BR112016017248B1 - ANTIBODY, HOST CELL, PHARMACEUTICAL COMPOSITION AND USE OF THE ANTIBODY - Google Patents

Abstract

ANTIBODY OR FRAGMENT THEREOF, LINKING AGENT, TRANSGENIC ANIMAL, HYBRIDOMA, VECTOR, PHARMACEUTICAL COMPOSITION, METHOD OF INDUCING SIGNALING SIMILAR TO FGF19 AND/OR FGF21, METHOD FOR ACTIVATING A KLOTHO BETA/FGF RECEPTOR COMPLEX, METHOD FOR IMPROVING METABOLISM GLUCOSE IN AN INDIVIDUAL, METHOD FOR DETECTING THE PRESENCE OF KLOTHO BETA, USE OF ANTIBODY OR FRAGMENT OF IT, USE OF PHARMACEUTICAL COMPOSITION, TREATMENT METHOD AND METHOD FOR IMPROVING METABOLIC PARAMETERS. The present invention provides binding proteins, such as antibodies, that bind to Klotho beta, including human Klotho beta, and methods for their use.

Description

CROSS REFERENCE FOR RELATED ORDERS

[0001] This application claims the benefit of priority of United States Provisional Application Serial No. 61/931,531, filed January 24, 2014, the entire contents of which are incorporated herein by reference.

FIELD

[0002] The present invention generally relates to binding proteins, such as antibodies, that bind to Klotho beta, including human Klotho beta, and methods for their use.

BACKGROUND

[0003] Klotho beta, which belongs to the Klotho family, is a type I single-pass membrane protein. Klotho beta has an extracellular domain consisting of two internal repeats that share homology with members of family 1 glycosidases but lack glycosidase catalytic activity. Klotho beta expression is mainly detected in the liver, pancreas and adipose tissue. Ito and colleagues reported that Klotho beta-deficient (KLB -/-) mice have elevated CYP7A1 and CY8B1 mRNA levels and exhibit increased bile acid synthesis and excretion (Ito et al, 2005, J Clin Invest 115: 2202-2208 ). Klotho beta forms a complex with fibroblast growth factor (FGF) receptors and functions as a coreceptor for FGFs, including FGF19 and FGF21.

[0004] Twenty-two members of the human FGF family have been identified and four tyrosine kinase receptors that bind FGF (FGFR1-FGFR4) have been identified. The interaction between FGF and its receptor results in the dimerization of FGFR, which allows the cytoplasmic domains of the receptor to transphosphorylate and become activated, which in turn leads to the phosphorylation and activation of downstream signaling molecules.

[0005] The high-affinity receptor for FGF19 is FGFR4 and the binding of FGF19 to FGFR4 is facilitated by Klotho beta. It has been reported that FGF19 transgenic mice have decreased adiposity, increased metabolic rate, reduced hepatic triglycerides, increased fatty acid oxidation, reduced glucose levels and increased insulin sensitivity (Tomlinson et al, 2002, Endocrinology 143: 1741-1747). Furthermore, these transgenic mice have not been reported to become obese or diabetic on a high-fat diet (Tomlinson et al, 2002, Endocrinology 143:1741-1747). It has also been reported that FGF19 treatment prevented or reversed diabetes in mice rendered obese by genetic ablation of brown adipose tissue or the genetic absence of leptin (Fu et al, 2004, Endocrinology 145:2594-2603).

[0006] FGF21 acts through the interaction of FGFRs and Klotho beta. FGFR1 is an abundant receptor in white adipose tissue and is most likely to be the major functional receptor for FGF21 in white adipose tissue. FGF21 expression is detected in the liver, thymus, adipose tissue, and islet beta cells of the pancreas. It has been reported that the interaction of FGF21 with the Klotho beta-FGFR complex stimulates glucose uptake, decreases glucagon secretion, improves insulin sensitivity and glucose disposal, promotes white adipose tissue in response to fasting, increases ketogenesis in the liver in response to fasting, it reduces plasma triglyceride levels and increases energy expenditure (Iglesias et al, 2012, European Journal of Endocrinology 167:301-309).

[0007] Since FGF19 and FGF21 require both FGFRs and Klotho beta for cell signaling, agents that mimic FGF19 and/or FGF21 may be desirable for their effects on either lipid metabolism or glucose metabolism. However, it is unclear what characteristics are necessary for an agent to confer cell signaling activity similar to that of FGF19 or FGF21.

SUMMARY

[0008] The present invention provides proteins that bind to Klotho beta, including binding proteins, such as antibodies, that bind to Klotho beta. Such binding proteins including antibodies can bind to a Klotho beta polypeptide, a Klotho beta fragment and/or a Klotho beta epitope. Such binding proteins, including antibodies, may be agonists (e.g., inducing FGF19 or FGF21-like signaling from an FGF receptor or activating a Klotho beta/FGF receptor complex).

[0009] The present invention also provides binding proteins, including antibodies or fragments thereof, that (i) bind to human Klotho beta, (ii) induce FGF19-like signaling and/or FGF21-like signaling, and ( iii) do not compete with FGF19 and/or FGF21 for interaction with Klotho beta.

[0010] In some embodiments, anti-Klotho beta antibodies are humanized antibodies that bind to a Klotho beta polypeptide, a Klotho beta fragment, or a Klotho beta epitope. In certain embodiments, an anti-Klotho beta antibody comprises a VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2 and/or VL CDR3 of a monoclonal antibody designated 5H23, 1C17, 1D19, 2L12, 3L3, 3N20, 4P5, 5C23, 5F7 or 1G19, as described herein, or a humanized variant thereof. In certain embodiments, an Anti-Klotho beta antibody may further comprise a VH FR1, VH FR2, VH FR3, VH FR4, VL FR1, VL FR2, VL FR3 and/or VL FR4 of a sequence of human immunoglobulin amino acids or a variant thereof.

[0011] In some embodiments, a binding protein (e.g., an anti-Klotho beta antibody) comprises six CDRs or fewer than six CDRs. In some embodiments, a binding protein (e.g., an Anti-Klotho beta antibody) comprises one, two, three, four, five or six CDRs selected from VH CDR1, VH CDR2, VH CDR3, VL CDR1, CDR2 of VL, and/or CDR3 of VL. In some embodiments, a binding protein (e.g., an Anti-Klotho beta antibody) comprises one, two, three, four, five, or six CDRs selected from VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2, and/or VL CDR3 of a monoclonal antibody designated as 5H23, 1C17, 1D19, 2L12, 3L3, 3N20, 4P5, 5C23, 5F7 or 1G19, as described herein, or a humanized variant thereof. In some embodiments, a binding protein (e.g., an Anti-Klotho beta antibody) further comprises a scaffold region or framework region, including a VH FR1, VH FR2, VH FR3, VH FR4, VL FR1 , FR2 of VL, FR3 of VL, and/or FR4 of VL of a human immunoglobulin amino acid sequence or a variant thereof.

[0012] In some embodiments, the antibody is a humanized antibody, a monoclonal antibody, a recombinant antibody, an antigen-binding fragment, or any combination thereof. In some embodiments, the antibody is a humanized monoclonal antibody, or antigen-binding fragment thereof, that binds to a Klotho beta polypeptide (e.g., a cell surface-expressed or soluble Klotho beta), a fragment of Klotho beta, or an epitope of Klotho beta.

[0013] The present invention also provides proteins, such as anti-Klotho beta antibodies, (i) that competitively block (e.g., in a dose-dependent manner) an Anti-Klotho beta antibody provided herein from binding to a Klotho beta polypeptide (e.g., a cell-surface expressed or soluble Klotho beta), a Klotho beta fragment, or a Klotho beta epitope and/or (ii) that bind to a Klotho beta epitope that is bound by an anti-Klotho beta antibody provided herein. In other embodiments, binding proteins, such as Anti-Klotho beta antibodies, competitively block (e.g., in a dose-dependent manner) the 5H23 or 1G19 monoclonal antibody described herein or a humanized variant thereof from binding to a Klotho beta polypeptide (e.g., a cell surface-expressed or soluble Klotho beta), a Klotho beta fragment, or a Klotho beta epitope. In other embodiments, binding proteins, such as anti-Klotho beta antibodies, bind to a Klotho beta epitope that is bound (e.g., recognized) by the monoclonal antibody 5H23, or 1G19 described herein or a humanized variant thereof.

[0014] The present invention also provides binding proteins, including antibodies or fragments thereof, which (i) bind to an epitope of human Klotho beta and cynomolgus monkey Klotho beta recognized by an antibody comprising a heavy chain variable region having the amino acid sequence of SEQ ID NO: 25 and a light chain variable region having the amino acid sequence of SEQ ID NO: 26; or (ii) compete for binding to human Klotho beta with an antibody comprising a heavy chain variable region having the amino acid sequence of SEQ ID NO: 25 and a light chain variable region having the amino acid sequence of SEQ ID NO: 26. In some embodiments, binding proteins, including antibodies or fragments thereof, are provided herein, which bind to a region, including an epitope, of human Klotho beta or cyno Klotho beta. In some embodiments, binding proteins, including antibodies or fragments thereof, bind to a region of human Klotho beta or cyno Klotho beta, including, for example, those that bind to: (i) a KLB2 domain of Klotho human beta comprising amino acid residues 509 to 1044 of SEQ ID NO: 297; (ii) a glycosyl hydrolase 1 region of a human Klotho beta KLB2 domain comprising amino acid residues 517-967 of SEQ ID NO: 297; (iii) a region of human Klotho beta comprising amino acid residues 657-703 of SEQ ID NO: 297; or (iv) a cyno beta Klotho region comprising amino acid residues 657-703 of SEQ ID NO: 299.

[0015] In some embodiments, binding proteins, including antibodies or fragments thereof, are provided herein that bind to a specific epitope of human Klotho beta, including, for example, those that bind to: (i) an epitope of Human beta Klotho comprising at least one of amino acid residues 657, 701 and/or 703 of human beta Klotho (SEQ ID NO: 297); (ii) a human Klotho beta epitope comprising at least amino acid residue 657 of SEQ ID NO: 297; (iii) a human Klotho beta epitope comprising at least amino acid residue 701 of SEQ ID NO: 297; (iv) a human Klotho beta epitope comprising at least amino acid residue 703 of SEQ ID NO: 297; (v) a human Klotho beta epitope comprising at least amino acid residues 657 and 701 of SEQ ID NO: 297; (vi) a human Klotho beta epitope comprising at least amino acid residues 657 and 703 of SEQ ID NO: 297; (vii) a human Klotho beta epitope comprising at least amino acid residues 701 and 703 of SEQ ID NO: 297; or (viii) a human Klotho beta epitope comprising at least amino acid residues 657, 701 and 703 of SEQ ID NO: 297. Such antibodies provided above may, in some embodiments, induce FGF19-like signaling and/or signaling similar to that of FGF21 or activate a Klotho beta/FGF receptor complex in a cell expressing human Klotho beta and an FGF receptor. Furthermore, in some embodiments, the antibody is a monoclonal antibody, e.g., a humanized, human, or chimeric antibody.

[0016] In some embodiments, binding proteins such as anti-Klotho beta antibodies provided herein are conjugated or recombinantly linked to a diagnostic agent, a detectable agent or therapeutic agent. In some aspects, the therapeutic agent is a drug, including one or more drugs, such as biguanides and sulfonylureas (e.g., metformin, tolbutamide, chlorpropamide, acetohexamide, tolazamide, glibenclamide, glyburide and glipizide), thiazolidinediones (e.g. , rosiglitazone, pioglitazone), GLP-1 analogues, PPAR gamma agonists (e.g. pioglitazone and rosiglitazone), dipeptidyl peptidase-4 (DPP-4) inhibitors (e.g. JANUVIN®, ONGLYZA®), formulations of bromocriptine and bile acid sequestrants (e.g., colesevelam), and insulin (e.g., bolus and basal analogues), alpha-glucosidase inhibitors (e.g., acarbose, roglibose), metformin (e.g., metformin hydrochloride) with or without a thiazolidinedione (TZD), SGLT-2 inhibitors, appetite suppression or weight loss drugs (e.g., Meridia®/sibutramine, Xenical®/ortistat). In some aspects, the detectable agent is a radioisotope, an enzyme, a fluorescent compound, a bioluminescent compound, or a chemiluminescent compound.

[0017] In certain embodiments, compositions comprising a binding protein such as an anti-Klotho beta antibody described herein are provided. Also provided herein are pharmaceutical compositions comprising a binding protein such as a Klotho beta antibody as described herein.

[0018] The present invention also provides isolated nucleic acid molecules that encode an immunoglobulin heavy chain, an immunoglobulin light chain, VH region, VL region, VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2 and/or VL CDR3 of binding proteins (e.g., anti-Klotho beta antibodies) that bind to a Klotho beta polypeptide, a Klotho beta polypeptide fragment, or a Klotho beta epitope. In some embodiments, the nucleic acid molecule encodes a VH region, VL region, VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2, and/or VL CDR3 of a monoclonal antibody designated as 5H23. , 1C17, 1D19, 2L12, 3L3, 3N20, 4P5, 5C23, 5F7 or 1G19, as described herein, or a humanized variant thereof. In some embodiments, the nucleic acid molecule further encodes a scaffold region or framework region, including FR1 of VH, FR2 of VH, FR3 of VH, FR4 of VH, FR1 of VL, FR2 of VL, FR3 of VL and/or FR4 of VL of a human immunoglobulin amino acid sequence or a variant thereof. Furthermore, provided herein are vectors and host cells comprising nucleic acid molecules encoding a binding protein such as an Anti-Klotho beta antibody, as well as methods of producing a binding protein such as an Anti-Klotho antibody. beta by culturing the host cells provided herein under conditions that promote the production of a binding protein such as an anti-Klotho beta antibody.

[0019] The present invention also provides methods of treating, preventing or alleviating a disease, disorder or condition (e.g., one or more symptoms) that include administering a therapeutically effective amount of a binding protein, such as an antibody Anti-Klotho beta is provided herein to an individual, including an individual in need thereof, thereby treating, preventing or alleviating the disease, disorder or condition. In some embodiments, the disease, disorder or condition is caused by, or otherwise associated with, Klotho beta, such as those related to FGF19-like and/or FGF21-like signaling in an individual. In certain embodiments, the disease is treatable by reducing blood glucose, insulin, or serum lipid levels (e.g., type 2 diabetes, obesity, dyslipidemia, NASH, cardiovascular disease, metabolic syndrome).

[0020] In some embodiments, the disease, disorder or condition is related to glucose metabolism or lipid metabolism. In some embodiments, the disease, disorder or condition is selected from the group of a hyperglycemic condition. (For example, diabetes, such as type I diabetes, type 2 diabetes, gestational diabetes, insulin resistance, hyperinsulinemia, glucose intolerance, metabolic syndrome, or obesity).

[0021] In some embodiments, the treatment, prevention or improvement methods include methods for improving glucose metabolism and/or methods for improving lipid metabolism. In some embodiments, the treatment, prevention, or enhancement methods result in reduced glucose levels (e.g., reduced blood glucose), increased insulin sensitivity, decreased insulin resistance, decreased glycogen, improved glucose tolerance. , improvement of glucose metabolism, improvement of homeostasis, improvement of pancreatic function, reduction of triglycerides, reduction of cholesterol, reduction of IDL, reduction of LDL, reduction of VLDL, reduction of blood pressure, reduction of internal thickening of a blood vessel and/ or decreased body mass or weight gain.

[0022] The present disclosure provides methods of treating a disease, disorder or condition associated with human FGF19 and/or human FGF21, which includes any disease, disorder or condition the onset of which in an individual (e.g., a patient) is caused by, at least in part, the induction of FGF19-like and/or FGF21-like signaling, which is initiated in vivo by the formation of a complex comprising FGFR1c, FGFR2c, FGFR3c or FGFR4 and Klotho beta and FGF19 or FGF21. The severity of the disease or condition can also be reduced by inducing FGF19-like and/or FGF21-like signaling. Examples of diseases and conditions that can be treated with binding proteins, such as anti-Klotho beta antibodies, include type 2 diabetes, obesity, dyslipidemia, NASH, cardiovascular disease, and metabolic syndrome.

[0023] As such, binding proteins such as Anti-Klotho beta antibodies described herein can be used to treat type 2 diabetes, obesity, dyslipidemia (e.g., hypertriglyceridemia), NASH, cardiovascular disease, and/or metabolic syndrome , as well as any disease, disorder or condition in which it is desirable to mimic or enhance the in vivo effects of FGF19 and/or FGF21, or may be used as a prophylactic treatment administered, for example, daily, weekly, biweekly, monthly, bimonthly, twice a year, etc., to prevent or reduce the frequency and/or severity of symptoms (e.g., elevated plasma glucose levels, elevated triglycerides and cholesterol levels), including, for example, to thereby provide a profile improved cardiovascular and/or glycemic risk factor. The present disclosure provides methods for improving metabolic parameters by administering to a subject a binding protein, including an antibody or a fragment thereof, as described herein or a pharmaceutical composition described herein, including, for example, wherein the improvement includes a decrease in body weight, body mass index, waist circumference, skinfold thickness, glucose, insulin and/or triglycerides.

[0024] The present invention also provides methods for inducing FGF19-like or FGF21-like signaling of cells that have cell surface expression of Klotho beta and one or more FGF receptors, such as FGFR1, FGFR2, FGFR3, or FGFR4 comprising contacting the cells with an effective amount of a binding protein (e.g., an antibody) that binds to Klotho beta as described herein. In some embodiments, the cell is an adipocyte or hepatocyte. In other embodiments, the cell is a cell transfected with a gene encoding Klotho beta and, optionally, a gene encoding an FGF receptor. Additional methods provided include the use of an anti-Klotho beta antibody as described herein, with activity to mediate FGF19-like and/or FGF21-like signaling effects.

[0025] The present invention also provides methods of modulating FGF19-like or FGF21-like signaling in an individual comprising administering an effective amount of a binding protein, such as an Anti-Klotho beta antibody as herein. described to an individual, including an individual in need thereof. In some embodiments, the modulation comprises FGF19-like activation. In some embodiments, the modulation comprises FGF21-like activation. In some embodiments, the modulation comprises increasing glucose metabolism (e.g., reducing glucose levels such as blood glucose levels).

[0026] The present description also provides methods for detecting Klotho beta in a sample comprising contacting the sample with a binding protein, such as an anti-Klotho beta antibody as described herein, which comprises a detectable agent. In certain embodiments, the sample comprises a cell that expresses Klotho beta on its surface.

[0027] The present invention also provides kits comprising a binding protein, such as an anti-Klotho beta antibody that binds to a Klotho beta polypeptide, a Klotho beta fragment or a Klotho beta epitope as described herein.

BRIEF DESCRIPTION OF FIGURES

[0028] Figure 1A-1B shows a sequence alignment of the heavy chain variable regions and light chain variable regions of the Anti-Klotho beta antibodies designated 5H23, 1C17, 1D19, 2L12, 3L3, 3N20, 4P5, 5C23, 5F7 and 1G19. CDR limits are indicated by Kabat, AbM, Chothia, Contact and IMGT numbering.

[0029] Figure 2A-1 and 2A-2 show sequence alignments of the heavy chain variable regions of anti-Klotho beta antibodies providing consensus CDR sequences. The top cluster consists of antibodies designated 5H23, 1D19, 2L12, 3L3, 4P5, 5C23, and 5F7. The bottom cluster consists of antibodies designated 1G19 and 1C17. The background pool consists only of the antibody designated 3N20. Variable residues are shown by "X". CDR limits are indicated by Kabat, AbM, Chothia, Contact and IMGT numbering.

[0030] Figure 2B-1 and 2B-2 show sequence alignments of the light chain variable regions of anti-Klotho beta antibodies providing consensus CDR sequences. The top cluster consists of antibodies designated 5H23, 1D19, 2L12, 3L3, 4P5, 5C23, and 5F7. The bottom cluster consists of antibodies designated 1G19 and 1C17. The background pool consists only of the antibody designated 3N20. Variable residues are shown by "X". CDR limits are indicated by Kabat, AbM, Chothia, Contact and IMGT numbering.

[0031] Figure 3A-1 and 3A-2 show an alignment of sequences of the heavy chain variable region of the anti-Klotho beta antibody designated 5H23 with the humanized sequences (vH1-vH9). Residues that are in bold indicate exemplary residues that have been modified from the original antibody. Residues that are bold and underlined indicate residues changed back to a rat residue.

[0032] Figure 3B shows an alignment of sequences of the light chain variable region of the anti-Klotho beta antibody designated 5H23 with the humanized sequences (vL1-vL5). Residues that are in bold indicate exemplary residues that have been modified. Residues that are bold and underlined indicate residues changed back to a rat residue.

[0033] Figure 3C-1 and 3C-2 show a sequence alignment of the light chain variable region of the anti-Klotho beta antibody designated 5H23 with the humanized sequences (v1-39a-v1-39p). Residues that are in bold indicate exemplary residues that have been modified.

[0034] Figure 3D-1 and 3D-2 show a sequence alignment of a light chain variable region of the anti-Klotho beta antibody designated 5H23 with various humanized sequences (v3-20a-v3-20j). Residues that are in bold indicate exemplary residues that have been modified.

[0035] Figure 4A-4C shows a sequence alignment between human, rat and chimeric Klotho beta polypeptides. The chMoHu chimeric polypeptide indicates mouse KLB(M1-F506)-human KLB (S509-S1044). The chHuMo chimeric polypeptide indicates human KLB (M1-F508)-mouse KLB (P507-S1043). Residues corresponding to mouse residues are in bold italics.

[0036] Figure 5A-5F shows a sequence alignment between Klotho beta polypeptides from several species described here.

[0037] Figure 6 shows a three-dimensional model of the three identified binding residues (dark spheres) in equivalent positions in human cytosolic beta-glucosidase. The structure shows the equivalent of Klotho-beta residues 521-963.

DETAILED DESCRIPTION

[0038] Binding proteins, such as antibodies that bind to Klotho beta, including human and/or cyno Klotho beta, are provided here. A unique property of such binding proteins, including the antibodies disclosed herein, is their agonistic nature, including the ability to mimic the in vivo effect of FGF19 and/or FGF21 and to induce FGF19-like signaling and/or similar signaling. that of FGF21. Most notably and specifically, some of the binding proteins such as antibodies to Klotho beta disclosed herein (i) bind human and Cyno Klotho beta, (ii) do not compete for binding with FGF19 and/or FGF21, and (iii) induce FGF19-like signaling and/or FGF21-like signaling, including, for example, in various in vitro cell-based assays. Such assays may include (1) an ELKluciferase reporter assay (see, for example, Example 4); (2) a cell assay mediated by recombinant FGF19 receptor for ERK-phosphorylation (see, for example, Example 4); and (3) a human adipocyte assay for ERK-phosphorylation (see, for example, Example 5). Binding proteins, such as anti-Klotho beta antibodies, as described herein, are therefore expected to exhibit in vivo activities that are consistent with the natural biological function of FGF19 and/or FGF21. This property makes the described binding proteins, including Anti-Klotho beta antibodies, viable therapies for the treatment of metabolic diseases (e.g., type 2 diabetes, obesity, dyslipidemia, NASH, cardiovascular disease, metabolic syndrome) and broadly any disease. , disorder or condition in which it is desirable to mimic or enhance the in vivo effects of FGF19 and/or FGF21.

[0039] The binding proteins, such as antibodies that bind to Klotho beta, that are provided here share the common characteristic of competing with each other for the binding of Klotho beta (see, for example, Example 3 describing antibodies in epitope bin of 5H23). This competitive inhibition indicates that each antibody binds to the same region of Klotho beta (e.g., the same epitope), thus ensuring similar effects. Anti-Klotho beta antibodies provided herein include humanized anti-Klotho beta antibodies, including humanized Anti-Klotho beta antibodies derived from, or based on, 5H23, 1C17, 1D19, 2L12, 3L3, 3N20, 4P5, 5C23, 5F7 and/or 1G19 having CDR sequence as described in Tables 1-10 or Figures 1-3, such as anti-Klotho beta antibodies, including humanized anti-Klotho beta antibodies, and binds to a specific domain of human Klotho beta (e.g., KL2 (residues S509-S1044); see Example 9). Furthermore, such binding can be largely attributed to the presence of particular amino acid residues within the KL2 region (e.g., H657, Y701 and R703), which comprise the epitope recognized by the anti-Klotho beta antibodies described herein. Taken together, the results described herein demonstrate that the effects observed for anti-Klotho beta antibody that is derived from, or based on, 5H23 or an antibody in the 5H23 epitope bin, including an antibody with one or more CDRs described in Tables 1-10 or Figures 1-3, can be extrapolated to other anti-Klotho beta antibodies described herein having the same or similar epitope specificity (e.g., the same or similar CDRs). For example, the in vitro activities of antibodies such as shown in Examples 4-7 and 9, as well as the in vivo effects demonstrated in Example 8 for an exemplary humanized anti-Klotho beta antibody, are representative of the activities and effects of anti-Klotho beta antibodies. Klotho beta described here.

[0040] In some embodiments of the present invention, binding proteins such as anti-Klotho beta antibodies may comprise immunoglobulin variable regions that comprise one or more complementary determining regions (CDRs) as described in Tables 1-10. In such binding proteins (e.g., anti-Klotho beta antibodies), the CDRs may be joined with one or more framework regions, which orient the CDR(s) in such a way that appropriate antigen-binding properties are achieved. of the CDR(s) are achieved. Such binding proteins, including anti-Klotho beta antibodies as described herein, may facilitate or enhance the interaction between FGFR1c and Klotho beta, and may induce FGF19-like and/or FGF21-like signaling.

General techniques

[0041] The techniques and procedures described or referenced herein include those that are generally well understood and/or commonly employed using conventional methodology by those skilled in the art, such as, for example, the widely used methodologies described in Sambrook et al., Molecular Cloning: A Laboratory Manual 3rd. edition (2001) Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y.; Current Protocols in Molecular Biology (F. M. Ausubel, et al. eds., (2003)); Therapeutic Monoclonal Antibodies: From Bench to Clinic, Z. An, ed, Wiley, Hoboken N.J. (2009); Monoclonal Antibodies: Methods and Protocols, M. Albitar, ed., Humana Press, Totawa, N.J. (2010); and Antibody Engineering, 2nd Ed., Vols 1 and 2, Kontermann and Dubel, eds., Springer-Verlag, Heidelberg, 2010.

TERMINOLOGY

[0042] Unless otherwise described, all technical and scientific terms used herein have the same meaning as is commonly understood by a person skilled in the art. For the purposes of interpreting this specification, the following description of terms will be applied and, whenever appropriate, terms used in the singular will also include the plural and vice versa. All patents, applications, published applications and other publications are incorporated herein by reference in their entirety. In the event that any description of the terms set forth conflicts with any document incorporated herein by reference, the term description set forth below shall control.

[0043] The term "Klotho beta" or "Klotho beta polypeptide" and similar terms, refers to a polypeptide ("polypeptide" and "protein" are used interchangeably herein) or any native Klotho beta from from any vertebrate source, including mammals such as primates (e.g., humans, cynomolgus monkey (Cyno)), dogs, and rodents (e.g., mice and rats) unless otherwise noted, and, in certain embodiments, included related Klotho beta polypeptides, including their SNP variants. Klotho beta comprises two domains, Klotho beta 1 (KLB1) and Klotho beta 2 (KLB2). Each Klotho beta domain comprises a glycosyl hydrolase 1 region. For example, the KLB1 domain of human Klotho beta comprises amino acid residues 1-508 with the glycosyl hydrolase 1 region comprising amino acid residues 77-508, and the KLB2 domain of Human Klotho beta comprises amino acid residues 509-1044 with the glycosyl hydrolase 1 region comprising amino acid residues 517-967. The amino acid sequence of human Klotho beta is given below: WPRLFP DGIVTVANAK GLQYYSTLLD ALVLRNIEPI VTLYHWDLPL 201 ALQEKYGGWK NDTIIDIFND YATYCFQMFG DRVKYWITIH NPYLVAWHGY 251 GTGMHAPGEK GNLAAVYTVG HNLIKAHSKV WHNYNTHFRP HQKGWLSITL 301 GSHWIEPNRS ENTMDIFKCQ QSMVSVLGWF ANPIHGDGDY PEGMRKKLFS 351 VLPIFSEAEK HEMRGTADFF AFSFGPNNFK PLNTMAKMGQ NVSLNLREAL 401 NWIKLEYNNP RILIAENGWF TDSRVKTEDT TAIYMMKNFL SQVLQAIRLD 451 EIRVFGYTAW SLLDGFEWQD AYTIRRGLFY V DFNSKQKER KPKSSAHYYK 501 51 LPEPLLHADG WLNPSTAEAF QAYAGLCFQE LGDLVKLWIT INEPNRLSDI 701 YNRSGNDTYG AAHNLLVAHA LAWRLYDRQF RPSQRGAVSL SLHADWAEPA 751 NPYADSHWRA AERFLQFEIA WFAEPLFKTG DYPAAMREYI ASKHRRGLSS 801 SALPRLTEAE RRLLKGTVDF CALNHFTTRF VMHEQLAGSR YDSDRDIQFL 851 QDITRLSSPT RLAVIPWGVR KLLRWVRRNY GDMDIYITAS GIDDQALEDD 901 RLRKYYLGKY LQEVLKAYLI DKVRIKGYYA FKLAEEKSKP RFGFFTSDFK 951 AKSSIQFYNK VISSRGFPFE NSSSRCSQTQ ENTECTVCLF LVQKKPLIFL 1001 GCCFFSTLVL LLSIAIFQRQ KRRKFWKAKN LQHIPLKKGK RVVS (SEQ ID NO: 297)

[0044] A human Klotho beta coding nucleic acid sequence is provided below: atgaagccaggctgtgcggcaggatctccagggaatgaatggattttcttcagcactgatga aataaccacacgctataggaatacaatgtccaacggggattgcaaagatctgtcatcctgt cagcacttattctgctacgagctgttactggattctctggagatggaagagctatat ggtct aaaaatcctaattttactccggtaaatgaaagtcagctgtttctctatgacactttccctaa aaactttttctggggtattgggactggagcattgcaagtggaagggagttggaagaaggatg gaaaaggaccttctatatgggatcatttcatccacacaccttaaaatgtcagcagcacg aatggttccagtga cagttatatttttctggaaaagacttatcagccctggattttatagg agtttctttttatcaattttcaatttcctggccaaggcttttccccgatggaatagtaacag ttgccaacgcaaaaggtctgcagtactacagtactcttctggacgctctagtgcttagaaac attgaacctatagttactttataccactgggatttgcctt tggcactacaagaaaatatgg ggggtggaaaaatgataccataatagatatcttcaatgactatgccacatactgtttccaga tgtttggggaccgtgtcaaatattggattacaattcacaacccatatctagtggcttggcat gggtatgggacaggtatgcatgcccctggagagaagggaaatttagcagctgtctacactgt gggacacaacttgatca aggctcactcgaaagtttggcataactacaacacacatttccgcc cacatcagaagggttggttatcgatcacgttgggatctcattggatcgagccaaaccggtcg gaaaacacgatggatatattcaaatgtcaacaatccatggtttctgtgcttggatggtttgc caaccctatccatggggatggcgactatccagaggggatgagaa agaagttgttctccgttc tacccattttctctgaagcagagaagcatgagatgagaggcacagctgatttctttgccttt tcttttggacccaacaacttcaagcccctaaacaccatggctaaaatgggacaaaatgtttc acttaatttaagagaagcgctgaactggattaaactggaatacaacaaccctcgaatcttga ttgctgaga atggctggttcacagacagtcgtgtgaaaacagaagacaccacggccatctac atgatgaagaatttcctcagccaggtgcttcaagcaataaggttagatgaaatacgagtgtt tggttatactgcctggtctctcctggatggctttgaatggcaggatgcttacaccatccgcc gaggattattttatgtggattttaacagtaaacagaaagagcgg aaacctaagtcttcagca cactactacaaacagatcatacgagaaaatggtttttctttaaaagagtccacgccagatgt gcagggccagtttccctgtgacttctcctggggtgtcactgaatctgttcttaagcccgagt ctgtggcttcgtccccacagttcagcgatcctcatctgtacgtgtggaacg ccactggcaac agactgttgcaccgagtggaaggggtgaggctgaaaacacgacccgctcaatgcacagattt tgtaaacatcaaaaacaacttgagatgttggcaagaatgaaagtcacccactacc ggtttg ctctggattgggcctcggtccttcccactggcaacctgtccgcggtgaaccgacaggccctg agg tactacaggtgcgtggtcagtgaggggctgaagcttggcatctccgcgatggtcaccct gtattatccgacccacgcccacctaggcctccccgagcctctgttgcatgccgacgggtggc tgaacccatcgacggccgaggccttccaggcctacgctgggctgtgcttccaggagctgggg gacctggtgaagctctggatcaccatcaacgagcctaaccggctaagtgacatctacaaccg ctctggcaacgacacctacggggcggcgcacaacctgctggtggcccacgccctggcctggc gcctctacgaccggcagttcaggccct cacagcgcggggccgtgtcgctgtcgctgcacgcg gactgggcggaacccgccaacccctatgctgactcgcactggagggcggccgagcgcttcct gcagttcgagatcgcctggttcgccgagccgctcttcaagaccggggactaccccgcggcca tgagggaatacattgcctccaagcaccgacgggggctt tccagctcggccctgccgcgcctc accgaggccgaaaggaggctgctcaagggcacggtcgacttctgcgcgctcaaccacttcac cactaggttcgtgatgcacgagcagctggccggcagccgctacgactcggacagggacatcc agtttctgcaggacatcacccgcctgagctcccccacgcgcctggctgt gattccctggggg gtgcgcaagctgctgcggtgggtccggaggaactacggcgacatggacatttacatcaccgc cagtggcatcgacgaccaggctctggaggatgaccggctccggaagtactacctagggaagt accttcaggaggtgctgaaagcatacctgattgataaagtcagaatcaaaggctattatgca ttcaaactggctgaagagaaat ctaaacccagatttggattcttcacatctgattttaaagc taaatcctcaatacaattttacaacaaagtgatcagcagcaggggcttcccttttgagaaca gtagttctagatgcagtcagacccaagaaaatacagagtgcactgtctgcttattccttgtg cagaagaaaccactgatattcctgggttgttgcttcttctccaccctggt tctactcttatc aattgccatttttcaaaggcagaagagaagaaagttttggaaagcaaaaaacttacaacaca taccattaaagaaaggcaagagagttgttagc (SEQ ID NO: 298)

