[0257] Table 127

[0258] Table 128

[0259] Table 129

[0260] Table 130

[0261] Table 131

[0262] Table 132

[0263] Table 133

[0264] Table 134

[0265] Table 135

[0266] Table 136

[0267] Table 137

[0268] Table 138

[0269] Table 139

[0270] Table 140

[0271] Table 141

[0272] Table 142

[0273] Table 143

[0274] Table 144

[0275] Table 145

[0276] Table 146

[0277] Table 147

[0278] Table 148

[0279] Table 149

[0280] Table 150

[0281] Table 151

[0282] Table 152

[0283] Table 153

[0284] Table 154

[0285] Table 155

[0286] Table 156

[0287] Table 157

[0288] Table 158

[0289] Table 159

[0290] Table 160

[0291] Table 161

[0292] Table 162

[0293] Table 163

[0294] Table 164

[0295] Table 165

[0296] Table 166

[0297] Table 167

[0298] Table 168

[0299] Table 169

[0300] Table 170

[0301] Table 171

[0302] Table 172

[0303] Table 173

[0304] Table 174

[0305] Table 175

[0306] Table 176

[0307] Table 177

[0308] Table 178

[0309] Table 179

[0310] Table 180

[0311] Table 181

[0312] Table 182

[0313] Table 183

[0314] Table 184

[0315] Table 185

[0316] Table 186

[0317] Table 187

[0318] Table 188

[0319] Table 189

[0320] Table 190

[0321] Table 191

[0322] Table 192

[0323] Table 193

[0324] Table 194

[0325] Table 195

[0326] Table 196

[0327] Table 197

[0328] Table 198

[0329] Table 199

[0330] Table 200

[0331] Table 201

[0332] Table 202

[0333] Table 203

[0334] Table 204

[0335] Table 205

[0336] Table 206

[0337] Table 207

[0338] Table 208

[0339] Table 209

[0340] Table 210

[0341] Table 211

[0342] Table 212

[0343] Table 213

[0344] Table 214

[0345] Table 215

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[0347] Table 217

[0348] Table 218

[0349] Table 219

[0350] Table 220

[0351] Table 221

[0352] Table 222

[0353] Table 223

[0354] Table 224

[0355] Table 225

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[0357] Table 227

[0358] Table 228

[0359] Table 229

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[0361] Table 231

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[0363] Table 233

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[0366] Table 236

[0367] Table 237

[0368] Table 238

[0369] Table 239

[0370] Table 240

[0371] Table 241

[0372] Table 242

[0373] Table 243

[0374] Table 244

[0375] Table 245

[0376] Table 246

[0377] Table 247

[0378] Table 248

[0379] Table 249

[0380] Table 250

[0381] Table 251

[0382] Table 252

[0383] Table 253

[0384] Table 254

[0385] Table 255

[0386] Table 256

[0387] Table 257

[0388] Table 258

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[0390] Table 260

[0391] Table 261

[0392] Table 262

[0393] Table 263

[0394] Table 264

[0395] Table 265

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[0397] Table 267

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[0399] Table 269

[0400] Table 270

[0401] Table 271

[0402] Table 272

[0403] Table 273

[0404] Table 274

[0405] Table 275

[0406] Table 276

[0407] Table 277

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[0410] Table 280

[0411] Table 281

[0412] Table 282

[0413] Table 283

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[0415] Table 285

[0416] Table 286

[0417] Table 287

[0418] Table 288

[0419] Table 289

[0420] Table 290

[0421] Table 291

[0422] Table 292

[0423] Table 293

[0424] Table 294

[0425] Table 295

[0426] Table 296

[0427] Table 297

[0428] Table 298

[0429] Table 299

[0430] Table 300

[0431]

Table 301

[0432]

Table 302

[0433]

Table 303

[0434] Table 304

[0435] Table 305

[0436] Table 306

[0437] Table 307

[0438] Table 308

[0439] Table 309

[0440] Table 310

[0441] Table 311

[0442] Table 312

[0443] Table 313

[0444] Table 314

[0445] Table 315

[0446] Table 316

[0447] Table 317

[0448] Table 318

[0449] Table 319

[0450] Table 320

[0451] Table 321

[0452] Table 322

[0453] Table 323

[0454] Table 324

[0455] Table 325

[0456] Table 326

[0457] Table 327

[0458] Table 328

[0459] Table 329

[0460] Table 330

[0461] Table 331

[0462] Table 332

[0463]

Table 333

[0464] Table 334

[0465] Table 335

[0466] Table 336

[0467] Table 337

[0468] Table 338

[0469] Table 339

[0470] Table 340

[0471] Table 341

[0472] Table 342

[0473] Table 343

[0474] Table 344

[0475] Table 345

[0476] Table 346

[0477] Table 347

[0478] Table 348

[0479] Table 349

[0480] Table 350

[0481] Table 351

[0482] Table 352

[0483] Table 353

[0484] Table 354

[0485] Table 355

[0486] Table 356

[0487] Table 357

[0488] Table 358

[0489] Table 359

[0490] Table 360

[0491] Table 361

[0492] Table 362

[0493] Table 363

[0494] Table 364

[0495] Table 365

[0496] Table 366

[0497] Table 367

[0498] Table 368

[0499] Table 369

[0500] Table 370

[0501] Table 371

[0502] Table 372

[0503] Table 373

[0504] Table 374

[0505] Table 375

[0506] Table 376

[0507] Table 377

[0508] Table 378

[0509] Table 379

[0510] Table 380

[0511] Table 381

[0512] Table 382

[0513] Table 383

[0514] Table 384

[0515] Table 385

[0516] Table 386

[0517] Table 387

[0518] Table 388

[0519] Table 389

[0520] Table 390

[0521] Table 391

[0522] Table 392

[0523]

Table 393

[0524] Table 394

[0525] Table 395

[0526] Table 396

[0527] Table 397

[0528] Table 398

[0529] Table 399

[0530] Table 400

[0531]

Table 401

[0532] Table 402

[0533] Table 403

[0534]

Table 404

[0535] [Table 405]

[0536] In one embodiment, the antibody or fragment thereof comprises a light chain variable region comprising a sequence selected from the group consisting of SEQ ID NOs: 1 to 157 and a heavy chain variable region comprising a sequence selected from the group consisting of SEQ ID NOs: 158 to 314, and the antibody or functional fragment thereof comprises an amino acid substitution of cysteine ​​or a non-standard amino acid at one or more conjugation sites selected from the group consisting of D70 of the antibody light chain for reference sequence SEQ ID NO: 455, E276 of the antibody heavy chain for reference sequence SEQ ID NO: 612, and T363 of the antibody heavy chain for reference sequence SEQ ID NO: 612.

[0537] In one embodiment, the antibody or fragment thereof comprises a light chain variable region comprising SEQ ID NO: 1 and a heavy chain variable region comprising SEQ ID NO: 158, a light chain variable region comprising SEQ ID NO: 2 and a heavy chain variable region comprising SEQ ID NO: 159, a light chain variable region comprising SEQ ID NO: 3 and a heavy chain variable region comprising SEQ ID NO: 160, a light chain variable region comprising SEQ ID NO: 4 and a heavy chain variable region comprising SEQ ID NO: 161, a light chain variable region comprising SEQ ID NO: 5 and a heavy chain variable region comprising SEQ ID NO: 162, a light chain variable region comprising SEQ ID NO: 6 and a heavy chain variable region comprising SEQ ID NO: 163, a light chain variable region comprising SEQ ID NO: 7 and a heavy chain variable region comprising SEQ ID NO: 164. a light chain variable region comprising SEQ ID NO: 8 and a heavy chain variable region comprising SEQ ID NO: 165; a light chain variable region comprising SEQ ID NO: 9 and a heavy chain variable region comprising SEQ ID NO: 166; a light chain variable region comprising SEQ ID NO: 10 and a heavy chain variable region comprising SEQ ID NO: 167; a light chain variable region comprising SEQ ID NO: 11 and a heavy chain variable region comprising SEQ ID NO: 168; a light chain variable region comprising SEQ ID NO: 12 and a heavy chain variable region comprising SEQ ID NO: 169; a light chain variable region comprising SEQ ID NO: 13 and a heavy chain variable region comprising SEQ ID NO: 170; a light chain variable region comprising SEQ ID NO: 14 and a heavy chain variable region comprising SEQ ID NO: 171; a light chain variable region comprising SEQ ID NO: 15 a light chain variable region comprising SEQ ID NO: 16 and a heavy chain variable region comprising SEQ ID NO: 173; a light chain variable region comprising SEQ ID NO: 17 and a heavy chain variable region comprising SEQ ID NO: 174; a light chain variable region comprising SEQ ID NO: 18 and a heavy chain variable region comprising SEQ ID NO: 175; a light chain variable region comprising SEQ ID NO: 19 and a heavy chain variable region comprising SEQ ID NO: 176; a light chain variable region comprising SEQ ID NO: 20 and a heavy chain variable region comprising SEQ ID NO: 177; a light chain variable region comprising SEQ ID NO: 21 and a heavy chain variable region comprising SEQ ID NO: 178; a light chain variable region comprising SEQ ID NO: 22 and a heavy chain variable region comprising SEQ ID NO: 1 a heavy chain variable region comprising SEQ ID NO: 79, a light chain variable region comprising SEQ ID NO: 23 and a heavy chain variable region comprising SEQ ID NO: 180, a light chain variable region comprising SEQ ID NO: 24 and a heavy chain variable region comprising SEQ ID NO: 181, a light chain variable region comprising SEQ ID NO: 25 and a heavy chain variable region comprising SEQ ID NO: 182, a light chain variable region comprising SEQ ID NO: 26 and a heavy chain variable region comprising SEQ ID NO: 183, a light chain variable region comprising SEQ ID NO: 27 and a heavy chain variable region comprising SEQ ID NO: 184, a light chain variable region comprising SEQ ID NO: 28 and a heavy chain variable region comprising SEQ ID NO: 185, a light chain variable region comprising SEQ ID NO: 29 and a heavy chain variable region comprising SEQ ID NO: 186,a light chain variable region comprising SEQ ID NO: 30 and a heavy chain variable region comprising SEQ ID NO: 187; a light chain variable region comprising SEQ ID NO: 31 and a heavy chain variable region comprising SEQ ID NO: 188; a light chain variable region comprising SEQ ID NO: 32 and a heavy chain variable region comprising SEQ ID NO: 189; a light chain variable region comprising SEQ ID NO: 33 and a heavy chain variable region comprising SEQ ID NO: 190; a light chain variable region comprising SEQ ID NO: 34 and a heavy chain variable region comprising SEQ ID NO: 191; a light chain variable region comprising SEQ ID NO: 35 and a heavy chain variable region comprising SEQ ID NO: 192; a light chain variable region comprising SEQ ID NO: 36 and a heavy chain variable region comprising SEQ ID NO: 193; a light chain variable region comprising SEQ ID NO: 38 and a heavy chain variable region comprising SEQ ID NO: 195; a light chain variable region comprising SEQ ID NO: 39 and a heavy chain variable region comprising SEQ ID NO: 196; a light chain variable region comprising SEQ ID NO: 40 and a heavy chain variable region comprising SEQ ID NO: 197; a light chain variable region comprising SEQ ID NO: 41 and a heavy chain variable region comprising SEQ ID NO: 198; a light chain variable region comprising SEQ ID NO: 42 and a heavy chain variable region comprising SEQ ID NO: 199; a light chain variable region comprising SEQ ID NO: 43 and a heavy chain variable region comprising SEQ ID NO: 200; a light chain variable region comprising SEQ ID NO: 44 and a heavy chain variable region comprising SEQ ID NO: 201, a light chain variable region comprising SEQ ID NO: 45 and a heavy chain variable region comprising SEQ ID NO: 202, a light chain variable region comprising SEQ ID NO: 46 and a heavy chain variable region comprising SEQ ID NO: 203, a light chain variable region comprising SEQ ID NO: 47 and a heavy chain variable region comprising SEQ ID NO: 204, a light chain variable region comprising SEQ ID NO: 48 and a heavy chain variable region comprising SEQ ID NO: 205, a light chain variable region comprising SEQ ID NO: 49 and a heavy chain variable region comprising SEQ ID NO: 206, a light chain variable region comprising SEQ ID NO: 50 and a heavy chain variable region comprising SEQ ID NO: 207, a light chain variable region comprising SEQ ID NO: 51 and SEQ ID NO: 208 a heavy chain variable region comprising SEQ ID NO: 209; a light chain variable region comprising SEQ ID NO: 53 and a heavy chain variable region comprising SEQ ID NO: 210; a light chain variable region comprising SEQ ID NO: 54 and a heavy chain variable region comprising SEQ ID NO: 211; a light chain variable region comprising SEQ ID NO: 55 and a heavy chain variable region comprising SEQ ID NO: 212; a light chain variable region comprising SEQ ID NO: 56 and a heavy chain variable region comprising SEQ ID NO: 213; a light chain variable region comprising SEQ ID NO: 57 and a heavy chain variable region comprising SEQ ID NO: 214; a light chain variable region comprising SEQ ID NO: 58 and a heavy chain variable region comprising SEQ ID NO: 215;a light chain variable region comprising SEQ ID NO: 59 and a heavy chain variable region comprising SEQ ID NO: 216; a light chain variable region comprising SEQ ID NO: 60 and a heavy chain variable region comprising SEQ ID NO: 217; a light chain variable region comprising SEQ ID NO: 61 and a heavy chain variable region comprising SEQ ID NO: 218; a light chain variable region comprising SEQ ID NO: 62 and a heavy chain variable region comprising SEQ ID NO: 219; a light chain variable region comprising SEQ ID NO: 63 and a heavy chain variable region comprising SEQ ID NO: 220; a light chain variable region comprising SEQ ID NO: 64 and a heavy chain variable region comprising SEQ ID NO: 221; a light chain variable region comprising SEQ ID NO: 65 and a heavy chain variable region comprising SEQ ID NO: 222; a light chain variable region comprising SEQ ID NO: 67 and a heavy chain variable region comprising SEQ ID NO: 224; a light chain variable region comprising SEQ ID NO: 68 and a heavy chain variable region comprising SEQ ID NO: 225; a light chain variable region comprising SEQ ID NO: 69 and a heavy chain variable region comprising SEQ ID NO: 226; a light chain variable region comprising SEQ ID NO: 70 and a heavy chain variable region comprising SEQ ID NO: 227; a light chain variable region comprising SEQ ID NO: 71 and a heavy chain variable region comprising SEQ ID NO: 228; a light chain variable region comprising SEQ ID NO: 72 and a heavy chain variable region comprising SEQ ID NO: 229; a light chain variable region comprising SEQ ID NO: 73; a heavy chain variable region comprising SEQ ID NO: 230, a light chain variable region comprising SEQ ID NO: 74 and a heavy chain variable region comprising SEQ ID NO: 231, a light chain variable region comprising SEQ ID NO: 75 and a heavy chain variable region comprising SEQ ID NO: 232, a light chain variable region comprising SEQ ID NO: 76 and a heavy chain variable region comprising SEQ ID NO: 233, a light chain variable region comprising SEQ ID NO: 77 and a heavy chain variable region comprising SEQ ID NO: 234, a light chain variable region comprising SEQ ID NO: 78 and a heavy chain variable region comprising SEQ ID NO: 235, a light chain variable region comprising SEQ ID NO: 79 and a heavy chain variable region comprising SEQ ID NO: 236, a light chain variable region comprising SEQ ID NO: 80 and SEQ ID NO: 237 a heavy chain variable region comprising SEQ ID NO: 238; a light chain variable region comprising SEQ ID NO: 82 and a heavy chain variable region comprising SEQ ID NO: 239; a light chain variable region comprising SEQ ID NO: 83 and a heavy chain variable region comprising SEQ ID NO: 240; a light chain variable region comprising SEQ ID NO: 84 and a heavy chain variable region comprising SEQ ID NO: 241; a light chain variable region comprising SEQ ID NO: 85 and a heavy chain variable region comprising SEQ ID NO: 242; a light chain variable region comprising SEQ ID NO: 86 and a heavy chain variable region comprising SEQ ID NO: 243; a light chain variable region comprising SEQ ID NO: 87 and a heavy chain variable region comprising SEQ ID NO: 244;a light chain variable region comprising SEQ ID NO: 88 and a heavy chain variable region comprising SEQ ID NO: 245, a light chain variable region comprising SEQ ID NO: 89 and a heavy chain variable region comprising SEQ ID NO: 246, a light chain variable region comprising SEQ ID NO: 90 and a heavy chain variable region comprising SEQ ID NO: 247, a light chain variable region comprising SEQ ID NO: 91 and a heavy chain variable region comprising SEQ ID NO: 248, a light chain variable region comprising SEQ ID NO: 92 and a heavy chain variable region comprising SEQ ID NO: 249, a light chain variable region comprising SEQ ID NO: 93 and a heavy chain variable region comprising SEQ ID NO: 250, a light chain variable region comprising SEQ ID NO: 94 and a heavy chain variable region comprising SEQ ID NO: 251, a light chain variable region comprising SEQ ID NO: 95 and a heavy chain variable region comprising SEQ ID NO: 252, a light chain variable region comprising SEQ ID NO: 96 and a heavy chain variable region comprising SEQ ID NO: 253, a light chain variable region comprising SEQ ID NO: 97 and a heavy chain variable region comprising SEQ ID NO: 254, a light chain variable region comprising SEQ ID NO: 98 and a heavy chain variable region comprising SEQ ID NO: 255, a light chain variable region comprising SEQ ID NO: 99 and a heavy chain variable region comprising SEQ ID NO: 256, a light chain variable region comprising SEQ ID NO: 100 and a heavy chain variable region comprising SEQ ID NO: 257, a light chain variable region comprising SEQ ID NO: 101 and a heavy chain variable region comprising SEQ ID NO: 258, a light chain variable region comprising SEQ ID NO: 10 a light chain variable region comprising SEQ ID NO: 102 and a heavy chain variable region comprising SEQ ID NO: 259, a light chain variable region comprising SEQ ID NO: 103 and a heavy chain variable region comprising SEQ ID NO: 260, a light chain variable region comprising SEQ ID NO: 104 and a heavy chain variable region comprising SEQ ID NO: 261, a light chain variable region comprising SEQ ID NO: 105 and a heavy chain variable region comprising SEQ ID NO: 262, a light chain variable region comprising SEQ ID NO: 106 and a heavy chain variable region comprising SEQ ID NO: 263, a light chain variable region comprising SEQ ID NO: 107 and a heavy chain variable region comprising SEQ ID NO: 264, a light chain variable region comprising SEQ ID NO: 108 and a heavy chain variable region comprising SEQ ID NO: 265, a light chain variable region comprising SEQ ID NO: 109 and a heavy chain variable region comprising SEQ ID NO: 266, a light chain variable region comprising SEQ ID NO: 110 and a heavy chain variable region comprising SEQ ID NO: 267, a light chain variable region comprising SEQ ID NO: 111 and a heavy chain variable region comprising SEQ ID NO: 268, a light chain variable region comprising SEQ ID NO: 112 and a heavy chain variable region comprising SEQ ID NO: 269, a light chain variable region comprising SEQ ID NO: 113 and a heavy chain variable region comprising SEQ ID NO: 270, a light chain variable region comprising SEQ ID NO: 114 and a heavy chain variable region comprising SEQ ID NO: 271, a light chain variable region comprising SEQ ID NO: 115 and a heavy chain variable region comprising SEQ ID NO: 272,a light chain variable region comprising SEQ ID NO: 116 and a heavy chain variable region comprising SEQ ID NO: 273, a light chain variable region comprising SEQ ID NO: 117 and a heavy chain variable region comprising SEQ ID NO: 274, a light chain variable region comprising SEQ ID NO: 118 and a heavy chain variable region comprising SEQ ID NO: 275, a light chain variable region comprising SEQ ID NO: 119 and a heavy chain variable region comprising SEQ ID NO: 276, a light chain variable region comprising SEQ ID NO: 120 and a heavy chain variable region comprising SEQ ID NO: 277, a light chain variable region comprising SEQ ID NO: 121 and a heavy chain variable region comprising SEQ ID NO: 278, a light chain variable region comprising SEQ ID NO: 122 and a heavy chain variable region comprising SEQ ID NO: 279, a light chain variable region comprising SEQ ID NO: 123 and a heavy chain variable region comprising SEQ ID NO: 280; a light chain variable region comprising SEQ ID NO: 124 and a heavy chain variable region comprising SEQ ID NO: 281; a light chain variable region comprising SEQ ID NO: 125 and a heavy chain variable region comprising SEQ ID NO: 282; a light chain variable region comprising SEQ ID NO: 126 and a heavy chain variable region comprising SEQ ID NO: 283; a light chain variable region comprising SEQ ID NO: 127 and a heavy chain variable region comprising SEQ ID NO: 284; a light chain variable region comprising SEQ ID NO: 128 and a heavy chain variable region comprising SEQ ID NO: 285; a light chain variable region comprising SEQ ID NO: 129 and a heavy chain variable region comprising SEQ ID NO: 286; a light chain variable region comprising SEQ ID NO: 130 and a heavy chain variable region comprising SEQ ID NO: 287, a light chain variable region comprising SEQ ID NO: 131 and a heavy chain variable region comprising SEQ ID NO: 288, a light chain variable region comprising SEQ ID NO: 132 and a heavy chain variable region comprising SEQ ID NO: 289, a light chain variable region comprising SEQ ID NO: 133 and a heavy chain variable region comprising SEQ ID NO: 290, a light chain variable region comprising SEQ ID NO: 134 and a heavy chain variable region comprising SEQ ID NO: 291, a light chain variable region comprising SEQ ID NO: 135 and a heavy chain variable region comprising SEQ ID NO: 292, a light chain variable region comprising SEQ ID NO: 136 and a heavy chain variable region comprising SEQ ID NO: 293, a light chain variable region comprising SEQ ID NO: 137 and a heavy chain variable region comprising SEQ ID NO: 294, a light chain variable region comprising SEQ ID NO: 138 and a heavy chain variable region comprising SEQ ID NO: 295, a light chain variable region comprising SEQ ID NO: 139 and a heavy chain variable region comprising SEQ ID NO: 296, a light chain variable region comprising SEQ ID NO: 140 and a heavy chain variable region comprising SEQ ID NO: 297, a light chain variable region comprising SEQ ID NO: 141 and a heavy chain variable region comprising SEQ ID NO: 298, a light chain variable region comprising SEQ ID NO: 142 and a heavy chain variable region comprising SEQ ID NO: 299, a light chain variable region comprising SEQ ID NO: 143 and a heavy chain variable region comprising SEQ ID NO: 300,a light chain variable region comprising SEQ ID NO: 144 and a heavy chain variable region comprising SEQ ID NO: 301; a light chain variable region comprising SEQ ID NO: 145 and a heavy chain variable region comprising SEQ ID NO: 302; a light chain variable region comprising SEQ ID NO: 146 and a heavy chain variable region comprising SEQ ID NO: 303; a light chain variable region comprising SEQ ID NO: 147 and a heavy chain variable region comprising SEQ ID NO: 304; a light chain variable region comprising SEQ ID NO: 148 and a heavy chain variable region comprising SEQ ID NO: 305; a light chain variable region comprising SEQ ID NO: 149 and a heavy chain variable region comprising SEQ ID NO: 306. a heavy chain variable region, a light chain variable region comprising SEQ ID NO: 150 and a heavy chain variable region comprising SEQ ID NO: 307, a light chain variable region comprising SEQ ID NO: 151 and a heavy chain variable region comprising SEQ ID NO: 308, a light chain variable region comprising SEQ ID NO: 152 and a heavy chain variable region comprising SEQ ID NO: 309, a light chain variable region comprising SEQ ID NO: 153 and a heavy chain variable region comprising SEQ ID NO: 310, a light chain variable region comprising SEQ ID NO: 154 and a heavy chain variable region comprising SEQ ID NO: 311, a light chain variable region comprising SEQ ID NO: 155 and a heavy chain variable region comprising SEQ ID NO: 312, a light chain variable region comprising SEQ ID NO: 156 and a heavy chain variable region comprising SEQ ID NO: and a heavy chain variable region comprising SEQ ID NO: 313, and a combination of a light chain variable region and a heavy chain variable region selected from the group consisting of a light chain variable region comprising SEQ ID NO: 157 and a heavy chain variable region comprising SEQ ID NO: 314, wherein the antibody or functional fragment thereof comprises an amino acid substitution of cysteine ​​or a non-canonical amino acid at one or more conjugation sites selected from the group consisting of D70 of the antibody light chain for reference sequence SEQ ID NO: 455, E276 of the antibody heavy chain for reference sequence SEQ ID NO: 612, and T363 of the antibody heavy chain for reference sequence SEQ ID NO: 612.