[0045] The amino acid sequence of Klotho beta from cynomolgus monkey (Cyno), scientific name Macaca fascicularis, is provided below: HTHLKN VSSTNGSSDS YIFLEKDLSA LDFIGVSFYQ 151 FSISWPRLFP DGIVTVANAK GLQYYNTLLD SLVLRNIEPI VTLYHWDLPL 201 ALQEKYGGWK NDTIIDIFND YATYCFQTFG DRVKYWITIH NPYLVAWHGY 251 GTGMHAPGEK GNLAAVYTVG HNLIKAHSKV WHNYNTHFRP HQKGWLSITL 301 GSHWIEPNRS ENTMDILKCQ QSMVSVLGWF HGDGDY PEGMKKKLLS 351 ILPLFSEAEK NEVRGTADFF AFSFGPNNFK PLNTMAKMGQ NVSLNLREAL 401 NWIKLEYNNP RILIAENGWF TDSHVKTEDT TAIYMMKNFL SQVLQAIRLD 451 EIRVFGYTAW SLLDGFEWQD AYTIRRGLFY VDFNSKQKER KPKSSAHYYK 501 QIIRENGFSL KEATPDVQGQ FPCDFSWGVT ESVLKPESVA SSPQFSDPYL 551 YVWNATGNRL LHRVEGVRLK TRPAQCTDFV NIKKQLEMLA RMS RA AERFLQFEIA WFAEPLFKTG DYPAAMREYI ASKHRRGLSS 801 SALPRLTEAE RRLLKGTVDF CALNHFTTRF VMHEQLAGSR YDSDRDIQFL 851 QDITRLSSPT RLAVIPWGVR KLLRWVRRNY GDMDIYITAS GIDDQALEDD 901 RLRKYYLEKY LQEVLKAYLI A FKLAEEKSKP RFGFFTSDFK 951 AKSSIQFYNK MISSSGFPSE NSSSRCSQTQ KNTECTVCLF LVQKKPLIFL 1001 GCCFFSTLVL LLSITIFHRQ KRRKFWKAKN LQHIPLKKGK RVLS (SEQ ID NO: 299)

[0046] A nucleic acid sequence encoding Klotho beta of cyno is provided below: atgaagcctggatgtgccgccggaagccccggcaacgagtggatcttcttcagcaccgacga gatcaccatccggtacagaaacaccatgagcaacggcggcctgcagcggagcgtgatcctgt ctgctctgaccctgctgagagccgtgaccggctt cagcggagatggcagagccgtgtggtcc aagaaccccaacttcacccccgtgaacgagagccagctgttcctgtacgataccttccccaa gaacttcttctggggcgtgggcacaggcgccctgcaggtggaaggatcctggaagaaggacg gcaagggccccagcatctgggaccactttgtgcacacccacctgaagaacgtgtcca gcacc aacggcagcagcgacagctacatctttctggaaaaggacctgagcgccctggacttcatcgg cgtgtccttctaccagttcagcatcagctggcccagactgttccccgacggcatcgtgacag tggccaatgccaagggcctgcagtactacaacaccctgctggacagcctggtgctgcggaac atcgagcccatcg tgaccctgtaccactgggacctgccactggctctgcaggagaaatacgg cggctggaagaacgacaccatcatcgacatcttcaacgactacgccacctactgcttccaga ccttcggcgacagagtgaagtactggatcacaatccacaacccctacctggtggcctggcac ggctatggcaccggaatgcatgcccctggcgagaagggaaatctggccgccg tgtacaccgt gggccacaacctgatcaaggcccacagcaaagtgtggcacaactacaatacccacttccggc cccaccagaagggctggctgtctatcacactgggcagccactggatcgagcctaaccgcagc gagaacaccatggacatcctgaagtgccagcagagcatggtgtccgtgctgggatggttcgc caaccccattcacggcga cggcgattaccccgagggcatgaagaagaagctgctgagcatcc tgcccctgttcagcgaggccgagaagaacgaagtgcggggcaccgccgatttcttcgccttt agcttcggccccaacaacttcaagcccctgaataccatggccaagatgggccagaatgtgtc cctgaacctgagagaggccctgaactggatcaagctggagtacaacaacccccggat cctga tcgccgagaacggctggttcaccgacagccacgtgaaaaccgaggacaccaccgccatctat atgatgaagaacttcctgagccaggtgctgcaggctatccggctggatgagatccgggtgtt cggctacaccgcctggtcactgctggacggcttcgaatggcaggacgcctacaccatcagac ggggcctgttctacgtgg acttcaacagcaagcagaaagagcggaagcccaagagcagcgcc cactactacaagcagatcatcagagagaatggcttcagcctgaaagaggccacccc cgacgt gcagggccagttcccttgtgatttctcttggggcgtgaccgagagcgtgctgaagcctgaaa gcgtggccagcagcccccagttcagcgacccttacctg tacgtgtggaacgccaccggcaac cggctgctgcatagagtggaaggcgtgcggctgaaaaccagacccgcccagtgcaccgactt cgtgaacatcaagaaacagctggaaatgctggcccggatgaaagtgacccactacagattcg ccctggactgggccagcgtgctgcctaccggaaatctgagcgccgtgaacaga caggccctg cggtactacagatgcgtggtgtccgagggcctgaagctgggcatcagcgccatggtcaccct gtactaccctacccacgcccacctgggactgcctgaacctctgctgcatgctggcggctggc tgaaccctagcaccgtggaagcctttcaggcctacgccgggctgtgcttccaggaactgggc gacctcgtgaagctgtggatcaccatcaacgagcccaacagactgagcgacatctacaacag aagcggcaacgacacctacggcgct gcccacaatctgctggtggctcatgccctggcttggc ggctgtacgacagacagttccggccttctcagcggggagccgtgtctctgtctctgcatgcc gattgggccgagcccgccaacccttacgccgactctcattggagagccgccgagcggttcct gcagttcgagatcgcttggtttgccgagcccctgtt caagaccggcgattaccctgccc ttctgcaggacatcacccggctgagcagccctacaagactggccgtgatcccttgggga gtgcggaagctgctgagatgggtgcgcagaaactacggcgacatggatatctacatcaccgc cagcggcatcgacgaccaggccctggaagatgaccggctgcggaagtactacctggaaagt acctgcaggaagtgctgaaggccta cctgatcgacaaagtgcggatcaagggctactacgcc ttcaagctggccgaggaaagagcaagcccagattcggcttcttcaccagcgacttcaaggc caagagcagcatccagttctacaacaagatgatcagcagcagcggcttccccagcgagaaca gcagctccagatgcagccagacccagaaaaacaccgagtgtaccgtgtg cctgttcctggtg cagaagaagcccctgatcttcctgggctgctgcttctttagcaccctggtgctgctgctgtc catcaccatcttccaccggcagaagcggagaaagttctggaaggccaaaaacctgcagcaca tccccctgaagaaaggcaagcgggtgctgagctga (SEQ ID NO: 300)

[0047] The amino acid sequence of rat Klotho beta homologue, scientific name Mus musculus, is given below: IFLEKDLLA LDFLGVSFYQ 151 FSISWPRLFP NGTVAAVNAQ GLRYYRALLD SLVLRNIEPI VTLYHWDLPL 201 TLQEEYGGWK NATMIDLFND YATYCFQTFG DRVKYWITIH NPYLVAWHGF 251 GTGMHAPGEK GNLTAVYTVG HNLIKAHSKV WHNYDKNFRP HQKGWLSITL 301 GSHWIEPNRT DNMEDVINCQ HSMSSVLGWF ANPIHGDGDY PEFMKTGAMI 351 PEFSEAEKEE VRGTADFFAF SFGPNNFRPS NTVVKMGQ NV SLNLRQVLNW 401 IKLEYDDPQI LISENGWFTD SYIKTEDTTA IYMMKNFLNQ VLQAIKFDEI 451 RVFGYTAWTL LDGFEWQDAY TTRRGLFYVD FNSEQKERKP KSSAHYYKQI 501 IQDNGFPLKE STPDMKGRFP CDFSWGVTES VLKPEFTVSS PQFTDPHLYV 55 1 WNVTGNRLLY RVEGVRLKTR PSQCTDYVSI KKRVEMLAKM KVTHYQFALD 601 WTSILPTGNL SKVNRQVLRY YRCVVSEGLK LGVFPMVTLY HPTHSHLGLP 651 LPLLSSGGWL NMNTAKAFQD YAELCFRELG DLVKLWITIN EPNRLSDMYN 701 RTSNDTYRAA HNLMIAHAQV WHLYDRQYRP VQHGAVSLSL HCDWAEPANP 751 FVDSHWKAAE RFLQFEIAWF ADPLFKTGDY PSVMKEYIAS KNQRGLSSSV 801 LPRFTAKESR LVKGTVDFYA LNHFTTRFVI HKQLNTNRSV ADRDVQFLQD 851 ITRLSSPSRL AVTPWGVRKL LAWIRRNYRD RDIYITANGI DDLALEDDQI 901 RKYYLEKYVQ EALKAYLIDK VKIKGYYAFK LTEEKSKPRF GFFTSDFRAK 951 SSVQFYSKLI SSSGLPA ENR SPACGQPAED TDCTICSFLV EKKPLIFFGC 1001 CFISTLAVLL SITVFHHQKR RKFQKARNLQ NIPLKKGHSR VFS (SEQ ID NO: 301)

[0048] A rat Klotho beta coding nucleic acid sequence is provided below: atgaagacaggctgtgcagcagggtctccggggaatgaatggattttcttcagctctgatga aagaaacacacgctctaggaaaacaatgtccaacagggcactgcaaagatctgccgtgctgt ctgcgtttgttctgctgcgagctgttaccggctt ctccggagacgggaaagcaatatgggat aaaaaacagtacgtgagtccggtaaacccaagtcagctgttcctctatgacactttccctaa aaacttttcctggggcgttgggaccggagcatttcaagtggaagggagttggaagacagatg gaagaggaccctcgatctgggatcggtacgtctactcacacctgagaggt gtcaacggcaca gacagatccactgacagttacatctttctggaaaaagacttgttggctctggattttttagg agtttctttttatcagttctcaatctcctggccacggttgtttcccaatggaacagtagcag cagtgaatgcgcaaggtctccggtactaccgtgcacttctggactcgctggtacttaggaat atcgagcc cattgttaccttgtaccattgggatttgcctctgacgctccaggaagaatatgg gggctggaaaaatgcaactatgatagatctcttcaacgactatgccacatactgcttccaga cctttggagaccgtgtcaaatattggattacaattcacaacccttaccttgttgcttggcat gggtttggcacaggtatgcatgcaccaggagagaagggaaatttaaca gctgtctacactgt gggacacaacctgatcaaggcacattcgaaagtgtggcataactacgacaaaaacttccgcc ctcatcagaagggttggctctccatcaccttggggtcccattggatagagccaaacagaaca gacaacatggaggacgtgatcaactgccagcactccatgtcctctgtgcttggatggttcgc caaccccatccacgggg acggcgactaccctgagttcatgaagacgggcgccatgatccccg agttctctgaggcagagaaggaggaggtgaggggcacggctgatttctttgccttttccttc gggcccaacaacttcaggccctcaaacaccgtggtgaaaatgggacaaaatgtatcactcaa cttaaggcaggtgctgaactggattaaactggaatacga tgaccctcaaatcttgatttcgg agaacggctggttcacagatagctatataaagacagaggacaccacggccatctacatgatg aagaatttcctaaaccaggttcttcaagcaataaaatttgatgaaatccgcgtgtttggtta tacggcctggactctcctggatggctttgagtggcaggatgcctatacgacccgacgagggc tg ttttatgtggactttaacagtgagcagaaagagaggaaacccaagtcctcggctcattac tacaagcagatcatacaagacaacggcttccctttgaaagagtccacgccagacatgaaggg tcggttcccctgtgatttctcttggggagtcactgagtctgttcttaagcccgagtttacgg tctcctccccgcagtttaccgatcctcacc tgtatgtgtggaatgtcactggcaacagattg ctctaccgagtggaaggggtaaggctgaaaacaagaccatcccagtgcacagattatgtgag catcaaaaaacgagttgaaatgttggcaaaaatgaaagtcacccactaccagtttgctctgg actggacctctatccttcccactggcaatctgtccaaagttaacaga caagtgttaaggtac tataggtgtgtggtgagcgaaggactgaagctgggcgtcttccccatggtgacgttgtacca cccaacccactcccatctcggcctccccctgccacttctgagcagtggggggtggctaaaca tgaacacagccaaggccttccaggactacgctgagctgtgcttccgggagttgggggacttg gtgaagctctggatcaccatcaatgagcctaacaggctgagtgacatgtacaaccgcacgag taatgacacctaccgtgcagcccacaacctgatgatcgcccatgcccaggtctggcacctct atgataggcagtataggccggtccagcatgg ggctgtgtcgctgtccttacattgcgactgg gcagaacctgccaacccctttgtggattcacactggaaggcagccgagcgcttcctccagtt tgagatcgcctggtttgcagatccgctcttcaagactggcgactatccatcggttatgaagg aatacatcgcctccaagaaccagcgagggctgtctagctcagtcctg a agctccttgcgtggatccggaggaactacagagacagggatatctacatcacagccaatgg catcgatgacctggctctagaggatgatcagatccgaaagtactacttggagaagtatgtcc aggaggctctgaaagcatatctcattgacaaggtcaaaatcaaaggctactatgcattcaaa ctgactgaagagaaatctaagcctagatttggatttttca cctctgacttcagagctaagtc ctctgtccagttttacagcaagctgatcagcagcagtggcctccccgctgagaacagaagtc ctgcgtgtggtcagcctgcggaagacacagactgcaccatttgctcatttctcgtggagaag aaaccactcatcttcttcggttgctgcttcatctccactctggctgtactgctat ccatcac cgtttttcatcatcaaaagagaagaaaattccagaaagcaaggaacttacaaaatataccat tgaagaaaggccacagcagagttttcagc (SEQ ID NO: 302)

The amino acid sequence of rat Klotho beta, scientific name Rattus norvegicus, is provided below: SISWPRLFPNGTVAAVNAKGLQYYRALLDSLVLRN IEPIVTLYHWDLPLTLQEEYGGWKNATMIDLFNDYATYCFQTFGDRVKYWITIHNPYLVAWH GFGTGMHAPGEKGNLTAVYTVGHNLIKAHSKVWHNYDKNFRPHQKGWLSITLGSHWIEPNRT ENMEDVINCQHSMSSVLGWFANPIHGDGDYPEFMKTSSVIPEFSEAE KEEVRGTADFFAFSF GPNNFRPSNTVVKMGQNVSLNLRQVLNWIKLEYDNPRILISENGWFTDSYIKTEDTTAIYMM KNFLNQVLQAIKFDEIQVFGYTAWTLLDGFEWQDAYTTRRGLFYVDFNSEQKERKPKSSAHY YKQIIQDNGFPLQESTPDMKGQFPCDFSWGVTESVLKPEFTVSSPQFTDPHLYVWNVT AEPANPYVESH WKAAERFLQFEIAWFADPLFKTGDYPLAMKEYIASKKQRGLSSSVLPRFTL KESRLVKGTIDFYALNHFTTRFVIHKQLNTNCSVADRDVQFLQDITRLSSPSRLAVTPWGMR KLLGWIRRNYRDMDIYVTANGIDDLALEDDQIRKYYLEKYVQEALKAYLIDKVKIKGYYAFK LTEEKSKPRFGFFTSDFKAK SSVQFYSKLISSSGFSSENRSPACGQPPEDTECAICSFLTQK KPLIFFGCCFISTLAALLSITIFHHRKRRKFQKARNLQNIPLKKGHSRVFS (SEQ ID NO:356)

[0050] A rat Klotho beta coding nucleic acid sequence is provided below: ATGAAGACAGGCTGTGCAGCAGGGTCTCCAGGGAATGAATGGGTTTTCTTCAGCTCTGATGA AAGAAGCACACGCTCTAGGAAAACAATGTCCAACGGAGCACTGCAAAGATCTGCCGTGCTGT CTGCATTGGTTCTGCTGCGAGCTGTTACCGGCTTCTCTGGAGACGGAAAAGCAATATGGGAT AAAAAACAATACGTGAG TCCGGTAAACCCAGGTCAGCTGTTCCTCTATGACACTTTCCCTAA AAACTTTTCCTGGGGCGTTGGGACCGGAGCATTTCAAGTGGAAGGGAGTTGGAAGGCAGATG GAAGAGGACCCTCGATCTGGGACCGTTATGTCGACTCACACCTGAGAGGTGTCAACAGCACA GACAGATCCACTGACAGTTATGTCTTTCTGGAAAAGGACTTGCTGGCTCTGGATTTTTTAGG AGTTTCTTTTTATCAGTTCTCAA TCTCCTGGCCGCGGTTGTTCCCCAACGGAACAGTAGCAG CTGTGAATGCAAAAGGTCTCCAGTACTACAGAGCACTTCTGGACTCGCTGGTACTTAGGAAT ATCGAACCCATTGTTACCTTATACCATTGGGATTTGCCTTTGACGCTACAGGAAGAATATGG GGGCTGGAAAAATGCAACTATGATAGATCTCTTCAATGACTATGCCACATACTGCTTCCAGA CCTTTGGAGACCGTGTCAAATATTGGATTACA ATTCACAACCCTTACCTCGTTGCTTGCAT GGGTTTGGCACAGGTATGCATGCGCCAGGAGAGAAGGGAAATTTAACAGCTGTCTACACTGT GGGACACAACCTGATCAAGGCGCATTCGAAAGTGTGGCATAACTACGACAAAAACTTCCGCC CTCATCAGAAGGGTTGGCTCTCCATCACCTTGGGGTCCCATTGGATAGAACCAAACAGAACA GAAAACATGGAGGACGTGATCAACTGCCAGCACTCCATGTCTTCTG TGCTCGGATGGTTTGC CAACCCCATCCACGGAGACGGCGACTACCCCGAGTTCATGAAGACGAGCTCCGTAATCCCTG AGTTCTCTGAGGCAGAGAAGGAGGAGGTGCGGGGCACTGCTGACTTCTTTGCCTTTTCCTTC GGGCCCAACAATTTCAGGCCCTCGAACACCGTGGTAAAAATGGGACAAAATGTATCACTCAA CTTAAGACAGGTGCTGAACTGGATTAACTAGAATATGACAACCCTCGAATCT TGATTTCGG AGAACGGCTGGTTCACAGATAGTTATATAAAGACGGAAGATACCACGGCCATCTACATGATG AAGAATTTCCTCAACCAGGTTCTTCAAGCAATAAAGTTTGATGAAATACAAGTGTTTGGTTA TACGGCTTGGACTCTCCTGGATGGCTTTGAGTGGCAGGATGCCTACACGACCCGACGAGGGC TGTTTTATGTGGACTTTAATAGTGAGCAGAAAGAGAGGAAACCCAAGTCCTCCGCTCATT AC TACAAACAGATTATACAAGACAACGGTTTCCCTTTGCAAGAATCCACACCAGACATGAAGGG TCAGTTTCCCTGTGACTTCTCCTGGGGAGTCACTGAGTCTGTTCTTAAGCCGGAGTTTACGG TGTCCTCCCCACAGTTTACTGATCCTCACCTGTATGTGTGGAATGTCACTGGCAACAGATTG CTATACCGAGTGGAAGGAGTCAGGCTAAAAACAAGACCGTCCCAATGCACAGATTATGTGAG CATCAA AAAACGAGTTGAAATGTTGGCCAAATGAAAGTCACCCACTACCAGTTTGCTCTGG ACTGGACCTCTATCCTCCCTACCGGAAATCTGTCTAAAATTAATAGACAAGTGTTGAGGTAC TATAGGTGTGTGGTGAGCGAAGGACTGAAGCTGGGCATCTCCCCTATGGTGACGTTGTACCA CCCGACCCACTCCCATCTAGGCCTCCCCATGCCACTTCTGAGCAGTGGGGATGGCTAAACA CCAACACAGCCAAGGCCTTCCAGGACTACGCAGGCCTGTGCTTCAAGGAGCTGGGGGACTTG GTAAAGCTCTGGATCACCATCAATGACCCAATAGGCTGAGTGACATGTACAACCGCACGAG TAACGACACCTACCGTGCGGCCCACAACCTGATGATCGCCCATGCCCAGGTCTGGCACCTCT ATGATAGGCAGTATAGGCCGGTCCAGCACGGGGCTGTGTCGCTGTCCTTACATTCCGACTGG ACCTGCCAACCCCTATGTGGAGTCTCACTGGAAGGCAGCCGAGCGCTTCCTCCAGTT TGAGATCGCCTGGTTTGCGGATCCACTCTTCAAGACTGGTGACTACCCGCTGGCCATGAAGG AATACATCGCCTCCAAGAAGCAGCGAGGGCTGTCTAGCTCAGTCCTGCCGCGCTTTACCTTG AAGGAGAGCAGGCTGGTGAAGGGGACCATCGACTTTTACGCACTGAACCACTTCACTACTAG ATTCGTGATACA CAAGCAGTTGAATACCAACTGCTCAGTGGCAGACAGGGACGTCCAGTTCC TGCAGGACATCACCCGCCTGAGCTCGCCCAGTCGCCTAGCCGTAACGCCCTGGGGAATGCGC AAGCTCCTTGGGTGGATCCGGAGGAACTACAGAGACATGGATATCTACGTCACAGCCAATGG CATTGATGATCTTGCTCTAGAGGACGATCAGATTAGAAAGTACTACTTGGAGAAGTACGTCC AGGAGGCTCTGAAAGC ATATCTGATTGACAAGGTCAAAATCAAAGGCTACTATGCATTCAAA CTGACTGAAGAGAAATCTAAGCCTAGATTTGGATTTTTCACATCTGACTTCAAAGCTAAATC TTCTGTACAGTTTTATAGCAAGCTGATCAGCAGCAGCGGCTTCTCCTCTGAGAACAGAAGTC CTGCCTGTGGTCAGCCTCCAGAAGACAGAATGCGCCATTTGCTCCTTCCTTACACAGAAG AAACCACTCATCTTCTTTGGTTG TTGCTTCATCTCCACTCTGGCTGCACTGCTATCAATCAC TATTTTTCATCATCGGAAGAGAAGAAAATTCCAGAAAGCAAGGAACTTACAAAATATACCAT TGAAGAAAGGGCACAGCAGAGTTTTTAGCTAA (SEQ ID NO:357)

[0051] The amino acid sequence of Hamster Klotho beta, scientific name Cricetulus griseus, is provided below: MKAGCAAGSPGNEWIFLSSYERNTRSKKTMSNRALQRSVVLSAFVLLRAVTGLSGDGKAIWD KKQYVSPVNASQLFLYDTFPKNFFWGVGTGAFQVEGNWQADGRGPSIWDRFIYTHLRDVSIT EKSADSYIFLEKDLLALDFLGVSFYQ FSISWPRLFPNGTVASVNAKGLQYYNKLLDSLILRN IEPVVTLYHWDLPLALQEDYGGWKNATMIDLFNDYATYCFQTFGDRVKYWITIHNPYLVAWH GFATGMHAPGETGNLTAVYIVGHNLIKAHSKVWHNYDKNFRPHQKGLLSITLGSHWIEPNKT ENMADTISCQHSMAFVLGWFANPIHADGDYPEFMKTLSTMPVFS EAEKEEVRGTADFFAFSF GPNNFRPSNTVVKMGQNVSLNLRQVLNWIKLEYDNPRILISENGWFTDSDIKTEDTTAIYMM KHFLNQVLQAIQFDEIRVFGYTAWSLLDGFEWQYAYTSRRGLFYVDFNSEQKERKPKTSAHY YKQIIQENGFPLKESTPDMQGQFPCDFSWGVTESVLKPEFMVSSPQFTDPHLY VWNATGNRL LQRVEGVRLKTKPSHCTDYVSIKKRVEMLAKMKVTHYQFALDWATILPTGNLSEVNRQVLRY YRCVVSEGLKLGVSPMVTLYHPTHSHLGLPEPLLNSGGWLNTYTAKAFQDYAGLCFQELGDL VKLWITINEPNRLSDMYNRTSNDTYRAAHNLMIAHAQVWRLYDRQYRPVQHGAVSLSLHSDW VE PANPYVDSHWKAAERFLLFEIAWFADPLFKTGDYPLAMKEYIASKNQQGLSRSVLPRFTP EESRLVKGTIDFYALNHFTTRFVIHKQLNSSRSMADRDVQFLQDITRLSSPSRLAVMPWGAR KLLGWIQRNYGDMDIYITANGIDDLALENDGIRKYYLEKYIQEALKAYLIDKVKIKGYYAFK LTEEKSKPRFG FFTSDFKAKSSVEFYSKLISRSGFPSETSNPACGQPPEDTDCTICSFFTQK KSLIFFGCCFISTLAVLLSITIFHHRKRRFHKSKNLENIPLKEGHSRVLS (SEQ ID NO: 408

[0052] A nucleic acid sequence coding for hamster Klotho beta is provided below: atgtccaacagggcactgcaaagatctgtcgtgctgtcagcgtttgttctgctgcgagctgt taccggattgtctggagacgggaaagcgatatgggataaaaaacagtacgtgagtccggtga atgcaagtcagctgtttctctatga cactttccctaaaaactttttctggggtgttggaact ggagcatttcaagtggaagggaattggcaggcagacggaagaggaccctcgatttgggatcg tttcatctacacacacctgagagatgtcagcatcacagagaaatccgccgacagttacattt ttctggaaaagatttgttggctctggatttttaggagttt ctttttatcagttctcaatc tcctggccacggttgttccccaatggaacagtagcatccgtgaatgcaaaaggtctccaata ctacaacaaacttctggactcgctgatacttaggaatattgagcccgttgttaccttatacc attgggatttgcctttggcgctacaggaagactatggggttggaaaaatgcaactatgata gatctcttca atgactatgccacatactgcttccagacctttggagaccgtgtcaagtattg gattacaattcacaacccttacctggttgcttggcatgggtttgccacaggtatgcatgcgc caggagagacgggaaatttaacagctgtctacattgtgggacacaacctgatcaaggctcat tcgaaagtgtggcataactacgacaaaaacttccgcccccatcaga agggtttgctgtccat taccttggggtcccactggatagaaccaaacaaaacagaaaacatggccgatacaatcagct gccagcactctatggcttttgtgcttgggtggtttgccaaccccatccatgcagacggcgac taccctgagttcatgaaaacattgtccaccatgccagtgttctctgaggcagagaaggagga ggtgagg ggcacagctgacttctttgccttttcctttgggcccaacaatttcaggccctcga acactgtagtgaaaatgggacaaaatgtatcactcaacttaagacaggtgctgaactggatt aaattagaatatgacaaccctcgaatcttgatttcggagaatggctggttcacagatagtga cataaagacagaggacaccacagccatctacatgatgaagcattt cctcaaccaggttcttc aagcaatacagtttgatgaaatacgagtgtttggttacacggcctggtctctcctggatggc tttgaatggcagtatgcctacacgtctcgccgaggactgttttatgtggactttaatagtga acagaaagaaaggaaacccaagacctcggcacattactacaaacagatcatacaagaaaatg gtt tccctttgaaagagtccacgccagacatgcagggtcagtttccctgtgacttctcctgg ggggtcaccgagtctgttcttaagccggagtttatggtttcctccccacagtttaccgaccc tcacctgtatgtgtggaatgccactggcaacagattgctacagcgagtagaaggagtaaggc taaaaacaaaaccatccc actgcacagactatgttagcatcaaaaaacgagttgagatgttg gccaaaatgaaagtcacccactaccagtttgctctggactgggccaccatccttcccactgg caatctgtctgaagttaatagacaagtactaaggtactataggtgtgtggtgagcgaaggac tgaagctgggcgtctctcccatggtgacgttgtaccacc cccacccactcccatctaggcctc cctgagccgcttcttaacagtgggggatggctaaacacttacaccgccaaggccttccagga ctacgcaggactgtgcttccaggaactaggggacttggtgaagctctggatcaccatc aatg agcctaataggctgagtgacatgtacaaccgcacgagtaatgacacctaccgtgcagcccat aacctgatgattgcccatgcccaggtctggcgtctctacgacaggcagtataggccagtcca gcatggagctgtgtcgctgtccctacattctgactgggtggaacctgccaacccctatgtgg actcacactggaaggcagcggagcgcttcctcctgtttgagatcgcctggttcgctgatccg ctcttcaagactggcgactatccactggccatgaaggagtacatcgcctccaagaaccagca agggctgtcccgctcagtcctgccgcgcttcaccccagaggagagcaggctggtgaagggca ccatcgacttctacgcactgaaccacttcactactaggttcgtgatacacaaacagctcaac agcagccgctctatggcagacagggacgtccagttcctgcaggacatcacccgcctgagctc gcccagccgcctggctgttatgccctggggagcacgcaagctgcttgggtggatccagagga actatggggacatggacatctacatcacagccaatggcatcgatgatctggctctggagaat gatgggatccgaaagtactacttggagaagtacatccaggaggctctgaaagcatacctcat tgacaaagtcaaaatcaaaggctattatgcattcaaactgactgaagagaaatctaagccta gatttggatttttcacatctgacttcaaagctaagtcatctgtagagttttatagcaagttg atcagcagaagtggcttcccctctgagactagcaatcccgcatgtggtcagcctccagaaga cacagactgcaccatctgctcatttttcactcagaagaaatctctgatcttctttggttgtt gcttcatctccactctggctgtactgctgtcaatcaccatttttcatcatcgaaagagaaga tttcataaatcaaagaacttagaaaatataccattgaaggaaggccacagtagagttcttag ctaa (SEQ ID NO: 409)

The amino acid sequence of rabbit Klotho beta, scientific name Oryctolagus cuniculus, is provided below: FPDGTVAVANAKGLQYYNRLLDSLLLRNI EPVVTLYHWDLPWALQEKYGGWKNETLIDLFNDYATYCFQTFGDRVKYWITIHNPYLVAWHG YGTGLHAPGEKGNVAAVYTVGHNLLKAHSKVWHNYNRNFRPHQKGWLSITLGSHWIEPNRAE SIVDILKCQQSMVSVLGWFANPIHGDGDYPEVMTKKLLSVLPAFSEAE KNEVRGTADFFAFS FGPNNFKPLNTMAKMGQNVSLNLRQVLNWIKLEYGNPRILIAENGWFTDSYVQTEDTTAIYM MKNFLNQVLQAIRLDGVRVFGYTAWSLLDGFEWQDAYNTRRGLFYVDFNSEQRERRPKSSAH YYKQVIGENGFTLREATPDLQGQFPCDFSWGVTESVLKPESVASSPQFSDPHLYVWNATG NL ERFLQFEIAWFAEPLFKTGDYPVAMREYIASKTRRGLSSSVLPRFS DAERRLVKGAADFYALNHFTTRFVMHEQQNGSRYDSDRDVQFLQDITRLASPSRLAVMPWGE GKLLRWMRNNYGDLDVYITANGIDDQALQNDQLRQYYLEKYVQEALKAYLIDKIKIKGYYAF KLTEEKSKPRFGFFTSDFKAKSSIQFY NKLITSNGFPSENGGPRCNQTQGNPECTVCLLLLQ KKPLIFFSCCFFCTLVLLSSITIFHRRKRRKFWKAKDLQHIPLKKGHKRVLS (SEQ ID NO: 410)