[0538] In one embodiment, the antibody or fragment thereof comprises a light chain variable region encoded by a polynucleotide sequence selected from the group consisting of SEQ ID NOs: 1571-1727 and a heavy chain variable region encoded by a polynucleotide sequence selected from the group consisting of SEQ ID NOs: 1728-1884, and the antibody or functional fragment thereof comprises an amino acid substitution of cysteine ​​or a non-standard amino acid at one or more conjugation sites selected from the group consisting of D70 of the antibody light chain for reference sequence SEQ ID NO: 455, E276 of the antibody heavy chain for reference sequence SEQ ID NO: 612, and T363 of the antibody heavy chain for reference sequence SEQ ID NO: 612.

[0539] In one embodiment, the antibody or fragment thereof comprises a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1571 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1728, a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1572 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1729, a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1573 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1730, a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1574 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1731, a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1575 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1732, a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1576 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1733 a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1734; a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1578 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1735; a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1579 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1736; a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1580 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1737; a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1581 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1738; a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1582 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1739;a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1583 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1740; a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1584 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1741; a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1585 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1742; a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1586 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1743; a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1587 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1744; a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1588 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1745; a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1589; a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1746 and a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1590 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1747; a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1591 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1748; a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1592 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1749; a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1593 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1750; a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1594 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1751; a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1595 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1752;a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1596 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1753; a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1597 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1754; a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1598 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1755; a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1599 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1756; a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1600 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1757; a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1601 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1758; a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1602; a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1759 and SEQ ID NO: 1759; a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1603 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1760; a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1604 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1761; a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1605 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1762; a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1606 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1763; a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1607 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1764; a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1608 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1765;a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1609 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1766; a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1610 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1767; a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1611 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1768; a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1612 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1769; a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1613 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1770; a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1614 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1771; a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1615; a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1772 and a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1616 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1773; a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1617 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1774; a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1618 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1775; a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1619 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1776; a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1620 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1777; a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1621 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1778;a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1622 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1779; a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1623 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1780; a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1624 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1781; a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1625 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1782; a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1626 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1783; a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1627 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1784; a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1628; a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1785, a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1629 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1786, a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1630 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1787, a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1631 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1788, a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1632 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1789, a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1633 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1790, a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1634 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1791,a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1635 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1792; a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1636 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1793; a polynucleotide comprising SEQ ID NO: 1637; a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1638 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1795; a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1639 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1796; a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1640 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1797; a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1641 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1798; a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1642 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1799; a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1643 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1800 a heavy chain variable region encoded by a polynucleotide sequence comprising: a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1644 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1801; a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1645 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1802; a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1646 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1803; a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1647 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1804; a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1648 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1805; a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1649 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1806;a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1650 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1807; a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1651 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1808; a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1652 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1809; a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1653 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1810; a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1654 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1811; a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1655 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1812; a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1656; a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1657 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1814; a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1658 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1815; a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1659 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1816; a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1660 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1817; a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1661 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1818; a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1662 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1819;a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1663 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1820; a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1664 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1821; a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1665 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1822; a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1666 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1823; a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1667 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1824; a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1668 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1825; a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1669; a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1673 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1830; a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1674 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1831; a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1675 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1832;a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1676 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1833; a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1677 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1834; a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1678 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1835; a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1679 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1836; a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1680 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1837; a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1681 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1838; a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1682; a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1687 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1844; a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1688 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1845; a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1689 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1846; a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1689 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1847; a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1689 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1846;a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1689 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1846; a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1690 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1847; a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1691 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1848; a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1692 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1849; a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1693 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1850; a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1694 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1851; a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1695; a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1696 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1853; a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1697 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1854; a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1698 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1855; a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1699 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1856; a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1700 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1857; a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1701 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1858;a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1702 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1859; a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1703 and a polynucleotide sequence comprising SEQ ID NO: 1860; a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1704 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1861; a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1705 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1862; a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1706 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1863; a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1707 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1864; a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1708 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1865; a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1709 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1866 a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1710 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1867; a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1711 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1868; a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1712 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1869; a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1713 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1870; a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1714 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1871; a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1715 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1872;a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO:1716 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO:1873; a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO:1717 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO:1874; a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO:1718 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO:1875; a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO:1719 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO:1876; a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO:1720 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO:1877; a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO:1721 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO:1878; a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO:1722. a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1726 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1883; a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1727 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1884; a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1728 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1885; a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1729 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1886; a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1729 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1887; a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1729 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1888; a light chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1729 and a heavy chain variable region encoded by a polynucleotide sequence comprising SEQ ID NO: 1889;The antibody light chain of the reference sequence SEQ ID NO: 455 comprises an amino acid substitution of cysteine ​​or a non-standard amino acid at one or more conjugation sites selected from the group consisting of D70, E276, and T363 of the antibody heavy chain of the reference sequence SEQ ID NO: 612.

[0540] The present invention relates to a composition comprising an anti-GIPR antigen-binding protein having at least one conjugation site. The conjugation site must be amenable to the conjugation of an additional functional moiety (e.g., a GLP-1 receptor agonist) through a defined conjugation chemistry via the side chain of an amino acid residue at the conjugation site. Achieving highly selective site-specific conjugation to an anti-GIPR antigen-binding protein according to the present invention requires consideration of various design criteria. First, a preferred conjugation or binding chemistry must be defined or specified. For example, a functional moiety such as a GLP-1 receptor agonist can be conjugated or bound to a selected conjugation site of an anti-GIPR antigen-binding protein through a combination of various conjugation chemistries known in the art. For example, maleimide-activated conjugation partners that target accessible cysteine ​​thiols on anti-GIPR antigen binding proteins are one embodiment, but many conjugation or linkage chemistries that target side chains of either standard or non-standard, e.g., unnatural, amino acids within the anti-GIPR antigen binding protein sequence can also be used in accordance with the present invention.

[0541] Chemical reactions for chemoselective conjugation include copper(I)-catalyzed azide-alkyne [3+2] dipolar cycloaddition, Staudinger ligation, other acyl transfer processes (S→N; X→N), oximation, hydrazone bond formation, and other suitable organic chemistry reactions, such as cross-coupling using water-soluble palladium catalysts. (For example, Bong et al., Chemoselective Pd(0)-catalyzed peptide coupling in water, Organic Letters 3(16):2509-11(2001); Dibowski et al., Bioconjugation of peptides by palladium-catalyzed CC cross-coupling in water,Angew.Chem.Int.Ed.37(4):476-78(1998);DeVasher et al.,Aqueous-phase,palladium-catalyzed cross-coupling of aryl bromides under mild conditions,using water-soluble,sterically demanding alkylphosphines,J.Org.Chem.69:7919-27(2004);Shaugnessy et al. al.,J.Org.Chem,2003,68,6767-6774;Prescher,JA and Bertozzi CR,Chemistry in living system, Nature Chemical Biology 1(1);13-21(2005)).

[0542] As noted above, conjugation (or covalent attachment) to the anti-GIPR antigen-binding protein is achieved through the side chain of an amino acid residue at a conjugation site, including, but not limited to, a cysteinyl residue. The amino acid residue at the selected internal conjugation site, e.g., a cysteinyl residue, may be an amino acid residue that occupies the same amino acid residue position in a native Fc domain sequence, or an amino acid residue that can be engineered into the Fc domain sequence by substitution or insertion.

[0543] Other examples of unnatural amino acid residues that may be particularly useful as conjugation sites in some embodiments of the processes and compositions of the invention include azide-containing amino acid residues, e.g., azidohomoalanine, p-azido-phenylalanine; keto-containing amino acid residues, e.g., p-acetyl-phenylalanine; alkyne-containing amino acid residues, e.g., p-ethynylphenylalanine, homopropargylglycine, p-(prop-2-ynyl)-tyrosine; alkene-containing amino acid residues, e.g., homoallylglycine; aryl halide-containing amino acid residues, e.g., p-iodophenylalanine, p-bromophenylalanine; and 1,2-aminothiol-containing amino acid residues.

[0544] Non-canonical amino acid residues can be incorporated by amino acid substitution or insertion. Non-canonical amino acid residues can be incorporated into peptides by chemical peptide synthesis rather than synthesis in a biological system, such as a recombinantly expressed cell, or those skilled in the art can use known techniques of protein engineering using recombinantly expressed cells. (See, e.g., Link et al., Non-canonical amino acids in protein engineering, Current Opinion in Biotechnology, 14(6):603-609 (2003); Schultz et al., In vivo incorporation of unnatural amino acids, U.S. Pat. No. 7,045,337.)

[0545] The selection of the location of the conjugation site within the entire anti-GIPR antigen-binding protein is another important aspect of selecting an internal conjugation site according to the present invention. Any exposed amino acid residue on the anti-GIPR antigen-binding protein can be a potentially useful conjugation site, and if it is not already present at the selected conjugation site in the anti-GIPR antigen-binding protein sequence, it can be mutated to cysteine ​​or some other reactive amino acid for site-selective conjugation. However, this approach does not take into account potential steric hindrances that may disrupt the activity of the conjugation partner or limit the reactivity of the genetically engineered mutation.

[0546] In one embodiment, the anti-GIPR antigen binding protein is an antibody or functional fragment thereof. In one embodiment, the anti-GIPR antibody or functional fragment thereof comprises an amino acid substitution of cysteine ​​or a non-canonical amino acid at one or more conjugation sites. These conjugation sites can be on essentially any residue of the antigen binding protein. In some embodiments, the conjugation site is located within the CL, CH1, CH2, or CH3 region of the antibody or functional fragment thereof. In some embodiments, the conjugation site is selected from the group consisting of D70 of the antibody light chain for reference sequence SEQ ID NO: 455, E276 of the antibody heavy chain for reference sequence SEQ ID NO: 612, and T363 of the antibody heavy chain for reference sequence SEQ ID NO: 612. For clarity, "D70 in the antibody light chain of reference sequence SEQ ID NO: 455" is the same substitution position as position D88 in AHo in the light chain of antibody 5G12.006 and position D70 in Kabat in the light chain of antibody 5G12.006, "E276 in the antibody heavy chain of reference sequence SEQ ID NO: 612" is the same substitution position as position E384 in AHo in the heavy chain of antibody 5G12.006 and position E285 in Kabat in the heavy chain of antibody 5G12.006, and "T363 in the antibody heavy chain of reference sequence SEQ ID NO: 612" is the same substitution position as position T487 in AHo in the heavy chain of antibody 5G12.006 and position T382 in Kabat in the heavy chain of antibody 5G12.006.