[0054] A rabbit Klotho beta coding nucleic acid sequence is provided below: tgaagccgtgataagacggtcccgcagttcgtggcaaatgaagccaggctgtgcggcaggat ctccagggaatgaatgggtttccttctgcaccgatgaaagaaacagacgctgtagggaaacg atgtccagcggacgcctgcggagatctgtcatgct gtcagccttcatcctgctgcgagccgt gactgggttccccggagacggaagagctgtatggtcgcaaaatcctaatttgagtccggtaa acgaaagtcagctgtttctctatgacactttcccaaaaaactttttctggggtgtggggact ggagccttccaagtggaagggagttggaagaaggatgggaaa ggactctctgtatgggatca tttcatcgctacacacctgaacgtcagcagccgcgatggctccagtgacagctacatttttt tggagaaagacttatcggcgctggattttttaggagtctctttttatcagttttcaatttcc tggccaagactgttcccggatggcacagtagcagtcgccaatgcaaaaggtctccagtacta taatcggctcctggactctctgctacttagaaacattgaacctgtagtcactttataccatt gggatctgccttgggcgctacaagaaaaatacggggggtggaaaaacgagacgttgattgat ttattcaatgactatgccacctactgtttccagacgtttggggaccgtgtcaaatactggat caccattcacaatccctatctggtggctt ggcatggctacgggacaggtctgcatgctccgg gagagaaggggaatgtggcagctgtctacactgtgggacacaacctgcttaaggctca ttca aaagtctggcacaactacaacaggaatttccgcccgcatcagaaaggctggctgtcgatcac gctgggatcccactggattgagccaaacagagcggaaagcatcgtggacatactcaagtgcc agcagtccatggtctcggtgctgggctggtttgccaacccgatccacggggacggggactac ccagaggtgatgacaaagaagctgctctccgtcctgcccgctttctcagaagcagagaagaa cgaggtacgaggcaccgcagacttctttgccttttcgtttggacccaacaacttcaagccct taaacaccatggctaaaatgg ggcagaatgtgtcactcaatctaagacaggtgctgaactgg attaaactggaatatggcaaccctcgaatcctgatcgctgagaacggctggttcacagacag ttacgtgcaaacagaagacaccacagccatctacatgatgaagaatttcctcaaccaggttc ttcaagcaataaggttggatggagtccgagtgtttggctacactgcct ggtctctcctggat ggcttcgaatggcaggacgcttacaacacccgccgtggactgttttatgtggacttcaacag cgaacagagagaaagaaggcccaagtcctcggcgcattactataaacaggtcataggagaaa acggcttcacgctcagagaggccaccccggatctgcaggggcagtttccctgtgacttctcc tggggcgtcacc gagtctgttcttaagcccgagtcggtggcttcctcgccacagttcagcga ccctcacctctacgtgtggaacgccactggcaaccgaatgcttcaccgggtggaaggggtga ggctgaaaacacggcccgctcagtgcacagatttcatcaccatcaagaaacaactcgagatg ttggcaagaatgaaagtcacccactt ccggtttgctctggactgggcctcggtccttcccac gggcaacctgtccgaggtgaaccgacaagccctgaggtactacaggtgtgtggtcaccgagg ggctgaagctcaacatctcgcccatggtcaccttgtactacccgacccatgcccacctgggc ctgcccgcgccgctgctgcacagcggggggtggctggacccatccacggccaaggccttccg cgactacgcagggctgtgcttccgggagctgggggacctggtgaagctctggatcaccatca acgagcccaaccggctgag cgacgtctacaaccgcaccagcaacgacacctaccaggccgcc cacaacctgctgatcgcgcacgcgctcgtgtggcacctgtacgaccgccagtaccggccgtc gcagcgcggggcgctgtcgctgtccctgcactcggactgggccgagcccgccaacccctacg tggcctcgcactggcaggcggccgagcg cttcctgcagttcgagattgcgtggttcgccgag cccctgttcaagaccggggactacccggtggccatgagggagtacatcgcctccaagacccg gcgcgggctctccagctccgtgctgccccgcttcagcgacgccgagcggcggctggtcaagg gcgccgccgacttctacgccctcaaccacttcaccaccaggt tcgtgatgcacgagcagcag aacggcagccgctacgactcggacagggacgtgcagttcctgcaggacatcacccgcctggc ctcacccagccgcctggccgtgatgccctggggcgagggcaagctgctgcggtggatgcgga acaactacggagacctggacgtctacatcacggccaatggcatcgacgaccaggccct gcag aacgaccagcttcgccagtactacctggagaagtacgtccaggaggctctgaaagcatatct gatagataaaataaaaatcaaaggctattatgcattcaaactgactgaagaaaaatctaaac ccaggtttggattcttcacctctgatttcaaagccaagtcttcaatacagttttacaaca aa ctaattaccagcaacggcttcccgtct gagaacggcggtcctagatgcaatcagactcaagg aaatcccgagtgcaccgtctgcttactcctcctgcagaagaagccgctgatattctttagct gctgcttcttctgcaccctggttctactctcatcaattaccatctttcacagacggaagaga agaaaattttggaaagcaaaggacttacaacacataccattaaagaaaggccacaagagagt c cttagctaaagtgaacttatttctctctgaagagtttagaaattcactccagttccatatg ctggtaacacaaaagacatacccgtattgtacacagagtatttgagatactgtgctaaccaa ggcgatgacaatcaaaacctctgccatgtggttgaatgcattttcccttaagcggtgacaat cagcgaactcagttcttggttctaa aggaggcttcgcactgccactaggctatgagtattac ctgacgcattgctttgtcaagtttgatgagctgtttcgcatcattctctagctttctttaga taccaatagctactatggtaaaagttgttttaaaagtcaaactctgtaaggcttcacagca gatttaaaactattctttacactggatctgtgattttgtcactc gtagcaaggtgctttccc cttttggtcctagtggctctcaaatagaaagcaaacacatcttagggtaatctacttatcta tagccaatcacagcactgacccacaactacacaaatccgttagctcttctccataaaacacc taattttgtgatcttttaagtaatctgaaatgtaaaagtatgacttccgtaacccatctcat ggaaagatcgactaa ggagagccatacccagctgtgaggacaatttagtcactaatctcacc gtactgcaacttcctcctttagagcaggcattccttaccatttttgtaagatgacatgattt agcatctagaacccctatctgcagtttctttctatggcttacctacatttcaagaatattga acggaaatttcagaaagatttccaagttttaaattgtgtactagcattagtgcat gatgaa atctcattttctttgctccatcctgcacaggatgtgaaacatccctctgtccagcaagtcca agctacctatattactcacttgatagtcaccatggttatccagctgttattacttgctcata cccaggtaacccttttttatttaatatagctccaaagtataagactagtgatgaaaaggag gtaagtcatcaaatatggaaggacagattaactct ggcactaagtgggaatgctgcaggttt tacaggaaaacaaaattcagtcagtggtttaaagcatcctctgaggtacctggggcacaatc tccacagataaggggaaagagcactgacaaagactaaacatcctaaaaagacgcaatgttct acttactggccatcagaataatggccaaaggaccctatacttgcttgctctctagccaagtt tcgct gcacataggtgtagaatgcagcgactgaccctggatgcgattcagaatgctgatctg agtgaactagttttttatacagcactttttaaagcctagaattcttccatctgaacttggga gttttgacttttttgaaattaattgtgcttaagatttattcagtgattctaaacactggagg tagaaaactgtatacccattatgcctattaatt tttcttgattagccaacatttaaataacc acaaagtggccagtcgttgtctttccctttcaggaatttaagtcaaaggatgctgctgcctg cgatgctggcacttcataggggtgacagtttgtgtccctgcggttccacttcctatccagct ccctgctaatggcttgggagagccctgcacccacatgggagacccaa aagcagatcctgctg ctttcagcctgctgcggccacttggagtatgaaccagtggatggaagatcaatgtctctccc aacaattctttgaataaattttttcaaaagtcaaaataaaattctccagctcaaaaagcttt agtagaaaacgatcctacattaaggcggttgtgattgtatcccaagtgcatctacgttacaa accaaattgagtatgcaattcagtatgctactagactataaggagaaaacagccaattcaaa gttaatttgctttctggttggccaaatgttttttttctct tcttgccaccactgttttacatgtactttagaagaaattttgactttttgcttcctttgaga aatcactattatcaaaggcaattcataattacaagtggtccattgtcttaagagctcaagat tatagcccttcaaacttgccaaactcctcaaatagtgaagctcctaacgaaggg tttacaac atcctgttccttaggggttatatttttaagtgactgtaatttacctaacaaatttaatctgg ctatctattggtaatacatgtaatattcaggtttatcataaacccacttaaaaactaaaggt taagtggaagttgctgcttttcaaagtaacaggcttctcaggggaaatatcaccttagcgt ccacctggtactacatctcgtgtatt cactgtaacccatctttccgaacatgtctgatatat atggaaacaacactagtgcttagcctctggaaatgaggccaggattttgtgattaaatgtct aatttattccaaataaactgatttacgccaata (SEQ ID NO: 411)

[0055] The amino acid sequence of dog Klotho beta, scientific name Canis lupus familiaris, is provided below: MKPGCAAGSPGNEWIFLSTDESNTHYRKTMCNHGLQRSVILSAFILLGAVPGFSGDGRAIWS KNPHFSPVNESQLFLYDTFPKNFFWGVGTGAFQVEGNWKTDGKGPSIWDHFIHTHLKNVNSM NSSSDSYIFLEKDLSALDFIGVSFYQFSISWPRL FPDGIAAVANAKGLQYYNSLLDALVLRN IEPIVTLYHWDLPALQEKYGGWKNETITDIFNDYATYCFQTFGDRVKYWITIHNPYLVAWH GYGTGMHAPGEKGNLAAVYTVGHNLIKAHSKVWHNYNTNFRPYQKGLLSITLGSHWIEPNRS ENMMDILKCQQSMVSVLGWFANPIHGNGDYPEVMKKKLLSTLPLFSEAEKNE VRGTADFFAF SFGPNNFKPQNTMAKMGQNVSLNLREVLNWIKLEYGNPRILIAENGWFTDSHVKTEDTTAIY MMKNFLNQVLQAIRFDEIQVFGYTAWSLLDGFEWQDAYSTRRGLFYVDFNSKQKERKPKSSA YYYKQIIQENGFTFKESTPDVQGQFPCDFSWGVTESVLKPKVVASSPQFSDPHLYVWNVT GN RLLHRVEGVRLKTRPAQCTDFVSIKRQLEMLARMNVTHYRFALDWPSILPTGNLSTVNRQAL RYYRCVVSESLKLSISPMVTLYYPTHAHLGLPSPLLHSGGWLNASTARAFQDYAGLCFQELG DLVKLWITINEPNRLSDVYSHTSSDTYRAAHNLLIAHALVWHLYDRRYRPAQRGAVSLSLHS DWAEPANPYADSHWKAAERFLQFE IAWFAEPLFKTGDYPPAMREYIASKNRQGLSRSTLPRF TDEERRLVKGAADFYALNHFTTRFVMHARQNGSRYDADRDVQFLQDITCLSSPSRLAVLPWG ERKVLRWIQKNYGDVDVYITASGIDDQSLENDELRKYYLEKYIQEALKAHLIDKVKVKGYYA FKLTEEKSKPRFGFFTSEFKAKSSVQLYNKLISNS GFPSENRSPRCSETQRNTECMVCLFLV QKKPLIFFSCCFFSTLVLLSSITILHKRKRRKIWKAKNLQHIPLKKSKNSLQS (SEQ ID NO: 412)

[0056] A dog Klotho beta coding nucleic acid sequence is provided below: acaatcacaagcttttactgaagcgttgataagacaggcgagcagttagtggcaaatgaagc caggctgtgcggctggatctccagggaatgaatggattttcctcagcaccgatgaaagcaac acacactataggaaacaatgtgcaaccacgggctacagagatctgt catcctgtcagcatt tattctcctaggagctgttcctggattctctggagacggaagagctatatggtctaaaaatc ctcattttagtccggtaaatgaaagtcagctgtttctctatgacacttttcctaaaaacttt ttttggggcgttgggactggagcatttcaagtggaagggaattggaagacagatggaaagg accctctatatgggatcatttcatccacacaccttaaaaatgtcaacagcatgaatagtt ccagtgacagttacatttttctggaaaagacctatcagccctggattttatcggagtttct ttttatcaattttcaatttcctggccaaggcttttccccgatggaatagcagcagttgccaa cgcaaaaggtctccagtactaca attctcttctcgatgctctagtacttaggaacattgaac ctatagttactttataccattgggatttgcctttggcactacaagaaaaatatggggggtgg aaaaatgaaaccataacggatatcttcaatgactatgccacctactgtttccagacgttcgg ggatcgtgtcaaatactggattacaattcacaatccatatctagttgcttggcatgggta tg ggacaggtatgcacgcgcctggagagaagggaaacttagcagctgtctacactgtgggacac aacctaatcaaggctcattcgaaagtttggcataactacaacaaatttccgcccatatca gaagggtttgttatcaatcacgttgggatcccattggattgaaccaaacagatcagaaaaca tggtttcagtgctcgggtggtttgccaacccc atccatgggaatggagactatccagaagtgatgaaaaagaagttgctctccactctacccct tttctctgaagcagagaagaatgaagtgaggggcacagctgacttctttgccttttcctttg gacccaacaatttcaagccccagaacaccatggctaaaatgggacaaaatgtg tcactcaat ttaagagaagtgctgaattggattaaactggaatatggcaacccccgaatcttgattgctga gaatggctggttcacagacagtcatgtgaaaacagaagataccacagccatttacatgatga agaatttcctcaaccaggttcttcaagcaataaggtttgacgaaatacaagtgtttggctac actgcttggtctctcctggatggct ttgaatggcaggatgcttactccactcgccgaggatt attttatgtggattttaatagtaaacaaaaagaaagaaagcccaagtcttcggcatattact ataaacagatcatacaagaaaatggttttactttcaaagagtccaccccagatgtgcagggt cagtttccctgtgacttctcatggggtgtcaccgaatctgtccttaagcccaaagtc gtggc ttcctccccacagttcagcgaccctcacctgtacgtgtggaatgtgacaggcaacagactgt tgcaccgagtggaaggggtgaggctgaagacacggccggctcaatgcacagatttt gtcagc atcaaaagacaacttgagatgttggcgaggatgaacgtcactcactacaggtttgctctgga ctggccct ccatccttcccaccggcaacctgtccacggttaaccgacaagccctgaggtact acaggtgtgtggtcagcgagtcgctgaagctcagcatctccccgatggtcacgctgtactac ccgacccacgcccacctgggcctcccctcgccgctgctgcacagcgggggctggctgaacgc gtccaccgcccgcgccttccaggactatgccgggctgtgcttccaggagctgggggacctgg tgaagctctgg atcaccatcaatgagcccaaccggctgagtgacgtctacagccacaccagc agcgacacctaccgggcagcgcacaacctgctgatcgcccacgccctggtgtggcacctgta cgaccggcggtaccggccggcgcagcgcggggccgtgtcgctgtccctgcactcggactggg cggagcccgccaacccctacgccgact cgcactggaaggcggccgagcgcttcctgcagttc gaaatcgcctggttcgccgagccgctcttcaagaccggggactacccgccggccatgaggga gtacatcgcctccaagaacaggcaggggctctcgcgctccaccctgccccgcttcaccgacg aggagaggaggctggtcaagggcgccgccgacttctacgcgctgaa ccacttcaccaccagg ttcgtgatgcacgcgcgccagaacggcagccgctacgacgcggaccgcgacgtccagttcct gcaggacatcacctgcctgagctcccccagccgcctggccgtcctgccctggggggagcgca aggtgctcaggtggatccagaagaactacggagacgtggacgtgtacatcacggccagtggc atcgatgaccagtctctggaaaatgatgagctcagaaaatacttggagaaatacatcca ggaggctctgaaagcacacctaattgataaagtcaaagtcaaaggctattatgcattcaaac tgactgaagaaaaatctaaacccagatttggattcttcacgtctgaattcaaagctaaatcc tcagttcagctttacaacaaactgatcagcaacag tggcttcccttctgagaacaggagtcc tagatgcagtgagactcaaagaaacacagagtgcatggtctgcttatttcttgtgcaaaaga aaccactgatattctttagttgttgcttcttctctaccctggttctactttcatcgattacc attcttcataagcgaaagagaagaaaatttggaaagcaaagaacttacaacatataccatt aaag tgaggccacagaaagttcttagtgaaactgatcctatttctgtctgcatgatagaaag tctaaaaattcactccagtcccaaatactggtaacatagaagacaatttgaaacactagtag taaccaaggtgatgacaatcaaggtctctgctgtgtggtccaaatgaattttccattaggtg ttgacatcactgaatacagtttttagatctgaagactaagat ctagagagtaagctaggatt atctgatacaatgcttcattaagtttaataagctgttatccatcattcttctctggcttcct tctagaaataccaatagctaattatagcaacttagaaaaaagtgcaacttttgctagactcc atagcagaaatctaaaactcttaacactggatattcagtgattattctatcacttctaacaa gatatacaatagggtaaatagtgctcctttatcatccattcc agcactttttttttccagcatagactcttaaacacattgatcctagtttttctcaatagaaa taaaaaatcatttagaaaacatggaattttgtgaggtctctccttgcattagatctgagttt tttttaaaaaaagacttaacttccataacccatctcatgggaagatca caggactaa gatt aaggagagttagacccatcaactgcctgaggagacagcactcaacctcacagtacagcaaat tccttgggacaaactgacagcaatcttccgcacttggattgttgaggcagcacacaagatct taacatacttaggaaagttaaatattctaaaaagatgtaaagttttattttattatcaagt cttcaaaggaccatattattccata agacttgctctctcctgagttccactcttctgacact atgtgtatatggggacactcaaactgcaccttgacattgcaactttggatacaattcagaat gtaaatgtttgaaggacttaaaactttctccactgcaccttttgaagctgggattaagtaaa tacgaactgggagtttgacttttttgaactctgtgcttgatttattcact gtattctaaatt ttaaggaaaacctgaatgtaaacccattcataccctttctttgggttagtaaacatttaacc acccatttca (SEQ ID NO: 413).

[0057] The amino acid sequence of the human/mouse Klotho beta chimeric protein (human KLB (M1-F508) - mouse KLB (P507-S1043)) is provided below: THLKNVSST NGSSDSYIFLEKDLSALDFIGVSFYQFSISWPRLFPDGIVTVANAKGLQYYSTLLDALVLRN IEPIVTLYHWDLPLALQEKYGGWKNDTIIDIFNDYATYCFQMFGDRVKYWITIHNPYLVAWH GYGTGMHAPGEKGNLAAVYTVGHNLIKAHSKVWHNYNTHFRPHQKGWLSITLGSHWIEPNRS ENTMDIFK CQQSMVSVLGWFANPIHGDGDYPEGMRKKLFSVLPIFSEAEKHEMRGTADFFAF SFGPNNFKPLNTMAKMGQNVSLNLREALNWIKLEYNNPRILIAENGWFTDSRVKTEDTTAIY MMKNFLSQVLQAIRLDEIRVFGYTAWSLLDGFEWQDAYTIRRGLFYVDFNSKQKERKPKSSA HYYKQIIRENGFPLKESTPDMKGRFPCDFS WGVTESVLKPEFTVSSPQFTDPHLYVWNVTGN RLLYRVEGVRLKTRPSQCTDYVSIKKRVEMLAKMKVTHYQFALDWTSILPTGNLSKVNRQVL RYYRCVVSEGLKLGVFPMVTLYHPTHSHLGLPLLSSGGWLNMNTAKAFQDYAELCFRELG DLVKLWITINEPNRLSDMYNRTSNDTYRAAHNLMIAHAQV WHLYDRQYRPVQHGAVSLSLHC DWAEPANPFVDSHWKAAERFLQFEIAWFADPLFKTGDYPSVMKEYIASKNQRGLSSSVLPRF TAKESRLVKGTVDFYALNHFTTRFVIHKQLNTNRSVADRDVQFLQDITRLSSPSRLAVTPWG VRKLLAWIRRNYRDRDIYITANGIDDLALEDDQIRKYYLEKYVQEALKAYLIDKVKIKGYYA FKLTEEKS KPRFGFFTSDFRAKSSVQFYSKLISSSGLPAENRSPACGQPAEDTDCTICSFLV EKKPLIFFGCCFISTLAVLLSITVFHHQKRRKFQKARNLQNIPLKKGHSRVFS (SEQ ID NO:374).

[0058] A nucleic acid sequence encoding the human/mouse beta Klotho chimeric protein is provided below: ATGAAGCCAGGCTGTGCGGCAGGATCTCCAGGGAATGAATGGATTTTCTTCAGCACTGATGA AATAACCACACGCTATAGGAATACAATGTCCAACGGGGGATTGCAAAGATCTGTCATCCTGT CAGCACTTATTCTGCTACGAGCTGTTACTGGATTCTCTGGAGATGGAAGAGCTATATGGTCT A AAAATCCTAATTTTACTCCGGTAATGAAAGTCAGCTGTTTCTCTATGACACTTTCCCTAA AAACTTTTTCTGGGGTATTGGGACTGGAGCATTGCAAGTGGAAGGGAGTTGGAAGAAGGATG GAAAAGGACCTTCTATATGGGATCATTTCATCCACACACCTTAAAAATGTCAGCAGCACG AATGGTTCCAGTGACAGTTATATTTTTCTGGAAAAAGACTTATCAGCCCTGGATTTTATAGG AGTTTCTTTTTATCA ATTTTCAATTTCCTGGCCAAGGCTTTTCCCCGATGGAATAGTAACAG TTGCCAACGCAAAAGGTCTGCAGTACTACAGTACTCTTCTGGACGCTCTAGTGCTTAGAAAC ATTGAACCTATAGTTACTTTATACCACTGGGATTTGCCTTTGGCACTACAAGAAAAATATGG GGGGTGGAAAAATGATACCATAATAGATATCTTCAATGACTATGCCACATACTGTTTCCAGA TGTTTGGGGACCGTGTCAAATATTGG ATTACAATTCACAACCCATATCTAGTGGCTTGGCAT GGGTATGGGACAGGTATGCATGCCCCTGGAGAGAAGGGAAATTTAGCAGCTGTCTACACTGT GGGACACAACTTGATCAAGGCTCACTCGAAAGTTTGGCATAACTACAACACACATTTCCGCC CACATCAGAAGGGTTGGTTATCGATCACGTTGGGATCTCATTGGATCGAGCCAAACCGGTCG GAAAACACGATGGATATATTCAAATGTCAACAATCCATGG TTTCTGTGCTTGGATGGTTTGC CAACCCTATCCATGGGGATGGCGACTATCCAGAGGGGATGAGAAAGAAGTTGTTCTCCGTTC TACCCATTTTCTCTGAAGCAGAGAAGCATGAGATGAGAGGCACAGCTGATTTCTTTGCCTTT TCTTTTGGACCCAACAACTTCAAGCCCCTAAACACCATGGCTAAAATGGGACAAAATGTTTC ACTTAATTTAAGAGAAGCGCTGAACTGGATTAACTGGAATACAACAACC CTCGAATCTTGA TTGCTGAGAATGGCTGGTTCACAGACAGTCGTGTGAAAACAGAAGACACCACGGCCATCTAC ATGATGAAGAATTTCCTCAGCCAGGTGCTTCAAGCAATAAGGTTAGATGAAATACGAGTGTT TGGTTATACTGCCTGGTCTCCTGGATGGCTTTGAATGGCAGGATGCTTACACCATCCGCC GAGGATTATTTTATGTGGATTTTAACAGTAAACAGAAAGAGCGGAAACCTAAGTCTT CAGCA CACTACTACAAACAGATCATACGAGAAAATGGTTTTCCTTTGAAAGAGTCCACGCCAGACAT GAAGGGTCGGTTCCCCTGTGATTTCTCTTGGGGAGTCACTGAGTCTGTTCTTAAGCCCGAGT TTACGGTCTCCTCCCCGCAGTTTACCGATCCTCACCTGTATGTGTGGAATGTCACTGGCAAC AGATTGCTCTACCGAGTGGAAGGGGTAAGGCTGAAAACAAGACCATCCCAGTGCACAGATTA TGTGAGCATCAAAAAACGAGTTGAAATGTTGGCAAAAATGAAAGTCACCCACTACCAGTTTG CTCTGGACTGGACCTCTATCCTTCCCACTGGCAATCTGTCCAAAGTTAACAGACAAGTGTTA AGGTACTATAGGTGTGTGGTGAGCGAAGGACTGAAGCTGGGCGTCTTCCCCATGGTGACGTT ACCCACTCCCATCTCGGCCTCCCCCTGCCACTTCTGAGCAGTGGGGGGTGGC TAAACATGAACACAGCCAAGGCCTTCCAGGACTACGCTGAGCTGTGCTTCCGGGAGTTGGGG GACTTGGTGAAGCTCTGGATCACCATCAATGAGCCTAACAGGCTGAGTGACATGTACAACCG CACGAGTAATGACACCTACCGTGCAGCCCACAACCTGATGATCGCCCATGCCCAGGTCTGGC CAGTATAGGCCGGTCCAGCATGGGGCTGTGTCGCTGTCCTTACATTGC GACTGGGCAGAACCTGCCAACCCCTTTGTGGATTCACACTGGAAGGCAGCCGAGCGCTTCCT CCAGTTTGAGATCGCCTGGTTTGCAGATCCGCTCTTCAAGACTGGCGACTATCCATCGGTTA TGAAGGAATACATCGCCTCCAAGAACCAGCGAGGGCTGTCTAGCTCAGTCCTGCCGCGCTTC ACCGCGAAGGAGAGCAGGCTG GTGAAGGGTACCGTCGACTTCTACGCACTGAACCACTTCAC TACGAGGTTCGTGATACACAAGCAGCTGAACACCAACCGCTCAGTTGCAGACAGGGACGTCC AGTTCCTGCAGGACATCACCCGCCTAAGCTCGCCCAGCCGCCTGGCTGTAACACCCTGGGGA GTGCGCAAGCTCCTTGCGTGGATCCGGAGGAACTACAGAGACAGGGATATCTACATCACAGC CAATGGCATCGATGACCTGGCTCTAGA GGATGATCAGATCCGAAAGTACTACTTGGAGAAGT ATGTCCAGGAGGCTCTGAAAGCATATCTCATTGACAAGGTCAAAATCAAAGGCTACTATGCA TTCAAACTGACTGAAGAGAAATCTAAGCCTAGATTTGGATTTTTCACCTCTGACTTCAGAGC TAAGTCCTCTGTCCAGTTTTACAGCAAGCTGATCAGCAGCAGTGGCCTCCCCGCTGAGAACA ACAGACTGCACCATTTGCTCATTTCTCGTG GAGAAGAAACCACTCATCTTCTTCGGTTGCTGCTTCATCTCCACTCTGGCTGTACTGCTATC CATCACCGTTTTTCATCATCAAAAGAGAAGAAAATTCCAGAAAGCAAGGAACTTACAAAATA TACCATTGAAGAAAGGCCACAGCAGAGTTTTCAGCTGA (SEQ ID NO:375)

[0059] The amino acid sequence of the rat/human Klotho beta chimeric protein (rat KLB (M1-F506) - human KLB (S509-S1044)) is provided below: WDRYVYSHLRGVNGT DRSTDSYIFLEKDLLALDFLGVSFYQFSISWPRLFPNGTVAAVNAQGLRYYRALLDSLVLRN IEPIVTLYHWDLPLTLQEEYGGWKNATMIDLFNDYATYCFQTFGDRVKYWITIHNPYLVAWH GFGTGMHAPGEKGNLTAVYTVGHNLIKAHSKVWHNYDKNFRPHQKGWLSITLGSHWIEPNRT DNMEDVINCQHSMSSVLGWFANPIHGDGDYPEFMKTGAMIPEFSEAEKEEVRGTADFFAFSF GPNNFRPSNTVVKMGQNVSLNLRQVLNWIKLEYDDPQILISENGWFTDSYIKTEDTTAIYMM KNFLNQVLQAIKFDEIRVFGYTAWTLLDGFEWQDAYTTRRGLFYVDFNSEQKERKPKSSAHY YKQIIQDNGFSLKEST PDVQGQFPCDFSWGVTESVLKPESVASSPQFSDPHLYVWNATGNRL LHRVEGVRLKTRPAQCTDFVNIKKQLEMLARMKVTHYRFALDWASVLPTGNLSAVNRQALRY YRCVVSEGLKLGISAMVTLYYPTHAHLGLPEPLLHADGWLNPSTAEAFQAYAGLCFQELGDL VKLWITINEPNRLSDIYNRSGNDTYGAAHNLLVAHALAWRLYDRQFRPSQRGAVSLSLHADW AEPANPYADSHWRAAERFLQFEIAWFAEPLFKTGDYPAAMREYIASKHRRGLSSSALPRLTE AERRLLKGTVDFCALNHFTTRFVMHEQLAGSRYDSDRDIQFLQDITRLSSPTRLAVIPWGVR KLLRWVRRNYGDMDIYITASGIDDQ ALEDDRLRKYYLGKYLQEVLKAYLIDKVRIKGYYAFK LAEEKSKPRFGFFTSDFKAKSSIQFYNKVISSRGFPFENSSSRCSQTQENTECTVCLFLVQK KPLIFLGCCFFSTLVLLLSIAIFQRQKRRKFWKAKNLQHIPLKKGKRVVS (SEQ ID NO:376)

[0060] A nucleic acid sequence encoding the rat/human Klotho beta chimeric protein is provided below: ATGAAGACAGGCTGTGCAGCAGGGTCTCCGGGGAATGAATGGATTTTCTTCAGCTCTGATGA AAGAAACACACGCTCTAGGAAAACAATGTCCAACAGGGCACTGCAAAGATCTGCCGTGCTGT CTGCGTTTGTTCTGCTGCGAGCTGTTACCGGCTTCTCCGGAGACGGGAAAGCAATATGGG AT AAAAAACAGTACGTGAGTCCGGTAAACCCAAGTCAGCTGTTCCTCTATGACACTTTCCCTAA AAACTTTTCCTGGGGCGTTGGGACCGGAGCATTTCAAGTGGAAGGGAGTTGGAAGACAGATG GAAGAGGACCCTCGATCTGGGATCGGTACGTCTACTCACACCTGAGAGGTGTCAACGGCACA GACAGATCCACTGACAGTTACATCTTTCTGGAAAAAGACTTGTTGGCTCTGGATTTTTTAGG AGTTTC TTTTTATCAGTTCTCAATCTCCTGGCCACGGTTGTTTCCCAATGGAACAGTAGCAG CAGTGAATGCGCAAGGTCTCCGGTACTACGTGCACTTCTGGACTCGCTGGTACTTAGGAAT ATCGAGCCCATTGTTACCTTGTACCATTGGGATTTGCCTCTGACGCTCCAGGAAGAATATGG GGGCTGGAAAAATGCAACTATGATAGATCTCTTCAACGACTATGCCACATACTGCTTCCAGA CCTTTGGAGACCGT GTCAAATATTGGATTACAATTCACAACCCTTACCTTGTTGCTTGGCAT GGGTTTGGCACAGGTATGCATGCACCAGGAGAGAAGGGAAATTTAACAGCTGTCTACACTGT GGGACACAACCTGATCAAGGCACATTCGAAAGTGTGGCATAACTACGACAAAAACTTCCGCC CTCATCAGAAGGGTTGGCTCTCCATCACCTTGGGGTCCCATTGGATAGAGCCAAACAGAACA GACAACATGGAGGACGTGATCAACTGCCA GCACTCCATGTCCTCTGTGCTTGGATGGTTCGC CAACCCCATCCACGGGGACGGCGACTACCCTGAGTTCATGAAGACGGGCGCCATGATCCCCG AGTTCTCTGAGGCAGAGAAGGAGGAGGTGAGGGGCACGGCTGATTTCTTTGCCTTTTTCCTTC GGGCCCAACAACTTCAGGCCCTCAAACACCGTGGTGAAAATGGGACAAAATGTATCACTCAA CTTAAGGCAGGTGCTGAACTGGATTAACTGGAATACGATGA CCCTCAAATCTTGATTTCGG AGAACGGCTGGTTCACAGATAGCTATATAAAGACAGAGGACACCACGGCCATCTACATGATG AAGAATTTCCTAAACCAGGTTCTTCAAGCAATAAAATTTGATGAAATCCGCGTGTTTGGTTA TACGGCCTGGACTCTCCTGGATGGCTTTGAGTGGCAGGATGCCTATACGACCCGACGAGGGC TGTTTTATGTGGACTTTAACAGTGAGCAGAAAGAGAGGAAACCCAAGTCC TCGGCTCATTAC TACAAGCAGATCATACAAGACAACGGCTTCTCTTTAAAAGAGTCCACGCCAGATGTGCAGGG CCAGTTTCCCTGTGACTTCTCCTGGGGTGTCACTGAATCTGTTCTTAAGCCCGAGTCTGTGG CTTCGTCCCCACAGTTCAGCGATCCTCATCTGTACGTGTGGAACGCCACTGGCAACAGACTG TTGCACCGAGTGGAAGGGGTGAGGCTGAAAACACGACCCGCTCAATGCACAGATTTTGTAAA CATCAAAAAACAACTTGAGATGTTGGCAAGAATGAAAGTCACCCACTACCGGTTTGCTCTGG ATTGGGCCTCGGTCCTTCCCACTGGCAACCTGTCCGCGGTGAACCGACAGGCCCTGAGGTAC TACAGGTGCGTGGTCAGTGAGGGGCTGAAGCTTGGCATCTCCGCGATGGTCACCCTGTATTA TCC GACCCACGCCCACCTAGGCCTCCCCGAGCCTCTGTTGCATGCCGACGGGTGGCTGAACC CATCGACGGCCGAGGCCTTCCAGGCCTACGCTGGGCTGTGCTTCCAGGAGCTGGGGGACCTG GTGAAGCTCTGGATCACCATCAACGAGCCTAACCGGCTAAGTGACATCTACAACCGCTCTGG CAACGACACCTACGGGGCGGCGCACAACCTGCTGGTGGCCCACGCCCTGGCCTGGCGCCTCT AC GACCGGCAGTTCAGGCCCTCACAGCGCGGGGCCGTGTCGCTGTCGCTGCACGCGGACTGG GCGGAACCCGCCAACCCCTATGCTGACTCGCACTGGAGGGCGGCCGAGCGCTTCCTGCAGTT CGAGATCGCCTGGTTCGCCGAGCCGCTCTTCAAGACCGGGGACTACCCCGCGGCCATGAGGG AATACATTGCCTCCAAGCACCGACGGGGGCTTTCCAGCTCGGCCCTGCCGCGCCTCACCGAG GCCG AAAGGAGGCTGCTCAAGGGCACGGTCGACTTCTGCGCGCTCAACCACTTCACCACTAG GTTCGTGATGCACGAGCAGCTGGCCGGCAGCCGCTACGACTCGGACAGGGACATCCAGTTTC TGCAGGACATCACCCGCCTGAGCTCCCCCACGCGCCTGGCTGTGATTCCCTGGGGGGTGCGC AAGCTGCTGCGGTGGGTCCGGAGGAACTACGGCGACATGGACATTTACATCACCGCCAGTGG CATCGACGACC AGGCTCTGGAGGATGACCGGCTCCGGAAGTACTACCTAGGGAAGTACCTTC AGGAGGTGCTGAAAGCATACCTGATTGATAAAGTCAGAATCAAAGGCTATTATGCATTCAAA CTGGCTGAAGAGAAATCTAAACCCAGATTTGGATTCTTCACATCTGATTTTAAAGCTAAATC CTCAATACAATTTTACAACAAAGTGATCAGCAGCAGGGGCTTCCCTTTTGAGAACAGTAGTT CTAGATGCAGTCAGACCCAA GAAAATACAGAGTGCACTGTCTGCTTATTCCTTGTGCAGAAG AAACCACTGATATTCCTGGGTTGTTGCTTCTTCTCCACCCTGGTTCTACTCTTATCAATTGC CATTTTTCAAAGGCAGAAGAGAAGAAAGTTTTGGAAAGCAAAAAACTTACAACACATACCAT TAAAGAAAGGCAAGAGAGTTGTTAGCTAG (SEQ ID NO:377)

[0061] Related Klotho beta polypeptides include allelic variants (e.g., SNP variants); splice variants; fragments; derivatives; substitution, deletion and insertion variants; fusion polypeptides; and preferably cross-species homologues which retain Klotho beta activity and/or are sufficient to generate an anti-Klotho beta immune response. As those skilled in the art will appreciate, an anti-Klotho beta antibody provided herein can bind to a Klotho beta polypeptide, a Klotho beta polypeptide fragment, a Klotho beta antigen, and/or a Klotho beta epitope. An epitope may be part of a larger Klotho beta antigen, which may be part of a larger Klotho beta polypeptide fragment, which, in turn, may be part of a larger Klotho beta polypeptide. Klotho beta can exist in a native or denatured form. The Klotho beta polypeptides described herein can be isolated from a variety of sources, such as from human tissue types or other sources, or prepared by recombinant or synthetic methods. A Klotho beta polypeptide may comprise a polypeptide having the same amino acid sequence as a corresponding naturally derived Klotho beta polypeptide. Klotho Beta polypeptides encompass truncated or secreted forms of a Klotho beta polypeptide (e.g., an extracellular domain sequence), variant forms (e.g., alternatively spliced forms), and allelic variants of the polypeptide. Orthologs to the Klotho beta polypeptide are also well known in the art.

[0062] The term "Klotho beta" encompasses "full-length", untransformed Klotho beta, as well as any form of Klotho beta that results from processing in the cell. The term also encompasses naturally occurring variants or mutations of Klotho beta (e.g., splice variants, allelic variants, SNP variants, and isoforms). The Klotho beta polypeptides described herein can be isolated from a variety of sources, such as from human tissue types or other sources, or prepared by recombinant or synthetic methods.

[0063] The terms "FGF19-like signaling" and "induces FGF19-like signaling", when applied to a binding protein such as an antibody that binds to Klotho beta of the present disclosure, means that the binding protein (e.g., antibody) mimics, or modulates, an in vivo biological effect induced by binding of (i) Klotho beta; (ii) FGFR1c, FGFR2c, FGFR3c, and FGFR4; or (iii) a complex comprising Klotho beta and one of FGFR1c, FGFR2c, FGFR3c, and FGFR4 and induces a biological response that would otherwise result from binding of FGF19 to (i) Klotho beta; (ii) FGFR1c, FGFR2c, FGFR3c or FGFR4; or (iii) a complex comprising Klotho beta and one of FGFR1c, FGFR2c, FGFR3c, and FGFR4 in vivo. When evaluating the binding and specificity of the Anti-Klotho beta antibody, e.g., an antibody or fragment thereof, which binds to Klotho beta (e.g., human Klotho beta), an antibody or fragment thereof is considered to induce a biological response when the answer is equal to or greater than 5%, and preferably equal to or greater than 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60 %, 65%, 70%, 75%, 80%, 85%, 90% or 95%, of the activity of a wild-type FGF19 standard comprising the mature form of SEQ ID NO: 304 (e.g., the mature form of human FGF19) and has the following properties: exhibiting an efficacy level equal to or greater than 5% of a FGF19 standard, with an EC50 equal to or less than 100 nM, e.g., 90 nM, 80 nM, 70 nM, 60 nM, 50 nM, 40 nM, 30 nM, 20 nM or 10 nM in (1) a recombinant FGF19 receptor-mediated luciferase reporter cell assay (see, for example, Example 4); (2) ERK-phosphorylation in a recombinant FGF19 receptor-mediated cellular assay (see, for example, Example 4); or (3) ERK-phosphorylation in human adipocytes (see, for example, Example 5).

[0064] The term "FGF19R" may refer to a multimeric receptor complex that FGF19 is known or suspected of forming in vivo. In various embodiments, FGF19R comprises (i) an FGFR, e.g., FGFR1c, FGFR2c, FGFR3c or FGFR4, and (ii) Klotho beta.

[0065] The terms "FGF21-like signaling" and "induces FGF21-like signaling", when applied to a binding protein such as an antibody that binds to Klotho beta of the present disclosure, means that the binding protein (e.g., antibody) mimics, or modulates, an in vivo biological effect induced by binding of (i) Klotho beta; (ii) FGFR1c, FGFR2c, FGFR3c, and FGFR4; or (iii) a complex comprising Klotho beta and one of FGFR1c, FGFR2c, FGFR3c, and FGFR4 and induces a biological response that would otherwise result from binding of FGF21 to (i) Klotho beta; (ii) FGFR1c, FGFR2c, FGFR3c or FGFR4; or (iii) a complex comprising Klotho beta and one of FGFR1c, FGFR2c, FGFR3c, and FGFR4 in vivo. When evaluating the binding and specificity of the Anti-Klotho beta antibody, e.g., an antibody or fragment thereof, which binds to Klotho beta (e.g., human Klotho beta), an antibody or fragment thereof is considered to induce a biological response when the answer is equal to or greater than 5%, and preferably equal to or greater than 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60 %, 65%, 70%, 75%, 80%, 85%, 90% or 95%, of the activity of a wild-type FGF21 standard comprising the mature form of SEQ ID NO: 306 or 429 (e.g., the mature form of human FGF21) and has the following properties: exhibiting an efficacy level equal to or greater than 5% of a standard FGF21, with an EC50 equal to or less than 100 nM, e.g. 90 nM, 80 nM, 70 nM , 60 nM, 50 nM, 40 nM, 30 nM, 20 nM or 10 nM in (1) a recombinant FGF21 receptor-mediated luciferase reporter cellular assay (see, for example, Example 4); (2) ERK-phosphorylation in a recombinant FGF21 receptor-mediated cellular assay (see, for example, Example 4); or (3) ERK-phosphorylation in human adipocytes (see, for example, Example 5).

[0066] The term "FGF21R" may refer to a multimeric receptor complex that FGF19 is known or suspected of forming in vivo. In various embodiments, FGF21R comprises (i) an FGFR, e.g., FGFR1c, FGFR2c, FGFR3c or FGFR4, and (ii) Klotho beta.

[0067] The term "binding protein" refers to a protein comprising a portion (e.g., one or more binding regions, such as CDRs) that binds to Klotho beta, including human and/or Klotho beta. cyno and, optionally, a framework portion (e.g., one or more framework regions) that allows the binding portion to adopt a conformation that promotes binding of the binding protein to a Klotho beta polypeptide, fragment or epitope. Examples of such binding proteins include antibodies, such as a human antibody, a humanized antibody; a chimeric antibody; a recombinant antibody; a single chain antibody; a diabody; a tribody; a tetrabody; a Fab fragment; an F(ab’)2; an IgD antibody; an IgE antibody; an IgM antibody; an IgG1 antibody; an IgG2 antibody; an IgG3 antibody; or an IgG4 antibody, and fragments thereof. The binding protein may comprise, for example, an alternative protein scaffold or artificial scaffold with grafted CDRs or CDR derivatives. Such scaffolds include, but are not limited to, antibody-derived scaffolds comprising mutations introduced to, for example, stabilize the three-dimensional structure of the binding protein, as well as entirely synthetic scaffolds comprising, for example, a biocompatible polymer. See, for example, Korndorfer et al., 2003, Proteins: Structure, Function, and Bioinformatics, 53(1):121-129 (2003); Roque et al., Biotechnol. Program. 20:639-654 (2004). Additionally, antibody peptide mimetics ("PAMs") can be used, as well as antibody mimetic-based scaffolds that utilize fibronectin components as a scaffold. In the context of the present disclosure, a binding protein is said to specifically bind or selectively bind to Klotho beta, for example, when the dissociation constant (KD) is < 10-8 M. The binding protein (e.g., antibodies ) can specifically bind to Klotho beta with high affinity when KD is < 10-9 M or KD is < 10-10 M. In some embodiments, binding proteins (e.g., antibodies) can bind to Klotho beta or a complex comprising FGFR1c and Klotho beta, including with a KD of between about 10-7 M and about 10-12 M and in other embodiments, binding proteins (e.g., antibodies) can bind with a KD of 1 -2x10-9 M.

[0068] The term "antibody" and "immunoglobulin" or "Ig" are used interchangeably herein, and is used in the broadest sense and specifically encompasses, for example, monoclonal individual Anti-Klotho beta antibodies (including agonist antibodies, antagonist antibodies , neutralizing antibodies, full-length or intact monoclonal antibodies), Anti-Klotho beta antibody compositions with polyepitopic or monoepitopic specificity, polyclonal or monovalent antibodies, multivalent antibodies, multispecific antibodies (e.g., bispecific antibodies, provided that they exhibit biological activity desired), formed from at least two intact antibodies, single-chain Anti-Klotho beta antibodies, and anti-Klotho-beta antibody fragments, as described below. An antibody may be human, humanized, chimeric and/or affinity matured, as well as an antibody from other species, e.g., mouse, rabbit, etc. The term "antibody" is intended to include a B cell polypeptide product within the immunoglobulin class of polypeptides that is capable of binding to a specific molecular antigen and is composed of two identical pairs of polypeptide chains, wherein each pair has a heavy chain (about 50-70 kDa) and a light chain (about 25 kDa) and each amino-terminal portion of each chain includes a variable region of about 100 to about 130 or more amino acids and each carboxy portion -terminal of each chain includes a constant region (See, Borrebaeck (ed.) (1995) Antibody Engineering, Second Ed., Oxford University Press.; Kuby (1997) Immunology, Third Ed., W.H. Freeman and Company, New York) . In specific embodiments, the specific molecular antigen can be bound by an antibody provided herein and includes a Klotho beta polypeptide, Klotho beta fragment or Klotho beta epitope. Antibodies include, but are not limited to, synthetic antibodies, monoclonal antibodies, recombinantly produced antibodies, multispecific antibodies (including bispecific antibodies), human antibodies, humanized antibodies, camelized antibodies, chimeric antibodies, intrabodies, anti-idiotypic antibodies (anti-Id ), and functional fragments (e.g., antigen-binding fragments such as Klotho beta-binding fragments) of any of the above, which refers to a portion of an antibody heavy or light chain polypeptide that retains some or all the binding activity of the antibody from which the fragment was derived. Examples of functional fragments (e.g., antigen-binding fragments, such as Klotho beta-binding fragments) include single-chain Fvs (scFv) (e.g., including monospecific, bispecific, etc.), Fab fragments, F(ab '), F(ab)2 fragments, F(ab')2 fragments, disulfide-linked Fv (sdFv), Fd fragments, Fv fragments, diabody, triabody, tetrabody and minibody. In particular, the antibodies provided herein include immunoglobulin molecules and immunologically active portions of immunoglobulin molecules, e.g., antigen-binding domains or molecules that contain an antigen-binding site that binds to a Klotho beta antigen (e.g. , one or more complementarity determining regions (CDRs) of an anti-Klotho beta antibody). Such antibody fragments can be found described in, for example, Harlow and Lane, Antibodies: A Laboratory Manual, Cold Spring Harbor Laboratory, New York (1989); Myers (ed.), Molec. Biology and Biotechnology: A Comprehensive Desk Reference, New York: VCH Publisher, Inc.; Huston et al., Cell Biophysics, 22:189-224 (1993); Plückthun and Skerra, Meth. Enzymol., 178:497-515 (1989) and in Day, E.D., Advanced Immunochemistry, Second Ed., Wiley-Liss, Inc., New York, NY (1990). The antibodies provided herein may be of any type (e.g., IgG, IgE, IgM, IgD, IgA, and IgY), any class (e.g., IgG1, IgG2, IgG3, IgG4, IgA1, and IgA2), or any subclass (e.g., example, IgG2a and IgG2b) of the immunoglobulin molecule. Anti-Klotho beta antibodies may be agonist antibodies or antagonist antibodies. Provided herein are Klotho beta agonist antibodies, including antibodies that induce FGF19-like signaling and/or FGF21-like signaling. Preferred Klotho beta agonist antibodies do not compete for the binding of FGF19 and/or FGF21 to an FGF receptor, including, for example, FGFR1c, FGFR2c, FGFR3c or FGFR4c.

[0069] The term "fibroblast growth factors" refers to a family of growth factors, including the twenty-two members of the human FGF family. The FGF19 subfamily of fibroblast growth factors consists of human FGF21, FGF23, and FGF19 and rat FGF15. The effects of FGF family members are the result of their heparin-dependent binding to one or more members of the FGF receptor tyrosine kinase (FGFR) family, which includes four members, each having a tyrosine kinase domain, FGFR1. , FGFR2, FGFR3 and FGFR4, as well as two splice variants each of FGFR1, FGFR2 and FGFR3. These splice variants, which occur in exon 3 of FGFR1, FGFR2, and FGFR3, are designated as "b" and "c" variants (e.g., FGFR1b, FGFR2b, FGFR3c, FGFR1c, FGFR2c, and FGFR3c, which are also known as FGFR1 (III)b, FGFR2(III)b, FGFR3(III)c, FGFR1(III)c, FGFR2(III)c and FGFR3(III)c, respectively). For example, FGF19 targets and has effects on both adipocytes and hepatocytes. Mice treated with recombinant human FGF19 (rhFGF19), despite being on a high-fat diet, show increased metabolic rate, increased lipid oxidation, a lower respiratory quotient, and weight loss. Furthermore, these mice had lower serum levels of leptin, insulin, cholesterol and triglycerides, and normal blood glucose levels, despite the high-fat diet and no decrease in appetite. Furthermore, obese mice that lacked leptin but included an FGF19 transgene showed weight loss, decreased cholesterol and triglycerides, and did not develop diabetes. Furthermore, obese diabetic mice that lacked leptin, when injected with rhFGF19, showed reversal of their metabolic characteristics, in the form of weight loss and reduced blood glucose. For example, FGF21 is primarily expressed by the liver and has metabolic effects similar to those of FGF19, such as increased metabolism through its effects on adipose tissue, weight loss, decreased blood glucose levels, and resistance to obesity and diabetes. FGF21 transgenic mice were also resistant to diet-induced obesity, and, in diabetic rodent models, administration of FGF21 reduced blood glucose and triglyceride levels. The metabolic effects of FGF19 and FGF21 occur through their binding to FGF receptors, including the FGFR1c, FGFR2c, and FGFR3c receptors, and required Klotho beta for binding. The bindings of FGF19 and FGF21 to FGFR1c and FGFR2c are significant. FGF19 has also been shown to have metabolic effects distinct from FGF21, including regulating bile production by the liver through its liver-specific effects, negatively regulating bile production in response to postprandial bile production, and mitogenic effects in the liver that are not observed in relation to FGF21. For example, FGF19 transgenic mice develop hepatic adenocarcinoma due to increased hepatocyte proliferation and dysplasia, and rhFGF19-treated mice exhibit hepatocyte proliferation of hepatocytes. These additional activities of FGF19 appear to be mediated through its binding to FGFR4. FGF19 can bind to FGFR4 in both a Klotho beta-dependent and Klotho beta-independent manner. Although FGF21 has also been shown to bind FGFR4 in a Klotho beta-dependent manner, efficient signaling has not previously been observed from FGF21 binding to FGFR4.

[0070] Binding proteins, such as anti-Klotho beta antibodies, as described herein, can induce FGF19-like signaling, as described herein. In vivo, the mature form of FGF19 is the active form of the molecule. A nucleic acid sequence encoding full-length FGF19 is provided below; the nucleotides encoding the signal sequence are underlined. atgcggagcgggtgtgtggtggtccacgtatggatcctggccggcctctggctggccgtggc cgggcgccccctcgccttctcggacgcggggccccacgtgcactacggctggggcgacccca tccgcctgcggcacctgtacacctccggcccccacgggctctccagctgcttcctgcgcatc cgtgccga cggcgtcgtggactgcgcgcggggccagagcgcgcacagtttgctggagatcaa ggcagtcgctctgcggaccgtggccatcaagggcgtgcacagcgtgcggtacctctgcatgg gcgccgacggcaagatgcaggggctgcttcagtactcggaggaagactgtgctttcgaggag cagatggctacaatgtgtaccgatccgagaagcaccgcctcccggtctccct gagcagtgccaaacagcggcagctgtacaagaacagaggctttcttccactctctcatttcc tgcccatgctgcccatggtcccagaggagcctgaggacctcaggggccacttggaatctgac atgttctcttcgcccctggagaccgacagcatggacccatttgg gcttgtcaccggactgga ggccgtgaggagtcccagctttgagaagtaa (SEQ ID NO:303)

[0071] A full-length amino acid sequence of FGF19 is provided; the amino acids that form the signal sequence are underlined: mrsgcvvvhvwilaglwlavagRPLAFS DAGPH VH YGWGDPIRLRHLYTSGPHGLSSCFLRI RADGVVDCARGQSAHSLLEIKAVALRTVAIKGVHSVRYLCMGADGKMQGLLQYSEEDCAFEE EIRPDGYNVYRSEKHRLPVSLSSAKQRQLYKNRGFLPLSHFLPMLPMVPEEP EDLRGHLESD MFSSPLETDSMDPFGLVTGLEAVRSPSFEK (SEQ ID NO:304)

[0072] Binding proteins, such as anti-Klotho beta antibodies, as described herein, can induce FGF21-like signaling, as described herein. In vivo, the Mature form of FGF21 is the active form of the molecule. A nucleic acid sequence encoding a full-length FGF21 is provided; the nucleotides encoding the signal sequence are underlined: atg gac tcg gac gag acc ggg ttc gag cac tca gga ctg tgg gtt tct gtg ctg gct ggt ctt ctg ctg gga gcc tgc cag gca cac ccc atc cct gac tcc agt cct ctc ctg caa ttc ggg ggc caa gtc cgg cag cgg tac ctc tac aca gat gat gcc cag cag aca gaa gcc cac ctg gag atc agg gag gat ggg acg gtg ggg ggc gct gct gac cag agc ccc gaa agt ctc ctg cag ctg aaa gcc ttg aag ccg gga gtt att caa atc ttg gga gtc aag aca tcc agg ttc ctg tgc cag cgg cca gat ggg gcc ctg tat gga tcg ctc cac ttt gac cct gag gcc tgc agc ttc cgg gag ctg ctt ctt gag gac gga tac aat gtt tac cag tcc gaa gcc cac ggc ctc ccg ctg cac ctg cca ggg aac aag tcc cca cac cgg gac cct gca ccc cga gga cca gct cgc ttc ctg cca cta cca ggc ctg ccc ccc gca ccc ccg gag cca ccc gga atc ctg gcc ccc cag ccc ccc gat gtg ggc tcc tcg gac cct ctg agc atg gtg gga cct tcc cag ggc cga agc ccc agc tac gct tcc tga (SEQ ID NO:305).

[0073] A full-length amino acid sequence of FGF21 is provided; The amino acids that form the signal sequence are underlined: (SEQ ID NO:306).

[0074] A nucleic acid sequence that also encodes full-length FGF21 is provided; the nucleotides encoding the signal sequence are underlined: atggactcggacgagaccgggttcgagcactcaggactgtgggtttctgtgctggctggtct tctgctgggagcctgccaggcaCACCCCATCCCTGAC TCCAGTCCTCTCCTGCAATTCGGGG GCCAAGTCCGGCAGCGGTACCTCTACAGATGATGCCCAGCAGACAGAAGCCCACCTGGAG ATCA GGGAGGATGGGACGGTGGGGCGCTGCTGACCAGAGCCCCGAAAGTCTCCTGCAGCT GAAAGCCTTGAAGCCGGGAGTTATTCAAATCTTGGGAGTCAAGACATCCAGGTTCCTGTGCC AGCGGCCAGATGGGGCCCTGTATGGATCGCTCCACTTTGACCCTGAGGCCTGCAGCTTCCGG GAGCTGCTTCTTGAGGACGGATACAATGTTTACCAGTCCGAAGCCCACGGCCTCCCGCTGCA CCTGCCAG GGAACAAGTCCCCACACCGGGACCCTGCACCCCGAGGACCAGCTCGCTTCCTGC CACTACCAGGCCTGCCCCCCGCACTCCCGGAGCCACCCGGAATCCTGGCCCCCCAGCCCCCC GATGTGGGCTCCTCGGACCCTCTGAGCATGGTGGGACCTTCCCAGGGCCGAAGCCCCAGCTA CGCTTCCTGA (SEQ ID NO:428).

[0075] An amino acid sequence that also encodes full-length FGF21 is provided; the amino acids encoding the signal sequence are underlined: mdsdetgfehsglwvsvlaglllgacqaHPIPDSSPLLQFGGQVRQRYLY T DDAQQTEAHLE IREDGTVGGAADQSPESLLQLKALKPGVIQILGVKTSRFLCQRPDGALYGSLHFDPEACSFR ELLLEDGYNVYQSEAHGLPLHLPGNKSPHRDPAPRGPARFLPLPGLPPALPEP PGILAPQPP DVGSSDPLSMVGPSQGRSPSYAS (SEQ ID NO:429)

[0076] Binding proteins, such as anti-Klotho beta antibodies, as described herein bind to Klotho beta alone or in complex with an FGF receptor, such as FGFR1c. A nucleic acid sequence encoding human FGFR1c (GenBank Accession Number NM 023110; also designated FGFRαIIIc) is provided below: atgtggagctggaagtgcctcctcttctgggctgtgctggtcacagcc acactctgcaccgctaggccgtccccgaccttgcctgaacaagcccag ccctggggagcccctgtggaagtggagtcct tcctggtccaccccggt gacctgctgcagcttcgctgtcggctgcgggacgatgtgcagagcatc aactggctgcgggacggggtgcagctggcggaaagcaaccgcacccg catcacaggggaggaggtggaggtgcaggactccgtgcccgcagact ccggcctctatgcttgcgtaaccagcagcccctcgg gcagtgacacca cctacttctccgtcaatgtttcagatgctctcccctcctcggaggatga tgatgatgatgatgactcctcttcagaggagaaagaaacagataaca ccaaaccaaaccgtatgcccgtagctccatattggacatcaccagaaa agatggaaagaaattgcatgcagtgccggctgccaagacagtgaag gcccttccagtgggacaccaaacccaacactgcgctggttg aaaaatggcaaagaattcaaacctgaccacagaattggaggctacaa ggtccgttatgccacctggagcatcataatggactctgtggtgccctc tgacaagggcaactacacctgcattgtggagaatgagtacggcagca tcaaccacacataccagctggatgtcgtggagcggtcc cctcaccggc ccatcctgcaagcagggttgcccgccaacaaaacagtggccctgggt agcaacgtggagttcatgtgtaaggtgtacagtgacccgcagccgcac atccagtggctaaagcacatcgaggtgaatgggagcaagattggccc agacaacctgccttatgtccagatcttgaagactgctggagttaatac caccgaca aagagatggaggtgcttcacttaagaaatgtctcctttga ggacgcaggggagtatacgtgcttggcgggtaactctatcggactctc ccatcactctgcatggttgaccgttctggaagccctggaagagaggcc ggcagtgatgacctcgcccctgtacctggagatcatcatctattgcac aggggccttcctcatctcctgcat ggtggggtcggtcatcgtctacaa gatgaagagtggtaccaagaagagtgacttccacagccagatggctg tgcacaagctggccaagagcatccctctgcgcagacaggtaacagtg tctgctgactccagtgcatccatgaactctggggttcttctggttcggc catcacggctctcctccagtgggactcccatgctagcaggggtctctg agtatgagcttcccgaagaccctcgct gggagctgcctcgggacagac tggtcttaggcaaacccctgggagagggctgctttgggcaggtggtgt tggcagaggctatcgggctggacaaggacaaacccaaccgtgtgacc aaagtggctgtgaagatgttgaagtcggacgcaacagagaaagactt gtcagacctgatctcagaaatggagatgatgaagatgatcgg gaagc ataagaatatcatcaacctgctgggggcctgcacgcaggatggtccct tgtatgtcatcgtggagtatgcctccaagggcaacctgcgggagtacc tgcaggcccggaggcccccagggctggaatactgctacaaccccagc cacaacccagaggagcagctctcctccaaggacctggtgtcctgcgcc taccaggtggcccgagg catggagtatctggcctccaagaagtgcata caccgagacctggcagccaggaatgtcctggtgacagaggacaatgt gatgaagatagcagactttggcctcgcacgggacattcaccacatcga ctactataaaaagacaaccaacggccgactgcctgtgaagtggatgg cacccgaggcattatttgaccggatctacacccaccagagtgatgtgt gg tctttcggggtgctcctgtgggagatcttcactctgggcggctcccc ataccccggtgtgcctgtggaggaacttttcaagctgctgaaggaggg tcaccgcatggacaagcccagtaactgcaccaacgagctgtacatgat gatgcgggactgctggcatgcagtgccctcacagagacccaccttcaa gcagctggtggaagacc tggaccgcatcgtggccttgacctccaacca ggagtacctggacctgtccatgcccctggaccagtactcccccagctt tcccgacacccggagctctacgtgctcctcaggggaggattccgtctt ctctcatgagccgctgcccgaggagccctgcctgccccgacacccagc ccagcttgccaatggcggactcaaacgccg ctga. (SEQ ID NO:307)

[0077] The amino acid sequence of human FGFR1c (GenBank Accession Number NP 075598) (also designated FGFRαIIIC) is provided below: MWSWKCLLFWAVLVTATLCTARPSPTLPEQAQPWGAPVEVESFLVHPG DLLQLRCRLRDDVQSINWLRDGVQLAESNRTRITGEEVEVQDSVPADSGL YACVTSSPSGSDTTYFSVNVSDALPSSEDDDDDDDS SSEEKETDNTKPNR MPVAPYWTSPEKMEKKLHAVPAAKTVKFKCPSSGTPNPTLRWLKNGKEF KPDHRIGGYKVRYATWSIIMDSVVPSDKGNYTCIVENEYGSINHTYQLDV VERSPHRPILQAGLPANKTVALGSNVEFMCKVYSDPQPHIQWLKHIEVNG SKIGPDNLPYVQILKTAGVNTTDKEMEVLHLRNVSFEDAGEY TCLAGNSIS KNIIN LLGACTQDGPLYVIVEYASKGNLREYLQARRPPGLEYCYNPSHNPEEQLS SKDLVSCAYQVARGMEYLASKKCIHRDLAARNVLVTEDNVMKIADFGL ARDIHHIDYYKKTTNGRLPVKWMAPEALFDRIYTHQSDVWSFGVLLWEI FTLGGSPYPGVPVEELFKLLKEGHRMDKPSNCTNELYMMMRDCWHAVP LVEDLDRIVALTSNQEYLDLSMPLDQYSPSFPDTRSSTCSSG EDSVFSHEPLPEEPCLPRHPAQLANGGLKRR. (SEQ ID NO:308)

[0078] Binding proteins, such as anti-Klotho beta antibodies, as described herein can bind to Klotho beta in complex with the extracellular portion of an FGF receptor such as FGFR1c. An example of an extracellular region of FGFR1c is: MWSWKCLLFWAVLVTATLCTARPSPTLPEQAQPWGAPVEVESFLVHPGDL LQLRCRLRDDVQSINWLRDGVQLAESNRTRITGEEVEVQDSVPADSGLYA CVTSSPSGSDTTYFSVNVSDALPSSEDDDDDDSSEEKETDNTKPNRMP VAPYWTSPEKMEKKLHAVPAAKTVKFKCPSSGTPNPTLR WLKNGKEFKPD HRIGGYKVRYATWSIIMDSVVPSDKGNYTCIVENEYGSINHTYQLDVVER SPHRPILQAGLPANKTVALGSNVEFMCKVYSDPQPHIQWLKHIEVNGSKI GPDNLPYVQILKTAGVNTTDKEMEVLHLRNVSFEDAGEYTCLAGNSIGLS HHSAWLTVLEALEERPAVMTSPLY. (SEQ ID NO:309)

[0079] An example of an extracellular region of FGFR1c (αIIIc) is: RPSPTLPEQAQPWGAPVEVESFLVHPGDLLQLRCRLRDDVQSINWLRDGVQLAESNRTRITG EEVEVQDSVPADSGLYACVTSSPSGSDTTYFSVNVSDALPSSEDDDDDDSSEEKETDNTK PNRMPVAPYWTSPEKMEKKLHAVPAAKTVKFKCPSSGTPNPTLRW LKNGKEFKPDHRIGGYK VRYATWSIIMDSVVPSDKGNYTCIVENEYGSINHTYQLDVVERSPHRPILQAGLPANKTVAL GSNVEFMCKVYSDPQPHIQWLKHIEVNGSKIGPDNLPYVQILKTAGVNTTDKEMEVLHLRNV SFEDAGEYTCLAGNSIGLSHHSAWLTVLEALEERPAVMTSPLYE. (SEQ ID NO:427)

[0080] An example of an extracellular region of FGFR1c (βIIIc) is: RPSPTLPEQDALPSEDDDDDDSSEEKETDNTKPNPVAPYWTSPEKMEKKLHAVPAAKTV KFKCPSSGTPNPTLRWLKNGKEFKPDHRIGGYKVRYATWSIIMDSVVPSDKGNYTCIVENEY GSINHTYQLDVVERSPHRPILQAGLPANKTVALGSNVEFMCKV YSDPQPHIQWLKHIEVNGS KIGPDNLPYVQILKTAGVNTTDKEMEVLHLRNVSFEDAGEYTCLAGNSIGLSHHSAWLTVLE ALEERPAVMTSPLYLE. (SEQ ID NO: 426)

[0081] As described herein, FGFR1c proteins may also include fragments. As used herein, the terms are used interchangeably to designate a receptor, in particular, and unless otherwise specified, a human receptor, which upon association with Klotho beta and FGF21 induces a signaling activity similar to that of FGF21.

[0082] The term FGFR1c also includes post-translational modifications of the amino acid sequence of FGFR1c, for example, possible N-linked glycosylation sites. Thus, antigen-binding proteins can bind to or be generated from glycosylated proteins in one or more of the positions.

[0083] Binding proteins, such as anti-Klotho beta antibodies, as described herein bind to Klotho beta alone or in complex with an FGF receptor, such as FGFR2c. A nucleic acid sequence encoding human FGFR2c is provided below: atggtcagctggggtcgtttcatctgcctggtcgtggtcaccatggcaaccttgtccctggc ccggccctccttcagtttagttgaggataccacattagagccagaagagccaccaaccaaat accaaatctctcaaccagaagtgtacgtggctgcgccaggggagtcgctagaggt gcgctgc ctgttgaaagatgccgccgtgatcagttggactaaggatggggtgcacttggggcccaacaa taggacagtgcttattggggagtacttgcagataaagggcgccacgcctagagactccggcc tctatgcttgtactgccagtaggactgtagacagtgaaacttggtacttcatggtgaatgtc acagatgccatctcat ccggagatgatgaggatgacaccgatggtgcggaagattttgtcag tgagaacagtaacaacaagagagcaccatactggaccaacacagaaaagatggaaaagcggc tccatgctgtgcctgcggccaacactgtcaagtttcgctgcccagccggggggaacccaatg ccaaccatgcggtggctgaaaaacgggaaggagtttaagca ggagcatcgcattggaggcta caaggtacgaaaccagcactggagcctcattatggaaagtgtggtcccatctgacaaggga a attatacctgtgtagtggagaatgaatacgggtccatcaatcacacgtaccacctggatgtt gtggagcgatcgcctcaccggcccatcctccaagccggactgccggcaaatgcctccacagt ggt cggaggagacgtagagtttgtctgcaaggtttacagtgatgcccagccccacatccagt ggatcaagcacgtggaaagaacggcagtaaatacgggcccgacgggctgccctacctcaag gttctcaaggccgccggtgttaacaccacggacaaagagattgaggttctctatattcgggaa atatacgtgcttggcgggtaattctattgggatatcct ttcactctgcatggttgacagttctgccagcgcctggaagagaaaaggagattacagcttcc ccagactacctggagatagccatttactgcataggggtcttcttaatcgcctgtatggtggt aacagtcatcctgtgccgaatgaagaacacgaccaagaagccagacttcagcagccag ccgg ctgtgcacaagctgaccaaacgtatccccctgcggagacaggtaacagtttcggctgagtcc agctcctccatgaactccaacaccccgctggtgaggataacaacacgcctctcttcaacggc agacacccccatgctggcaggggtctccgagtatgaacttccagaggacccaaaatgggagt ttccaagagataagctgacactgggcaagcccc tgggagaaggttgctttgggcaagtggtc atggcggaagcagtgggaattgacaaagacaagcccaaggaggcggtcaccgtggccgtgaa gatgttgaaagatgatgccacagagaaagacctttctgatctggtgtcagagatggagatga tgaagatgattgggaaacacaagaatacataaatcttcttggagcctgcacacagg atggg cctctctatgtcatagttgagtatgcctctaaaggcaacctccgagaatacctccgagcccg gaggccacccgggatggagtactcctatgacattaaccgtgttcctgaggagcagatgacct tcaaggacttggtgtcatgcacctaccagctggccagaggcatggagtacttggcttcccaa aaatgtattcatcgagatttagca gccagaaatgttttggtaacagaaaacaatgtgatgaa aatagcagactttggactcgccagagatatcaacaatatagactattacaaaaagaccacca atgggcggcttccagtcaagtggatggctccagaagccctgtttgatagagtatacactcat cagagtgatgtctggtccttcggggtgttaatgtgggagatcttcactttagggggctc gcc ctacccagggattcccgtggaggaactttttaagctgctgaaggaaggacacagaatggata agccagccaactgcaccaacgaactgtacatgatgatgagggactgttggcatgcagtgccc tcccagagaccaacgttcaagcagttggtagaagacttggatcgaattctcactctcacaac caatgaggaatacttggacctcagccaacctctcgaacagtattcacctagttaccctgaca caagaagttcttgttcttcaggagatgattctgttttttctccagaccccatgccttacgaa ccatgccttcctcagtatccacacataaacggcag tgttaaaacatga (SEQ ID NO:310)

[0084] The amino acid sequence of human FGFR2c is provided below; the amino acids that make up the signal sequence are underlined: mvswgrficlvvvtmatlslaRPSFSLVEDTTLEPEEPPTKYQISQPEVYVAAPGE SLEVRC LLKDAAVISWTKDGVHLGPNNRTVLIGEYLQIKGATPRDSGLYACTASRTVDSETWYFMVNV TDAISSGDDEDDTDGAEDFVSENSNNKRAPYWTNTEKMEKRLHAVPAANTVKFRCPAGGN PM PTMRWLKNGKEFKQEHRIGGYKVRNQHWSLIMESVVPSDKGNYTCVVENEYGSINHTYHLDV VERSPHRPILQAGLPANASTVVGGDVEFVCKVYSDAQPHIQWIKHVEKNGSKYGPDGLPYLK VLKAAGVNTTDKEIEVLYIRNVTFEDAGEYTCLAGNSIGISFHSAWLTVLPAPGREKEITAS PDYLEIAIYCIGVFL IACMVVTVILCRMKNTTKKPDFSSQPAVHKLTKRIPLRRQVTVSAES SSSMNSNTPLVRITTRLSSTADTPMLAGVSEYELPEDPKWEFPRDKLTLGKPLGEGCFGQVV MAEAVGIDKDKPKEAVTVAVKMLKDDATEKDLSDLVSEMEMMKMIGKHKNIINLLGACTQDG PLYVIVEYASKGNLREYLRARRPPGMEYSYDINRVPEE QMTFKDLVSCTYQLARGMEYLASQ KCIHRDLAARNVLVTENNVMKIADFGLARDINNIDYYKKTTNGRLPVKWMAPEALFDRVYTH QSDVWSFGVLMWEIFTLGGSPYPGIPVEELFKLLKEGHRMDKPANCTNELYMMMRDCWHAVP SQRPTFKQLVEDLDRILTLTNEEYLDLSQPLEQYSPSYPDTRSSCSSGDDSVFSPDPMPYE PC LPQYPHINGSVKT (SEQ ID NO:311)

[0085] Binding proteins, such as anti-Klotho beta antibodies, as described herein, bind to Klotho beta alone or in complex with an FGF receptor, such as FGFR3c. A nucleic acid sequence encoding human FGFR3c (GenBank Accession Number NP 000133) is provided below: atgggcgcccctgcctgcgccctcgcgctctgcgtggccgtggccatcgtggccggcgcctc ctcggagtccttggggacggagcagcgcgtcgtggggcgagcggcagaagtcccgggcccag agcc cggccagcaggagcagttggtcttcggcagcggggatgctgtggagctgagctgtccc ccgcccgggggtggtcccatggggcccactgtctgggtcaaggatggcacagggctggtgcc ctcggagcgtgtcctggtggggccccagcggctgcaggtgctgaatgcctcccacgaggact ccggggcctacagctg ccggcagcggctcacgcagcgcgtactgtgccacttcagtgtgcgg gtgacagacgctccatcctcgggagatgacgaagacggggaggacgaggctgaggacacagg tgtggacacaggggccccttactggacacggcccgagcggatggacaagaagctgctggccg tgccggccgccaacaccgtccgcttccgctgcccagccg ctggcaaccccactccctccatc tcctggctgaagaacggcagggagttccgcggcgagcaccgcattggaggcatcaagctgcg gcatcagcagtggagcctggtcatggaaagcgtggtgccctcggaccgcggcaactacacct gcgtcgtggagaacaagtttggcagcatccggcagacgtacacgctggacgtgctggag cgc tccccgcaccggcccatcctgcaggcggggctgccggccaaccagacggcggtgctgggcag cgacgtggagttccactgcaaggtgtacagtgacgcacagccccacatccagtggctcaagc acgtggaggtgaatggcagcaaggtgggcccggacggcacaccctacgttaccgtgctcaag acggcgggcgc taacaccaccgacaaggagctagaggttctctccttgcacaacgtcacctt tgaggacgccggggagtacacctgcctggcgggcaattctattgggttttctcatcactctg cgtggctggtggtgctgccagccgaggaggagctggtggaggctgacgaggcgggcagtgtg tatgcaggcatcctcagctacggggtgggctt cttcctgttcatcctggtggtggcggctgt gacgctctgccgcctgcgcagcccccccaagaaaggcctgggctcccccaccgtgcacaaga tctcccgcttcccgctcaagcgacaggtgtccctggagtccaacgcgtccatgagctccaac acaccactggtgcgcatcgcaaggctgtcctcaggggagggccccac gctggccaatgtctc cgagctcgagctgcctgccgaccccaaatgggagctgtctcgggcccggctgaccctgggca agccccttggggagggctgcttcggccaggtggtcatggcggaggccatcggcattgacaag gaccgggccgccaagcctgtcaccgtagccgtgaagatgctgaaagacgatgccactgacaa ggacctg tcggacctggtgtctgagatggagatgatgaagatgatcgggaaacacaaaaaca tcatcaacctgctgggcgcctgcacgcagggcgggcccctgtacgtgctggtggagtacgcg gccaagggtaacctgcgggagtttctgcgggcgcggcggcccccgggcctggactactcctt cgacacctgcaagccg cccgaggagcagctcaccttcaaggacctggtgtcctgtgcctacc aggtggcccggggcatggagtacttggcctcccagaagtgcatccacagggacctggctgcc cgcaatgtgctggtgaccgaggacaacgtgatgaagatcgcagacttcgggctggcccggga cgtgcacaacctcgactactacaagaagacaaccaacggccggctgcccgtgaagtggatgg cgcctgaggccttgtttgaccgag tctacactcaccagagtgacgtctggtcctttggggtc ctgctctgggagatcttcacgctggggggctccccgtaccccggcatccctgtggaggagct cttcaagctgctgaaggagggccaccgcatggacaagcccgccaactgcac acacgacctgt acatgatcatgcgggagtgctggcatgccgcgccctcccagaggccca ccttcaagcagctg gtggaggacctggaccgtgtccttaccgtgacgtccaccgacgagtacctggacctgtcggc gcctttcgagcagtactccccgggtggccaggacacccccagctccagctcctcaggggacg actccgtgtttgcccacgacctgctgcccccggccccacccagcagtgggggctcgcggac g tga (SEQ ID NO:312)

[0086] The amino acid sequences of human FGFR3c are provided below; The amino acids that form the signal sequence are underlined: GEHRIGGIKLRHQQWSLVMESVVPSDRGNYTCVVENKFGSIRQTYTLDVLER SPHRPILQAGLPANQTAVLGSDVEFHCKVYSDAQPHIQWLKHVEVNGSKVGPDGTPYVTVLK TAGANTTDKELEVLSLHNVTFEDAGEYTCLAGNSIGFSHHSAWLVVLPAEEELVEADEAGSV YAGILSYGVGFFLFILVVAAVTLCRLRSPPKKGLG SPTVHKISRFPLKRQVSLESNASMSSN TPLVRIARLSSGEGPTLANVSELELPADPKWELSRARLTLGKPLGEGCFGQVVMAEAIGIDK DRAAKPVTVAVKMLKDDATDKDLSDLVSEMEMMKMIGKHKNIINLLGACTQGGPLYVLVEYA AKGNLREFLRARRPPGLDYSFDTCKPPEEQLTFKDLVSCAYQVARGMEYLASQKCIHR DLAA RNVLVTEDNVMKIADFGLARDVHNLDYYKKTTNGRLPVKWMAPEALFDRVYTHQSDVWSFGV LLWEIFTLGGSPYPGIPVEELFKLLKEGHRMDKPANCTHDLYMIMRECWHAAPSQRPTFKQL VEDLDRVLTVTSTDEYLDLSAPFEQYSPGGQDTPSSSSSGDDSVFAHDLLPPAPPSSGGSRT (SEQ ID NO:313)

[0087] Binding proteins, such as anti-Klotho beta antibodies, as described herein, bind to Klotho beta alone or in complex with an FGF receptor, such as FGFR4. A nucleic acid sequence encoding human FGFR4 is provided below: atgcggctgctgctggccctgttgggggtcctgctgagtgtgcctgggcctccagtcttgtc cctggaggcctctgaggaagtggagcttgagccctgcctggctcccagcctggagcagcaag agcaggagctgacagtagcccttgggcagcctgtgcgtct gtgctgtgggcgggctgagcgt ggtggccactggtacaaggagggcagtcgcctggcacctgctggccgtgtacggggctggag gggccgcctagagattgccagcttcctacctgaggatgctggccgctacctctgcctggcac gaggctccatgatcgtcctgcagaatctcaccttgattacaggtgactccttgacct ccagc aacgatgatgaggaccccaagtcccatagggacccctcgaataggcacagttacccccagca agcaccctactggacacacccccagcgcatggagaagaaactgcatgcagtacctgcgggga acaccgtcaagttccgctgtccagctgcaggcaaccccacgcccaccatccgctggcttaag gatggacaggcctttcatggggagaaccgcat tggaggcattcggctgcgccatcagcactg gagtctcgtgatggagagcgtggtgccctcggaccgcggcacatacacctgcctggtagaga acgctgtgggcagcatccgctataactacctgctagatgtgctggagcggtccccgcaccgg cccatcctgcaggccgggctcccggccaacaccacagccgtggtgggcagc gacgtggagct gctgtgcaaggtgtacagcgatgcccagccccacatccagtggctgaagcacatcgtcatca acggcagcagcttcggagccgacggtttcccctatgtgcaagtcctaaagactgcagacatc aatagctcagaggtggaggtcctgtacctgcggaacgtgtcagccgaggacgcaggcgagta ca cctgcctcgcaggcaattccatcggcctctcctaccagtctgcctggctcacggtgctgc cagaggaggaccccacatggaccgcagcagcgcccgaggccaggtatacggacatcatcctg tacgcgtcgggctccctggccttggctgtgctcctgctgctggccgggctgtatcgagggca ggcgctccacggccggcacc cccgcccgcccgccactgtgcagaagctctcccgcttccctc tggcccgacagttctccctggagtcaggctcttccggcaagtcaagctcatccctggtacga ggcgtgcgtctctcctccagcggccccgccttgctcgccggcctcgtgagtctagatctacc tctcgacccactatgggagttcccccgggaca ggctggtgcttgggaagcccctaggcgagg gctgctttggccaggtagtacgtgcagaggcctttggcatggaccctgcccggcctgaccaa gccagcactgtggccgtcaagatgctcaaagacaacgcctctgacaaggacctggccgacct ggtctcggagatggaggtgatgaagctgatcggccgacacaagaacatcatcaacctgct TG cagtatctggagtcccggaagtgtatccaccgggacctggctgcccgcaatgtgctggtg actgaggacaatgtgatgaagattgctgactttgggctggcccgcggcgtccaccacattga ctactataagaaaaccagcaacggccgcctgcctgtgaagtggatggcgcc cgaggccttgt ttgaccgggtgtacacaca ccagagtgacgtgtggtcttttgggatcctgctatgggagatc ttcaccctcgggggctccccgtatcctggcatcccggtggaggagctgttctcgctgctgcg ggagggacatcggatggaccgacccccacactgccccccagagctgtacgggctgatgcgtg agtgctggcacgcagcgccctcccaga ggcctaccttcaagcagctggtggaggcgctggac aaggtcctgctggccgtctctgaggagtacctcgacctccgcctgaccttcggaccctattc cccctctggtggggacgccagcagcacctgctcctccagcgattctgtcttcagccacgacc ccctgccattgggatccagctccttccccttcgggtctggggtgcagacatga (SEQ ID NO:314)

[0088] The amino acid sequence of human FGFR4 (GenBank Accession Number NP. 002002.3) is provided below; the amino acids that form the signal sequence are underlined: mrlllallgvllsvpgppvlsLEASEEVELEPCLAPSLEQQEQELTVALGQPVRLCCGRAER GGHWYKEGSRLAPAGRVRGWRGRLEIASFLPEDAGRYLCLARGSMIVLQNLTLITGDSLTSS NDDEDPKSHRDPSNRHSYPQQAPYWTHPQRMEKKLHAVPAGNTVKFRCPAAGNPTPTIRWLK DGQAFHGENRIGGIRLRHQHWSLVMESVVPSDRGTYTCLVENAVGSIRYNYLLDVLERSPHR PILQAGLPANTTAVVGSDVELLCKVYSDAQPHIQWLKHIVINGSSFGADGFPYVQVLKTADI NSSEVEVLYLRNVSAEDAGEYTCLAGNSIGLSYQSAWLTVLPEEDPTWTAAAPEARYTDIIL YASGSLALAVLLLLAGLYRGQALHGRHPRPPA Tv VMKIADFGLARGVHHIDYYKKTSNGRLPVKWMAPEALFDRVYTHQSDVWSFGILLWEI FTLGGSPYPGIPVEELFSLLREGHRMDRPPHCPPELYGLMRECWHAAPSQRPTFKQLVEALD KVLLAVSEEYLDLRLTFGPYSPSGGDASSTCSSSDSVFSHDPLPLGSSSFPFGSGVQT (SEQ ID NO:315)

[0089] An "antigen" is a predetermined antigen to which an antibody can selectively bind. A target antigen may be a polypeptide, carbohydrate, nucleic acid, lipid, hapten, or other naturally occurring or synthetic compound. Preferably, the target antigen is a polypeptide.

[0090] The term "antigen-binding fragment", "antigen-binding domain", "antigen-binding region", and similar terms refer to the portion of an antibody comprising amino acid residues that interact with a antigen and confer on the binding agent its specificity and affinity for the antigen (e.g., complementarity determining regions (CDRs)).

[0091] The terms "alloy" or "bond" refer to an interaction between molecules, including, for example, to form a complex. The interactions may be, for example, non-covalent interactions, including hydrogen bonds, ionic bonds, hydrophobic interactions, and/or van der Waals. A complex may also include the bond of two or more molecules held together by covalent or noncovalent bonds, interactions, or forces. The strength of the total noncovalent interactions between a single antigen-binding site of an antibody and a single epitope of a target molecule, such as Klotho beta, is the affinity of the antibody or functional fragment for that epitope. The ratio of association (k1) to dissociation (k-1) of an antibody to a monovalent antigen (k1/k-1) is the association constant K, which is a measure of affinity. The value of K varies for different antibody and antigen complexes and depends on both k1 and k-1. The association constant K for an antibody provided herein can be determined using any method provided herein or any other method well known to those skilled in the art. The affinity of a binding site does not always reflect the true strength of the interaction between an antibody and an antigen. When complex antigens containing multiple repeating antigenic determinants, such as a polyvalent Klotho beta, come into contact with antibodies containing multiple binding sites, the interaction of the antibody with the antigen at one site will increase the likelihood of a reaction at a second site. . The resistance of such multiple interactions between an antibody and the multivalent antigen is called avidity. The avidity of an antibody may be a better measure of its binding capacity, which is the affinity of its individual binding sites. For example, high avidity may compensate for low affinity as is sometimes found for pentameric IgM antibodies, which may have a lower affinity than IgG, but the high avidity of IgM, resulting from its polyvalence, allows it to bind to the antigen. effectively.

[0092] The terms "antibodies that specifically bind to Klotho beta," "antibodies that specifically bind to an epitope of Klotho beta," and similar terms are also used interchangeably herein and refer to antibodies that bind specifically to a Klotho beta polypeptide, such as a Klotho beta antigen, or fragment, or epitope (e.g., human Klotho beta such as a human Klotho beta polypeptide, antigen, or epitope). An antibody that specifically binds to Klotho beta, (e.g., human Klotho beta) can bind to the extracellular domain or peptide derived from the extracellular domain of Klotho beta. An antibody that specifically binds to a Klotho beta antigen (e.g., human Klotho beta) may be cross-reactive with related antigens (e.g., Cyno Klotho beta). In certain embodiments, an antibody that specifically binds to a Klotho beta antigen does not cross-react with other antigens. An antibody that specifically binds to a Klotho beta antigen can be identified, for example, by immunoassays, BIAcore, or other techniques known to those skilled in the art. An antibody specifically binds to a Klotho beta antigen when it binds to a Klotho beta antigen with greater affinity than to any antigen in cross-reactive assays, as determined using experimental techniques such as radioimmunoassays (RIA) and immunosorbent assays. enzymatic (ELISA). Typically, a specific or selective reaction will be at least twice the background signal or noise and may be more than 10 times the background. See, for example, Paul, ed., 1989, Fundamental Immunology Second Edition, Raven Press, New York, on pages 332 to 336 for a discussion of antibody specificity. An antibody that "binds" an antigen of interest (e.g., a target antigen, such as Klotho beta) is one that binds to the antigen with sufficient affinity for the antibody to be useful as a therapeutic agent in targeting a cell or tissue. that expresses the antigen, and does not significantly cross-react with other proteins. In such embodiments, the extent of binding of the antibody to a "non-target" protein will be less than about 10% of the binding of the antibody to its particular target protein, e.g., as determined by fluorescence activated cell (FACS) or analysis. radioimmunoprecipitation (RIA). With respect to the binding of an antibody to a target molecule, the term "specific binding" or "specifically binds to" or is "specific for" a particular polypeptide or an epitope on a particular polypeptide target means binding that is measurably different of a non-specific interaction. Specific binding can be measured, for example, by determining the binding of a molecule compared to the binding of a control molecule, which is generally a molecule of similar structure that has no binding activity. For example, specific binding can be determined by competition with a control molecule that is similar to the target, e.g., an excess of unlabeled target. In this case, specific binding is indicated if the binding of the labeled target to a probe is competitively inhibited by excess unlabeled target. The term "specific binding" or "specifically binds to" or is "specific for" a particular polypeptide or an epitope on a particular polypeptide target as used herein may be displayed, for example, by a molecule having a Kd for the target of at least about 104 M, alternatively at least about 10-5 M, alternatively at least about 10-6 M, alternatively at least about 10-7 M, alternatively at least about 10-8 M, alternatively at least about 10-9 M, alternatively at least about 10-10 M, alternatively at least about 10-11 M, alternatively at least about 10-12 M or greater. In one embodiment, the term "specific binding" refers to a binding in which the molecule binds to a particular polypeptide or epitope on a particular polypeptide without substantially binding to any other polypeptide or polypeptide epitope. In certain embodiments, an antibody that binds to Klotho beta has a dissociation constant (Kd) of less than or equal to 10 nM, 5 nM, 4 nM, 3 nM, 2 nM, 1 nM, 0.9 nM, 0.8 nM, 0.7 nM, 0.6 nM, 0.5 nM, 0.4 nM, 0.3 nM, 0.2 nM, or 0.1 nM. The lower KD, the greater the affinity of the anti-Klotho beta antibody. In certain embodiments, the anti-Klotho beta antibody binds to an epitope of Klotho beta that is conserved among Klotho beta from different species (e.g., between human and Cyno Klotho beta).

[0093] The term "compete" when used in the context of Anti-Klotho beta antibodies (e.g., agonist antibodies and binding proteins that bind to (i) Klotho beta; or (ii) a complex comprising Klotho beta and one of FGFR1c, FGFR2c, FGFR3c, and FGFR4) competing for the same epitope or binding site on a target means competition between As determined by an assay in which the antibody (or binding fragment) thereof under study prevents or inhibits specific binding of a reference molecule (e.g., a reference ligand, or reference antigen-binding protein, such as a reference antibody) to a common antigen (e.g., Klotho beta or a fragment thereof). Numerous types of competitive binding assays can be used to determine whether a test antibody competes with a reference antibody for binding to Klotho beta (e.g., human Klotho beta). Examples of assays that may be employed include direct or indirect solid-phase radioimmunoassay (RIA), direct or indirect solid-phase enzyme immunoassay (EIA), sandwich competition assay (see, e.g., Stahli et al., (1983) Methods in Enzymology 9:242-253); Direct solid phase biotin-avidin EIA (see, e.g., Kirkland et al, (1986) J. Immunol. 137:3614-3619), direct solid phase labeled assay, direct solid phase labeled sandwich assay (see , e.g., Harlow and Lane, (1988) Antibodies, A Laboratory manual, Cold Spring Harbor Press); solid phase direct label RIA using 1-125 labeling (see, for example, Morel et al, (1988) Molec Immunol 25:7-15); Direct solid phase biotin-avidin EIA (see, for example, Cheung, et al, (1990) Virology 176:546-552); and direct labeled RIA (Moldenhauer et al, (1990) Scand J. Immunol 32:77-82). Typically, such an assay involves the use of a purified antigen (e.g., Klotho beta, such as human Klotho beta) bound to a solid surface or cells carrying both an unlabeled test antigen-binding protein (e.g., Antibody antibody). - test Klotho beta) or a labeled reference antigen-binding protein (e.g. reference anti-Klotho beta antibody). Competitive inhibition can be measured by determining the amount of tracer 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. Antibodies identified by competition assay (competing antibodies) include antibodies that bind to the same epitope as the reference antibody and/or antibodies that bind to an adjacent epitope sufficiently close to the epitope bound by reference antibodies for steric hindrance to occur. Additional details on methods for determining competitive binding are described herein. Normally, when a competing antibody protein is present in excess, it will inhibit the specific binding of a reference antibody to a common antigen by at least 23%, e.g. 40%, 45%, 50%, 55%, 60%, 65%, 70% or 75%. In some examples, binding is inhibited by at least 80%, 85%, 90%, 95%, 96% or 97%, 98%, 99% or more.

[0094] The term "Anti-Klotho beta antibody" or "an antibody that binds to Klotho beta" includes an antibody that is capable of binding to Klotho beta with sufficient affinity for the antibody to be useful as a diagnostic agent and/or or therapeutic to target Klotho beta. Preferably, the extent of binding of an Anti-Klotho beta antibody to an unrelated non-Klotho beta protein is less than about 10% of the antibody's binding to Klotho beta as measured, for example, by analysis of cells activated by fluorescence (FACS) or an immunoassay such as a radioimmunoassay (RIA). An antibody that "specifically binds to" or is "specific for" Klotho beta is illustrated above. In certain embodiments, an antibody that binds to Klotho beta as described herein has a dissociation constant (Kd) of less than or equal to 10 nM, 9 nM, 8 nM, 7 nM, 6 nM, 5 nM, 4 nM, 0.9 nM, 0.8 nM, 0.7 nM, 0.6 nM, 0.5 nM, 0.4 nM, 0.3 nM, 0.2 nM, or 0.1 nM, and /or is greater than or equal to 0.1 nM. In certain embodiments, the anti-Klotho beta antibody binds to an epitope of Klotho beta that is conserved among Klotho beta from different species (e.g., between human and Cyno Klotho beta).

[0095] An "isolated" antibody is substantially free of cellular material or other contaminating proteins of the cellular or tissue source and/or other contaminating components from which the antibody is derived, or substantially free of chemical precursors or other chemicals when chemically synthesized. The language "substantially free of cellular material" includes preparations of an antibody in which the antibody is separated from the cellular components of the cells from which it is isolated or recombinantly produced. Thus, an antibody that is substantially free of cellular material includes antibody preparations having less than about 30%, 25%, 20%, 15%, 10%, 5%, or 1% (by dry weight) heterologous protein. (also referred to herein as a "contaminant protein"). In certain embodiments, when the antibody is produced recombinantly, it is substantially free of culture medium, e.g., the culture medium represents less than about 20%, 15%, 10%, 5%, or 1% of the volume of the protein preparation. In certain embodiments, when the antibody is produced by chemical synthesis, it is substantially free of chemical precursors or other chemicals, for example, it is separated from chemical precursors or other chemicals that are involved in the synthesis of the protein. Therefore, such antibody preparations contain less than about 30%, 25%, 20%, 15%, 10%, 5%, or 1% (by dry weight) chemical precursors or other compounds other than the antibody of interest. . Contaminating components may also include, but are not limited to, materials that would interfere with therapeutic uses for the antibody, and may include enzymes, hormones, and other proteinaceous or non-proteinaceous solutes. In certain embodiments, the antibody will be purified (1) to greater than 95% by weight of antibody as determined by the Lowry method (Lowry et al. J. Bio. Chem. 193: 265-275, 1951), such as 96 %, 97%, 98%, or 99%, by weight, (2) to a degree sufficient to obtain at least 15 residues of the N-terminal or internal amino acid sequence by use of a spinning cup sequencer, or (3) to homogeneity by SDS-PAGE under reducing or non-reducing conditions using Coomassie blue or, preferably, silver staining. Isolated antibody includes antibody in situ within recombinant cells since at least one component of the antibody's natural environment will not be present. Typically, however, the isolated antibody will be prepared by at least one purification step. In specific embodiments, the antibodies provided herein are isolated.

[0096] A 4-chain antibody unit is a heterotetrameric glycoprotein composed of two identical light (L) chains and two identical heavy (H) chains. In the case of IgGs, the 4-chain unit is generally about 150,000 daltons. Each L chain is linked to one H chain by a covalent disulfide bond, while the two H chains are linked to each other by one or more disulfide bonds depending on the isotype of the H chain. Each H and L chain also have intrachain disulfide bridges regularly. spaced. Each H chain has, at the N-terminus, a variable domain (VH) followed by three constant domains (CH) for each of the α and y chains and four CH domains for the μ and ε isotypes. Each L chain has at the N-terminal end, a variable domain (VL) followed by a constant domain (CL) at its other end. VL is aligned with VH and CL is aligned with the first constant domain of the heavy chain (CH1). Particular amino acid residues are believed to form an interface between the light chain and heavy chain variable domains. The pairing of a VH and VL together forms a single antigen-binding site. For the structure and properties of the different classes of antibodies, see, for example, Basic and Clinical Immunology, 8th edition, Daniel P. Stites, Abba I. Terr, and Tristram G. Parslow (eds.), Appleton & Lange, Norwalk, CT , 1994, page 71 and Chapter 6.

[0097] The term "variable region" or "variable domain" refers to a portion of the heavy or light chains of an antibody that is generally located at the amino-terminus of the light or heavy chain and has a length of about 120 to 130 amino acids in the heavy chain and about 100 to 110 amino acids in the light chain, and are used in the binding and specificity of each particular antibody for its particular antigen. The heavy chain variable region may be referred to as "VH". The light chain variable region may be referred to as "VL". The term "variable" refers to the fact that certain segments of the variable regions differ extensively in sequence among antibodies. The V region mediates antigen binding and defines the specificity of a particular antibody for its particular antigen. However, the variability is not uniformly distributed across the entire 110 amino acid length of the variable regions. Instead, V regions consist of less variable (e.g., relatively invariant) stretches called structural regions (FRs) of about 15-30 amino acids separated by shorter regions of greater variability (e.g., extreme variability) called "regions." hypervariables" that are each about 9-12 amino acids long. The variable regions of the heavy and light chains each comprise four FRs, largely adopting a β-sheet configuration, connected by three hypervariable regions, which form loops linking to, and in some cases, forming part of, the β-sheet structure. The hypervariable regions on each chain are held together in close proximity by the FRs and, with the hypervariable regions on the other chain, contribute to the formation of the antigen-binding site of antibodies (see, e.g., Kabat et al., Sequences of Proteins of Immunological Interest, 5th Ed. Public Health Service, National Institutes of Health, Bethesda, MD, 1991)). Constant regions are not directly involved in the binding of an antibody to an antigen, but exhibit various effector functions, such as antibody participation in antibody-dependent cellular cytotoxicity (ADCC) and complement-dependent cytotoxicity (CDC). The variable regions differ extensively in sequence between different antibodies. Sequence variability is concentrated in the CDRs, while the less variable portions in the variable region are referred to as framework regions (FR). The CDRs of the light and heavy chains are mainly responsible for the interaction of the antibody with the antigen. In specific embodiments, the variable region is a human variable region.

[0098] The term "variable region residue numbering as in Kabat" or "amino acid position numbering as in Kabat", and variations thereof, refers to the numbering system used for heavy chain variable regions or variable regions light chain antibody assembly in Kabat et al., Sequences of Proteins of Immunological Interest, 5th Ed. Public Health Service, National Institutes of Health, Bethesda, MD. (1991). Using this numbering system, the actual linear amino acid sequence may contain fewer or additional amino acids corresponding to a shortening of, or insertion of, an FR or CDR of the variable domain. For example, a heavy chain variable domain may include a single amino acid insertion (residue 52a according to Kabat) after residue 52 of H2 and inserted residues (e.g., residues 82a, 82b, and 82c, etc., according to Kabat) after heavy chain FR residue 82. The Kabat numbering of residues can be determined for a given antibody by aligning regions of antibody sequence homology with a numbered "standard" Kabat sequence. The Kabat numbering system is generally used when referring to a residue in the variable domain (approximately residues 1-107 of the light chain and residues 1113 of the heavy chain) (e.g., Kabat et al., Sequences of Immunological Interest. 5th Ed. Public Health Service, National Institutes of Health, Bethesda, Md. (1991)). The "EU numbering system" or "EU index" is generally used when referring to a residue in an immunoglobulin heavy chain constant region (e.g., the EU index reported in Kabat et al., supra). The "EU index as in Kabat" refers to the numbering of the residues of the human EU IgG 1 antibody. Other numbering systems have been described, including, for example, by AbM, Chothia, Contact, IMGT and Ahon. Various number systems are illustrated in Figures 1-3.

[0099] An "intact" antibody is one that comprises an antigen-binding site as well as a CL and at least heavy chain constant regions, CH1, CH2 and CH3. Constant regions may include human constant regions or variant amino acid sequences. Preferably, an intact antibody has one or more effector functions.