[0547] 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 that are 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, except that one or both of the domains differ 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, and each such sequence difference is independently either a single amino acid deletion, insertion, or substitution, and such deletions, insertions, and / or substitutions result in no more than 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15 amino acid changes 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 with the N-terminal methionine deleted. 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 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.

[0548] 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.

[0549] Other antigen binding proteins provided are variants of antibodies formed by the combination of heavy and light chains shown in Table 3, and comprise a light chain and / or a heavy chain, each having 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 comprise at least one heavy chain and one light chain, while in other cases, the variant form comprises two identical light chains and two identical heavy chains.

[0550] Various combinations of heavy chain variable regions may be combined with any of various combinations of light chain variable regions.

[0551] In additional embodiments, the isolated antigen binding protein provided herein is a human antibody comprising a sequence set forth in Table 3, and is of the IgG1, IgG2, IgG3, or IgG4 type.

[0552] 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 contain both a CDRH3 and a CDRL3. Specific light chain and heavy chain CDRs are identified in Table 4A and Table 4B, respectively.

[0553] 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., U.S. 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.

[0554] The structure and properties of CDRs in naturally occurring antibodies are described above. Briefly, in conventional antibodies, the CDRs are incorporated into frameworks in the heavy and light chain variable regions, where they constitute the regions responsible for antigen binding and recognition. The variable regions comprise at least three heavy chain or light chain CDRs (see Kabat et al., 1991, supra, "Sequences of Proteins of Immunological Interest," Public Health Service NIH, Bethesda, MD; 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 conventional antibody structure, as well as be incorporated into a variety of other polypeptide structures described herein.

[0555] In one embodiment, the antibody or fragment thereof comprises CDRL1, CDRL2, CDRL3, CDRH1, CDRH2 and CDRH3, and the antibody or functional fragment thereof comprises an amino acid substitution of cysteine ​​or a non-canonical amino acid at one or more conjugation sites selected from the group consisting of D70 of the antibody light chain for reference sequence SEQ ID NO: 455, E276 of the antibody heavy chain for reference sequence SEQ ID NO: 612 and T363 of the antibody heavy chain for reference sequence SEQ ID NO: 612. In one embodiment, the antibody or fragment thereof comprises CDRL1, CDRL2, CDRL3, CDRH1, CDRH2, and CDRH3, wherein each of CDRL1, CDRL2, CDRL3, CDRH1, CDRH2, and CDRH3 is selected from the group consisting of SEQ ID NO:629, SEQ ID NO:786, SEQ ID NO:943, SEQ ID NO:1100, SEQ ID NO:1257, and SEQ ID NO:1414; SEQ ID NO:630, SEQ ID NO:787, SEQ ID NO:944, SEQ ID NO:1101, SEQ ID NO:1258, and SEQ ID NO:1415; SEQ ID NO:631, SEQ ID NO:788, SEQ ID NO:945, SEQ ID NO:1102, SEQ ID NO:1259, and SEQ ID NO:1416; SEQ ID NO:632, SEQ ID NO:789, SEQ ID NO:946, SEQ ID NO:1103, SEQ ID NO:1260, and SEQ ID NO:1417; SEQ ID NO:633, SEQ ID NO:790, SEQ ID NO:947, SEQ ID NO:1104, SEQ ID NO:1261, and SEQ ID NO:1418; SEQ ID NO:634, SEQ ID NO:791, SEQ ID NO:948, SEQ ID NO:1105, SEQ ID NO:1262 and SEQ ID NO:1419, SEQ ID NO:635, SEQ ID NO:792, SEQ ID NO:949, SEQ ID NO:1106, SEQ ID NO:1263 and SEQ ID NO:1420, SEQ ID NO:636, SEQ ID NO:793, SEQ ID NO:950, SEQ ID NO:1107, SEQ ID NO:1264 and SEQ ID NO:1421, SEQ ID NO:637, SEQ ID NO:794, SEQ ID NO:951, SEQ ID NO:1108, SEQ ID NO:1265 and SEQ ID NO:1422 and SEQ ID NO: 638, SEQ ID NO: 795, SEQ ID NO: 952, SEQ ID NO: 1109, SEQ ID NO: 1266 and SEQ ID NO: 1423, and SEQ ID NO: 639, SEQ ID NO: 796, SEQ ID NO: 953, SEQ ID NO: 1110, SEQ ID NO: 1267 and SEQ ID NO: 1424, and SEQ ID NO: 640, SEQ ID NO: 797, SEQ ID NO: 954, SEQ ID NO: 1111, SEQ ID NO: 1268 and SEQ ID NO: 1425, and SEQ ID NO: 641, SEQ ID NO: 798, SEQ ID NO: 955, SEQ ID NO: 1112,SEQ ID NO: 1269 and SEQ ID NO: 1426, SEQ ID NO: 642, SEQ ID NO: 799, SEQ ID NO: 956, SEQ ID NO: 1113, SEQ ID NO: 1270 and SEQ ID NO: 1427, SEQ ID NO: 643, SEQ ID NO: 800, SEQ ID NO: 957, SEQ ID NO: 1114, SEQ ID NO: 1271 and SEQ ID NO: 1428, SEQ ID NO: 644, SEQ ID NO: 801, SEQ ID NO: 958, SEQ ID NO: 1115, SEQ ID NO: 1272 and SEQ ID NO: 1429, SEQ ID NO: 645, SEQ ID NO: 802, SEQ ID NO: 959, SEQ ID NO: 1116, SEQ ID NO: 1273 and SEQ ID NO: 1430, SEQ ID NO: 646, SEQ ID NO: 803, SEQ ID NO: Sequence number 960, sequence number 1117, sequence number 1274, and sequence number 1431; sequence number 647, sequence number 804, sequence number 961, sequence number 1118, sequence number 1275, and sequence number 1432; sequence number 648, sequence number 805, sequence number 962, sequence number 1119, sequence number 1276, and sequence number 1433; sequence number 649, sequence number 806, sequence number 963, sequence number 1120, sequence number 1277, and sequence number 1434; sequence number 650, sequence number 807, sequence number 964, sequence number 1121, sequence number 1278, and sequence number 1435; sequence number SEQ ID NO: 651, SEQ ID NO: 808, SEQ ID NO: 965, SEQ ID NO: 1122, SEQ ID NO: 1279 and SEQ ID NO: 1436, SEQ ID NO: 652, SEQ ID NO: 809, SEQ ID NO: 966, SEQ ID NO: 1123, SEQ ID NO: 1280 and SEQ ID NO: 1437, SEQ ID NO: 653, SEQ ID NO: 810, SEQ ID NO: 967, SEQ ID NO: 1124, SEQ ID NO: 1281 and SEQ ID NO: 1438, SEQ ID NO: 654, SEQ ID NO: 811, SEQ ID NO: 968, SEQ ID NO: 1125, SEQ ID NO: 1282 and SEQ ID NO: 1439, SEQ ID NO: 655, SEQ ID NO: 812, SEQ ID NO: 969, SEQ ID NO: 1126, SEQ ID NO: 12 83 and SEQ ID NO: 1440, SEQ ID NO: 656, SEQ ID NO: 813, SEQ ID NO: 970, SEQ ID NO: 1127, SEQ ID NO: 1284 and SEQ ID NO: 1441, SEQ ID NO: 657, SEQ ID NO: 814, SEQ ID NO: 971, SEQ ID NO: 1128, SEQ ID NO: 1285 and SEQ ID NO: 1442, SEQ ID NO: 658, SEQ ID NO: 815, SEQ ID NO: 972, SEQ ID NO: 1129, SEQ ID NO: 1286 and SEQ ID NO: 1443, SEQ ID NO: 659, SEQ ID NO: 816, SEQ ID NO: 973, SEQ ID NO: 1130, SEQ ID NO: 1287 and SEQ ID NO: 1444, SEQ ID NO: 660, SEQ ID NO: 817, SEQ ID NO: 974,SEQ ID NO: 1131, SEQ ID NO: 1288 and SEQ ID NO: 1445, SEQ ID NO: 661, SEQ ID NO: 818, SEQ ID NO: 975, SEQ ID NO: 1132, SEQ ID NO: 1289 and SEQ ID NO: 1446, SEQ ID NO: 662, SEQ ID NO: 819, SEQ ID NO: 976, SEQ ID NO: 1133, SEQ ID NO: 1290 and SEQ ID NO: 1447, SEQ ID NO: 663, SEQ ID NO: 820, SEQ ID NO: 977, SEQ ID NO: 1134, SEQ ID NO: 1291 and SEQ ID NO: 1448, SEQ ID NO: 664, SEQ ID NO: 821, SEQ ID NO: 978, SEQ ID NO: 1135, SEQ ID NO: 1292 and SEQ ID NO: 1449, SEQ ID NO: 665, SEQ ID NO: 822, SEQ ID NO: 979, SEQ ID NO: 1136, SEQ ID NO: 1293 and SEQ ID NO: 1450, SEQ ID NO: 666, SEQ ID NO: 823, SEQ ID NO: 980, SEQ ID NO: 1137, SEQ ID NO: 1294 and SEQ ID NO: 1451, SEQ ID NO: 667, SEQ ID NO: 824, SEQ ID NO: 981, SEQ ID NO: 1138, SEQ ID NO: 1295 and SEQ ID NO: 1452, SEQ ID NO: 668, SEQ ID NO: 825, SEQ ID NO: 982, SEQ ID NO: 1139, SEQ ID NO: 1296 and SEQ ID NO: 1453, SEQ ID NO: 669, SEQ ID NO: 826, SEQ ID NO: 983, SEQ ID NO: 1140, SEQ ID NO: 1297 and SEQ ID NO: No. 1454, SEQ ID NO: 670, SEQ ID NO: 827, SEQ ID NO: 984, SEQ ID NO: 1141, SEQ ID NO: 1298 and SEQ ID NO: 1455, SEQ ID NO: 671, SEQ ID NO: 828, SEQ ID NO: 985, SEQ ID NO: 1142, SEQ ID NO: 1299 and SEQ ID NO: 1456, SEQ ID NO: 672, SEQ ID NO: 829, SEQ ID NO: 986, SEQ ID NO: 1143, SEQ ID NO: 1300 and SEQ ID NO: 1457, SEQ ID NO: 673, SEQ ID NO: 830, SEQ ID NO: 987, SEQ ID NO: 1144, SEQ ID NO: 1301 and SEQ ID NO: 1458, SEQ ID NO: 674, SEQ ID NO: 831, SEQ ID NO: 988, SEQ ID NO: 114 5, SEQ ID NO: 1302 and SEQ ID NO: 1459, SEQ ID NO: 675, SEQ ID NO: 832, SEQ ID NO: 989, SEQ ID NO: 1146, SEQ ID NO: 1303 and SEQ ID NO: 1460, SEQ ID NO: 676, SEQ ID NO: 833, SEQ ID NO: 990, SEQ ID NO: 1147, SEQ ID NO: 1304 and SEQ ID NO: 1461, SEQ ID NO: 677, SEQ ID NO: 834, SEQ ID NO: 991, SEQ ID NO: 1148, SEQ ID NO: 1305 and SEQ ID NO: 1462, SEQ ID NO: 678, SEQ ID NO: 835, SEQ ID NO: 992, SEQ ID NO: 1149, SEQ ID NO: 1306 and SEQ ID NO: 1463, SEQ ID NO: 679, SEQ ID NO: 836,SEQ ID NO:993, SEQ ID NO:1150, SEQ ID NO:1307 and SEQ ID NO:1464, SEQ ID NO:680, SEQ ID NO:837, SEQ ID NO:994, SEQ ID NO:1151, SEQ ID NO:1308 and SEQ ID NO:1465, SEQ ID NO:681, SEQ ID NO:838, SEQ ID NO:995, SEQ ID NO:1152, SEQ ID NO:1309 and SEQ ID NO:1466, SEQ ID NO:682, SEQ ID NO:839, SEQ ID NO:996, SEQ ID NO:1153, SEQ ID NO:1310 and SEQ ID NO:1467, SEQ ID NO:683, SEQ ID NO:840, SEQ ID NO:997, SEQ ID NO:1154, SEQ ID NO:1311 and SEQ ID NO:14 68, SEQ ID NO: 684, SEQ ID NO: 841, SEQ ID NO: 998, SEQ ID NO: 1155, SEQ ID NO: 1312 and SEQ ID NO: 1469, SEQ ID NO: 685, SEQ ID NO: 842, SEQ ID NO: 999, SEQ ID NO: 1156, SEQ ID NO: 1313 and SEQ ID NO: 1470, SEQ ID NO: 686, SEQ ID NO: 843, SEQ ID NO: 1000, SEQ ID NO: 1157, SEQ ID NO: 1314 and SEQ ID NO: 1471, SEQ ID NO: 687, SEQ ID NO: 844, SEQ ID NO: 1001, SEQ ID NO: 1158, SEQ ID NO: 1315 and SEQ ID NO: 1472, SEQ ID NO: 688, SEQ ID NO: 845, SEQ ID NO: 1002, SEQ ID NO: No. 1159, SEQ ID NO: 1316 and SEQ ID NO: 1473, SEQ ID NO: 689, SEQ ID NO: 846, SEQ ID NO: 1003, SEQ ID NO: 1160, SEQ ID NO: 1317 and SEQ ID NO: 1474, SEQ ID NO: 690, SEQ ID NO: 847, SEQ ID NO: 1004, SEQ ID NO: 1161, SEQ ID NO: 1318 and SEQ ID NO: 1475, SEQ ID NO: 691, SEQ ID NO: 848, SEQ ID NO: 1005, SEQ ID NO: 1162, SEQ ID NO: 1319 and SEQ ID NO: 1476, SEQ ID NO: 692, SEQ ID NO: 849, SEQ ID NO: 1006, SEQ ID NO: 1163, SEQ ID NO: 1320 