[0100] "Antibody fragments" comprise a portion of an intact antibody, preferably the antigen-binding or variable region of the intact antibody. Examples of antibody fragments include, without limitation, Fab, Fab', F(ab')2, and Fv fragments; diabodies and di-diabodies (see, for example, Holliger, P. et al., (1993) Proc. Natl. Acad. Sci. 90:6444-8; Lu, D. et al., (2005) J. Biol Chem. 280:19665-72; Hudson et al., Nat. Med. 9:129-134 (2003); single-chain antibody molecules (see, for example, US Patent Nos. 4,946,778; 5,260,203; 5,482,858 and 5,476,786); dual variable domain antibodies (see, for example, US Patent No. 7,612,181); single variable domain antibodies (SdAbs) (see, for example, Woolven et al, Immunogenetics 50:98-101, 1999; Streltsov et al, Proc Natl Acad Sci USA 101:12444-12449, 2004); and multispecific antibodies formed from antibody fragments.

[0101] A "functional fragment" or "binding fragment" or "antigen-binding fragment" of a therapeutic antibody will exhibit at least one, if not part or all, of the biological functions attributed to the intact antibody, the function comprising at least minus binding to the target antigen, (e.g., a Klotho beta-binding fragment or fragment that binds to Klotho beta).

[0102] The term "fusion protein" as used herein refers to a polypeptide that comprises an amino acid sequence of an antibody and an amino acid sequence of a heterologous polypeptide or protein (e.g., a polypeptide or protein that is not normally part of the antibody (e.g., a non-anti-Klotho beta antigen-binding antibody). The term "fusion", when used in relation to Klotho beta or an anti-Klotho beta antibody, refers to the union of a peptide or polypeptide, or fragment, variant and/or derivative thereof, with a heterologous peptide or polypeptide. In certain embodiments, the fusion protein retains the biological activity of the Klotho beta or anti-Klotho beta antibody. In certain embodiments, the fusion protein comprises a Klotho beta antibody VH region, VL region, VH CDR (one, two, or three VH CDRs), and/or VL CDR (one, two, or three Klotho beta antibody CDRs). VL), wherein the fusion protein binds to a Klotho beta epitope, a Klotho beta fragment and/or a Klotho beta polypeptide.

[0103] The term "heavy chain", when used in reference to an antibody, refers to a polypeptide chain of about 50-70 kDa, wherein the amino-terminal portion includes a variable region of about 120 to 130 or more amino acids and a carboxy-terminal portion that includes a constant region. The constant region can be one of five distinct types, (e.g., isotypes) referred to as alpha (α), delta (δ), epsilon (ε), gamma (Y), and mu (μ), based on the amino acid sequence of the heavy chain constant region. The different heavy chains differ in size: α, δ, and Y contain about 450 amino acids, while μ and ε contain about 550 amino acids. When combined with a light chain, these distinct types of heavy chains give rise to five well-known classes (e.g., isotypes) of antibodies, IgA, IgD, IgE, IgG, and IgM, respectively, including four subclasses of IgG, namely IgG1, IgG2, IgG3 and IgG4. The heavy chain may be a human heavy chain.

[0104] The term "light chain", when used in reference to an antibody, refers to a polypeptide chain of about 25 kDa, wherein the amino-terminal portion includes a variable region of about 100 to about 110 or more amino acids and a carboxy-terminal portion that includes a constant region. The approximate length of a light chain is 211 to 217 amino acids. There are two distinct types, called kappa (K) and lambda (À) based on the amino acid sequence of the constant domains. Light chain amino acid sequences are well known in the art. The light chain may be a human light chain.

[0105] The term "host", as used herein, refers to an animal, such as a mammal (e.g., a human).

[0106] The term "host cell", as used herein, refers to a particular individual cell that can be transfected with a nucleic acid molecule and the progeny or potential progeny of such cell. The progeny of such a cell may not be identical to the parental cell transfected with the nucleic acid molecule due to mutations or environmental influences that may occur in successive generations or integrations of the nucleic acid molecule into the host cell genome.

[0107] The term "monoclonal antibody" as used herein refers to an antibody obtained from a population of substantially homogeneous antibodies, for example, individual antibodies constituting the population are identical except for possible naturally occurring mutations that may be present. present in smaller amounts, and each monoclonal antibody typically recognizes a single epitope on the antigen. In specific embodiments, a "monoclonal antibody" as used herein is an antibody produced by a single hybridoma or other cell, wherein the antibody binds only to one epitope of Klotho beta, as determined, for example, by ELISA or other antigen binding or competitive binding assay known in the art. The term "monoclonal antibody" is not limited to any particular method of making the antibody. For example, monoclonal antibodies useful in the present invention can be prepared by the hybridoma methodology first described by Kohler et al, Nature, 256:495 (1975), or can be produced using recombinant DNA methods in bacterial, eukaryotic animal, or plant cells. (see, eg, US Patent No. 4,816,567). "Monoclonal antibodies" can also be isolated from phage antibody libraries using the techniques described in Clackson et al, Nature, 352:624-628 (1991) and Marks et al, J. Mol. Biol., 222:581- 597 (1991), for example. Other methods for preparing clonal cell lines and monoclonal antibodies expressed in this way are well known in the art (see, for example, Chapter 11 in: Short Protocols in Molecular Biology, (2002) 5th Ed., Ausubel et al. , eds., John Wiley and Sons, New York). Examples of methods of producing monoclonal antibodies are provided in the examples presented here.

[0108] The term "native", when used in connection with biological materials such as nucleic acid molecules, polypeptides, host cells and the like, refers to those that are found in nature and not manipulated, modified and/or altered ( e.g., isolated, purified, selected) by a human being.

[0109] Antibodies provided herein may include "chimeric" antibodies in which a portion of the heavy and/or light chain is identical or homologous to corresponding sequences in antibodies derived from a particular corresponding species or belonging to a particular class of antibody or subclass, while the remainder of the chain(s) is identical or homologous to corresponding sequences in antibodies derived from another species or belonging to another class or subclass of antibodies, as well as fragments of such antibodies, provided they exhibit the desired biological activity ( see US Patent No. 4816567; and Morrison et al, Proc Natl Acad Sci USA, 81: 6851-6855 (1984)).

[0110] "Humanized" forms of non-human (e.g., murine) antibodies are chimeric antibodies that include human immunoglobulins (e.g., receptor antibody) in which native CDR residues are replaced with residues from the corresponding CDR of a non-human species (e.g., donor antibody) such as mouse, rat, rabbit or non-human primate, having desired specificity, affinity and capacity. In some cases, one or more residues in the FR region of the human immunoglobulin are replaced by corresponding non-human residues. Furthermore, humanized antibodies may comprise residues that are not found in the recipient antibody or the donor antibody. These modifications are made to further refine the performance of the antibody. A humanized heavy or light chain antibody may comprise substantially all of at least one or more variable regions, wherein all or substantially all of the CDRs correspond to those of a non-human immunoglobulin and all or substantially all of the FRs are those of a sequence of human immunoglobulin. In certain embodiments, the humanized antibody will comprise at least a portion of an immunoglobulin constant region (Fc), typically that of a human immunoglobulin. For more details, see, Jones et al., Nature, 321:522-525 (1986); Riechmann et al., Nature, 332:323-329 (1988); and Presta, Curr. Op. Struct. Biol., 2:593-596 (1992); Carter et al., Proc. Natl. Acd. Sci. USA 89:4285-4289 (1992); and US Patent Nos: 6,800,738 (issued Oct. 5, 2004), 6,719,971 (issued Sept. 27, 2005), 6,639,055 (issued Oct. 28, 2003), 6,407,213 (issued June 18, 2003), 2002), and 6,054,297 (issued April 25, 2000).

[0111] A "human antibody" is one that has an amino acid sequence that corresponds to that of an antibody produced by a human and/or was produced using any of the techniques for producing human antibodies as disclosed herein. This definition of a human antibody specifically excludes a humanized antibody comprising non-human antigen-binding residues. Human antibodies can be produced using various techniques known in the art, including phage display libraries (Hoogenboom and Winter, J. Mol Biol, 227:381 (1991); Marks et al, J. Mol Biol, 222:581 (1991) and yeast display libraries (Chao et al, Nature Protocols. 1:755-768 (2006)). Also available for the preparation of human monoclonal antibodies are the methods described in Cole et al, Monoclonal Antibodies and Cancer Therapy. , Alan R. Liss, p. 77 (1985); Boerner et al., J. Immunol., 147(1):86-95 (1991). :368-74 (2001). Human antibodies can be prepared by administering the antigen to a transgenic animal that has been engineered to produce such antibodies in response to antigenic challenge, but whose endogenous loci have been inactivated, e.g., mice (see, for example, , Jakobovits, A., Curr. Biotechnol. 1997, 8(4):455-8; and Pat. US Nos. 6075181 and 6150584 on XENOMOUSETM technology). See also, for example, Li et al., Proc. Natl. Academic. Sci. USA, 103: 3557-3562 (2006) on human antibodies generated through a human B cell hybridoma technology.

[0112] A "CDR" refers to one of three hypervariable regions (H1, H2 or H3) within the non-framework region of the immunoglobulin (Ig or antibody) VH β-sheet framework, or one of three hypervariable regions (L1, L2 or L3) within the non-framework region of the VL β-sheet framework of the antibody. Therefore, CDRs are variable region sequences interspersed within the framework region sequences. CDR regions are well known to those skilled in the art and have been defined by, for example, Kabat as regions of greatest hypervariability within antibody variable (V) domains (Kabat et al, J. Biol Chem 252:6609-6616 (1977) ; Kabat, Adv Prot Chem 32:1-75 (1978)). CDR region sequences were also structurally defined by Chothia as those residues that are not part of the conserved β-sheet framework, and thus are capable of adapting different conformations (Chothia and Lesk, J. Mol Biol 196:901-917 (1987) ). Both terms are well known in the art. Sequences of the CDR region were also defined by ABM, Contact and IMGT. Sequences of the CDR region are illustrated in Figures 1-3. The positions of the CDRs within a canonical antibody variable region were determined by comparison of numerous structures (Al-Lazikani et al., J. Mol. Biol. 273:927948 (1997); Morea et al., Methods 20:267- 279 (2000)). Since the number of residues of a hypervariable region varies in different antibodies, additional residues relative to canonical positions are conventionally numbered with a, b, c, and so on alongside the number of residues in the canonical variable region numbering scheme (Al -Lazikani et al, supra (1997)). Such similar nomenclature is well known to those skilled in the art.

[0113] The term "hypervariable region", "HVR", or "HV", when used herein, refers to regions of an antibody variable region that are hypervariable in sequence and/or form structurally defined loops. In general, antibodies comprise six hypervariable regions; three in VH (H1, H2, H3), and three in VL (L1, L2, L3). A number of delimitations of the hypervariable region are in use and are covered here. Kabat's complementarity determining regions (CDRs) are based on sequence variability and are the most commonly used (see, e.g., Kabat et al., Sequences of Proteins of Immunological Interest, 5th Ed. Public Health Service, National Institutes of Health, Bethesda, MD (1991)). Chothia refers instead to the location of structural loops (see, e.g., Chothia and Lesk, J. Mol Biol 196:901-917 (1987)). The end of the Chothia CDR-H1 handle when numbered using the Kabat numbering convention varies between H32 and H34 depending on the length of the handle (this is due to the fact that the Kabat numbering scheme places the inserts at H35A and H35B; if neither 35A nor 35B are present, the loop ends in 32; if at least 35A is present, the loop ends in 33; if both 35A and 35B are present, the loop ends in 34). The AbM hypervariable regions represent a compromise between the CDRs of Kabat and structural loops of Chothia, and are used by Oxford Molecular's AbM antibody modeling software (see, for example, Martin, in Antibody Engineering, Vol. 2, Chapter 3, Springer Verlag). The "contact" hypervariable regions are based on an analysis of the available complex crystal structures. The residues of each of these hypervariable regions or CDRs are indicated below.

[0114] Recently, a universal numbering system has been developed and widely adopted, Immunogenetics (IMGT) Information System® (Lafranc et al, Dev Comp Immunol 27 (1):55-77 (2003)). IMGT is an integrated information system specialized in immunoglobulins (IG), T cell receptors (TR) and major histocompatibility complex (MHC) of humans and other vertebrates. Here, CDRs are referred to in terms of both their amino acid sequence and location within the light or heavy chain. Because the "location" of CDRs within the structure of the immunoglobulin variable domain is conserved between species and present in structures called loops, when using numbering systems that align variable domain sequences according to structural features, CDRs and residues of framework are readily identified. This information can be used in grafting and replacing immunoglobulin CDR residues from one species to an acceptor framework from, typically, a human antibody. An additional numbering system (AHon) was developed by Honegger and Plückthun, J. Mol. Biol. 309: 657-670 (2001). The correspondence between the numbering system, including, for example, Kabat numbering and the unique IMGT numbering system, is well known to one skilled in the art (see, for example, Kabat, supra; Chothia and Lesk, supra; Martin , supra; Lefranc et al., supra) and is also illustrated in Figures 1-3. An exemplary system, shown here, combines Kabat and Chothia.

[0115] Hypervariable regions may comprise "extended hypervariable regions" as follows: 24-36 or 24-34 (L1), 46-56 or 50-56 (L2) and 89-97 or 89-96 (L3) in VL and 26-35 or 26-35A (H1), 50-65 or 49-65 (H2) and 93-102, 94-102, or 95-102 (H3) in VH. As used herein, the terms "HVR" and "CDR" are used interchangeably.

[0116] The term "constant region" or "constant domain" refers to a carboxy-terminal portion of the light and heavy chain that is not directly involved in antibody binding to antigen, but exhibits various effector functions, such as interaction with the Fc receptor. The terms refer to the portion of an immunoglobulin molecule having a more conserved amino acid sequence in relation to the other portion of the immunoglobulin, the variable region, which contains the antigen-binding site. The constant region may contain the CH1, CH2 and CH3 regions of the heavy chain and the CL region of the light chain.

[0117] The term "framework" or "FR" residues are those variable region residues that flank the CDRs. FR residues are present, for example, in chimeric, humanized, human antibodies, domain antibodies, diabodies, linear antibodies, and bispecific antibodies. FR residues are those variable domain residues that are different from hypervariable region residues or CDR residues.

[0118] An "affinity matured" antibody is one with one or more changes (e.g., amino acid sequence variations, including changes, additions and/or deletions) in one or more HVRs thereof that result in an improvement in the affinity of the antibody. antibody to the antigen, compared to a parent antibody that does not have this change(s). Preferred affinity matured antibodies will have nanomolar or even picomolar affinities for the target antigen. Affinity matured antibodies are produced using procedures known in the art. For review, see Hudson and Souriau, Nature Medicine 9:129-134 (2003); Hoogenboom, Nature Biotechnol. 23: 1105-1116 (2005); Quiroz and Sinclair, Quiroz and Sinclair, Revista Ingeneria Biomedia 4:39-51 (2010).

[0119] A "blocking" or "antagonist" antibody is one that inhibits or reduces the biological activity of the antigen it binds. For example, blocking antibodies or antagonist antibodies can substantially or completely inhibit the biological activity of the antigen.

[0120] An "agonist antibody" is an antibody that triggers a response, for example, one that mimics at least one of the functional activities of a polypeptide of interest (e.g., FGF19 or FGF21). An agonist antibody comprises an antibody that is a ligand mimetic, for example, wherein a ligand binds to a cell surface receptor and the binding induces cell signaling or activities through an intercellular cell signaling pathway and wherein the antibody induces similar cell signaling or activation.

[0121] A Klotho beta "agonist" refers to a molecule that is capable of activating or otherwise increasing one or more of the biological activities of Klotho beta, such as in a cell that expresses Klotho beta and an FGF receptor . In some embodiments, a Klotho beta agonist (e.g., an agonist antibody, as described herein) may, for example, act to activate or otherwise enhance cellular activation and/or signaling pathways of a cell expressing a Klotho protein. beta and an FGF receptor, thereby increasing a Klotho beta-mediated biological activity of the cell relative to the Klotho beta-mediated biological activity in the absence of agonist. In some embodiments the antibodies provided herein are anti-Klotho beta agonist antibodies, including antibodies that induce FGF19-like signaling and/or FGF21-like signaling.

[0122] "Binding affinity" generally refers to the strength of the sum total of noncovalent interactions between a single binding site of a molecule (e.g., a binding protein such as an antibody) and its binding partner (e.g. an antigen). Unless otherwise indicated, as used herein, "binding affinity" refers to the intrinsic binding affinity that reflects a 1:1 interaction between members of a binding pair (e.g., antibody and antigen). The affinity of an X-binding molecule for its Y-binding partner can generally be represented by the dissociation constant (KD). Affinity can be measured by common methods known in the art, including those described herein. Low-affinity antibodies generally bind to the antigen slowly and tend to dissociate easily, while high-affinity antibodies generally bind to the antigen faster and tend to remain bound longer. A variety of methods of measuring binding affinity are known in the art, any of which can be used for the purposes of the present disclosure. Specific illustrative embodiments include the following. In one embodiment, "KD" or "KD value" may be measured by assays known in the art, for example by a binding assay. KD can be measured in a radioactively labeled antigen binding assay (RIA), for example, performed with the Fab version of an antibody of interest and its antigen (Chen, et al, (1999) J. Mol Biol 293:865- 881). KD or KD value can also be measured using surface plasmon resonance assays by Biacore, using, for example, a BIAcoreTM-2000 or a BIAcoreTM-3000 (BIAcore, Inc., Piscataway, NJ), or by biolayer interferometry. using, for example, the OctetQK384 system (ForteBio, Menlo Park, CA). An "on-rate" or "association rate" or "rate to association" or "kon" can also be determined with the same surface plasmon resonance or biolayer interferometry techniques described above, using, for example, a BIAcoreTM-2000 or a BIAcoreTM-3000 (BIAcore, Inc., Piscataway, NJ), or the OctetQK384 system (ForteBio, Menlo Park, CA).

[0123] The phrase "substantially similar" or "substantially the same" denotes a sufficiently high degree of similarity between two numerical values (e.g., one associated with an antibody of the present disclosure and the other associated with a reference antibody) so as to that one skilled in the art would consider the difference between the two values to be of little or no statistical and/or biological significance in the context of the biological characteristic measured by the values (e.g., KD values). For example, the difference between two values may be less than about 50%, less than about 40%, less than about 30%, less than about 20%, less than about 10%, less than about 5%, as a function of the value for the reference antibody.

[0124] The phrase "substantially reduced," or "substantially different," as used herein, denotes a sufficiently high degree of difference between two numerical values (e.g., one associated with an antibody of the present disclosure and the other associated with a reference antibody) such that one skilled in the art would consider the difference between the two values to be of statistical significance in the context of the biological characteristic measured by the values. For example, the difference between said two values may preferably be greater than about 10%, greater than about 20%, greater than about 30%, greater than about 40%, greater than about 50% as a function of the reference antibody value.

[0125] Antibody "effector functions" refer to the biological activities attributable to the Fc region (e.g., a native sequence Fc region or variant amino acid sequence Fc region) of an antibody and vary with the antibody isotype. Examples of antibody effector functions include: C1q binding and complement-dependent cytotoxicity; binding to the Fc receptor; antibody-dependent cell-mediated cytotoxicity (ADCC); phagocytosis; downregulation of cell surface receptors (e.g., B cell receptor); and the activation of B cells.

[0126] The term "Fc region" is used herein to define a C-terminal region of an immunoglobulin heavy chain, including, for example, native sequence Fc regions, recombinant Fc regions, and variant Fc regions. Although the boundaries of the Fc region of an immunoglobulin heavy chain can vary, the Fc region of the human IgG heavy chain is often defined to stretch from an amino acid residue at position Cys226, or Pro230, to its carboxyl terminus. The C-terminal lysine (residue 447 according to the EU numbering system) of the Fc region can be removed, for example, during production or purification of the antibody, or by recombinant engineering of the nucleic acid encoding an antibody heavy chain. . Accordingly, an intact antibody composition may comprise populations of antibodies with all K447 residues removed, populations of antibodies with no K447 residues removed, and antibody populations having a mixture of antibodies with and without the K447 residue.

[0127] A "functional Fc region" has an "effector function" of a native sequence Fc region. Examples of “effector functions” include C1q binding; complement-dependent cytotoxicity (CDC); binding to the Fc receptor; antibody-dependent cell-mediated cytotoxicity (ADCC); phagocytosis; downregulation of cell surface receptors (e.g. B cell receptor; BCR), etc. Such effector functions generally require that the Fc region be combined with a binding region or binding domain (e.g., an antibody variable region or domain) and can be assessed using various assays as disclosed.

[0128] A "native sequence Fc region" comprises an amino acid sequence identical to the amino acid sequence of an Fc region found in nature, and not manipulated, modified and/or altered (e.g., isolated, purified, selected, including or combining with other sequences, such as variable region sequences) from a human. Human native sequence Fc regions include a human native Fc sequence region of IgG1 (non-A and A allotypes); native human Fc sequence region of IgG2; native human IgG3 Fc sequence region; and the Fc region of human IgG4 native sequence, as well as its naturally occurring variants.

[0129] A "variant Fc region" comprises an amino acid sequence that differs from that of a native sequence Fc region by virtue of at least one amino acid modification, (e.g., substitution, addition or deletion) preferably one or more amino acid substitutions. Preferably, the variant Fc region has at least one amino acid substitution compared to a native sequence Fc region or the Fc region of a parent polypeptide, e.g., from about one to about ten amino acid substitutions, and from preferably from about one to about five amino acid substitutions in a native sequence Fc region or in the Fc region of the parent polypeptide. The variant Fc region herein preferably will have at least about 80% homology to a native sequence Fc region and/or to an Fc region of a parent polypeptide, and more preferably at least about 90% homology thereto, e.g. example, at least about 95% homology with it. For example, a variant with two amino acid changes to alanine at two positions in the Fc sequence of human IgG1 is shown in bold in the amino acid sequence given below: ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGL YSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPALAGGPSVF LFPPKPKDTLMISRTPEVT CVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVV SVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSL TCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSV MHEALHNHYTQKSLSLS PGK (SEQ ID NO: 316) A variant sequence can be used in humanized heavy chain constructs, as shown below for a humanized 5H23-VH3 (see, for example, Example 7) designated 5H23(vH3)-hIgG1(E233A) (L235A), as provided below; the amino acids that make up the signal sequence are underlined and the variable region sequence is in bold: mdmrvpaqllgllllwlrgarcQVQLQQSGAEVKKPGASVKVSCKAS GYTFTSYDINWVRQA PGQGLEWIGWIYPGDGSTKYNEKFKGKATITRDTSASTAYMELSSLRSEDTAVYFCARSDYY GSRSFAYWGQGTLVTVSSASTKGPS VFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSG ALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKT HTCPPCPAPALAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVH NAKTKPREEQYNSTYRVVSVLTVLHQDWLNGK EYKCKVSNKALPAPIEKTISKAKGQPREPQ VYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSK LTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK (SEQ ID NO: 317)

[0130] A "light chain constant region" includes kappa and lambda constant regions. An exemplary kappa constant region is provided below: RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSK DSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 318) Such a kappa constant region sequence can be used in humanized light chain constructs such as shown below for a humanized 5H23-VL2 ( see, for example, Example 7), as given below; the amino acids that make up the signal sequence are underlined and the variable region sequence is in bold: mdmrvpaqllgllllwlrgarcDIVMTQSPDSLAVSLGERATINCRASKSVSTSGYVYMHWY QQKPGQPPKLLIYLASYLESGVPDRFSGSGSGTDFTLTISSVQAEDVAVYYCQHSRDLTFPF GGGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVC LLNNFYPREAKVQWKVDNALQSGNSQ ESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 319)

[0131] The term "variant" when used in relation to Klotho beta or an Anti-Klotho beta antibody may refer to a peptide or polypeptide comprising one or more (such as, for example, about 1 to about 25 , about 1 to about 20, about 1 to about 15, about 1 to about 10, or about 1 to about 5) amino acid sequence substitutions, deletions and/or additions, compared to a native or unmodified Klotho beta sequence. For example, a Klotho beta variant may result from one or more (such as, for example, about 1 to about 25, about 1 to about 20, about 1 to about 15, about 1 to about of 10, or about 1 to about 5) changes to a native Klotho beta amino acid sequence. Also by way of example, a variant of an Anti-Klotho beta antibody may result from one or more (such as, for example, about 1 to about 25, about 1 to about 20, about 1 to about 15, about 1 to about 10, or about 1 to about 5) changes to an amino acid sequence of a native or previously unmodified anti-Klotho beta antibody. Variants may be naturally occurring, such as allelic or splice variants, or they may be artificially constructed. Polypeptide variants can be prepared from the corresponding nucleic acid molecules that encode the variants. In specific embodiments, the Klotho beta variant or anti-Klotho beta antibody variant at least retains the functional activity of Klotho beta or the anti-Klotho beta antibody, respectively. In specific embodiments, an Anti-Klotho beta antibody variant binds to Klotho beta and/or is an agonist at Klotho beta activity. In specific embodiments, an anti-Klotho beta antibody variant binds to Klotho beta and/or is an agonist at Klotho beta activity. In certain embodiments, the variant is encoded by a variant single nucleotide polymorphism (SNP) of a nucleic acid molecule encoding VH or VL regions or subregions of Klotho beta or anti-Klotho beta antibodies, such as one or more CDRs.

[0132] The term "vector" refers to a substance that is used to carry or include a nucleic acid sequence, including, for example, in order to introduce a nucleic acid sequence into a host cell. Applicable vectors for use include, for example, expression vectors, plasmids, phage vectors, viral vectors, episomes and artificial chromosomes, which may include selection sequences or markers operable for stable integration into the chromosome of a host cell. Furthermore, vectors may include one or more selectable marker genes and appropriate expression control sequences. Selectable marker genes that can be included, for example, provide resistance to antibiotics or toxins, complement auxotrophic deficiencies, or provide critical nutrients not in the culture medium. Expression control sequences may include constitutive and inducible promoters, transcription enhancers, transcription terminators, and the like, which are well known in the art. When two or more nucleic acid molecules are to be co-expressed (e.g., both an antibody heavy and light chain or antibody VH and VL), both nucleic acid molecules can be inserted, for example, into a single vector expression vectors or in separate expression vectors. For a single expression vector, the encoding nucleic acids can be operably linked to a common expression control sequence or linked to different expression control sequences, such as an inducible promoter and a constitutive promoter. The introduction of nucleic acid molecules into a host cell can be confirmed using methods well known in the art. Such methods include, for example, nucleic acid analysis, such as Northern hybridizations or polymerase chain reaction (PCR) amplification of mRNA, or immunoblotting for expression of gene products, or other analytical methods suitable for testing expression. of an introduced nucleic acid sequence or its corresponding gene product. It is understood by those skilled in the art that nucleic acid molecules are expressed in an amount sufficient to produce a desired product (e.g., an Anti-Klotho beta antibody as described herein), and it is further understood that expression levels may be optimized to obtain sufficient expression using methods well known in the art.

[0133] "Antibody-dependent cell-mediated cytotoxicity" or "ADCC" refers to a form of cytotoxicity in which secreted Ig binds to Fc receptors (FcRs) present on certain cytotoxic cells (e.g., natural killer cells ( NK), neutrophils, and macrophages) enable these cytotoxic effector cells to specifically bind to a target cell carrying the antigen and subsequently kill the target cell with cytotoxins. Antibodies "arm" cytotoxic cells and are absolutely necessary for such killing. The primary cells to mediate ADCC, NK cells, express FcyRIII only, while monocytes express FCYRI, FCYRII, and FCYRIII. Expression of FcR in hematopoietic cells is known (see, for example, Table 3, page 464, Ravetch and Kinet, Annu Rev. Immunol. 9:457-92 (1991)). To evaluate the ADCC activity of a molecule of interest, an in vitro ADCC assay, (see, for example, US Patent No. 5500362 or 5821337) can be performed. Useful effector cells for such assays include peripheral blood mononuclear cells (PBMC) and natural killer (NK) cells. Alternatively, or additionally, the ADCC activity of the molecule of interest can be assessed in vivo, for example in an animal model (see, for example, Clynes et al (USA) 95:652-656 (1998)). Antibodies with little or no ADCC activity can be selected for use.

[0134] "Fc receptor" or "FcR" describes a receptor that binds to the Fc region of an antibody. The preferred FcR is a native sequence human FcR. Furthermore, a preferred FcR is one that binds to an IgG antibody (e.g., a gamma receptor) and includes receptors from the FcYRI, FCYRII and FCYRIII subclasses, including allelic variants and alternatively spliced forms of these receptors. FCYRII receptors include FCYRIIA (an "activating receptor") and FCYRIIB (an "inhibiting receptor"), which have similar amino acid sequences that differ primarily in their cytoplasmic domains (see, e.g., Daeron, Annu. Rev. Immunol 15:203-234 (1997)). FcRs are known (see, for example, Ravetch and Kinet, Annu. Rev. Immunol. 9:457-492 (1991); Capel et al., Immunomethods 4:25-34 (1994); and de Haas et al. , J. Lab. Clin. 126:330-41 (1995)). Other FcRs, including those to be identified in the future, are encompassed by the term "FcR" herein. The term also includes the neonatal receptor, FcRn, which is responsible for the transfer of maternal IgG to the fetus (see, for example, Guyer et al, J. Immunol 117:587 (1976) and Kim et al, J. Immunol. 24 :249 (1994)). Antibody variants with enhanced or decreased binding to FcR have been described (see, for example, WO 2000/42072; US Patent Nos. 7,183,387, 7,332,581 and 7,335,742; Shields et al, J. Biol Chem 9(2): 6591-6604 (2001)).

[0135] "Complement-dependent cytotoxicity" or "CDC" refers to the lysis of a target cell in the presence of complement. Activation of the classical complement pathway is initiated by the binding of the first component of the complement system (C1q) to antibodies (of the appropriate subclass) that are bound to its cognate antigen. To assess complement activation, a CDC assay, (see, for example, Gazzano-Santoro et al, J. Immunol Methods 202:163 (1996)), can be performed. Polypeptide variants with altered Fc region amino acid sequences (polypeptides with a variant Fc region) and increased or reduced C1q binding capacity have been described, (see, for example, US Patent No. 6,194,551, WO 1999/ 51642, Idusogie et al., J. Immunol 164: 4178-4184 (2000)). Antibodies with little or no CDC activity can be selected for use.

[0136] A Klotho beta polypeptide "extracellular domain" or "ECD" refers to a form of the Klotho beta polypeptide that is essentially free of the transmembrane and cytoplasmic domains. For example, a Klotho beta polypeptide ECD may have less than 1% of these transmembrane and/or cytoplasmic domains and, preferably, may have less than 0.5% of such domains. The term "identity" refers to a relationship between the sequences of two or more polypeptide molecules or two or more nucleic acid molecules, as determined by alignment and comparison of the sequences. "Percent identity" means the percentage of identical residues among the amino acids or nucleotides in the molecules being compared and is calculated based on the size of the smallest of the molecules being compared. For these calculations, gaps in the alignments (if any) must be addressed by a particular mathematical model or computer program (e.g., an "algorithm"). Methods that can be used to calculate the identity of aligned nucleic acids or polypeptides include those described in Computational Molecular Biology, (Lesk, A. M., ed.), (1988) New York: Oxford University Press; Biocomputing Informatics and Genome Projects, (Smith, D. W., ed.), 1993, New York: Academic Press; Computer Analysis of Sequence Data, Part I, (Griffin, A. M., and Griffin, H. G., eds.), 1994, New Jersey: Humana Press; von Heinje, G., (1987) Sequence Analysis in Molecular Biology, New York: Academic Press; Sequence Analysis Primer, (Gribskov, M. and Devereux, J., eds.), 1991, New York: M. Stockton Press; and Carillo et al., (1988) SIAM J. Applied Math. 48:1073.

[0137] For the calculation of percentage identity, the sequences to be compared can be aligned in a way that gives them the greatest correspondence between the sequences. One computer program that can be 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, Wis. .). The GAP computer algorithm typically aligns two polypeptides or polynucleotides for which percent sequence identity must be determined. Sequences can be aligned for optimal matching of their respective amino acids or nucleotides (the "combined span", as determined by the algorithm). A large gap penalty (which is calculated as 3 times the average diagonal, where the "average diagonal" is the average of the diagonal of the comparison matrix being used; the "diagonal" is the score or number assigned to each perfect amino acid match by the particular comparison matrix) and a gap extension penalty (which is usually 1/10 times the gap opening penalty), as well as a comparison matrix such as PAM 250 or BLOSUM 62 are used in together with the algorithm. In certain embodiments, a standard comparison matrix (see, Dayhoff et al, (1978) Atlas of Protein Sequence and Structure 5:345-352 for the PAM 250 comparison matrix; Henikoff et al, (1992) Proc Natl Acad. Sci . USA 89:10915-10919 for the BLOSUM 62 comparison matrix is also used by the algorithm.