and SEQ ID NO: 1477, SEQ ID NO: 693, SEQ ID NO:850, SEQ ID NO:1007, SEQ ID NO:1164, SEQ ID NO:1321 and SEQ ID NO:1478, SEQ ID NO:694, SEQ ID NO:851, SEQ ID NO:1008, SEQ ID NO:1165, SEQ ID NO:1322 and SEQ ID NO:1479, SEQ ID NO:695, SEQ ID NO:852, SEQ ID NO:1009, SEQ ID NO:1166, SEQ ID NO:1323 and SEQ ID NO:1480, SEQ ID NO:696, SEQ ID NO:853, SEQ ID NO:1010, SEQ ID NO:1167, SEQ ID NO:1324 and SEQ ID NO:1481, SEQ ID NO:697, SEQ ID NO:854, SEQ ID NO:1011, SEQ ID NO:1168,SEQ ID NO: 1325 and SEQ ID NO: 1482, SEQ ID NO: 698, SEQ ID NO: 855, SEQ ID NO: 1012, SEQ ID NO: 1169, SEQ ID NO: 1326 and SEQ ID NO: 1483, SEQ ID NO: 699, SEQ ID NO: 856, SEQ ID NO: 1013, SEQ ID NO: 1170, SEQ ID NO: 1327 and SEQ ID NO: 1484, SEQ ID NO: 700, SEQ ID NO: 857, SEQ ID NO: 1014, SEQ ID NO: 1171, SEQ ID NO: 1328 and SEQ ID NO: 1485, SEQ ID NO: 701, SEQ ID NO: 858, SEQ ID NO: 1015, SEQ ID NO: 1172, SEQ ID NO: 1329 and SEQ ID NO: 1486, SEQ ID NO:702, SEQ ID NO:859, SEQ ID NO:1016, SEQ ID NO:1173, SEQ ID NO:1330 and SEQ ID NO:1487, SEQ ID NO:703, SEQ ID NO:860, SEQ ID NO:1017, SEQ ID NO:1174, SEQ ID NO:1331 and SEQ ID NO:1488, SEQ ID NO:704, SEQ ID NO:861, SEQ ID NO:1018, SEQ ID NO:1175, SEQ ID NO:1332 and SEQ ID NO:1489, SEQ ID NO:705, SEQ ID NO:862, SEQ ID NO:1019, SEQ ID NO:1176, SEQ ID NO:1333 and SEQ ID NO:1490, SEQ ID NO:706, SEQ ID NO:863, SEQ ID NO:1 020, SEQ ID NO: 1177, SEQ ID NO: 1334 and SEQ ID NO: 1491, SEQ ID NO: 707, SEQ ID NO: 864, SEQ ID NO: 1021, SEQ ID NO: 1178, SEQ ID NO: 1335 and SEQ ID NO: 1492, SEQ ID NO: 708, SEQ ID NO: 865, SEQ ID NO: 1022, SEQ ID NO: 1179, SEQ ID NO: 1336 and SEQ ID NO: 1493, SEQ ID NO: 709, SEQ ID NO: 866, SEQ ID NO: 1023, SEQ ID NO: 1180, SEQ ID NO: 1337 and SEQ ID NO: 1494, SEQ ID NO: 710, SEQ ID NO: 867, SEQ ID NO: 1024, SEQ ID NO: 1181, SEQ ID NO: 133 8 and SEQ ID NO:1495, SEQ ID NO:711, SEQ ID NO:868, SEQ ID NO:1025, SEQ ID NO:1182, SEQ ID NO:1339 and SEQ ID NO:1496, SEQ ID NO:712, SEQ ID NO:869, SEQ ID NO:1026, SEQ ID NO:1183, SEQ ID NO:1340 and SEQ ID NO:1497, SEQ ID NO:713, SEQ ID NO:870, SEQ ID NO:1027, SEQ ID NO:1184, SEQ ID NO:1341 and SEQ ID NO:1498, SEQ ID NO:714, SEQ ID NO:871, SEQ ID NO:1028, SEQ ID NO:1185, SEQ ID NO:1342 and SEQ ID NO:1499, SEQ ID NO:715, SEQ ID NO: 872, SEQ ID NO: 1029, SEQ ID NO: 1186, SEQ ID NO: 1343 and SEQ ID NO: 1500, SEQ ID NO: 716, SEQ ID NO: 873, SEQ ID NO: 1030, SEQ ID NO: 1187, SEQ ID NO: 1344 and SEQ ID NO: 1501, SEQ ID NO: 717, SEQ ID NO: 874, SEQ ID NO: 1031, SEQ ID NO: 1188, SEQ ID NO: 1345 and SEQ ID NO: 1502, SEQ ID NO: 718, SEQ ID NO: 875, SEQ ID NO: 1032, SEQ ID NO: 1189, SEQ ID NO: 1346 and SEQ ID NO: 1503, SEQ ID NO: 719, SEQ ID NO: 876, SEQ ID NO: 1033, SEQ ID NO: 1190, SEQ ID NO: 134 7 and SEQ ID NO:1504, SEQ ID NO:720, SEQ ID NO:877, SEQ ID NO:1034, SEQ ID NO:1191, SEQ ID NO:1348 and SEQ ID NO:1505, SEQ ID NO:721, SEQ ID NO:878, SEQ ID NO:1035, SEQ ID NO:1192, SEQ ID NO:1349 and SEQ ID NO:1506, SEQ ID NO:722, SEQ ID NO:879, SEQ ID NO:1036, SEQ ID NO:1193, SEQ ID NO:1350 and SEQ ID NO:1507, SEQ ID NO:723, SEQ ID NO:880, SEQ ID NO:1037, SEQ ID NO:1194, SEQ ID NO:1351 and SEQ ID NO:1508, SEQ ID NO:724, SEQ ID NO:881, SEQ ID NO:1 038, SEQ ID NO: 1195, SEQ ID NO: 1352 and SEQ ID NO: 1509, SEQ ID NO: 725, SEQ ID NO: 882, SEQ ID NO: 1039, SEQ ID NO: 1196, SEQ ID NO: 1353 and SEQ ID NO: 1510, SEQ ID NO: 726, SEQ ID NO: 883, SEQ ID NO: 1040, SEQ ID NO: 1197, SEQ ID NO: 1354 and SEQ ID NO: 1511, SEQ ID NO: 727, SEQ ID NO: 884, SEQ ID NO: 1041, SEQ ID NO: 1198, SEQ ID NO: 1355 and SEQ ID NO: 1512, SEQ ID NO: 728, SEQ ID NO: 885, SEQ ID NO: 1042, SEQ ID NO: 1199, SEQ ID NO: 1356 and SEQ ID NO: 1513, SEQ ID NO: 729, SEQ ID NO: 886, SEQ ID NO: 1043, SEQ ID NO: 1200, SEQ ID NO: 1357 and SEQ ID NO: 1514, SEQ ID NO: 730, SEQ ID NO: 887, SEQ ID NO: 1044, SEQ ID NO: 1201, SEQ ID NO: 1358 and SEQ ID NO: 1515, SEQ ID NO: 731, SEQ ID NO: 888, SEQ ID NO: 1045, SEQ ID NO: 1202, SEQ ID NO: 1359 and SEQ ID NO: 1516, SEQ ID NO: 732, SEQ ID NO: 889, SEQ ID NO: 1046, SEQ ID NO: 1203, SEQ ID NO: 1360 and SEQ ID NO: 1517, SEQ ID NO: 733, SEQ ID NO: 890, SEQ ID NO: 1047, SEQ ID NO: 1204,SEQ ID NO: 1361 and SEQ ID NO: 1518, SEQ ID NO: 734, SEQ ID NO: 891, SEQ ID NO: 1048, SEQ ID NO: 1205, SEQ ID NO: 1362 and SEQ ID NO: 1519, SEQ ID NO: 735, SEQ ID NO: 892, SEQ ID NO: 1049, SEQ ID NO: 1206, SEQ ID NO: 1363 and SEQ ID NO: 1520, SEQ ID NO: 736, SEQ ID NO: 893, SEQ ID NO: 1050, SEQ ID NO: 1207, SEQ ID NO: 1364 and SEQ ID NO: 1521, SEQ ID NO: 737, SEQ ID NO: 894, SEQ ID NO: 1051, SEQ ID NO: 1208, SEQ ID NO: 1365 and SEQ ID NO: 1522, SEQ ID NO: 738, SEQ ID NO: 89 5, SEQ ID NO: 1052, SEQ ID NO: 1209, SEQ ID NO: 1366 and SEQ ID NO: 1523, SEQ ID NO: 739, SEQ ID NO: 896, SEQ ID NO: 1053, SEQ ID NO: 1210, SEQ ID NO: 1367 and SEQ ID NO: 1524, SEQ ID NO: 740, SEQ ID NO: 897, SEQ ID NO: 1054, SEQ ID NO: 1211, SEQ ID NO: 1368 and SEQ ID NO: 1525, SEQ ID NO: 741, SEQ ID NO: 898, SEQ ID NO: 1055, SEQ ID NO: 1212, SEQ ID NO: 1369 and SEQ ID NO: 1526, SEQ ID NO: 742, SEQ ID NO: 899, SEQ ID NO: 1056, SEQ ID NO: 1213, SEQ ID NO: 1370 and SEQ ID NO: 1527, SEQ ID NO:743, SEQ ID NO:900, SEQ ID NO:1057, SEQ ID NO:1214, SEQ ID NO:1371 and SEQ ID NO:1528, SEQ ID NO:744, SEQ ID NO:901, SEQ ID NO:1058, SEQ ID NO:1215, SEQ ID NO:1372 and SEQ ID NO:1529, SEQ ID NO:745, SEQ ID NO:902, SEQ ID NO:1059, SEQ ID NO:1216, SEQ ID NO:1373 and SEQ ID NO:1530, SEQ ID NO:746, SEQ ID NO:903, SEQ ID NO:1060, SEQ ID NO:1217, SEQ ID NO:1374 and SEQ ID NO:1531, SEQ ID NO:747, SEQ ID NO:904, SEQ ID NO:1061, SEQ ID NO: No. 1218, SEQ ID NO: 1375 and SEQ ID NO: 1532, SEQ ID NO: 748, SEQ ID NO: 905, SEQ ID NO: 1062, SEQ ID NO: 1219, SEQ ID NO: 1376 and SEQ ID NO: 1533, SEQ ID NO: 749, SEQ ID NO: 906, SEQ ID NO: 1063, SEQ ID NO: 1220, SEQ ID NO: 1377 and SEQ ID NO: 1534, SEQ ID NO: 750, SEQ ID NO: 907, SEQ ID NO: 1064, SEQ ID NO: 1221, SEQ ID NO: 1378 and SEQ ID NO: 1535, SEQ ID NO: 751, SEQ ID NO: 908, SEQ ID NO: 1065, SEQ ID NO: 1222, SEQ ID NO: 1379 and SEQ ID NO: 1536, SEQ ID NO: 752,SEQ ID NO: 909, SEQ ID NO: 1066, SEQ ID NO: 1223, SEQ ID NO: 1380 and SEQ ID NO: 1537, SEQ ID NO: 753, SEQ ID NO: 910, SEQ ID NO: 1067, SEQ ID NO: 1224, SEQ ID NO: 1381 and SEQ ID NO: 1538, SEQ ID NO: 754, SEQ ID NO: 911, SEQ ID NO: 1068, SEQ ID NO: 1225, SEQ ID NO: 1382 and SEQ ID NO: 1539, SEQ ID NO: 755, SEQ ID NO: 912, SEQ ID NO: 1069, SEQ ID NO: 1226, SEQ ID NO: 1383 and SEQ ID NO: 1540, SEQ ID NO: 756, SEQ ID NO: 913, SEQ ID NO: 1070, SEQ ID NO: 1227, SEQ ID NO: 138 4 and SEQ ID NO:1541, SEQ ID NO:757, SEQ ID NO:914, SEQ ID NO:1071, SEQ ID NO:1228, SEQ ID NO:1385 and SEQ ID NO:1542, SEQ ID NO:758, SEQ ID NO:915, SEQ ID NO:1072, SEQ ID NO:1229, SEQ ID NO:1386 and SEQ ID NO:1543, SEQ ID NO:759, SEQ ID NO:916, SEQ ID NO:1073, SEQ ID NO:1230, SEQ ID NO:1387 and SEQ ID NO:1544, SEQ ID NO:760, SEQ ID NO:917, SEQ ID NO:1074, SEQ ID NO:1231, SEQ ID NO:1388 and SEQ ID NO:1545, SEQ ID NO:761, SEQ ID NO:918, SEQ ID NO:1 075, SEQ ID NO: 1232, SEQ ID NO: 1389 and SEQ ID NO: 1546, SEQ ID NO: 762, SEQ ID NO: 919, SEQ ID NO: 1076, SEQ ID NO: 1233, SEQ ID NO: 1390 and SEQ ID NO: 1547, SEQ ID NO: 763, SEQ ID NO: 920, SEQ ID NO: 1077, SEQ ID NO: 1234, SEQ ID NO: 1391 and SEQ ID NO: 1548, SEQ ID NO: 764, SEQ ID NO: 921, SEQ ID NO: 1078, SEQ ID NO: 1235, SEQ ID NO: 1392 and SEQ ID NO: 1549, SEQ ID NO: 765, SEQ ID NO: 922, SEQ ID NO: 1079, SEQ ID NO: 1236, SEQ ID NO: 1393 and SEQ ID NO: 1550, SEQ ID NO: 766, SEQ ID NO: 923, SEQ ID NO: 1080, SEQ ID NO: 1237, SEQ ID NO: 1394 and SEQ ID NO: 1551, SEQ ID NO: 767, SEQ ID NO: 924, SEQ ID NO: 1081, SEQ ID NO: 1238, SEQ ID NO: 1395 and SEQ ID NO: 1552, SEQ ID NO: 768, SEQ ID NO: 925, SEQ ID NO: 1082, SEQ ID NO: 1239, SEQ ID NO: 1396 and SEQ ID NO: 1553, SEQ ID NO: 769, SEQ ID NO: 926, SEQ ID NO: 1083, SEQ ID NO: 1240, SEQ ID NO: 1397 and SEQ ID NO: 1554, SEQ ID NO: 770, SEQ ID NO: 927, SEQ ID NO: 1084, SEQ ID NO: 1241,SEQ ID NO: 1398 and SEQ ID NO: 1555, SEQ ID NO: 771, SEQ ID NO: 928, SEQ ID NO: 1085, SEQ ID NO: 1242, SEQ ID NO: 1399 and SEQ ID NO: 1556, SEQ ID NO: 772, SEQ ID NO: 929, SEQ ID NO: 1086, SEQ ID NO: 1243, SEQ ID NO: 1400 and SEQ ID NO: 1557, SEQ ID NO: 773, SEQ ID NO: 930, SEQ ID NO: 1087, SEQ ID NO: 1244, SEQ ID NO: 1401 and SEQ ID NO: 1558, SEQ ID NO: 774, SEQ ID NO: 931, SEQ ID NO: 1088, SEQ ID NO: 1245, SEQ ID NO: 1402 and SEQ ID NO: 1559, SEQ ID NO: 775, SEQ ID NO: 9 32, SEQ ID NO: 1089, SEQ ID NO: 1246, SEQ ID NO: 1403 and SEQ ID NO: 1560, SEQ ID NO: 776, SEQ ID NO: 933, SEQ ID NO: 1090, SEQ ID NO: 1247, SEQ ID NO: 1404 and SEQ ID NO: 1561, SEQ ID NO: 777, SEQ ID NO: 934, SEQ ID NO: 1091, SEQ ID NO: 1248, SEQ ID NO: 1405 and SEQ ID NO: 1562, SEQ ID NO: 778, SEQ ID NO: 935, SEQ ID NO: 1092, SEQ ID NO: 1249, SEQ ID NO: 1406 and SEQ ID NO: 1563, SEQ ID NO: 779, SEQ ID NO: 936, SEQ ID NO: 1093, SEQ ID NO: 1250, SEQ ID NO: 1407 and SEQ ID NO: No. 1564, SEQ ID NO: 780, SEQ ID NO: 937, SEQ ID NO: 1094, SEQ ID NO: 1251, SEQ ID NO: 1408 and SEQ ID NO: 1565, SEQ ID NO: 781, SEQ ID NO: 938, SEQ ID NO: 1095, SEQ ID NO: 1252, SEQ ID NO: 1409 and SEQ ID NO: 1566, SEQ ID NO: 782, SEQ ID NO: 939, SEQ ID NO: 1096, SEQ ID NO: 1253, SEQ ID NO: 1410 and SEQ ID NO: 1567, SEQ ID NO: 783, SEQ ID NO: 940, SEQ ID NO: 1097, SEQ ID NO: 1254, SEQ ID NO: 1411 and SEQ ID NO: 1568, SEQ ID NO: 784, SEQ ID NO: 941, SEQ ID NO: 1098, SEQ ID NO: SEQ ID NO: 1255, SEQ ID NO: 1412 and SEQ ID NO: 1569, and SEQ ID NO: 785, SEQ ID NO: 942, SEQ ID NO: 1099, SEQ ID NO: 1256, SEQ ID NO: 1413 and SEQ ID NO: 1570, wherein the antibody or functional fragment thereof comprises an amino acid substitution of cysteine ​​or a non-canonical amino acid at one or more conjugation sites selected from the group consisting of D70 of the antibody light chain relative to reference sequence SEQ ID NO: 455, E276 of the antibody heavy chain relative to reference sequence SEQ ID NO: 612, and T363 of the antibody heavy chain relative to reference sequence SEQ ID NO: 612.

[0556] In one embodiment, the antibody or fragment thereof comprises CDRL1, CDRL2, CDRL3, CDRH1, CDRH2 and CDRH3, and the antibody or functional fragment thereof comprises an amino acid substitution of cysteine ​​or a non-canonical amino acid at one or more conjugation sites selected from the group consisting of D70 of the antibody light chain relative to reference sequence SEQ ID NO: 455, E276 of the antibody heavy chain relative to reference sequence SEQ ID NO: 612, and T363 of the antibody heavy chain relative to reference sequence SEQ ID NO: 612, encoded by a polynucleotide. In one embodiment, the antibody or functional fragment thereof comprises CDRL1, CDRL2, CDRL3, CDRH1, CDRH2, and CDRH3, wherein the antibody or functional fragment thereof comprises an amino acid substitution of a cysteine ​​or a non-canonical amino acid at one or more conjugation sites selected from the group consisting of D70 in the antibody light chain relative to reference sequence SEQ ID NO:455, E276 in the antibody heavy chain relative to reference sequence SEQ ID NO:612, and T363 in the antibody heavy chain relative to reference sequence SEQ ID NO:612, and wherein each CDRL1, CDRL2, CDRL3, CDRH1, CDRH2, and CDRH3 is selected from the group consisting of SEQ ID NO:2199, SEQ ID NO:2356, SEQ ID NO:2513, SEQ ID NO:2670, SEQ ID NO:2827, and SEQ ID NO:2984, and SEQ ID NO:2200, SEQ ID NO:2357, SEQ ID NO:2514, SEQ ID NO:2671, SEQ ID NO:2828, and SEQ ID NO:2985, and SEQ ID NO:2201, SEQ ID NO:2357, SEQ ID NO:2514, SEQ ID NO:2671, SEQ ID NO:2828, and SEQ ID NO:2985, respectively. Sequence number 2358, sequence number 2515, sequence number 2672, sequence number 2829 and sequence number 2986, sequence number 2202, sequence number 2359, sequence number 2516, sequence number 2673, sequence number 2830 and sequence number 2987, sequence number 2203, sequence number 2360, sequence number 2517, sequence number 2674, sequence number 2831 and sequence number 2988, sequence number 2204, sequence number 2361, sequence number 2518, sequence number 26 75, SEQ ID NO: 2832 and SEQ ID NO: 2989, SEQ ID NO: 2205, SEQ ID NO: 2362, SEQ ID NO: 2519, SEQ ID NO: 2676, SEQ ID NO: 2833 and SEQ ID NO: 2990, SEQ ID NO: 2206, SEQ ID NO: 2363, SEQ ID NO: 2520, SEQ ID NO: 2677, SEQ ID NO: 2834 and SEQ ID NO: 2991, SEQ ID NO: 2207, SEQ ID NO: 2364, SEQ ID NO: 2521, SEQ ID NO: 2678, SEQ ID NO: 2835 and SEQ ID NO: 2992,SEQ ID NO: 2208, SEQ ID NO: 2365, SEQ ID NO: 2522, SEQ ID NO: 2679, SEQ ID NO: 2836 and SEQ ID NO: 2993, SEQ ID NO: 2209, SEQ ID NO: 2366, SEQ ID NO: 2523, SEQ ID NO: 2680, SEQ ID NO: 2837 and SEQ ID NO: 2994, SEQ ID NO: 2210, SEQ ID NO: 2367, SEQ ID NO: 2524, SEQ ID NO: 2681, SEQ ID NO: 2838 and SEQ ID NO: 2995, SEQ ID NO: 2211, SEQ ID NO: 2368, SEQ ID NO: 2525, SEQ ID NO: 2682, SEQ ID NO: 2839 and SEQ ID NO: 2996, SEQ ID NO: 2212, SEQ ID NO: 2369, SEQ ID NO: 2 526, SEQ ID NO: 2683, SEQ ID NO: 2840 and SEQ ID NO: 2997, SEQ ID NO: 2213, SEQ ID NO: 2370, SEQ ID NO: 2527, SEQ ID NO: 2684, SEQ ID NO: 2841 and SEQ ID NO: 2998, SEQ ID NO: 2214, SEQ ID NO: 2371, SEQ ID NO: 2528, SEQ ID NO: 2685, SEQ ID NO: 2842 and SEQ ID NO: 2999, SEQ ID NO: 2215, SEQ ID NO: 2372, SEQ ID NO: 2529, SEQ ID NO: 2686, SEQ ID NO: 2843 and SEQ ID NO: 3000, SEQ ID NO: 2216, SEQ ID NO: 2373, SEQ ID NO: 2530, SEQ ID NO: 2687, SEQ ID NO: 2844 and SEQ ID NO:3001, SEQ ID NO:2217, SEQ ID NO:2374, SEQ ID NO:2531, SEQ ID NO:2688, SEQ ID NO:2845 and SEQ ID NO:3002, SEQ ID NO:2218, SEQ ID NO:2375, SEQ ID NO:2532, SEQ ID NO:2689, SEQ ID NO:2846 and SEQ ID NO:3003, SEQ ID NO:2219, SEQ ID NO:2376, SEQ ID NO:2533, SEQ ID NO:2690, SEQ ID NO:2847 and SEQ ID NO:3004, SEQ ID NO:2220, SEQ ID NO:2377, SEQ ID NO:2534, SEQ ID NO:2691, SEQ ID NO:2848 and SEQ ID NO:3005, SEQ ID NO:2221, SEQ ID NO: 2378, SEQ ID NO: 2535, SEQ ID NO: 2692, SEQ ID NO: 2849 and SEQ ID NO: 3006, SEQ ID NO: 2222, SEQ ID NO: 2379, SEQ ID NO: 2536, SEQ ID NO: 2693, SEQ ID NO: 2850 and SEQ ID NO: 3007, SEQ ID NO: 2223, SEQ ID NO: 2380, SEQ ID NO: 2537, SEQ ID NO: 2694, SEQ ID NO: 2851 and SEQ ID NO: 3008, SEQ ID NO: 2224, SEQ ID NO: 2381, SEQ ID NO: 2538, SEQ ID NO: 2695, SEQ ID NO: 2852 and SEQ ID NO: 3009, SEQ ID NO: 2225, SEQ ID NO: 2382, SEQ ID NO: 2539, SEQ ID NO: 2696,SEQ ID NO:2853 and SEQ ID NO:3010, SEQ ID NO:2226, SEQ ID NO:2383, SEQ ID NO:2540, SEQ ID NO:2697, SEQ ID NO:2854 and SEQ ID NO:3011, SEQ ID NO:2227, SEQ ID NO:2384, SEQ ID NO:2541, SEQ ID NO:2698, SEQ ID NO:2855 and SEQ ID NO:3012, SEQ ID NO:2228, SEQ ID NO:2385, SEQ ID NO:2542, SEQ ID NO:2699, SEQ ID NO:2856 and SEQ ID NO:3013, SEQ ID NO:2229, SEQ ID NO:2386, SEQ ID NO:2543, SEQ ID NO:2700, SEQ ID NO:2857 and SEQ ID NO:3014, SEQ ID NO:2 230, SEQ ID NO: 2387, SEQ ID NO: 2544, SEQ ID NO: 2701, SEQ ID NO: 2858 and SEQ ID NO: 3015, SEQ ID NO: 2231, SEQ ID NO: 2388, SEQ ID NO: 2545, SEQ ID NO: 2702, SEQ ID NO: 2859 and SEQ ID NO: 3016, SEQ ID NO: 2232, SEQ ID NO: 2389, SEQ ID NO: 2546, SEQ ID NO: 2703, SEQ ID NO: 2860 and SEQ ID NO: 3017, SEQ ID NO: 2233, SEQ ID NO: 2390, SEQ ID NO: 2547, SEQ ID NO: 2704, SEQ ID NO: 2861 and SEQ ID NO: 3018, SEQ ID NO: 2234, SEQ ID NO: 2391, SEQ ID NO: 2548, SEQ ID NO: SEQ ID NO:2705, SEQ ID NO:2862 and SEQ ID NO:3019, SEQ ID NO:2235, SEQ ID NO:2392, SEQ ID NO:2549, SEQ ID NO:2706, SEQ ID NO:2863 and SEQ ID NO:3020, SEQ ID NO:2236, SEQ ID NO:2393, SEQ ID NO:2550, SEQ ID NO:2707, SEQ ID NO:2864 and SEQ ID NO:3021, SEQ ID NO:2237, SEQ ID NO:2394, SEQ ID NO:2551, SEQ ID NO:2708, SEQ ID NO:2865 and SEQ ID NO:3022, SEQ ID NO:2238, SEQ ID NO:2395, SEQ ID NO:2552, SEQ ID NO:2709, SEQ ID NO:2866 and SEQ ID NO:3023, Sequence number 2239, sequence number 2396, sequence number 2553, sequence number 2710, sequence number 2867 and sequence number 3024, sequence number 2240, sequence number 2397, sequence number 2554, sequence number 2711, sequence number 2868 and sequence number 3025, sequence number 2241, sequence number 2398, sequence number 2555, sequence number 2712, sequence number 2869 and sequence number 3026, sequence number 2242, sequence number 2399, sequence number 2556, sequence number 2713, sequence number 2870 and sequence number 3027, sequence number 2243, sequence number 2400, sequence number 2557,SEQ ID NO: 2714, SEQ ID NO: 2871 and SEQ ID NO: 3028, SEQ ID NO: 2244, SEQ ID NO: 2401, SEQ ID NO: 2558, SEQ ID NO: 2715, SEQ ID NO: 2872 and SEQ ID NO: 3029, SEQ ID NO: 2245, SEQ ID NO: 2402, SEQ ID NO: 2559, SEQ ID NO: 2716, SEQ ID NO: 2873 and SEQ ID NO: 3030, SEQ ID NO: 2246, SEQ ID NO: 2403, SEQ ID NO: 2560, SEQ ID NO: 2717, SEQ ID NO: 2874 and SEQ ID NO: 3031, SEQ ID NO: 2247, SEQ ID NO: 2404, SEQ ID NO: 2561, SEQ ID NO: 2718, SEQ ID NO: 2875 and SEQ ID NO: 30 32, SEQ ID NO: 2248, SEQ ID NO: 2405, SEQ ID NO: 2562, SEQ ID NO: 2719, SEQ ID NO: 2876 and SEQ ID NO: 3033, SEQ ID NO: 2249, SEQ ID NO: 2406, SEQ ID NO: 2563, SEQ ID NO: 2720, SEQ ID NO: 2877 and SEQ ID NO: 3034, SEQ ID NO: 2250, SEQ ID NO: 2407, SEQ ID NO: 2564, SEQ ID NO: 2721, SEQ ID NO: 2878 and SEQ ID NO: 3035, SEQ ID NO: 2251, SEQ ID NO: 2408, SEQ ID NO: 2565, SEQ ID NO: 2722, SEQ ID NO: 2879 and SEQ ID NO: 3036, SEQ ID NO: 2252, SEQ ID NO: 2409, SEQ ID NO: 2566, SEQ ID NO: 2723, SEQ ID NO: 2880 and SEQ ID NO: 3037, SEQ ID NO: 2253, SEQ ID NO: 2410, SEQ ID NO: 2567, SEQ ID NO: 2724, SEQ ID NO: 2881 and SEQ ID NO: 3038, SEQ ID NO: 2254, SEQ ID NO: 2411, SEQ ID NO: 2568, SEQ ID NO: 2725, SEQ ID NO: 2882 and SEQ ID NO: 3039, SEQ ID NO: 2255, SEQ ID NO: 2412, SEQ ID NO: 2569, SEQ ID NO: 2726, SEQ ID NO: 2883 and SEQ ID NO: 3040, SEQ ID NO: 2256, SEQ ID NO: 2413, SEQ ID NO: 2570, SEQ ID NO: 2727, SEQ ID NO: 2884 and SEQ ID NO: Number 3041, SEQ ID NO: 2257, SEQ ID NO: 2414, SEQ ID NO: 2571, SEQ ID NO: 2728, SEQ ID NO: 2885 and SEQ ID NO: 3042, SEQ ID NO: 2258, SEQ ID NO: 2415, SEQ ID NO: 2572, SEQ ID NO: 2729, SEQ ID NO: 2886 and SEQ ID NO: 3043, SEQ ID NO: 2259, SEQ ID NO: 2416, SEQ ID NO: 2573, SEQ ID NO: 2730, SEQ ID NO: 2887 and SEQ ID NO: 3044, SEQ ID NO: 2260, SEQ ID NO: 2417, SEQ ID NO: 2574, SEQ ID NO: 2731, SEQ ID NO: 2888 and SEQ ID NO: 3045, SEQ ID NO: 2261, SEQ ID NO: 2418,SEQ ID NO: 2575, SEQ ID NO: 2732, SEQ ID NO: 2889 and SEQ ID NO: 3046, SEQ ID NO: 2262, SEQ ID NO: 2419, SEQ ID NO: 2576, SEQ ID NO: 2733, SEQ ID NO: 2890 and SEQ ID NO: 3047, SEQ ID NO: 2263, SEQ ID NO: 2420, SEQ ID NO: 2577, SEQ ID NO: 2734, SEQ ID NO: 2891 and SEQ ID NO: 3048, SEQ ID NO: 2264, SEQ ID NO: 2421, SEQ ID NO: 2578, SEQ ID NO: 2735, SEQ ID NO: 2892 and SEQ ID NO: 3049, SEQ ID NO: 2265, SEQ ID NO: 2422, SEQ ID NO: 2579, SEQ ID NO: 2736, SEQ ID NO: 289 3 and SEQ ID NO: 3050, SEQ ID NO: 2266, SEQ ID NO: 2423, SEQ ID NO: 2580, SEQ ID NO: 2737, SEQ ID NO: 2894 and SEQ ID NO: 3051, SEQ ID NO: 2267, SEQ ID NO: 2424, SEQ ID NO: 2581, SEQ ID NO: 2738, SEQ ID NO: 2895 and SEQ ID NO: 3052, SEQ ID NO: 2268, SEQ ID NO: 2425, SEQ ID NO: 2582, SEQ ID NO: 2739, SEQ ID NO: 2896 and SEQ ID NO: 3053, SEQ ID NO: 2269, SEQ ID NO: 2426, SEQ ID NO: 2583, SEQ ID NO: 2740, SEQ ID NO: 2897 and SEQ ID NO: 3054, SEQ ID NO: 2270, SEQ ID NO: 2427, SEQ ID NO: 2584, SEQ ID NO: 2741, SEQ ID NO: 2898 and SEQ ID NO: 3055, SEQ ID NO: 2271, SEQ ID NO: 2428, SEQ ID NO: 2585, SEQ ID NO: 2742, SEQ ID NO: 2899 and SEQ ID NO: 3056, SEQ ID NO: 2272, SEQ ID NO: 2429, SEQ ID NO: 2586, SEQ ID NO: 2743, SEQ ID NO: 2900 and SEQ ID NO: 3057, SEQ ID NO: 2273, SEQ ID NO: 2430, SEQ ID NO: 2587, SEQ ID NO: 2744, SEQ ID NO: 2901 and SEQ ID NO: 3058, SEQ ID NO: 2274, SEQ ID NO: 2431, SEQ ID NO: 2588, SEQ ID NO: 2745, SEQ ID NO: No. 2902 and SEQ ID NO: 3059, SEQ ID NO: 2275, SEQ ID NO: 2432, SEQ ID NO: 2589, SEQ ID NO: 2746, SEQ ID NO: 2903 and SEQ ID NO: 3060, SEQ ID NO: 2276, SEQ ID NO: 2433, SEQ ID NO: 2590, SEQ ID NO: 2747, SEQ ID NO: 2904 and SEQ ID NO: 3061, SEQ ID NO: 2277, SEQ ID NO: 2434, SEQ ID NO: 2591, SEQ ID NO: 2748, SEQ ID NO: 2905 and SEQ ID NO: 3062, SEQ ID NO: 2278, SEQ ID NO: 2435, SEQ ID NO: 2592, SEQ ID NO: 2749, SEQ ID NO: 2906 and SEQ ID NO: 3063, SEQ ID NO: 2279,SEQ ID NO: 2436, SEQ ID NO: 2593, SEQ ID NO: 2750, SEQ ID NO: 2907 and SEQ ID NO: 3064, SEQ ID NO: 2280, SEQ ID NO: 2437, SEQ ID NO: 2594, SEQ ID NO: 2751, SEQ ID NO: 2908 and SEQ ID NO: 3065, SEQ ID NO: 2281, SEQ ID NO: 2438, SEQ ID NO: 2595, SEQ ID NO: 2752, sequences, SEQ ID NO: 2909 and SEQ ID NO: 3066, SEQ ID NO: 2282, SEQ ID NO: 2439, SEQ ID NO: 2596, SEQ ID NO: 2753, SEQ ID NO: 2910 and SEQ ID NO: 3067, SEQ ID NO: 2283, SEQ ID NO: 2440, SEQ ID NO: 2597, SEQ ID NO: 2754, SEQ ID NO: 2911 and SEQ ID NO: 3068, SEQ ID NO: 2284, SEQ ID NO: 2441, SEQ ID NO: 2598, SEQ ID NO: 2755, SEQ ID NO: 2912 and SEQ ID NO: 3069, SEQ ID NO: 2285, SEQ ID NO: 2442, SEQ ID NO: 2599, SEQ ID NO: 2756, SEQ ID NO: 2913 and SEQ ID NO: 3070, SEQ ID NO: 228 6, SEQ ID NO:2443, SEQ ID NO:2600, SEQ ID NO:2757, SEQ ID NO:2914 and SEQ ID NO:3071, SEQ ID NO:2287, SEQ ID NO:2444, SEQ ID NO:2601, SEQ ID NO:2758, SEQ ID NO:2915 and SEQ ID NO:3072, SEQ ID NO:2288, SEQ ID NO:2445, SEQ ID NO:2602, SEQ ID NO:2759, SEQ ID NO:2916 and SEQ ID NO:3073, SEQ ID NO:2289, SEQ ID NO:2446, SEQ ID NO:2603, SEQ ID NO:2760, SEQ ID NO:2917 and SEQ ID NO:3074, SEQ ID NO:2290, SEQ ID NO:2447, SEQ ID NO:2604, SEQ ID NO:2 761, SEQ ID NO: 2918 and SEQ ID NO: 3075, SEQ ID NO: 2291, SEQ ID NO: 2448, SEQ ID NO: 2605, SEQ ID NO: 2762, SEQ ID NO: 2919 and SEQ ID NO: 3076, SEQ ID NO: 2292, SEQ ID NO: 2449, SEQ ID NO: 2606, SEQ ID NO: 2763, SEQ ID NO: 2920 and SEQ ID NO: 3077, SEQ ID NO: 2293, SEQ ID NO: 2450, SEQ ID NO: 2607, SEQ ID NO: 2764, SEQ ID NO: 2921 and SEQ ID NO: 3078, SEQ ID NO: 2294, SEQ ID NO: 2451, SEQ ID NO: 2608, SEQ ID NO: 2765, SEQ ID NO: 2922 and SEQ ID NO: 3079, Sequence number 2295, sequence number 2452, sequence number 2609, sequence number 2766, sequence number 2923 and sequence number 3080, sequence number 2296, sequence number 2453, sequence number 2610, sequence number 2767, sequence number 2924 and sequence number 3081, sequence number 2297, sequence number 2454, sequence number 2611, sequence number 2768, sequence number 2925 and sequence number 3082, sequence number 2298, sequence number 2455, sequence number 2612, sequence number 2769, sequence number 2926 and sequence number 3083, sequence number 2299, sequence number 2456, sequence number 2613,SEQ ID NO: 2770, SEQ ID NO: 2927 and SEQ ID NO: 3084, SEQ ID NO: 2300, SEQ ID NO: 2457, SEQ ID NO: 2614, SEQ ID NO: 2771, SEQ ID NO: 2928 and SEQ ID NO: 3085, SEQ ID NO: 2301, SEQ ID NO: 2458, SEQ ID NO: 2615, SEQ ID NO: 2772, SEQ ID NO: 2929 and SEQ ID NO: 3086, SEQ ID NO: 2302, SEQ ID NO: 2459, SEQ ID NO: 2616, SEQ ID NO: 2773, SEQ ID NO: 2930 and SEQ ID NO: 3087, SEQ ID NO: 2303, SEQ ID NO: 2460, SEQ ID NO: 2617, SEQ ID NO: 2774, SEQ ID NO: 2931 and SEQ ID NO: 30 88, SEQ ID NO: 2304, SEQ ID NO: 2461, SEQ ID NO: 2618, SEQ ID NO: 2775, SEQ ID NO: 2932 and SEQ ID NO: 3089, SEQ ID NO: 2305, SEQ ID NO: 2462, SEQ ID NO: 2619, SEQ ID NO: 2776, SEQ ID NO: 2933 and SEQ ID NO: 3090, SEQ ID NO: 2306, SEQ ID NO: 2463, SEQ ID NO: 2620, SEQ ID NO: 2777, SEQ ID NO: 2934 and SEQ ID NO: 3091, SEQ ID NO: 2307, SEQ ID NO: 2464, SEQ ID NO: 2621, SEQ ID NO: 2778, SEQ ID NO: 2935 and SEQ ID NO: 3092, SEQ ID NO: 2308, SEQ ID NO: 2465, SEQ ID NO: 2622, SEQ ID NO: 2779, SEQ ID NO: 2936 and SEQ ID NO: 3093, SEQ ID NO: 2309, SEQ ID NO: 2466, SEQ ID NO: 2623, SEQ ID NO: 2780, SEQ ID NO: 2937 and SEQ ID NO: 3094, SEQ ID NO: 2310, SEQ ID NO: 2467, SEQ ID NO: 2624, SEQ ID NO: 2781, SEQ ID NO: 2938 and SEQ ID NO: 3095, SEQ ID NO: 2311, SEQ ID NO: 2468, SEQ ID NO: 2625, SEQ ID NO: 2782, SEQ ID NO: 2939 and SEQ ID NO: 3096, SEQ ID NO: 2312, SEQ ID NO: 2469, SEQ ID NO: 2626, SEQ ID NO: 2783, SEQ ID NO: 2940 and SEQ ID NO: Number 3097, SEQ ID NO: 2313, SEQ ID NO: 2470, SEQ ID NO: 2627, SEQ ID NO: 2784, SEQ ID NO: 2941 and SEQ ID NO: 3098, SEQ ID NO: 2314, SEQ ID NO: 2471, SEQ ID NO: 2628, SEQ ID NO: 2785, SEQ ID NO: 2942 and SEQ ID NO: 3099, SEQ ID NO: 2315, SEQ ID NO: 2472, SEQ ID NO: 2629, SEQ ID NO: 2786, SEQ ID NO: 2943 and SEQ ID NO: 3100, SEQ ID NO: 2316, SEQ ID NO: 2473, SEQ ID NO: 2630, SEQ ID NO: 2787, SEQ ID NO: 2944 and SEQ ID NO: 3101, SEQ ID NO: 2317, SEQ ID NO: 2474,SEQ ID NO: 2631, SEQ ID NO: 2788, SEQ ID NO: 2945 and SEQ ID NO: 3102, SEQ ID NO: 2318, SEQ ID NO: 2475, SEQ ID NO: 2632, SEQ ID NO: 2789, SEQ ID NO: 2946 and SEQ ID NO: 3103, SEQ ID NO: 2319, SEQ ID NO: 2476, SEQ ID NO: 2633, SEQ ID NO: 2790, SEQ ID NO: 2947 and SEQ ID NO: 3104, SEQ ID NO: 2320, SEQ ID NO: 2477, SEQ ID NO: 2634, SEQ ID NO: 2791, SEQ ID NO: 2948 and SEQ ID NO: 3105, SEQ ID NO: 2321, SEQ ID NO: 2478, SEQ ID NO: 2635, SEQ ID NO: 2792, SEQ ID NO: 294 9 and SEQ ID NO:3106, SEQ ID NO:2322, SEQ ID NO:2479, SEQ ID NO:2636, SEQ ID NO:2793, SEQ ID NO:2950 and SEQ ID NO:3107, SEQ ID NO:2323, SEQ ID NO:2480, SEQ ID NO:2637, SEQ ID NO:2794, SEQ ID NO:2951 and SEQ ID NO:3108, SEQ ID NO:2324, SEQ ID NO:2481, SEQ ID NO:2638, SEQ ID NO:2795, SEQ ID NO:2952 and SEQ ID NO:3109, SEQ ID NO:2325, SEQ ID NO:2482, SEQ ID NO:2639, SEQ ID NO:2796, SEQ ID NO:2953 and SEQ ID NO:3110, SEQ ID NO:2326, SEQ ID NO: 2483, SEQ ID NO: 2640, SEQ ID NO: 2797, SEQ ID NO: 2954 and SEQ ID NO: 3111, SEQ ID NO: 2327, SEQ ID NO: 2484, SEQ ID NO: 2641, SEQ ID NO: 2798, SEQ ID NO: 2955 and SEQ ID NO: 3112, SEQ ID NO: 2328, SEQ ID NO: 2485, SEQ ID NO: 2642, SEQ ID NO: 2799, SEQ ID NO: 2956 and SEQ ID NO: 3113, SEQ ID NO: 2329, SEQ ID NO: 2486, SEQ ID NO: 2643, SEQ ID NO: 2800, SEQ ID NO: 2957 and SEQ ID NO: 3114, SEQ ID NO: 2330, SEQ ID NO: 2487, SEQ ID NO: 2644, SEQ ID NO: 2801, SEQ ID NO: No. 2958 and SEQ ID NO: 3115, SEQ ID NO: 2331, SEQ ID NO: 2488, SEQ ID NO: 2645, SEQ ID NO: 2802, SEQ ID NO: 2959 and SEQ ID NO: 3116, SEQ ID NO: 2332, SEQ ID NO: 2489, SEQ ID NO: 2646, SEQ ID NO: 2803, SEQ ID NO: 2960 and SEQ ID NO: 3117, SEQ ID NO: 2333, SEQ ID NO: 2490, SEQ ID NO: 2647, SEQ ID NO: 2804, SEQ ID NO: 2961 and SEQ ID NO: 3118, SEQ ID NO: 2334, SEQ ID NO: 2491, SEQ ID NO: 2648, SEQ ID NO: 2805, SEQ ID NO: 2962 and SEQ ID NO: 3119, SEQ ID NO: 2335,SEQ ID NO: 2492, SEQ ID NO: 2649, SEQ ID NO: 2806, SEQ ID NO: 2963 and SEQ ID NO: 3120, SEQ ID NO: 2336, SEQ ID NO: 2493, SEQ ID NO: 2650, SEQ ID NO: 2807, SEQ ID NO: 2964 and SEQ ID NO: 3121, SEQ ID NO: 2337, SEQ ID NO: 2494, SEQ ID NO: 2651, SEQ ID NO: 2808, SEQ ID NO: 2965 and SEQ ID NO: 3122, SEQ ID NO: 2338, SEQ ID NO: 2495, SEQ ID NO: 2652, SEQ ID NO: 2809, SEQ ID NO: 2966 and SEQ ID NO: 3123, SEQ ID NO: 2339, SEQ ID NO: 2496, SEQ ID NO: 2653, SEQ ID NO: 281 0, SEQ ID NO: 2967 and SEQ ID NO: 3124, SEQ ID NO: 2340, SEQ ID NO: 2497, SEQ ID NO: 2654, SEQ ID NO: 2811, SEQ ID NO: 2968 and SEQ ID NO: 3125, SEQ ID NO: 2341, SEQ ID NO: 2498, SEQ ID NO: 2655, SEQ ID NO: 2812, SEQ ID NO: 2969 and SEQ ID NO: 3126, SEQ ID NO: 2342, SEQ ID NO: 2499, SEQ ID NO: 2656, SEQ ID NO: 2813, SEQ ID NO: 2970 and SEQ ID NO: 3127, SEQ ID NO: 2343, SEQ ID NO: 2500, SEQ ID NO: 2657, SEQ ID NO: 2814, SEQ ID NO: 2971 and SEQ ID NO: 3128, SEQ ID NO: 2344, SEQ ID NO: 2501, SEQ ID NO: 2658, SEQ ID NO: 2815, SEQ ID NO: 2972 ​​and SEQ ID NO: 3129, SEQ ID NO: 2345, SEQ ID NO: 2502, SEQ ID NO: 2659, SEQ ID NO: 2816, SEQ ID NO: 2973 and SEQ ID NO: 3130, SEQ ID NO: 2346, SEQ ID NO: 2503, SEQ ID NO: 266, SEQ ID NO: 2817, SEQ ID NO: 2974 and SEQ ID NO: 3131, SEQ ID NO: 2347, SEQ ID NO: 2504, SEQ ID NO: 2661, SEQ ID NO: 2818, SEQ ID NO: 2975 and SEQ ID NO: 3132, SEQ ID NO: 2348, SEQ ID NO: 2505, SEQ ID NO: 2662, SEQ ID NO: No. 2819, SEQ ID NO: 2976 and SEQ ID NO: 3133, SEQ ID NO: 2349, SEQ ID NO: 2506, SEQ ID NO: 2663, SEQ ID NO: 2820, SEQ ID NO: 2977 and SEQ ID NO: 3134, SEQ ID NO: 2350, SEQ ID NO: 2507, SEQ ID NO: 2664, SEQ ID NO: 2821, SEQ ID NO: 2978 and SEQ ID NO: 3135, SEQ ID NO: 2351, SEQ ID NO: 2508, SEQ ID NO: 2665, SEQ ID NO: 2822, SEQ ID NO: 2979 and SEQ ID NO: 3136, SEQ ID NO: 2352, SEQ ID NO: 2509, SEQ ID NO: 2666, SEQ ID NO: 2823, SEQ ID NO: 2980 and SEQ ID NO: 3137,Encoded by a sequence selected from the group consisting of SEQ ID NO: 2353, SEQ ID NO: 2510, SEQ ID NO: 2667, SEQ ID NO: 2824, SEQ ID NO: 2981 and SEQ ID NO: 3138, SEQ ID NO: 2354, SEQ ID NO: 2511, SEQ ID NO: 2668, SEQ ID NO: 2825, SEQ ID NO: 2982 and SEQ ID NO: 3139, SEQ ID NO: 2355, SEQ ID NO: 2512, SEQ ID NO: 2669, SEQ ID NO: 2826, SEQ ID NO: 2983 and SEQ ID NO: 3140.