[0138] Exemplary parameters for determining the percentage identity of polypeptides or nucleotide sequences using the GAP program are as follows: (i) Algorithm: Needleman et al, 1970, J. Mol. Biol. 48:443-453; (ii) Comparison matrix:. BLOSUM 62 by Henikoff et al, 1992, supra; (iii) Gap Penalty: 12 (but with no penalty for final gaps) (iv) Gap Length Penalty: 4; and (v) Similarity threshold: 0.

[0139] Certain alignment schemes for aligning two amino acid sequences may result in matching only a small region of the two sequences, and this small aligned region may have very high sequence identity, although there is no significant relationship between the two amino acid sequences. full length. Therefore, the selected alignment method (e.g., the GAP program) can be adjusted if desired to result in an alignment that spans a series of amino acids, e.g., at least 50 contiguous amino acids of the target polypeptide.

[0140] "Percent amino acid sequence identity (%)" with respect to a reference polypeptide sequence is defined as the percentage of amino acid residues in a candidate sequence that are identical to amino acid residues in the reference polypeptide sequence, after align sequences and introduce gaps if necessary to achieve maximum percent sequence identity, and not considering any conservative substitutions as part of sequence identity. Alignment for purposes of determining percent amino acid sequence identity can be accomplished in a variety of ways that are within the skill of the art, for example, using publicly available computer programs such as BLAST, BLAST-2, ALIGN. or Megaalign software (DNASTAR). Those skilled in the art can determine appropriate parameters for sequence alignment, including any algorithms necessary to achieve maximum alignment over the full length of the sequences being compared.

[0141] A "modification" of an amino acid residue/position refers to a change of a primary amino acid sequence compared to a starting amino acid sequence, wherein the change results from a sequence change involving said amino acid residue/positions. For example, typical modifications include replacing the residue with another amino acid (e.g., a conservative or non-conservative substitution), insertion of one or more (e.g., generally less than 5, 4, or 3) amino acids adjacent to said residue/ position, and/or elimination of said waste/position.

[0142] An "epitope" is the location on the surface of an antigen molecule to which a single antibody molecule binds, such as a region located on the surface of an antigen, such as a Klotho beta polypeptide, a polypeptide fragment of Klotho beta or a Klotho beta epitope, which is capable of being bound to one or more antigen-binding regions of an antibody, and which has antigenic or immunogenic activity in an animal, such as a mammal (e.g., a human ), which is capable of triggering an immune response. An epitope that has immunogenic activity is a portion of a polypeptide that induces an antibody response in an animal. An epitope that has antigenic activity is a portion of a polypeptide to which an antibody binds, as determined by any method well known in the art, including, for example, by an immunoassay. Antigenic epitopes do not necessarily have to be immunogenic. Epitopes generally consist of chemically active surface clusters of molecules such as amino acids or sugar side chains, and have specific three-dimensional structural features as well as specific charge characteristics. The term "epitope" specifically includes linear epitopes and conformational epitopes. A region of a polypeptide that contributes to an epitope may be contiguous amino acids of the polypeptide or the epitope may join from two or more non-contiguous regions of the polypeptide. The epitope may or may not be a three-dimensional surface feature of the antigen. In certain embodiments, a Klotho beta epitope is a three-dimensional surface feature of a Klotho beta polypeptide. In other embodiments, a Klotho beta epitope is a linear function of a Klotho beta polypeptide. Generally an antigen has several or many different epitopes and can react with many different antibodies.

[0143] An antibody binds to "an epitope" or "essentially the same epitope" or "the same epitope" as a reference antibody, when the two antibodies recognize identical, overlapping or adjacent epitopes in three-dimensional space. The most widely used and rapid methods for determining whether two antibodies bind to identical, overlapping, or adjacent epitopes in three-dimensional space are competition assays, which can be configured in a number of different formats, for example, using a labeled antigen or antibody. marked. In some assays, the antigen is immobilized on a 96-well plate, or expressed on the surface of cells, and the ability of unlabeled antibodies to block the binding of labeled antibodies is measured using radioactive, fluorescent, or enzymatic labels.

[0144] "Epitope mapping" is the process of identifying the binding sites, or epitopes, of antibodies on their target antigens. Antibody epitopes can be linear epitopes or conformational epitopes. Linear epitopes are formed by a continuous sequence of amino acids in a protein. Conformational epitopes are formed from amino acids that are discontinuous in the protein sequence, but are brought together by folding the protein into its three-dimensional structure. Induced epitopes are formed when the three-dimensional structure of the protein is in an altered conformation, such as following activation or the binding of another protein or a ligand (e.g., the binding of Klotho beta to an FGR receptor such as FGRFR1c, FGFR2c , FGFR3c, or FGFR4c.

[0145] "Epitope Binning" is the process of grouping antibodies based on the epitopes they recognize. More particularly, epitope binning comprises methods and systems for discriminating the epitope recognition properties of different antibodies, using competitive assays coupled with computational processes to cluster antibodies based on their epitope recognition properties and identify antibodies having epitope specificities. different connections.

[0146] A "Klotho beta-mediated disease" and "Klotho beta-mediated disorder" and "Klotho beta-mediated condition" are used interchangeably and refer to any disease, disorder or condition that is completely or partially caused by , is either the result of Klotho beta or the interaction of a Klotho beta with an FGF receptor such as FGFR1c, FGFR2c, FGFR3c, or FGFR4 and/or alternatively any disease, disorder, or condition in which it is desirable to mimic or enhance the effects in live of FGF19 and/or FGF21.

[0147] The term "therapeutically effective amount" as used herein refers to the amount of an agent (e.g., an antibody described herein or any other agent described in the present invention) that is sufficient to reduce and/or improve the severity and /or the duration of a given disease, disorder or condition, and/or a related symptom. A therapeutically effective amount of an agent, including a therapeutic agent, may be an amount necessary to (i) reduce or ameliorate the advancement or progression of a particular disease, disorder or condition, (ii) reduce or ameliorate the recurrence, development or onset of a given disease, disorder or conditions, and/or (iii) to improve or enhance the prophylactic or therapeutic effect of another therapy (e.g., a therapy other than administration of an antibody provided herein). A "therapeutically effective amount" of a substance/molecule/agent of the present disclosure (e.g., an Anti-Klotho beta antibody) may vary according to factors such as the disease state, age, sex, and weight of the individual, and the ability of the substance/molecule/agent to elicit a desired response in the individual. A therapeutically effective amount encompasses an amount in which any toxic or harmful effects of the substance/molecule/agent are outweighed by the therapeutically beneficial effects. In certain embodiments, the term "therapeutically effective amount" refers to an amount of an antibody or other agent (e.g., or drug) effective to "treat" a disease, disorder, or condition in an individual or mammal.

[0148] An "effective amount" is generally an amount sufficient to reduce the severity and/or frequency of symptoms, eliminate the symptoms and/or underlying cause, prevent the occurrence of symptoms and/or their underlying cause, and/or improve or correct harm that results from, or is associated with, a disease, disorder or condition, including, for example, diabetes, obesity, dyslipidemia, cardiovascular disease, metabolic syndrome or broadly any disease, disorder or condition in which it is desirable to imitate or enhance the in vivo effects of FGF19 and/or FGF21. In some embodiments, the effective amount is a therapeutically effective amount or a prophylactically effective amount. A "therapeutically effective amount" is an amount sufficient to cure a disease, disorder, or condition (e.g., type 2 diabetes, obesity, dyslipidemia, NASH, cardiovascular disease, metabolic syndrome, or broadly any disease, disorder, or condition in which it is desirable to mimic or enhance the in vivo effects of FGF19 and/or FGF21) or symptoms, in particular a disease, disorder or condition, or the symptoms associated with such a disease, disorder or condition, or otherwise prevent, impede, delay or reverse the progression of the disease, disorder or condition, or any other undesirable symptom associated with such a disease, disorder or condition, in any form whatsoever. A "prophylactically effective amount" is an amount of a pharmaceutical composition that, when administered to an individual, has the intended prophylactic effect, e.g., preventing or delaying the onset (or recurrence) of diabetes, obesity, or dyslipidemia, or reducing the likelihood of the onset (or recurrence) of a disease, disorder or condition or associated symptom(s), including, for example, diabetes, obesity, dyslipidemia, cardiovascular disease, metabolic syndrome or broadly any disease, disorder or condition in which it is It is desirable to mimic or enhance the in vivo effects of FGF19 and/or FGF21 or associated symptoms. Complete therapeutic or prophylactic effect does not necessarily occur upon administration of one dose, and may occur only after administration of a series of doses. Thus, a therapeutically or prophylactically effective amount can be administered in one or more administrations.

[0149] A "prophylactically effective amount" refers to an effective amount, in dosages and for periods of time necessary, to achieve the desired prophylactic result. Typically, but not necessarily, since a prophylactic dose is used in individuals prior to or at an early stage of a disease, disorder or condition, a prophylactically effective amount may be less than a therapeutically effective amount.

[0150] "Chronic" administration refers to the administration of the agent(s) in a continuous mode (e.g., over a period of time, such as days, weeks, months, or years), as opposed to an acute mode , in order to maintain the initial therapeutic effect (activity) for a prolonged period of time. "Intermittent" administration is treatment that is not carried out consecutively without interruption, but is instead cyclical in nature.

[0151] Administration "in combination with" one or more additional therapeutic agents includes simultaneous (e.g., concurrent) and consecutive administration in any order. The term "in combination" in the context of administering other therapies (e.g., other agents) includes the use of more than one therapy (e.g., one agent). The use of the term "in combination" does not restrict the order in which therapies are administered to an individual. A first therapy (e.g., agent) may be administered earlier (e.g., 1 minute, 15 minutes, 30 minutes, 45 minutes, 1 hour, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 7 hours, 8 hours, 12 hours, 24 hours, 48 hours, 72 hours, 96 hours, 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 8 weeks, 9 weeks, 10 weeks, 11 weeks, or 12 weeks), simultaneously, or later (e.g. 1 minute, 15 minutes, 30 minutes, 45 minutes, 1 hour, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 7 hours, 8 hours, 12 hours, 24 hours, 48 hours, 72 hours, 96 hours, 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks, 9 weeks, 10 weeks, 11 weeks or 12 weeks) administration of a second therapy (e.g., agent) to an individual who has, has, or is susceptible to a Klotho beta-mediated disease.

[0152] Any additional therapy (e.g., agent) can be administered in any order with the other additional therapies (e.g., agents). In certain embodiments, antibodies may be administered in combination with one or more therapies, such as agents (e.g., therapies, including agents, other than antibodies that are currently administered) to prevent, treat, manage, and/or or improve a Klotho beta-mediated disease. Non-limiting examples of therapies (e.g., agents) that may be administered in combination with an antibody include, for example, analgesic agents, anesthetic agents, antibiotics or immunomodulatory agents, or any other agent listed in the United States Pharmacopeia and/or Physician's Desk Reference. Examples of agents useful in combination therapy include, but are not limited to, the following: nonsteroidal anti-inflammatory drugs (NSAIDs) such as aspirin, ibuprofen, and other propionic acid derivatives (alminoprofen, benoxaprofen, bucloxic acid, carprofen, fenbufen, fenoprofen, fluprofen, flurbiprofen, indoprofen, ketoprofen, miroprofen, naproxen, oxaprozin, pirprofen, pranoprofen, suprofen, tiaprofenic acid, and tioxaprofen), acetic acid derivatives (indomethacin, acemetacin, alclofenac, clidanac, diclofenac, fenclofenac, fenclozic acid, fentiazac , fuirofenac, ibufenac, isoxepac, oxpinac, sulindac, tiopinac, tolmetin, zidomethacin and zomepirac), fenamic acid derivatives (flufenamic acid, meclofenamic acid, mefenamic acid, niflumic acid and tolfenamic acid), biphenylcarboxylic acid derivatives (diflunisal and flufenisal) , oxicams (isoxicam, piroxicam, sudoxicam and tenoxicam), salicylates (acetyl salicylic acid, sulfasalazine) and pyrazolones (apazone, bezpiperilone, feprazone, mofebutazone, oxyphenbutazone, phenylbutazone). Other combinations include cyclooxygenase-2 (COX-2) inhibitors. Other combination agents include steroids such as prednisolone, prednisone, methylprednisolone, betamethasone, dexamethasone, or hydrocortisone. Such a combination may be particularly advantageous, since one or more side effects of the steroid may be reduced or even eliminated by gradually decreasing the dose of steroid required in treating patients in combination with the antibodies of the present invention. Additional examples of agents for combinations include cytokine suppressive anti-inflammatory drugs (CSAIDs); antibodies or antagonists of other human cytokines or growth factors, e.g., TNF, LT, IL-1β, IL-2, IL-6, IL-7, IL-8, IL-15, IL-16, IL-18 , EMAP-II, GM-CSF, FGF, or PDGF. Combinations of agents may include TNF antagonists such as chimeric, humanized or human TNF antibodies, REMICADE, anti-TNF antibody fragments (e.g., CDP870), and soluble p55 or p75 TNF receptors, derivatives thereof, p75TNFRIgG (Enbrel ®) or p55TNFR1gG (LENERCEPT®), soluble IL-13 receptor (sIL-13), and also TNFα-converting enzyme (TACE) inhibitors; Similarly, IL-1 inhibitors (e.g., interleukin-1 converting enzyme inhibitors) may be effective. Other combinations include interleukin-11, anti-P7s and p-selectin glycoprotein ligand (PSGL). Other examples of agents useful in combination therapy include interferon-β1a (AVONEX); interferon-β1b (BETASERON®); Copaxone; hyperbaric oxygen therapy; intravenous immunoglobulin; clabribine; and antibodies to or antagonists of other human cytokines or growth factors (e.g., antibodies to CD40 and CD80 ligand).

[0153] "Carriers" as used herein include pharmaceutically acceptable carriers, excipients or stabilizers that are non-toxic to the cell or mammal exposed to them, in the dosages and concentrations employed. Often, the physiologically acceptable carrier is a pH-buffered aqueous solution. Examples of physiologically acceptable carriers include buffers such as phosphate, citrate, and other organic acids; antioxidants including ascorbic acid; low molecular weight polypeptides (e.g., less than about 10 amino acid residues); proteins, such as serum albumin, gelatin, or immunoglobulins; hydrophilic polymers such as polyvinylpyrrolidone; amino acids, such as glycine, glutamine, asparagine, arginine or lysine; monosaccharides, disaccharides, and other carbohydrates including glucose, mannose or dextrins; chelating agents such as EDTA; , polyethylene glycol (PEG), and PLURONICS™. The term "carrier" may also refer to a diluent, adjuvant (e.g., Freund's adjuvant (complete or incomplete)), excipient or vehicle with which the therapeutic is administered. Carriers, including pharmaceutical carriers, may be sterile liquids, such as water and oils, including those of petroleum, animal, vegetable or synthetic origin, such as peanut oil, soybean oil, mineral oil, sesame oil and the like. Water is an exemplary carrier when a composition (e.g., a pharmaceutical composition) is administered intravenously. Saline solutions and aqueous solutions of dextrose and glycerol can also be used as liquid carriers, particularly for injectable solutions. Suitable excipients (e.g. pharmaceutical excipients) include starch, glucose, lactose, sucrose, gelatin, malt, rice, flour, chalk, silica gel, sodium stearate, glycerol monostearate, talc, sodium chloride, dry skimmed milk, glycerol, propylene, glycol, water, ethanol and the like. The composition, if desired, may also contain minor amounts of wetting or emulsifying agents, or pH buffering agents. The compositions may take the form of solutions, suspensions, emulsions, tablets, pills, capsules, powders, sustained-release formulations and the like. Oral compositions, including formulations, may include standard carriers such as pharmaceutical grades of mannitol, lactose, starch, magnesium stearate, sodium saccharin, cellulose, magnesium carbonate, etc. Examples of suitable pharmaceutical carriers are described in Remington's Pharmaceutical Sciences (1990) Mack Publishing Co., Easton, PA. The compositions, including pharmaceutical compounds, may contain a prophylactically or therapeutically effective amount of an Anti-Klotho beta antibody, for example, in isolated or purified form, together with a suitable amount of carrier so as to provide the form for administration appropriate to the individual. (e.g. patient). The formulation must be suitable for the method of administration.

[0154] The term "pharmaceutically acceptable" as used herein means being approved by a federal government or state government regulatory agency or listed in the United States Pharmacopeia, European Pharmacopoeia, or other generally recognized pharmacopoeia for use in animals, and more particularly in humans.

[0155] The term "pharmaceutical formulation" refers to a preparation that is such that the biological activity of the active ingredient (e.g., an Anti-Klotho beta antibody) is effective, and that does not contain any additional components. that are unacceptably toxic to the individual to whom the formulation would be administered. Such a formulation may be sterile.

[0156] A "sterile" formulation is aseptic or free of all living microorganisms and their spores.

[0157] "Polyclonal antibodies", as used herein, refer to a population of antibodies generated in an immunogenic response to a protein having many epitopes and thus includes a variety of different antibodies directed to the same epitope and to different epitopes within of the protein. Methods for producing polyclonal antibodies are known in the art (See, for example, Chapter 11 in: Short Protocols in Molecular Biology, (2002) 5th Ed., Ausubel et al., eds., John Wiley and Sons, New York ).

[0158] An "isolated nucleic acid" is a nucleic acid, for example, an RNA, DNA, or a mixed polymer, that is substantially separated from other DNA sequences in the genome, as well as proteins or complexes, such as ribosomes and polymerases , which naturally accompany a native sequence. An "isolated" nucleic acid molecule is one that is separated from other nucleic acid molecules that are present in the natural source of the nucleic acid molecule. Furthermore, an "isolated" nucleic acid molecule, such as a cDNA molecule, may be substantially free of other cellular material, or culture medium when produced by recombinant techniques, or substantially free of chemical precursors or other chemicals when synthesized. chemically. In a specific embodiment, one or more nucleic acid molecules encoding an antibody as described herein are isolated or purified. The term encompasses nucleic acid sequences that have been removed from their naturally occurring environment, and includes recombinant or cloned DNA isolates and chemically synthesized analogues or biologically synthesized analogues by heterologous systems. A substantially pure molecule may include isolated forms of the molecule.

[0159] "Polynucleotide", or "nucleic acid", as used interchangeably herein, refers to nucleotide polymers of any length and includes DNA and RNA. Nucleotides can be deoxyribonucleotides, ribonucleotides, modified nucleotides or bases, and/or their analogues or any substrate that can be incorporated into a polymer by DNA or RNA polymerase or by a synthesis reaction. A polynucleotide may comprise modified nucleotides, such as methylated nucleotides and analogues thereof. "Oligonucleotide" as used herein generally refers to short, generally single-stranded and generally synthetic polynucleotides, which are generally, but not necessarily, less than about 200 nucleotides in length. The terms "oligonucleotide" and "polynucleotide" are not mutually exclusive. The above description for polynucleotides is equally and fully applicable to oligonucleotides. A cell that produces an anti-Klotho beta antibody of the present invention can include a progenitor hybridoma cell, as well as bacterial and eukaryotic host cells into which the nucleic acid encoding the antibodies has been introduced. Suitable host cells are described below.

[0160] Unless otherwise specified, the left end of any single-stranded polynucleotide sequence disclosed herein is the 5' end; the leftward direction of double-stranded polynucleotide sequences is referred to as the 5' direction. The direction of addition from 5' to 3' of nascent RNA transcripts is termed the direction of transcription; the sequence regions in the DNA chain having the same sequence as the RNA transcript that are 5' to 5' end of the RNA transcript are called "upstream sequences"; The sequence regions in the DNA chain having the same sequence as the RNA transcript that are 3' to 3' end of the RNA transcript are called "downstream sequences".

[0161] The term "package leaflet" is used to refer to the instructions usually included in commercial packages of therapeutic products, which contain information about the indications, use, dosage, administration, contraindications and/or warnings regarding the use of such therapeutic products.

[0162] The terms "prevent", "preventing", and "prevention" refer to total or partial inhibition of the development, recurrence, appearance or spread of a Klotho beta-mediated disease and/or symptom related thereto, resulting from administering a therapy or a combination of therapies provided herein (e.g., a combination of therapeutic or prophylactic agents, such as an antibody provided herein).

[0163] The term "prophylactic agent" refers to any agent that can totally or partially inhibit the development, recurrence, appearance or spread of a Klotho beta-mediated disease and/or symptom related thereto in an individual. In certain embodiments, the term "prophylactic agent" refers to an anti-Klotho beta antibody as described herein. In certain other embodiments, the term "prophylactic agent" refers to an agent other than an anti-Klotho beta antibody as described herein. In certain embodiments, a prophylactic agent is an agent that is known to be useful or has been or is being used to prevent a Klotho beta-mediated disease, disorder or condition, and/or a symptom related thereto or prevent the onset, development , progression and/or severity of a Klotho beta-mediated disease, disorder or condition, and/or a related symptom. In specific embodiments, the prophylactic agent is a humanized anti-Klotho beta antibody, such as a humanized anti-Klotho beta monoclonal antibody.

[0164] In certain embodiments, a "prophylactically effective serum titer" is the serum titer in an individual, preferably a human, that fully or partially inhibits the development, recurrence, onset or spread of a Klotho beta-mediated disease, disorder or condition and/or symptom related to it in the individual.

[0165] In certain embodiments, a "therapeutically effective serum titer" is the serum titer in an individual, preferably a human, that reduces the severity, duration and/or symptoms associated with a Klotho beta-mediated disease, disorder or condition, in the individual.

[0166] The term "recombinant antibody" refers to an antibody that is prepared, expressed, created or isolated by recombinant means. Recombinant antibodies can be antibodies expressed using a recombinant expression vector transfected into a host cell, antibodies isolated from a combinatorial library of antibodies and recombinants, antibodies isolated from an animal (e.g., a mouse or a cow) that is transgenic and/or transchromosomal to human immunoglobulin genes (see, e.g., Taylor, L. D. et al. (1992) Nucl. Acids Res. 20:6287-6295) or antibodies prepared, expressed, raised or isolated by any other means involving splicing of immunoglobulin gene sequences to other DNA sequences. Such recombinant antibodies may have variable and constant regions, including those derived from human germline immunoglobulin sequences (see Kabat, E. A. et al. (1991) Sequences of Proteins of Immunological Interest, Fifth Edition, U.S. Department of Health and Human Services, NIH Publication No. 91-3242). In certain embodiments, however, such recombinant antibodies can be subjected to in vitro mutagenesis (or, when an animal transgenic for human Ig sequences is used, in vivo somatic mutagenesis) and, therefore, the amino acid sequences of the VH and VL regions of the Recombinant antibodies are sequences that, although derived from and related to human germline VH and VL sequences, may not naturally exist in the germline repertoire of human antibodies in vivo.

[0167] The term "serum titer" refers to an average serum titer in an individual from multiple samples (e.g., at one time present or multiple time points) or in a population of at least 10, such like at least 20, or at least 40 individuals, up to about 100, 1000 or more.

[0168] The term "side effects" includes the undesirable and/or negative effects of a therapy (e.g., a prophylactic or therapeutic agent). Unwanted effects are not necessarily adverse. An adverse effect of a therapy (e.g., a prophylactic or therapeutic agent) may be harmful or uncomfortable or risky. Examples of side effects include diarrhea, cough, gastroenteritis, wheezing, nausea, vomiting, anorexia, abdominal cramps, fever, pain, loss of body weight, dehydration, alopecia, dyspnea, insomnia, dizziness, mucositis, muscle and nerve effects, fatigue , dry mouth and loss of appetite, rashes or swelling at the injection site, flu-like symptoms such as fever, chills and fatigue, digestive system problems and allergic reactions. Additional undesirable effects experienced by patients are numerous and known in the art. Many are described in Physician's Desk Reference (Ed. 68, 2014).

[0169] The terms "individual" and "patient" may be used interchangeably. As used herein, in certain embodiments, an individual is a mammal, such as a non-primate (e.g., cows, pigs, horses, cats, dogs, rats, etc.) or a primate (e.g., monkey and human). ). In specific embodiments, the individual is a human being. In one embodiment, the subject is a mammal (e.g., a human) having a Klotho beta-mediated disease, disorder or condition. In another embodiment, the subject is a mammal (e.g., a human) at risk of developing a Klotho beta-mediated disease, disorder or condition.

[0170] The term "substantially all" refers to at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80 %, at least about 85%, at least about 90%, at least about 95%, at least about 98%, at least about 99%, or about 100%.

[0171] The term "therapeutic agent" refers to any agent that can be used in the treatment, prevention or relief of a disease, disorder or condition, including in the treatment, prevention or relief of one or more symptoms of a disease mediated by Klotho beta, disorder or condition and/or a related symptom. In certain embodiments, a therapeutic agent refers to an anti-Klotho beta antibody as described herein. In certain other embodiments, a therapeutic agent refers to an agent other than an antibody provided herein. In certain embodiments, a therapeutic agent is an agent that is known to be useful for, or has been or is being used for the treatment, prevention, or relief of one or more symptoms of a Klotho beta-mediated disease, disorder or condition, or a related symptom.

[0172] The combination of therapies (e.g., use of agents, including therapeutic agents) may be more effective than the additive effects of any two or more therapies alone (e.g., synergistic). A synergistic effect is unexpected and cannot be predicted. For example, a synergistic effect of a combination of therapeutic agents allows for the use of lower doses of one or more of the agents and/or less frequent administration of the agents to an individual with a Klotho beta-mediated disease. The ability to use lower dosages of therapies and/or administer the therapies less frequently reduces the toxicity associated with administering the therapies to an individual, without reducing the effectiveness of the therapies for preventing, treating, or alleviating one or more symptoms. of a disease mediated by Klotho beta. Furthermore, a synergistic effect may result in improved efficacy of therapies for the prevention, treatment or alleviation of one or more symptoms of a Klotho beta-mediated disease. Finally, the synergistic effect of a combination of therapies (e.g., therapeutic agents) can avoid or reduce unwanted or adverse side effects associated with the use of either therapy alone.

[0173] The term "therapy" refers to any protocol, method and/or agent that can be used in the prevention, administration, treatment and/or improvement of a Klotho beta-mediated disease, disorder or condition. In certain embodiments, the terms "therapies" and "therapy" refer to a biological therapy, supportive therapy, and/or other therapies useful in the prevention, administration, treatment, and/or amelioration of a Klotho beta-mediated disease, disorder or condition, known to one skilled in the art, such as medical personnel.

[0174] The term "detectable probe" refers to a composition that provides a detectable signal. The term includes, without limitation, any fluorophore, chromophore, radioactive label, enzyme, antibody, or antibody fragment, and the like, which provides a detectable signal through its activity.

[0175] The term "diagnostic agent" designates a substance administered to an individual that assists in diagnosing a disease, disorder or conditions. Such substances can be used to reveal, identify and/or define the location of a disease-causing process. In certain embodiments, a diagnostic agent includes a substance that is conjugated to an anti-Klotho beta antibody as described herein, which, when administered to an individual or contacted with a sample from an individual assists in the diagnosis of a Klotho-mediated disease. beta.

[0176] The term "detectable agent" refers to a substance that can be used to determine the existence or presence of a desired molecule, such as an anti-Klotho beta antibody as described herein, in a sample or object. A detectable agent may be a substance that is capable of being visualized or a substance that is otherwise capable of being determined and/or measured (e.g., by quantification).

[0177] The term "encode" or grammatical equivalents thereof as used in reference to the nucleic acid molecule refers to a nucleic acid molecule in its native state or when manipulated by methods well known to those skilled in the art which can be transcribed to produce mRNA, which is then translated into a polypeptide and/or a fragment thereof. The antisense strand is the complement of such a nucleic acid molecule, and the coding sequence can be deduced from it.

[0178] The term "excipient" refers to an inert substance, which is commonly used as a diluent, carrier, preservative, binding agent, or stabilizing agent, and includes, but is not limited to, proteins (e.g., serum albumin, etc.), amino acids (e.g. aspartic acid, glutamic acid, lysine, arginine, glycine, histidine, etc.), fatty acids and phospholipids (e.g. alkyl sulfonates, caprylate, etc.), surfactants (e.g. SDS, polysorbate, nonionic surfactant, etc.), saccharides (e.g. sucrose, maltose, trehalose, etc.) and polyols (e.g. mannitol, sorbitol, etc.). See, also, Remington's Pharmaceutical Sciences (1990) Mack Publishing Co., Easton, PA, which is incorporated herein by reference in its entirety.

[0179] In the context of a peptide or polypeptide, the term "fragment" as used herein refers to a peptide or polypeptide that comprises less than the full-length amino acid sequence. Such a fragment may arise, for example, from an amino-terminal truncation, a carboxy-terminal truncation, and/or an internal deletion of a residue(s) from the amino acid sequence. Fragments may, for example, result from alternative RNA splicing or in vivo protease activity. In certain embodiments, Klotho beta fragments include polypeptides that comprise an amino acid sequence of at least 5 contiguous amino acid residues, at least 10 contiguous amino acid residues, at least 15 contiguous amino acid residues, at least 20 contiguous amino acid residues, at least at least 25 contiguous amino acid residues, at least 40 contiguous amino acid residues, at least 50 contiguous amino acid residues, at least 60 contiguous amino acid residues, at least 70 contiguous amino acid residues, at least 80 contiguous amino acid residues, at least 90 contiguous amino acid residues at least 100 contiguous amino acid residues at least 125 contiguous amino acid residues with at least 150 contiguous amino acid residues at least 175 contiguous amino acid residues at least 200 contiguous amino acid residues at least 250, at least 300, at least 350, at least 400, at least 450, at least 500, at least 550, at least 600, at least 650, at least 700, at least 750, at least 800, at least 850, at least 900, or at least 950 contiguous amino acid residues of the amino acid sequence of a Klotho beta polypeptide or an antibody that binds to a Klotho beta polypeptide. In a specific embodiment, a fragment of a Klotho beta polypeptide or an antibody that binds a Klotho beta antigen retains at least one, at least two, or at least three or more functions of the polypeptide or antibody.

[0180] The terms "administer," "administration," and "management" refer to the beneficial effects that an individual derives from a therapy (e.g., a prophylactic or therapeutic agent), which does not result in a cure of the disease. In certain embodiments, the subject is administered one or more therapies (e.g., prophylactic or therapeutic agents, such as an antibody provided herein) to "manage" a Klotho beta-mediated disease, one or more of its symptoms, as well as to prevent the progression or worsening of the disease.

[0181] The terms "about" or "approximately" mean within 20%, within 15%, within 10%, within 9%, within 8%, within 7%, within 6%, within 5%, within 4%, within 3%, within 2%, or within 1% or less of a given value or range.

[0182] "Administer" or "administration" refers to the act of injecting or otherwise physically delivering a substance as it exists outside the body (e.g., an Anti-Klotho beta antibody as described herein) into a patient, such as by mucosal, intradermal, intravenous, intramuscular delivery and/or any other physical delivery method described herein or known in the art. When a disease, disorder or condition, or a symptom thereof is being treated, administration of the substance typically occurs after the onset of the disease, disorder or condition, or its symptoms. When a disease, disorder or condition or its symptoms are being prevented, administration of the substance normally occurs before the onset of the disease, disorder or condition or its symptoms.

[0183] In the context of a polypeptide, the term "analog" as used herein refers to a polypeptide that has a similar or identical function to that of a Klotho beta polypeptide, a fragment of a Klotho beta polypeptide, or a Anti-Klotho beta antibody, but does not necessarily comprise an identical or similar amino acid sequence of a Klotho beta polypeptide, a fragment of a Klotho beta polypeptide, or an Anti-Klotho beta antibody, or have an identical or similar structure of an Klotho beta polypeptide, a fragment of a Klotho beta polypeptide, or an anti-Klotho beta antibody. A polypeptide having a similar amino acid sequence refers to a polypeptide that satisfies at least one of the following characteristics: (a) a polypeptide having an amino acid sequence that is at least 30%, at least 35%, at least 40% , at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90% , at least 95%, or at least 99% identical to the amino acid sequence of a Klotho beta polypeptide (e.g., SEQ ID NO: 297, a fragment of a Klotho beta polypeptide, or an Anti-Klotho beta antibody described herein (b) a polypeptide encoded by a nucleotide sequence that hybridizes under stringent conditions to a nucleotide sequence encoding a Klotho beta polypeptide, a fragment of a Klotho beta polypeptide, or an Anti-Klotho beta antibody (or region of VH or VL thereof) described herein of at least 5 amino acid residues, at least 10 amino acid residues, at least 15 amino acid residues, at least 20 amino acid residues, at least 25 amino acid residues, at least 40 amino acid residues , at least 50 amino acid residues, at least 60 amino acid residues, at least 70 amino acid residues, at least 80 amino acid residues, at least 90 amino acid residues, at least 100 amino acid residues, at least 125 amino acid residues, or at least 150 amino acid residues (see, for example, Sambrook et al. (2001) Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY; Maniatis et al. (1982) Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Press, Cold Spring Harbor, NY); and (c) a polypeptide encoded by a nucleotide sequence that is at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identical to the nucleotide sequence encoding a Klotho polypeptide beta, a fragment of a Klotho beta polypeptide, or an anti-Klotho beta antibody (or VH or VL region thereof) described herein. A polypeptide with a structure similar to a Klotho beta polypeptide, a fragment of a Klotho beta polypeptide, or an Anti-Klotho beta antibody described herein refers to a polypeptide that has a secondary, tertiary, or quaternary structure similar to a Klotho beta polypeptide, a fragment of a Klotho beta, or a Klotho beta antibody described herein. The structure of a polypeptide can be determined by methods known to those skilled in the art, including but not limited to, X-ray crystallography, nuclear magnetic resonance, and crystallographic electron microscopy.