[0557] In another aspect, the antigen binding protein comprises one, two, three, four, five, or six variant forms of the CDRs set out in Table 4A and Table 4B, which 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 Table 4A and Table 4B, respectively. Some antigen binding proteins comprise one, two, three, four, five, or six CDRs set out in Table 4A and Table 4B, which 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.

[0558] 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 Table 4A and Table 4B. In another aspect, the antigen binding protein comprises one, two, three, four, five, or six variant forms of the CDRs set forth in one of the rows directed to the antibodies of Table 4A and Table 4B, wherein the CDRs each 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 Table 4A and Table 4B. 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 those 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 aspect, the antigen binding protein comprises all six CDRs set forth in the rows of Table 4A and Table 4B, and 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.

[0559] In one embodiment, the antibody or fragment thereof comprises a light chain comprising a sequence selected from the group consisting of SEQ ID NOs: 472-628 and a heavy chain comprising a sequence selected from the group consisting of SEQ ID NOs: 472-628, and the antibody or functional fragment thereof comprises an amino acid substitution of cysteine ​​or a non-standard amino acid at one or more conjugation sites selected from the group consisting of D70 of the antibody light chain for reference sequence SEQ ID NO: 455, E276 of the antibody heavy chain for reference sequence SEQ ID NO: 612, and T363 of the antibody heavy chain for reference sequence SEQ ID NO: 612. In one embodiment, the antibody or fragment thereof comprises a light chain comprising SEQ ID NO: 315 and a heavy chain comprising SEQ ID NO: 472, a light chain comprising SEQ ID NO: 316 and a heavy chain comprising SEQ ID NO: 473, a light chain comprising SEQ ID NO: 317 and a heavy chain comprising SEQ ID NO: 474, a light chain comprising SEQ ID NO: 318 and a heavy chain comprising SEQ ID NO: 475, a light chain comprising SEQ ID NO: 319 and a heavy chain comprising SEQ ID NO: 476, a light chain comprising SEQ ID NO: 320 and a heavy chain comprising SEQ ID NO: 477, a light chain comprising SEQ ID NO: 321 and a heavy chain comprising SEQ ID NO: 478, a light chain comprising SEQ ID NO: 322 and a heavy chain comprising SEQ ID NO: 479, a light chain comprising SEQ ID NO: 323 and a heavy chain comprising SEQ ID NO: 480, a light chain comprising SEQ ID NO: 324 and a heavy chain comprising SEQ ID NO: 481, a light chain comprising SEQ ID NO: 325 and a heavy chain comprising SEQ ID NO: 482, a light chain comprising SEQ ID NO: 326 and a heavy chain comprising SEQ ID NO: 483, a light chain comprising SEQ ID NO: 327 and a heavy chain comprising SEQ ID NO: 484, a light chain comprising SEQ ID NO: 328 and a heavy chain comprising SEQ ID NO: 485, a light chain comprising SEQ ID NO: 329 and a heavy chain comprising SEQ ID NO: 490, a light chain comprising SEQ ID NO: 491 and a heavy chain comprising SEQ ID NO: 492, a light chain comprising SEQ ID NO: 493 and a heavy chain comprising SEQ ID NO: 494, a light chain comprising SEQ ID NO: 495 and a heavy chain comprising SEQ ID NO: 4 a light chain comprising SEQ ID NO: 328 and a heavy chain comprising SEQ ID NO: 485, a light chain comprising SEQ ID NO: 329 and a heavy chain comprising SEQ ID NO: 486, a light chain comprising SEQ ID NO: 330 and a heavy chain comprising SEQ ID NO: 487, a light chain comprising SEQ ID NO: 331 and a heavy chain comprising SEQ ID NO: 488, a light chain comprising SEQ ID NO: 332 and a heavy chain comprising SEQ ID NO: 489, a light chain comprising SEQ ID NO: 333 and a heavy chain comprising SEQ ID NO: 490, a light chain comprising SEQ ID NO: 334 and a heavy chain comprising SEQ ID NO: 491, a light chain comprising SEQ ID NO: 335 and a heavy chain comprising SEQ ID NO: 492, a light chain comprising SEQ ID NO: 336 and a heavy chain comprising SEQ ID NO: 493, a light chain comprising SEQ ID NO: 337 and a heavy chain comprising SEQ ID NO: 494, a light chain comprising SEQ ID NO: 338 and a heavy chain comprising SEQ ID NO: 495, a light chain comprising SEQ ID NO: 339 and a heavy chain comprising SEQ ID NO: 496, a light chain comprising SEQ ID NO: 340 and a heavy chain comprising SEQ ID NO: 497, a light chain comprising SEQ ID NO: 341 and a heavy chain comprising SEQ ID NO: 498.a light chain comprising SEQ ID NO: 342 and a heavy chain comprising SEQ ID NO: 499, a light chain comprising SEQ ID NO: 343 and a heavy chain comprising SEQ ID NO: 500, a light chain comprising SEQ ID NO: 344 and a heavy chain comprising SEQ ID NO: 501, a light chain comprising SEQ ID NO: 345 and a heavy chain comprising SEQ ID NO: 502, a light chain comprising SEQ ID NO: 346 and a heavy chain comprising SEQ ID NO: 503, a light chain comprising SEQ ID NO: 347 and a heavy chain comprising SEQ ID NO: 504, a light chain comprising SEQ ID NO: 348 and a heavy chain comprising SEQ ID NO: 505, a light chain comprising SEQ ID NO: 349 and a heavy chain comprising SEQ ID NO: 506, a light chain comprising SEQ ID NO: 350 and a heavy chain comprising SEQ ID NO: 507, a light chain comprising SEQ ID NO: 35 1 and a heavy chain comprising SEQ ID NO: 508, a light chain comprising SEQ ID NO: 352 and a heavy chain comprising SEQ ID NO: 509, a light chain comprising SEQ ID NO: 353 and a heavy chain comprising SEQ ID NO: 510, a light chain comprising SEQ ID NO: 354 and a heavy chain comprising SEQ ID NO: 511, a light chain comprising SEQ ID NO: 355 and a heavy chain comprising SEQ ID NO: 512, a light chain comprising SEQ ID NO: 356 and a heavy chain comprising SEQ ID NO: 513, a light chain comprising SEQ ID NO: 357 and a heavy chain comprising SEQ ID NO: 514, a light chain comprising SEQ ID NO: 358 and a heavy chain comprising SEQ ID NO: 515, a light chain comprising SEQ ID NO: 359 and a heavy chain comprising SEQ ID NO: 516, a light chain comprising SEQ ID NO: 360, and and a heavy chain comprising SEQ ID NO: 517, a light chain comprising SEQ ID NO: 361 and a heavy chain comprising SEQ ID NO: 518, a light chain comprising SEQ ID NO: 362 and a heavy chain comprising SEQ ID NO: 519, a light chain comprising SEQ ID NO: 363 and a heavy chain comprising SEQ ID NO: 520, a light chain comprising SEQ ID NO: 364 and a heavy chain comprising SEQ ID NO: 521, a light chain comprising SEQ ID NO: 365 and a heavy chain comprising SEQ ID NO: 522, a light chain comprising SEQ ID NO: 366 and a heavy chain comprising SEQ ID NO: 523, a light chain comprising SEQ ID NO: 367 and a heavy chain comprising SEQ ID NO: 524, a light chain comprising SEQ ID NO: 368 and a heavy chain comprising SEQ ID NO: 525, a light chain comprising SEQ ID NO: 369 and a heavy chain comprising SEQ ID NO: 52 a heavy chain comprising SEQ ID NO: 6, a light chain comprising SEQ ID NO: 370 and a heavy chain comprising SEQ ID NO: 527, a light chain comprising SEQ ID NO: 371 and a heavy chain comprising SEQ ID NO: 528, a light chain comprising SEQ ID NO: 372 and a heavy chain comprising SEQ ID NO: 529, a light chain comprising SEQ ID NO: 373 and a heavy chain comprising SEQ ID NO: 530, a light chain comprising SEQ ID NO: 374 and a heavy chain comprising SEQ ID NO: 531, a light chain comprising SEQ ID NO: 375 and a heavy chain comprising SEQ ID NO: 532, a light chain comprising SEQ ID NO: 376 and a heavy chain comprising SEQ ID NO: 533, a light chain comprising SEQ ID NO: 377 and a heavy chain comprising SEQ ID NO: 534, a light chain comprising SEQ ID NO: 378 and a heavy chain comprising SEQ ID NO: 535,a light chain comprising SEQ ID NO: 379 and a heavy chain comprising SEQ ID NO: 536, a light chain comprising SEQ ID NO: 380 and a heavy chain comprising SEQ ID NO: 537, a light chain comprising SEQ ID NO: 381 and a heavy chain comprising SEQ ID NO: 538, a light chain comprising SEQ ID NO: 382 and a heavy chain comprising SEQ ID NO: 539, a light chain comprising SEQ ID NO: 383 and a heavy chain comprising SEQ ID NO: 540, a light chain comprising SEQ ID NO: 384 and a heavy chain comprising SEQ ID NO: 541, a light chain comprising SEQ ID NO: 385 and a heavy chain comprising SEQ ID NO: 542, a light chain comprising SEQ ID NO: 386 and a heavy chain comprising SEQ ID NO: 543, a light chain comprising SEQ ID NO: 387 and a heavy chain comprising SEQ ID NO: 544, a light chain comprising SEQ ID NO: 38 a light chain comprising SEQ ID NO: 394 and a heavy chain comprising SEQ ID NO: 551; a light chain comprising SEQ ID NO: 395 and a heavy chain comprising SEQ ID NO: 552; a light chain comprising SEQ ID NO: 396 and a heavy chain comprising SEQ ID NO: 553; a light chain comprising SEQ ID NO: 397; and a heavy chain comprising SEQ ID NO: 554, a light chain comprising SEQ ID NO: 398 and a heavy chain comprising SEQ ID NO: 555, a light chain comprising SEQ ID NO: 399 and a heavy chain comprising SEQ ID NO: 556, a light chain comprising SEQ ID NO: 400 and a heavy chain comprising SEQ ID NO: 557, a light chain comprising SEQ ID NO: 401 and a heavy chain comprising SEQ ID NO: 558, a light chain comprising SEQ ID NO: 402 and a heavy chain comprising SEQ ID NO: 559, a light chain comprising SEQ ID NO: 403 and a heavy chain comprising SEQ ID NO: 560, a light chain comprising SEQ ID NO: 404 and a heavy chain comprising SEQ ID NO: 561, a light chain comprising SEQ ID NO: 405 and a heavy chain comprising SEQ ID NO: 562, a light chain comprising SEQ ID NO: 406 and a heavy chain comprising SEQ ID NO: 56 a heavy chain comprising SEQ ID NO: 3, a light chain comprising SEQ ID NO: 407 and a heavy chain comprising SEQ ID NO: 564, a light chain comprising SEQ ID NO: 408 and a heavy chain comprising SEQ ID NO: 565, a light chain comprising SEQ ID NO: 409 and a heavy chain comprising SEQ ID NO: 566, a light chain comprising SEQ ID NO: 410 and a heavy chain comprising SEQ ID NO: 567, a light chain comprising SEQ ID NO: 411 and a heavy chain comprising SEQ ID NO: 568, a light chain comprising SEQ ID NO: 412 and a heavy chain comprising SEQ ID NO: 569, a light chain comprising SEQ ID NO: 413 and a heavy chain comprising SEQ ID NO: 570, a light chain comprising SEQ ID NO: 414 and a heavy chain comprising SEQ ID NO: 571, a light chain comprising SEQ ID NO: 415 and a heavy chain comprising SEQ ID NO: 572,a light chain comprising SEQ ID NO: 416 and a heavy chain comprising SEQ ID NO: 573, a light chain comprising SEQ ID NO: 417 and a heavy chain comprising SEQ ID NO: 574, a light chain comprising SEQ ID NO: 418 and a heavy chain comprising SEQ ID NO: 575, a light chain comprising SEQ ID NO: 419 and a heavy chain comprising SEQ ID NO: 576, a light chain comprising SEQ ID NO: 420 and a heavy chain comprising SEQ ID NO: 577, a light chain comprising SEQ ID NO: 421 and a heavy chain comprising SEQ ID NO: 578, a light chain comprising SEQ ID NO: 422 and a heavy chain comprising SEQ ID NO: 579, a light chain comprising SEQ ID NO: 423 and a heavy chain comprising SEQ ID NO: 580, a light chain comprising SEQ ID NO: 424 and a heavy chain comprising SEQ ID NO: 581, a light chain comprising SEQ ID NO: 42 a light chain comprising SEQ ID NO: 5 and a heavy chain comprising SEQ ID NO: 582, a light chain comprising SEQ ID NO: 426 and a heavy chain comprising SEQ ID NO: 583, a light chain comprising SEQ ID NO: 427 and a heavy chain comprising SEQ ID NO: 584, a light chain comprising SEQ ID NO: 428 and a heavy chain comprising SEQ ID NO: 585, a light chain comprising SEQ ID NO: 429 and a heavy chain comprising SEQ ID NO: 586, a light chain comprising SEQ ID NO: 430 and a heavy chain comprising SEQ ID NO: 587, a light chain comprising SEQ ID NO: 431 and a heavy chain comprising SEQ ID NO: 588, a light chain comprising SEQ ID NO: 432 and a heavy chain comprising SEQ ID NO: 589, a light chain comprising SEQ ID NO: 433 and a heavy chain comprising SEQ ID NO: 590, a light chain comprising SEQ ID NO: 434, and and a heavy chain comprising SEQ ID NO:591, a light chain comprising SEQ ID NO:435 and a heavy chain comprising SEQ ID NO:592, a light chain comprising SEQ ID NO:436 and a heavy chain comprising SEQ ID NO:593, a light chain comprising SEQ ID NO:437 and a heavy chain comprising SEQ ID NO:594, a light chain comprising SEQ ID NO:438 and a heavy chain comprising SEQ ID NO:595, a light chain comprising SEQ ID NO:439 and a heavy chain comprising SEQ ID NO:596, a light chain comprising SEQ ID NO:440 and a heavy chain comprising SEQ ID NO:597, a light chain comprising SEQ ID NO:441 and a heavy chain comprising SEQ ID NO:598, a light chain comprising SEQ ID NO:442 and a heavy chain comprising SEQ ID NO:599, a light chain comprising SEQ ID NO:443 and a heavy chain comprising SEQ ID NO:60 a heavy chain comprising SEQ ID NO: 0, a light chain comprising SEQ ID NO: 444 and a heavy chain comprising SEQ ID NO: 601, a light chain comprising SEQ ID NO: 445 and a heavy chain comprising SEQ ID NO: 602, a light chain comprising SEQ ID NO: 446 and a heavy chain comprising SEQ ID NO: 603, a light chain comprising SEQ ID NO: 447 and a heavy chain comprising SEQ ID NO: 604, a light chain comprising SEQ ID NO: 448 and a heavy chain comprising SEQ ID NO: 605, a light chain comprising SEQ ID NO: 449 and a heavy chain comprising SEQ ID NO: 606, a light chain comprising SEQ ID NO: 450 and a heavy chain comprising SEQ ID NO: 607, a light chain comprising SEQ ID NO: 451 and a heavy chain comprising SEQ ID NO: 608, a light chain comprising SEQ ID NO: 452 and a heavy chain comprising SEQ ID NO: 609,a light chain comprising SEQ ID NO: 453 and a heavy chain comprising SEQ ID NO: 610, a light chain comprising SEQ ID NO: 454 and a heavy chain comprising SEQ ID NO: 611, a light chain comprising SEQ ID NO: 455 and a heavy chain comprising SEQ ID NO: 612, a light chain comprising SEQ ID NO: 456 and a heavy chain comprising SEQ ID NO: 613, a light chain comprising SEQ ID NO: 457 and a heavy chain comprising SEQ ID NO: 614, a light chain comprising SEQ ID NO: 458 and a heavy chain comprising SEQ ID NO: 615, a light chain comprising SEQ ID NO: 459 and a heavy chain comprising SEQ ID NO: 616, a light chain comprising SEQ ID NO: 460 and a heavy chain comprising SEQ ID NO: 617, a light chain comprising SEQ ID NO: 461 and a heavy chain comprising SEQ ID NO: 618, a light chain comprising SEQ ID NO: 462 and a heavy chain comprising SEQ ID NO: 619, a light chain comprising SEQ ID NO: 463 and a heavy chain comprising SEQ ID NO: 620, a light chain comprising SEQ ID NO: 464 and a heavy chain comprising SEQ ID NO: 621, a light chain comprising SEQ ID NO: 465 and SEQ ID NO: 622 a light chain comprising SEQ ID NO: 466 and a heavy chain comprising SEQ ID NO: 623, a light chain comprising SEQ ID NO: 467 and a heavy chain comprising SEQ ID NO: 624, a light chain comprising SEQ ID NO: 468 and a heavy chain comprising SEQ ID NO: 625, a light chain comprising SEQ ID NO: 469 and a heavy chain comprising SEQ ID NO: 626, a light chain comprising SEQ ID NO: 470 and a heavy chain comprising SEQ ID NO: 627, and a light chain comprising SEQ ID NO: 471 and a heavy chain comprising SEQ ID NO: 628, wherein the antibody or functional fragment thereof comprises an amino acid substitution of cysteine ​​or a non-standard amino acid at one or more conjugation sites selected from the group consisting of D70 of the antibody light chain relative to reference sequence SEQ ID NO: 455, E276 of the antibody heavy chain relative to reference sequence SEQ ID NO: 612, and T363 of the antibody heavy chain relative to reference sequence SEQ ID NO: 612.

[0560] In one embodiment, the antibody or fragment thereof comprises a light chain encoded by a polynucleotide sequence selected from the group consisting of SEQ ID NOs: 1885-2014 and a heavy chain comprising a sequence selected from the group consisting of SEQ ID NOs: 2042-2198, wherein the antibody or functional fragment thereof comprises an amino acid substitution of cysteine ​​or a non-standard amino acid at one or more conjugation sites selected from the group consisting of D70 of the antibody light chain relative to reference sequence SEQ ID NO: 455, E276 of the antibody heavy chain relative to reference sequence SEQ ID NO: 612, and T363 of the antibody heavy chain relative to reference sequence SEQ ID NO: 612. In one embodiment, the antibody or fragment thereof comprises a combination of a light chain and a heavy chain, and the antibody or functional fragment thereof comprises a light chain encoded by a polynucleotide sequence comprising SEQ ID NO: 1885 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO: 2042, a light chain encoded by a polynucleotide sequence comprising SEQ ID NO: 1886 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO: 2043, a light chain encoded by a polynucleotide sequence comprising SEQ ID NO: 1887 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO: 2044, a light chain encoded by a polynucleotide sequence comprising SEQ ID NO: 1888 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO: 2045, a light chain encoded by a polynucleotide sequence comprising SEQ ID NO: 1889 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO: 2046. a heavy chain encoded by a polynucleotide sequence comprising: a light chain encoded by a polynucleotide sequence comprising SEQ ID NO: 1890 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO: 2047; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO: 1891 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO: 2048; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO: 1892 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO: 2049; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO: 1893 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO: 2050; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO: 1894 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO: 2051;a light chain encoded by a polynucleotide sequence comprising SEQ ID NO: 1895 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO: 2052; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO: 1896 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO: 2053; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO: 1897 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO: 2054; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO: 1898 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO: 2055; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO: 1899 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO: 2056; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO: 1900 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO: 2057; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO: 1901 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO: 2058; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:1902 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:2059; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:1903 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:2060; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:1904 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:2061; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:1905 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:2062; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:1906 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:2063; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:1907 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:2064; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:1908 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:2065;a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:1909 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:2066; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:1910 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:2067; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:1911 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:2068; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:1912 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:2069; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:1913 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:2070; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:1914 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:2071; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:1915 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:2072; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:1916 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:2073; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:1917 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:2074; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:1918 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:2075; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:1919 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:2076; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:1920 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:2077; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:1921 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:2078; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:1922 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:2079;a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:1923 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:2080; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:1924 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:2081; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:1925 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:2082; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:1926 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:2083; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:1927 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:2084; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:1928 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:2085; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:1929 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:2086; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:1930 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:2087; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:1931 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:2088; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:1932 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:2089; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:1933 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:2090; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:1934 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:2091; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:1935 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:2092; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:1936 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:2093;a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:1937 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:2094; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:1938 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:2095; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:1939 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:2096; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:1940 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:2097; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:1941 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:2098; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:1942 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:2099; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:1943 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:2100; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:1944 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:2101; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:1945 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:2102; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:1946 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:2103; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:1947 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:2104; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:1948 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:2105; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:1949 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:2106; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:1950 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:2107;a light chain encoded by a polynucleotide sequence comprising SEQ ID NO: 1951 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO: 2108; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO: 1952 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO: 2109; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO: 1953 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO: 2110; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO: 1954 and a polynucleotide sequence comprising SEQ ID NO: 2111; a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO: 1955 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO: 2112, a light chain encoded by a polynucleotide sequence comprising SEQ ID NO: 1956 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO: 2113, a light chain encoded by a polynucleotide sequence comprising SEQ ID NO: 1957 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO: 2114, a light chain encoded by a polynucleotide sequence comprising SEQ ID NO: 1958 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO: 2115, a light chain encoded by a polynucleotide sequence comprising SEQ ID NO: 1959 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO: 2116, a light chain encoded by a polynucleotide sequence comprising SEQ ID NO: 1960 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO: 2117, a light chain encoded by a polynucleotide sequence comprising SEQ ID NO: 1961 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO: 2118 a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO: 1962 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO: 2119; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO: 1963 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO: 2120; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO: 1964 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO: 2121; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO: 1965 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO: 2122; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO: 1966 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO: 2123; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO: 1967 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO: 2124; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO: 1968 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO: 2125;a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:1969 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:2126; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:1970 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:2127; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:1971 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:2128; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:1972 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:2129; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:1973 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:2130; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:1974 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:2131; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:1975 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:2132; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:1976 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:2133; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:1977 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:2134; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:1978 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:2135; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:1979 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:2136; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:1980 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:2137; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:1981 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:2138; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:1982 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:2139;a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:1983 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:2140; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:1984 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:2141; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:1985 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:2142; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:1986 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:2143; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:1987 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:2144; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:1988 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:2145; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:1989 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:2146; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:1990 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:2147; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:1991 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:2148; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:1992 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:2149; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:1993 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:2150; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:1994 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:2151; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:1995 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:2152; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:1996 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:2153;a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:1997 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:2154; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:1998 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:2155; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:1999 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:2156; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:2000 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:2157; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:2001 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:2158; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:2002 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:2159; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:2003 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:2160; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:2004 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:2161; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:2005 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:2162; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:2006 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:2163; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:2007 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:2164; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:2008 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:2165; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:2009 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:2166; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:2010 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:2167;a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:2011 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:2168; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:2012 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:2169; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:2013 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:2170; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:2014 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:2171; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:2015 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:2172; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:2016 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:2173; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:2017 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:2174; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:2018 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:2175; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:2019 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:2176; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:2020 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:2177; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:2021 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:2178; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:2022 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:2179; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:2023 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:2180; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:2024 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:2181;a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:2025 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:2182; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:2026 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:2183; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:2027 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:2184; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:2028 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:2185; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:2029 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:2186; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:2030 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:2187; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:2031 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:2188; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:2032 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:2189; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:2033 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:2190; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:2034 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO:2191; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO:2035 and a polynucleotide sequence comprising SEQ ID NO:2192 a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO: 2193; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO: 2037 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO: 2194; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO: 2038 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO: 2195; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO: 2039 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO: 2196; a light chain encoded by a polynucleotide sequence comprising SEQ ID NO: 2040 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO: 2197; and a light chain encoded by a polynucleotide sequence comprising SEQ ID NO: 2041 and a heavy chain encoded by a polynucleotide sequence comprising SEQ ID NO: 2198.and comprising an amino acid substitution of cysteine ​​or a non-standard amino acid at one or more conjugation sites selected from the group consisting of E276 of the antibody heavy chain relative to the reference sequence SEQ ID NO: 612 and T363 of the antibody heavy chain relative to the reference sequence SEQ ID NO: 612.

[0561] 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 that are 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, except 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 resulting in no more than 1, 2 or less, 3 or less, 4 or less, 5 or less, 6 or less, 7 or less, 8 or less, 9 or less, 10 or less, 11 or less, 12 or less, 13 or less, 14 or less or 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 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 differs from the sequences specified 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.

[0562] In another embodiment, the antigen binding protein consists solely of a light chain polypeptide or a heavy chain polypeptide as shown in Table 5.

[0563] 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.

[0564] 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.

[0565] 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: 3141). Such antigen binding proteins may bind to the same epitope as one of the antigen binding proteins described herein, or may bind to 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, antigen binding proteins provided include: An antibody that competes with any of the antibodies listed in Table 4A and Table 4B and has all six CDRs listed therein; competes with an antibody having the VH and VL listed in Table 3 against any of the antibodies listed in Table 3; or Competes with an antibody having two light chains and two heavy chains specified against any of the antibodies in Table 5 Includes:

[0566] The antigen-binding proteins provided include monoclonal antibodies that bind to GIPR. Monoclonal antibodies can be produced using any method known in the art, for example, by immortalizing spleen cells collected from transgenic animals after the completion of the immunization schedule. Spleen cells can be immortalized using any method known in the art, for example, by fusing spleen cells with myeloma cells to produce hybridomas. 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 specific selection media that support the growth of only 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, while 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.

[0567] In some cases, hybridoma cell lines are produced by immunizing an animal (e.g., a transgenic animal having human immunoglobulin sequences) with a GIPR immunogen, collecting spleen cells from the immunized animal, generating hybridoma cells by fusing the collected spleen cells with a myeloma cell line, establishing hybridoma cell lines from the hybridoma cells, and identifying hybridoma cell lines that produce antibodies that bind to a GIPR polypeptide. Such hybridoma cell lines and the anti-GIPR monoclonal antibodies they produce are embodiments of the present application.

[0568] Monoclonal antibodies secreted by hybridoma cell lines 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.

[0569] Chimeric and humanized antibodies based on the aforementioned sequences are also provided. Monoclonal antibodies intended for use as therapeutic agents may be modified in various ways before use. One example is a chimeric antibody, which is an antibody composed of protein segments from different antibodies 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 to or homologous to corresponding sequences in antibodies from a particular species or belonging to a particular antibody class or subclass, while the remainder of the chain is identical to or homologous to corresponding sequences in antibodies from another species or belonging to another antibody class or subclass. For methods related to chimeric antibodies, see, e.g., U.S. Pat. No. 4,816,567 and Morrison et al., 1985, Proc. Natl. Acad. Sci. USA 81:6851-6855, both of which are incorporated herein by reference. CDR grafting is described, for example, in US Pat. Nos. 6,180,370, 5,693,762, 5,693,761, 5,585,089 and 5,530,101.

[0570] Generally, 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 is identical to 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 derived from, 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.

[0571] One useful form of chimeric antibody is a "humanized" antibody. Generally, humanized antibodies are generated from monoclonal antibodies initially produced in non-human animals. Specific 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 typically originate from non-antigen-recognizing portions of the antibody. Humanization can be performed using various methods, for example, by substituting at least a portion of a rodent variable region for the corresponding region of a human antibody (see, e.g., U.S. Pat. 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).

[0572] 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 antibodies of the same or different lineages. For example, the heavy and light chain variable regions V H 1. V H 2. V H 3. V H 4. V H 5. V H6. 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 Two CDRs can be grafted onto consensus human FRs. To create consensus human FRs, consensus amino acid sequences 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 a GIPR antibody are not exchanged, and the remaining FR amino acids are exchanged. A "rare amino acid" is a specific amino acid that occurs at a position not normally found in an FR. Alternatively, a variable region grafted from one heavy or light chain can be used with a constant region that is different from the constant region of that particular heavy or light chain disclosed herein. In other embodiments, the grafted variable region is part of a single-chain Fv antibody.

[0573] In certain embodiments, constant regions from a species other than human can be used in conjunction with human variable regions to generate hybrid antibodies.

[0574] 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 achieving the generation of fully human antibodies is the "humanization" of the mouse humoral immune system. The introduction of human immunoglobulin (Ig) loci into mice in which the endogenous Ig genes have been inactivated 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 the immunogenic and allergic responses that can occur when mouse mAbs or mouse-derived mAbs are administered to humans as therapeutic agents.

[0575] Fully human antibodies can be produced by immunizing transgenic animals (usually mice) that lack endogenous immunoglobulin production and have the capacity to produce a repertoire of human antibodies. Antigens of interest typically contain six or more consecutive amino acids and are optionally conjugated to a carrier such as a hapten. See, e.g., Jakobovits et al., 1993, Proc. Natl. Acad. Sci. USA 90:2551-2555; Jakobovits et al., 1993, Nature 362:255-258; and Bruggermann et al., 1993, Year in Immunol. 7:33. In one example of such a method, a transgenic animal is generated by disabling the endogenous mouse immunoglobulin loci encoding 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 crossbred to produce animals with all of the desired immune system modifications. When administered an immunogen, these transgenic animals produce antibodies immunospecific for the immunogen, but these antibodies 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 546073B1 and EP 546073A1.

[0576] The transgenic mice, referred to herein as "HuMab" mice, contain human immunoglobulin gene minilocuses encoding unrearranged human heavy chain (mu and gamma) and kappa light chain immunoglobulin sequences, along 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 entirety 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 Nos. 5,545,807, WO 93 / 1227, WO 92 / 22646, and WO 92 / 03918, the disclosures of all of which are incorporated herein by reference in their entirety and for all purposes. Techniques utilizing the production of human antibodies in such transgenic mice are also disclosed in WO 98 / 24893 and Mendez et al., 1997, Nature Genetics 15:146-156, which are incorporated herein by reference. For example, the HCo7 and HCo12 transgenic mouse strains can be used to generate human monoclonal antibodies against GIPR. Further details regarding the production of human antibodies using transgenic mice are provided below.

[0577] 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 collected from cultured hybridoma cells.

[0578] Fully human antibodies can also be obtained from phage display libraries (as 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 WO 99 / 10494, incorporated herein by reference.

[0579] 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.

[0580] In one embodiment, for example, the antigen binding proteins are mutant forms of the antigen binding proteins 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.

[0581] 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 influence chain orientation: Gly, Pro, and 6) Aromatics: Trp, Tyr, Phe.

[0582] Conservative amino acid substitutions may involve the exchange of one member of such a class for 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.

[0583] Non-conservative substitutions may involve exchanging 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 non-homologous regions of the molecule.

[0584] According to certain embodiments, when making such changes, the hydropathic index of amino acids may be taken into consideration. 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).

[0585] The importance of hydropathic profiles in conferring interactive biological function to proteins is understood in the art (see, e.g., Kyte et al., 1982, J. Mol. Biol. 157:105-131). It is known that certain amino acids can be substituted for other amino acids having 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 is included, and in other aspects, within ±0.5 is included.

[0586] 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 use of the resulting biologically functional protein or peptide is, as in the present case, in an immunological embodiment. In certain embodiments, the greatest local average hydrophilicity of a protein, as governed by the hydrophilicity of its adjacent amino acids, correlates with its immunogenicity and antigen binding or immunogenicity, i.e., a biological property of the protein.

[0587] The following amino acid residues have been assigned the following 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 changes are made based on similar hydrophilicity values, substitutions of amino acids whose hydrophilicity values ​​are within ±2 of each other 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."

[0588] Table 6 shows exemplary conservative amino acid substitutions.

[0589] [Table 406]

[0590] Those skilled in the art will be able to determine suitable variants of the polypeptides provided herein using well-known techniques. By targeting regions not believed to be important for activity, those skilled in the art will be able to identify suitable regions of the molecule that can be altered without impairing activity. Those skilled in the art will also be able to identify residues and portions of the molecule that are conserved among similar polypeptides. In additional embodiments, regions that may not be important for biological activity or structure may be subject to conservative amino acid substitutions without impairing biological activity or adversely affecting polypeptide structure.

[0591] Furthermore, one skilled in the art can evaluate structure-function studies that identify residues important for activity or structure in similar polypeptides. By considering such comparisons, one can predict 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 such predicted important amino acid residues.

[0592] Those skilled in the art can also analyze the three-dimensional structure of similar polypeptides and the amino acid sequence associated with that structure. Taking such information into account, those skilled in the art can predict the alignment of the amino acid residues of an antibody with respect to its three-dimensional structure. Because 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 fundamental 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 at which additional substitutions, either alone or in combination with other mutations, should be avoided.

[0593] 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 assist 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. 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).

[0594] 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).

[0595] 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 for forming protein complexes, (4) altered binding affinity for a ligand or antigen, and / or (5) conferring or modifying other physicochemical or functional properties on such polypeptides. For example, single or multiple amino acid substitutions (in certain embodiments, conservative amino acid substitutions) may be made in naturally occurring sequences. Substitutions may be made in portions of antibodies located outside of domains that form intermolecular contacts. 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, W.H. 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.

[0596] Preferred antibody variants additionally include cysteine ​​variants, in which one or more cysteine ​​residues in the parent or native amino acid sequence have been deleted or substituted with another amino acid (e.g., serine). Cysteine ​​variants are particularly useful when the antibody must be refolded into a biologically active conformation. Cysteine ​​variants may have fewer cysteine ​​residues compared to the native antibody, and typically have a number that minimizes interactions resulting from unpaired cysteines.

[0597] The disclosed heavy and light chains, variable region domains, and CDRs can be used to prepare polypeptides comprising an antigen-binding region capable of specifically binding to GIPR. For example, one or more CDRs can be incorporated into a molecule (e.g., a polypeptide) either covalently or noncovalently to prepare an immunoadhesin. The CDRs can be incorporated into the immunoadhesin as part of a larger polypeptide chain, can be covalently linked to another polypeptide chain in the immunoadhesin, or can be incorporated noncovalently into the immunoadhesin. The CDRs enable the immunoadhesin to specifically bind to a particular antigen of interest (e.g., a GIPR polypeptide or an epitope thereof).

[0598] 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, peptidomimetics are proteins structurally similar to antibodies that exhibit a desired biological activity (such as the ability to specifically bind to GIPR herein), but which have one or more peptide bonds optionally replaced by a bond selected from -CHNH-, -CHS-, -CH-CH-, -CH-CH- (cis and trans), -COCH-, -CH(OH)CH-, and CHSO-, 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 proteins with improved stability. Additionally, controlled peptides containing the consensus sequence or substantially identical consensus sequence variants 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.

[0599] 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 on the antibody or fragment, such as increased half-life, for 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 pharmaceutically 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 pegylated single-chain polypeptides described herein. 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.

[0600] Other derivatives include covalent or aggregate conjugation of the GIPR antigen-binding protein with other proteins or polypeptides, such as by expression of a recombinant fusion protein containing a heterologous polypeptide fused to the N- 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 also 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, 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 to which a specific monoclonal antibody (mAb) can reversibly bind, 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).

[0601] 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. Various methods for labeling proteins are known in the art and may be used as appropriate.

[0602] 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.

[0603] Generally, labels are divided into various classes depending on the assay in which they are to be detected: a) isotopic labels (which may 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 for labeling proteins are known in the art.

[0604] Particular labels include optical dyes, which include, but are not limited to, chromophores, phosphors, and fluorophores, the latter of which are often specific. Fluorophores can be either "small molecule" fluorophores or proteinaceous fluorophores.

[0605] By "fluorescent label" is intended any molecule that can 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 Fluor 488, Alexa Fluor 546, Alexa Fluor 568, Alexa Fluor 594, Alexa Fluor 633, Alexa Fluor 660, Alexa Fluor 680), 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.

[0606] Suitable proteinaceous fluorescent labels include, but are not limited to, green fluorescent protein (EGFP), including Renilla, Ptilosarcus, or Aequorea GFP (Chalfie et al., 1994, Science 263:802-805), EGFP (Clontech Labs., Inc., Genbank accession number U55762), blue fluorescent protein (BFP, Quantum Biotechnologies, Inc., Quebec, Canada; Stauber, 1998, Biotechniques 24:462-471; 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., 1994, J. Immunol. 150:5408-5417), and β-galactosidase (Nolan et al., 1994, 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. Pat. 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).

[0607] 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. Nucleic acids can 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 can include one or more additional sequences, e.g., regulatory sequences, and / or can be part of a longer nucleic acid, e.g., a vector. Nucleic acids can be single- or double-stranded and can comprise RNA and / or DNA nucleotides, and artificial variants thereof (e.g., peptide nucleic acids). Any of the variable regions provided herein can 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.

[0608] Nucleic acids encoding a particular antigen-binding protein or portion thereof (e.g., a 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 approach, 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.

[0609] In one embodiment, a nucleic acid that hybridizes to another nucleic acid under specific hybridization conditions is further provided. Nucleic acid hybridization methods are well known in the art. See, for example, 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 containing 5x sodium chloride / sodium citrate (SSC), 0.5% SDS, 1.0 mM EDTA (pH 8.0), a hybridization buffer containing about 50% formamide, 6x SSC, and a hybridization temperature of about 55°C (or other similar hybridization solutions, such as those containing about 50% formamide, are used at a hybridization temperature of 42°C), and washing conditions of 0.5x SSC, 0.1% SDS at 60°C. Stringent hybridization conditions include hybridization in 6×SSC at 45° C., followed by one or more washes in 0.1×SSC, 0.2% SDS at 68° C. Moreover, one of skill in the art can manipulate hybridization and / or wash conditions to increase or decrease the stringency of hybridization, such that nucleic acids containing nucleotide sequences 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.

[0610] Basic parameters influencing the selection of hybridization conditions and guidance for devising appropriate conditions are provided, for example, by Sambrook, Fritsch, and Maniatis (2001, supra, 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.

[0611] Changes can be introduced by mutagenesis into a nucleic acid, which results in a change 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, for example, a site-directed mutagenesis protocol is used to alter one or more specific amino acid residues. In another embodiment, for example, a random mutagenesis protocol is used to alter one or more randomly selected residues. Regardless of how it is performed, the mutant polypeptides can be expressed and screened for desired properties.

[0612] 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 made. 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, a mutation can alter the biological activity quantitatively or qualitatively. 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 altered in amino acid sequence by mutagenesis using molecular biology techniques well established in the art.

[0613] In another aspect, nucleic acid molecules suitable for use as primers or hybridization probes for the detection of nucleic acid sequences are provided. The nucleic acid molecules 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.

[0614] 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 encoding a single polypeptide. The probe can contain a labeling group, such as a radioisotope, a fluorescent compound, an enzyme, or an enzyme cofactor. Such probes can be used to identify cells expressing the polypeptide.

[0615] 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 in a host cell. The recombinant expression vector comprises one or more regulatory sequences selected based on the host cell to be used for expression, and these 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., the SV40 early gene enhancer, the Rous sarcoma virus promoter, and the 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 specific treatments or conditions (e.g., the metallothionin promoter in mammalian cells, and tetracycline (tet)- and / or streptomycin-responsive promoters in both prokaryotic and eukaryotic systems (see ibid.). It will be appreciated by those skilled in the art that the design of the expression vector can depend on such factors as the choice of the host cell to be transformed, the expression level 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.

[0616] 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. Stable transfection of mammalian cells depends on the expression vector and gene transfer technique used, and it is known that only a small fraction of cells may integrate 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 that have incorporated the selectable marker gene will survive, while other cells die).

[0617] Also provided herein are expression systems and constructs in the form of plasmids, expression vectors, transcription cassettes, or expression cassettes that comprise at least one of the above-described polynucleotides, as well as host cells containing such expression systems or constructs.

[0618] 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).

[0619] 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 polynucleotides 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, such as those exemplified by U.S. Pat. Nos. 4,399,216; 4,912,040; 4,740,461; and 4,959,455. The optimal transformation procedure used will depend on the type of host cell 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 polynucleotides in liposomes, mixing nucleic acids with positively charged lipids, and direct microinjection of DNA into the nucleus.

[0620] Recombinant expression constructs typically include a nucleic acid molecule encoding a polypeptide comprising one or more of the following: one or more of the CDRs provided herein, a light chain constant region, a light...

Claims

1. A composition for reducing weight in a human subject, comprising an intact human or humanized IgG antibody that specifically binds to the N-terminal extracellular domain of the human gastric inhibitory peptide receptor (GIPR) and a glucagon-like polypeptide-1 (GLP-1) receptor agonist, wherein the GLP-1 receptor agonist is conjugated to the antibody via a cysteine ​​at one or more conjugation sites of the antibody.

2. The composition described in claim 1, for parenteral administration.

3. The composition described in claim 1, for subcutaneous administration.

4. A pharmaceutical composition for the treatment of type 2 diabetes in a human subject comprising an intact human or humanized IgG antibody that specifically binds to the N-terminal extracellular domain of the human gastric inhibitory peptide receptor (GIPR) and a glucagon-like polypeptide-1 (GLP-1) receptor agonist, wherein the GLP-1 receptor agonist is conjugated to the antibody via a cysteine ​​at one or more conjugation sites of the antibody.

5. The pharmaceutical composition of claim 4, for parenteral administration.

6. The pharmaceutical composition of claim 4, for subcutaneous administration.

7. A composition for reducing weight gain in a human subject, comprising an intact human or humanized IgG antibody that specifically binds to the N-terminal extracellular domain of the human gastric inhibitory peptide receptor (GIPR) and a glucagon-like polypeptide-1 (GLP-1) receptor agonist, wherein the GLP-1 receptor agonist is conjugated to the antibody via a cysteine ​​at one or more conjugation sites of the antibody.

8. The composition described in claim 7, for parenteral administration.

9. The composition described in claim 7, for subcutaneous administration.