[0184] The term "composition" is intended to encompass a product that contains the specified ingredients (e.g., an antibody provided herein) in, optionally, the specified amounts, as well as any product that results, directly or indirectly, from the combination of the specified ingredients in, optionally, the specified values.

[0185] In the context of a polypeptide, the term "derivative" as used herein refers to a polypeptide comprising an amino acid sequence of a Klotho beta polypeptide, a fragment of a Klotho beta polypeptide, or an antibody which binds to a Klotho beta polypeptide that has been altered by the introduction of amino acid residue substitutions, deletions or additions. The term "derivative" as used herein also refers to a Klotho beta polypeptide, a fragment of a Klotho beta polypeptide, or an antibody that binds to a Klotho beta polypeptide that has been chemically modified, by example, by covalently binding any type of molecule to the polypeptide. For example, but not by way of limitation, a Klotho beta polypeptide, a fragment of a Klotho beta polypeptide, or a Klotho beta antibody may be chemically modified, for example, by glycosylation, acetylation, pegylation, phosphorylation, amidation, derivatization by known protecting/blocking groups, proteolytic cleavage, binding to a cellular ligand or other protein, etc. Derivatives are modified in a way that is different from that occurring naturally or from peptides or polypeptides, either in the type or location of the attached molecules. Derivatives also include the deletion of one or more chemical groups that are naturally present in the peptide or polypeptide. A derivative of a Klotho beta polypeptide, a fragment of a Klotho beta polypeptide, or a Klotho beta antibody can be chemically modified by chemical modifications using techniques known to those skilled in the art, including, but not limited to, cleavage. specific chemistry, acetylation, formulation, metabolic synthesis of tunicamycin, etc. Furthermore, a derivative of a Klotho beta polypeptide, a fragment of a Klotho beta polypeptide, or a Klotho beta antibody may contain one or more non-classical amino acids. A polypeptide derivative has a function similar or identical to that of a Klotho beta polypeptide, a fragment of a Klotho beta polypeptide, or a Klotho beta antibody described herein.

COMPOSITIONS AND METHODS FOR THEIR MANUFACTURE

[0186] Binding proteins, such as antibodies that bind to Klotho beta (e.g., human and/or Cyno Klotho beta) are provided. The antibodies of the present disclosure are useful, for example, for the diagnosis or treatment of diseases, disorders or conditions associated with expression of Klotho beta. In certain embodiments, the antibodies of the present invention are useful for the diagnosis or treatment of a disease, disorder or condition, such as type 2 diabetes, obesity, dyslipidemia, NASH, cardiovascular disease, metabolic syndrome or broadly any disease, disorder or condition in that it is desirable to mimic or enhance the in vivo effects of FGF19 and/or FGF21.

[0187] Antibodies are provided here that bind to a Klotho beta polypeptide, a Klotho beta polypeptide fragment, Klotho beta peptide, or a Klotho beta epitope. In some embodiments, anti-Klotho beta antibodies bind to the extracellular domain (ECD) of Klotho beta. Also provided are antibodies that competitively block an anti-Klotho beta antibody provided herein from binding to a Klotho beta polypeptide. The anti-Klotho beta antibodies provided herein can also be conjugated or recombinantly fused to a diagnostic agent, a detectable agent or a therapeutic agent. Compositions comprising a Klotho beta antibody are further provided.

[0188] Also provided herein are isolated nucleic acid molecules encoding an immunoglobulin heavy chain, light chain, VH region, VL region, VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2 and /or VL CDR3 of anti-Klotho beta antibodies that bind to a Klotho beta polypeptide, a Klotho beta polypeptide fragment, a Klotho beta peptide or a Klotho beta epitope. Vectors and host cells comprising nucleic acid molecules encoding anti-Klotho beta antibodies that bind to a Klotho beta polypeptide, a Klotho beta polypeptide fragment, a Klotho beta peptide or a Klotho beta epitope are further provided. Also provided are methods of preparing antibodies that bind to a Klotho beta polypeptide, a Klotho beta polypeptide fragment, a Klotho beta peptide, or a Klotho beta epitope.

[0189] Methods of using anti-Klotho beta antibodies are provided. The methods include treating, preventing or alleviating a disease, disorder or condition, including treating, preventing or alleviating one or more symptoms of a disease, disorder or condition in an individual. Non-limiting examples of diseases, disorders or conditions include glucose utilization disorders and the sequelae associated therewith, including diabetes mellitus (type I and type 2), gestational diabetes, hyperglycemia, insulin resistance, abnormal glucose metabolism, " prediabetes" (impaired fasting glucose (IFG) or impaired glucose tolerance (IGT)), or other physiological disorders associated with, or resulting from, a hyperglycemic condition, including, for example, histopathological changes such as cell destruction β of the pancreas. For example, individuals with diseases, disorders or conditions in need of treatment may have a fasting plasma glucose (FPG) level greater than about 100 mg/dl. Other hyperglycemic-related disorders include kidney damage (e.g., tubule damage or nephropathy), liver degeneration, eye damage (e.g., diabetic retinopathy or cataracts), and diabetic foot disorders. Other diseases, disorders or conditions include dyslipidemias and their sequelae, such as, for example, atherosclerosis, coronary artery disease, cerebrovascular diseases and the like or other of the diseases, disorders or conditions that may be associated with metabolic syndrome, such as such as obesity and high body mass (including its co-morbid conditions such as, but not limited to, non-alcoholic fatty liver disease (NAFLD), non-alcoholic steatohepatitis (NASH), and polycystic ovarian syndrome (PCOS)) , or thrombosis, hypercoagulability and prothrombotic states (arterial and venous), hypertension, cardiovascular diseases, stroke and heart failure. These diseases, disorders, or conditions include atherosclerosis, chronic inflammatory bowel diseases (e.g., Crohn's disease and ulcerative colitis), asthma, lupus erythematosus, arthritis, or other rheumatic inflammatory disorders. Other diseases, disorders or conditions include fat cell tumors, lipomatous carcinomas, including, for example, liposarcoma, solid tumors, and neoplasms. Other diseases, disorders or conditions include neurodegenerative diseases and/or demyelinating diseases of the central and peripheral nervous systems and/or neurological diseases involving neuro-inflammatory processes and/or other peripheral neuropathies, including Alzheimer's disease, multiple sclerosis, Parkinson's disease , progressive multifocal leukoencephalopathy and Guillain-Barre syndrome. Other diseases, disorders or conditions include dermatological and skin disorders and/or disorders of wound healing processes, including erythematous-scaly dermatoses. Other diseases, disorders, or conditions include syndrome X, osteoarthritis, and acute respiratory distress syndrome. As used herein, the term "hyperglycemic" or "hyperglycemia" when used in reference to a disease, disorder or condition of an individual refers to a transient or chronic abnormally high level of glucose present in the blood of an individual. The disease, disorder, or condition may be caused by a delay in glucose metabolism or absorption such that the individual experiences glucose intolerance or a high glucose state not normally found in normal individuals (e.g., in intolerant prediabetic individuals). glucose levels at risk of developing diabetes or in diabetic individuals). For example, fasting plasma glucose (FPG) levels for normoglycemia may be less than about 100 mg/dl, for impaired glucose metabolism between about 100 and 126 mg/dl, and for diabetics greater than about 126 mg/dl. Prevention methods (e.g., in individuals predisposed to have a particular disorder(s), refer to delaying, slowing, or inhibiting the progression of, the onset of, or treating (e.g., ameliorating) obesity or a undesirable body mass (e.g., a greater than normal body mass index, or "BMI" relative to an appropriate matched individual of comparable age, sex, race, etc.). Methods of treating obesity or undesirable body mass (including conditions co-morbid with obesity, e.g., obstructive sleep apnea, arthritis, cancer (e.g., breast, endometrial, and colon), gallstones or hyperglycemia, include the contact or administration of a binding protein, such as an anti-Klotho beta antibody as described herein, in an amount effective to treat obesity or an undesirable body mass. For example, an individual may have a mass index. greater than 25, e.g., 25-30, 30-35, 35-40, or greater than 40. Prevention methods (e.g., in individuals predisposed to having a particular disorder(s)) refer to whether to delay, slow down or inhibit the progression of, the initiation of, or treatment of undesirable or abnormally elevated serum/plasma LDL, VLDL, triglycerides or cholesterol levels, which, alone or in combination, can lead to, e.g. , the formation of plaque, narrowing or blocking of blood vessels and increased risk of hypertension, stroke and coronary artery disease. Such diseases, disorders or conditions may be due to, for example, genetic predisposition or diet.

Anti-Klotho beta antibodies

[0190] In one embodiment, the present invention provides anti-Klotho beta antibodies that may find use in the present invention as therapeutic agents. Examples of antibodies include polyclonal, monoclonal, humanized, human, bispecific and heteroconjugate antibodies, as well as variants thereof having improved affinity or other properties.

[0191] In some embodiments, provided herein are antibodies that bind to Klotho beta, including a Klotho beta polypeptide, a Klotho beta polypeptide fragment, a Klotho beta peptide or a Klotho beta epitope. In some embodiments, anti-Klotho beta antibodies are humanized antibodies (e.g., comprising human constant regions) that bind to Klotho beta, including Klotho beta polypeptide, a Klotho beta polypeptide fragment, a Klotho beta peptide, or an epitope. from Klotho beta.

[0192] In certain embodiments, the anti-Klotho beta antibody comprises a VH region, VL region, VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2 and/or VL CDR3 of either of the murine monoclonal antibodies described herein, such as an amino acid sequence represented in Tables 1-10. Therefore, in some embodiments, the isolated antibody or functional fragment thereof provided herein comprises one, two and/or three heavy chain CDRs and/or one, two and/or three light chain CDRs from: (a) the antibody designated 5H23; (b) the antibody designated 1C17; (c) the antibody designated 1D19; (d) the antibody designated 2L12; (e) the antibody designated 3L3; (f) the antibody designated 3N20; (g) the antibody designated 4P5; (h) the antibody designated 5C23; (i) the antibody designated 5F7; (j) the antibody designated 1G19, as shown in Tables 1-10.

[0193] The antibody designated 5H23 comprises a VH sequence that is SEQ ID NO: 25 and a VL sequence that is SEQ ID NO: 26.

[0194] The antibody designated 1C17 comprises a VH sequence that is SEQ ID NO: 51 and a VL sequence that is SEQ ID NO: 52.

[0195] The antibody designated 1D19 comprises a VH sequence that is SEQ ID NO: 77 and a VL sequence that is SEQ ID NO: 78.

[0196] The antibody designated 2L12 comprises a VH sequence that is SEQ ID NO: 103 and a VL sequence that is SEQ ID NO: 104.

[0197] The antibody designated 3L3 comprises a VH sequence that is SEQ ID NO: 129 and a VL sequence that is SEQ ID NO: 130.

[0198] The antibody designated 3N20 comprises a VH sequence that is SEQ ID NO: 155 and a VL sequence that is SEQ ID NO: 156.

[0199] The antibody designated 4P5 comprises a VH sequence that is SEQ ID NO: 181 and a VL sequence that is SEQ ID NO: 182.

[0200] The antibody designated 5C23 comprises a VH sequence that is SEQ ID NO: 207 and a VL sequence that is SEQ ID NO: 208.

[0201] The antibody designated 5F7 comprises a VH sequence that is SEQ ID NO: 233 and a VL sequence that is SEQ ID NO: 234.

[0202] The antibody designated IG19 comprises a VH sequence that is SEQ ID NO: 259 and a VL sequence that is SEQ ID NO: 260. Table 1: CDR sequences of Antibody 5H23 Table 2: 1C17 Antibody CDR Sequences Table 3: 1D19 Antibody CDR Sequences Table 4: 2L12 Antibody CDR Sequences Table 5: 3L3 Antibody CDR Sequences Table 6: 3N20 Antibody CDR Sequences Table 7: 4P5 Antibody CDR Sequences Table 8: 5C23 Antibody CDR Sequences Table 9: 5F7 Antibody CDR Sequences Table 10: 1G19 Antibody CDR Sequences

[0203] In some embodiments, the antibodies provided herein comprise a VH region or VH domain. In other embodiments, the antibodies provided herein comprise a VL region or VL chain. In some embodiments, the antibodies provided herein have a combination of (i) a VH domain or VH region; and/or (ii) a VL domain or VL region.

[0204] In some embodiments, an antibody provided herein comprises or consists of six CDRs, e.g., VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2, and/or VL CDR3 identified in Tables 1-10. In some embodiments, an antibody provided herein may comprise fewer than six CDRs. In some embodiments, the antibody comprises or consists of one, two, three, four, or five CDRs selected from the group consisting of VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2, and /or VL CDR3 identified in Tables 1-10. In some embodiments, the antibody comprises or consists of one, two, three, four, or five CDRs selected from the group consisting of VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2, and /or murine monoclonal antibody VL CDR3 selected from the group consisting of: (a) the antibody designated 5H23; (b) the antibody designated 1C17; (c) the antibody designated 1D19; (d) the antibody designated 2L12; (e) the antibody designated 3L3; (f) the antibody designated 3N20; (g) the antibody designated 4P5; (h) the antibody designated 5C23; (i) the antibody designated 5F7; (j) the antibody designated 1G19; described here. Therefore, in some embodiments, the antibody comprises or consists of one, two, three, four, or five CDRs of any of VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2, and/or VL CDR3 identified in Tables 1-10.

[0205] In some embodiments, the antibodies provided herein comprise one or more (e.g., one, two, or three) VH CDRs listed in Tables 1-10. In other embodiments, the antibodies provided herein comprise one or more (e.g., one, two, or three) VL CDRs listed in Tables 1-10. In still other embodiments, the antibodies provided herein comprise one or more (e.g., one, two, or three) VH CDRs listed in Tables 1-10 and one or more VL CDRs listed in Tables 1-10. Therefore, in some embodiments, the antibodies comprise a VH CDR1 having an amino acid sequence of any one of SEQ ID NOS: 1, 7, 12, 13, 18, 27, 33, 38, 39, 44, 53, 59 , 64, 65, 70, 79, 85, 90, 91, 96, 105, 111, 116, 117, 122, 131, 137, 142, 143, 148, 157, 163, 168, 169, 174, 183, 189 , 194, 195, 200, 209, 215, 220, 221, 226, 235, 241, 246, 247, and 252. In some embodiments, the antibodies comprise a VH CDR2 having an amino acid sequence of any one of SEQ ID NOS: 2, 8, 14, 19, 24, 28, 34, 40, 45, 50, 54, 60, 66, 71, 76, 80, 86, 92, 97, 102, 106, 112, 118, 123, 128, 132, 138, 144, 149, 154, 158, 164, 170, 175, 180, 184, 190, 196, 201, 206, 210, 216, 222, 227, 232, 236, 242, 248, 3, and 258. In some embodiments, the antibodies comprise a VH CDR3 having an amino acid sequence of any one of SEQ ID NOS: 3, 9, 15, 20, 29, 35, 41, 46, 55, 61, 67, 72 , 81, 87, 93, 98, 107, 113, 119, 124, 133, 139, 145, 150, 159, 165, 171, 176, 185, 191, 197, 202, 211, 217, 223, 228, 237 , 243, 249, and 254. In some embodiments, the antibodies comprise a VH CDR1 and/or a VH CDR2 and/or a VH CDR3 independently selected from a VH CDR1, VH CDR2, VH CDR3 as illustrated in any of the amino acid sequences illustrated in Tables 1-10. In some embodiments, the antibodies comprise a VL CDR1 having the amino acid sequence of any one of SEQ ID NOS: 4, 10, 16, 21, 30, 36, 42, 47, 56, 62, 68, 73, 82, 88, 94, 99, 108, 114, 120, 125, 134, 140, 146, 151, 160, 166, 172, 177, 186, 192, 198, 203, 212, 218, 224, 229, 238, 244, 250, and 255. In another embodiment, the antibodies comprise a VL CDR2 having the amino acid sequence of any of SEQ ID NOS: 5, 11, 22, 31, 37, 48, 57, 63, 74, 83, 89, 100, 109, 115, 126, 135, 141, 152, 161, 167, 178, 187, 193, 204, 213, 219, 230, 239, 245, and 256. In some embodiments, the antibodies comprise a CDR3 of VL having the amino acid sequence of any of SEQ ID NOS: 6, 17, 23, 32, 43, 49, 58, 69, 75, 84, 95, 101, 110, 121, 127, 136, 147, 153 , 162, 173, 179, 188, 199, 205, 214, 225, 231, 240, 251, and 257. In some embodiments, the antibodies comprise a VL CDR1 and/or a VL CDR2 and/or a VL CDR3. VL independently selected from a VL CDR1, VL CDR2, VL CDR3 as illustrated in any of the amino acid sequences illustrated in Tables 1-10.

[0206] In some embodiments, the antibodies provided herein comprise a heavy chain variable region (VH) comprising: (1) a VH CDR1 having an amino acid sequence selected from the group consisting of: (i) SEQ ID NO :1, 27, 53, 79, 105, 131, 157, 183, 209, and or 235, (ii) SEQ ID NO:7, 33, 59, 85, 111, 137, 163, 189, 215 or 241, (iii) SEQ ID NO:12, 38, 64, 90, 116, 142, 168, 194, 220 or 246, (iv) SEQ ID NO:13, 39, 65, 91, 117, 143, 169, 195, 221 or 247, and (v) SEQ ID NO: 18, 44, 70, 96, 122, 148, 174, 200, 226 or 252; (2) a VH CDR2 having an amino acid sequence selected from the group consisting of: (i) SEQ ID NO: 2, 28, 54, 80, 106, 132, 158, 184, 210, and or 236, (ii) SEQ ID NO:8, 34, 60, 86, 112, 138, 164, 190, 216 or 242, (iii) SEQ ID NO:14, 40, 66, 92, 118, 144, 170, 196, 222 or 248, (iv) SEQ ID NO:19, 45, 71, 97, 123, 149, 175, 201, 227 or 253, and (v) SEQ ID NO:24, 50, 76, 102, 128, 154 , 180, 206, 232 or 258; and (3) a VH CDR3 having an amino acid sequence selected from the group consisting of: (i) SEQ ID NO: 3, 29, 55, 81, 107, 133, 159, 185, 211, and or 237 , (ii) SEQ ID NO:9, 35, 61, 87, 113, 139, 165, 191, 217 or 243, (iii) SEQ ID NO:15, 41, 67, 93, 119, 145, 171, 197 , 223 or 249, and (iv) SEQ ID NO: 20, 46, 72, 98, 124, 150, 176, 202, 228 or 254; and/or a light chain (VL) variable region comprising: (1) a VL CDR1 having an amino acid sequence selected from the group consisting of: (i) SEQ ID NO:4, 30, 56, 82, 108, 134, 160, 186, 212, and or 238, (ii) SEQ ID NO: 10, 36, 52, 88, 114, 140, 166, 192, 218 or 244, (iii) SEQ ID NO: 16, 42, 68, 94, 120, 146, 172, 198, 224 or 250, and (iv) SEQ ID NO: 21, 47, 73, 99, 125, 151, 177, 203, 229 or 255; (2) a VL CDR2 having an amino acid sequence selected from the group consisting of: (i) SEQ ID NO: 5, 31, 57, 83, 109, 135, 161, 187, 213, and or 239, (ii) SEQ ID NO:11, 37, 63, 89, 115, 141, 167, 193, 219 or 245, and (iii) SEQ ID NO:22, 48, 74, 100, 126, 152, 178, 204 , 230 or 256; and (3) a VL CDR3 having an amino acid sequence selected from the group consisting of: (i) SEQ ID NO: 6, 32, 58, 84, 110, 136, 162, 188, 214, and or 240 , (ii) SEQ ID NO:17, 43, 69, 95, 121, 147, 173, 199, 225 or 251, and (iii) SEQ ID NO:23, 49, 75, 101, 127, 153, 179, 205, 231 or 257.

[0207] In some embodiments, the antibodies provided herein comprise a heavy chain variable region (VH) comprising: (1) a VH CDR1 having an amino acid sequence selected from the group consisting of: (i) SEQ ID NO :1, 27, 53, 79, 105, 131, 157, 183, 209, and or 235, (ii) SEQ ID NO:7, 33, 59, 85, 111, 137, 163, 189, 215 or 241, (iii) SEQ ID NO:12, 38, 64, 90, 116, 142, 168, 194, 220 or 246, (iv) SEQ ID NO:13, 39, 65, 91, 117, 143, 169, 195, 221 or 247, and (v) SEQ ID NO: 18, 44, 70, 96, 122, 148, 174, 200, 226 or 252; (2) a VH CDR2 having an amino acid sequence selected from the group consisting of: (i) SEQ ID NO: 2, 28, 54, 80, 106, 132, 158, 184, 210, and or 236, (ii) SEQ ID NO:8, 34, 60, 86, 112, 138, 164, 190, 216 or 242, (iii) SEQ ID NO:14, 40, 66, 92, 118, 144, 170, 196, 222 or 248, (iv) SEQ ID NO:19, 45, 71, 97, 123, 149, 175, 201, 227 or 253, and (v) SEQ ID NO:24, 50, 76, 102, 128, 154 , 180, 206, 232 or 258; and (3) a VH CDR3 having an amino acid sequence selected from the group consisting of: (i) SEQ ID NO: 3, 29, 55, 81, 107, 133, 159, 185, 211, and or 237 , (ii) SEQ ID NO:9, 35, 61, 87, 113, 139, 165, 191, 217 or 243, (iii) SEQ ID NO:15, 41, 67, 93, 119, 145, 171, 197 , 223 or 249, and (iv) SEQ ID NO: 20, 46, 72, 98, 124, 150, 176, 202, 228 or 254.

[0208] In some embodiments, the antibodies provided herein comprise a light chain variable region (VL) comprising: (1) a VL CDR1 having an amino acid sequence selected from the group consisting of: (i) SEQ ID NO :4, 30, 56, 82, 108, 134, 160, 186, 212, and or 238, (ii) SEQ ID NO: 10, 36, 52, 88, 114, 140, 166, 192, 218 or 244, (iii) SEQ ID NO:16, 42, 68, 94, 120, 146, 172, 198, 224 or 250, and (iv) SEQ ID NO:21, 47, 73, 99, 125, 151, 177, 203 , 229 or 255; (2) a VL CDR2 having an amino acid sequence selected from the group consisting of: (i) SEQ ID NO: 5, 31, 57, 83, 109, 135, 161, 187, 213, and or 239, (ii) SEQ ID NO:11, 37, 63, 89, 115, 141, 167, 193, 219 or 245, and (iii) SEQ ID NO:22, 48, 74, 100, 126, 152, 178, 204 , 230 or 256; and (3) a VL CDR3 having an amino acid sequence selected from the group consisting of: (i) SEQ ID NO: 6, 32, 58, 84, 110, 136, 162, 188, 214, and or 240 , (ii) SEQ ID NO:17, 43, 69, 95, 121, 147, 173, 199, 225 or 251, and (iii) SEQ ID NO:23, 49, 75, 101, 127, 153, 179, 205, 231 or 257.

[0209] Also provided herein are antibodies comprising one or more VH CDRs and one or more (e.g., one, two or three) VL CDRs listed in Tables 1-10. In particular, provided herein is an antibody comprising a VH CDR1 (SEQ ID NOS: 1, 7, 12, 13, 18, 27, 33, 38, 39, 44, 53, 59, 64, 65, 70, 79, 85, 90, 91, 96, 105, 111, 116, 117, 122, 131, 137, 142, 143, 148, 157, 163, 168, 169, 174, 183, 189, 194, 195, 200, 209, 215, 220, 221, 226, 235, 241, 246, 247, and 252.) and a VL CDR1 (SEQ ID NOS: 4, 10, 16, 21, 30, 36, 42, 47, 56, 62, 68, 73, 82, 88, 94, 99, 108, 114, 120, 125, 134, 140, 146, 151, 160, 166, 172, 177, 186, 192, 198, 203, 212, 218, 224, 229, 238, 244, 250, and 255); a VH CDR1 (SEQ ID NOS: 1, 7, 12, 13, 18, 27, 33, 38, 39, 44, 53, 59, 64, 65, 70, 79, 85, 90, 91, 96, 105 , 111, 116, 117, 122, 131, 137, 142, 143, 148, 157, 163, 168, 169, 174, 183, 189, 194, 195, 200, 209, 215, 220, 221, 226, 235 , 241, 246, 247, and 252); a VL CDR2 (SEQ ID NOS: 5, 11, 22, 31, 37, 48, 57, 63, 74, 83, 89, 100, 109, 115, 126, 135, 141, 152, 161, 167, 178 , 187, 193, 204, 213, 219, 230, 239, 245, and 256) and a VH CDR1 (SEQ ID NOS 1, 7, 12, 13, 18, 27, 33, 38, 39, 44, 53 , 59, 64, 65, 70, 79, 85, 90, 91, 96, 105, 111, 116, 117, 122, 131, 137, 142, 143, 148, 157, 163, 168, 169, 174, 183 , 189, 194, 195, 200, 209, 215, 220, 221, 226, 235, 241, 246, 247, and 252) VL CDR3 (SEQ ID NOS: 6, 17, 23, 32, 43, 49, 58, 69, 75, 84, 95, 101, 110, 121, 127, 136, 147, 153, 162, 173, 179, 188, 199, 205, 214, 225, 231, 240, 251, and 257) and a VH CDR2 (SEQ ID NOS: 2, 8, 14, 19, 24, 28, 34, 40, 45, 50, 54, 60, 66, 71, 76, 80, 86, 92, 97, 102, 106 , 112, 118, 123, 128, 132, 138, 144, 149, 154, 158, 164, 170, 175, 180, 184, 190, 196, 201, 206, 210, 216, 222, 227, 232, 236 , 242, 248, 253, and 258); a VL CDR1 (SEQ ID NOS: 4, 10, 16, 21, 30, 36, 42, 47, 56, 62, 68, 73, 82, 88, 94, 99, 108, 114, 120, 125, 134 , 140, 146, 151, 160, 166, 172, 177, 186, 192, 198, 203, 212, 218, 224, 229, 238, 244, 250, and 255) and a VH CDR2 (SEQ ID NOS: 2, 8, 14, 19, 24, 28, 34, 40, 45, 50, 54, 60, 66, 71, 76, 80, 86, 92, 97, 102, 106, 112, 118, 123, 128, 132, 138, 144, 149, 154, 158, 164, 170, 175, 180, 184, 190, 196, 201, 206, 210, 216, 222, 227, 232, 236, 242, 248, 253, and 2 58 ); and a CDR3 of VL (SEQ ID NOS: 6, 17, 23, 32, 43, 49, 58, 69, 75, 84, 95, 101, 110, 121, 127, 136, 147, 153, 162, 173, 179, 188, 199, 205, 214, 225, 231, 240, 251, and 257); a VH CDR3 (SEQ ID NOS: 3, 9, 15, 20, 29, 35, 41, 46, 55, 61, 67, 72, 81, 87, 93, 98, 107, 113, 119, 124, 133 , 139, 145, 150, 159, 165, 171, 176, 185, 191, 197, 202, 211, 217, 223, 228, 237, 243, 249, and 254) and a VL CDR1 (SEQ ID NOS: 4, 10, 16, 21, 30, 36, 42, 47, 56, 62, 68, 73, 82, 88, 94, 99, 108, 114, 120, 125, 134, 140, 146, 151, 160, 166, 172, 177, 186, 192, 198, 203, 212, 218, 224, 229, 238, 244, 250, and 255); a VH CDR3 (SEQ ID NOS: 3, 9, 15, 20, 29, 35, 41, 46, 55, 61, 67, 72, 81, 87, 93, 98, 107, 113, 119, 124, 133 , 139, 145, 150, 159, 165, 171, 176, 185, 191, 197, 202, 211, 217, 223, 228, 237, 243, 249, and 254) and a VL CDR2 (SEQ ID NOS: 5, 11, 22, 31, 37, 48, 57, 63, 74, 83, 89, 100, 109, 115, 126, 135, 141, 152, 161, 167, 178, 187, 193, 204, 213, 219, 230, 239, 245, and 256); a VH CDR3 (SEQ ID NOS: 3, 9, 15, 20, 29, 35, 41, 46, 55, 61, 67, 72, 81, 87, 93, 98, 107, 113, 119, 124, 133 , 139, 145, 150, 159, 165, 171, 176, 185, 191, 197, 202, 211, 217, 223, 228, 237, 243, 249, and 254) and a VL CDR3 (SEQ ID NOS: 6, 17, 23, 32, 43, 49, 58, 69, 75, 84, 95, 101, 110, 121, 127, 136, 147, 153, 162, 173, 179, 188, 199, 205, 214, 225, 231, 240, 251, and 257); a VH CDR1 (SEQ ID NOS: 1, 7, 12, 13, 18, 27, 33, 38, 39, 44, 53, 59, 64, 65, 70, 79, 85, 90, 91, 96, 105 , 111, 116, 117, 122, 131, 137, 142, 143, 148, 157, 163, 168, 169, 174, 183, 189, 194, 195, 200, 209, 215, 220, 221, 226, 235 , 241, 246, 247, and 252), a VH CDR2 (SEQ ID NOS: 2, 8, 14, 19, 24, 28, 34, 40, 45, 50, 54, 60, 66, 71, 76, 80, 86, 92, 97, 102, 106, 112, 118, 123, 128, 132, 138, 144, 149, 154, 158, 164, 170, 175, 180, 184, 190, 196, 201, 206, 210, 216, 222, 227, 232, 236, 242, 248, 253, and 258) and a VL CDR1 (SEQ ID NOS: 4, 10, 16, 21, 30, 36, 42, 47, 56, 62 , 68, 73, 82, 88, 94, 99, 108, 114, 120, 125, 134, 140, 146, 151, 160, 166, 172, 177, 186, 192, 198, 203, 212, 218, 224 , 229, 238, 244, 250, and 255); a VH CDR1 (SEQ ID NOS: 1, 7, 12, 13, 18, 27, 33, 38, 39, 44, 53, 59, 64, 65, 70, 79, 85, 90, 91, 96, 105 , 111, 116, 117, 122, 131, 137, 142, 143, 148, 157, 163, 168, 169, 174, 183, 189, 194, 195, 200, 209, 215, 220, 221, 226, 235 , 241, 246, 247, and 252), a VH CDR2 (SEQ ID NOS: 2, 8, 14, 19, 24, 28, 34, 40, 45, 50, 54, 60, 66, 71, 76, 80, 86, 92, 97, 102, 106, 112, 118, 123, 128, 132, 138, 144, 149, 154, 158, 164, 170, 175, 180, 184, 190, 196, 201, 206, 210, 216, 222, 227, 232, 236, 242, 248, 253, and 258) and a VL CDR2 (SEQ ID NOS: 5, 11, 22, 31, 37, 48, 57, 63, 74, 83 , 89, 100, 109, 115, 126, 135, 141, 152, 161, 167, 178, 187, 193, 204, 213, 219, 230, 239, 245, and 256); a VH CDR1 (SEQ ID NOS: 1, 7, 12, 13, 18, 27, 33, 38, 39, 44, 53, 59, 64, 65, 70, 79, 85, 90, 91, 96, 105, 111, 116, 117, 122, 131, 137, 142, 143, 148, 157, 163, 168, 169, 174, 183, 189, 194, 195, 200, 209, 215, 220, 221, 6, 235, 241, 246, 247, and 252), a VH CDR2 (SEQ ID NOS: 2, 8, 14, 19, 24, 28, 34, 40, 45, 50, 54, 60, 66, 71, 76 , 80, 86, 92, 97, 102, 106, 112, 118, 123, 128, 132, 138, 144, 149, 154, 158, 164, 170, 175, 180, 184, 190, 196, 201, 206 , 210, 216, 222, 227, 232, 236, 242, 248, 253, and 258) and a VL CDR3 (SEQ ID NOS: 6, 17, 23, 32, 43, 49, 58, 69, 75, 84, 95, 101, 110, 121, 127, 136, 147, 153, 162, 173, 179, 188, 199, 205, 214, 225, 231, 240, 251, and 257); a VH CDR2 (SEQ ID NOS: 2, 8, 14, 19, 24, 28, 34, 40, 45, 50, 54, 60, 66, 71, 76, 80, 86, 92, 97, 102, 106 , 112, 118, 123, 128, 132, 138, 144, 149, 154, 158, 164, 170, 175, 180, 184, 190, 196, 201, 206, 210, 216, 222, 227, 232, 236 , 242, 248, 253, and 258), a VH CDR3 (SEQ ID NOS: 3, 9, 15, 20, 29, 35, 41, 46, 55, 61, 67, 72, 81, 87, 93, 98, 107, 113, 119, 124, 133, 139, 145, 150, 159, 165, 171, 176, 185, 191, 197, 202, 211, 217, 223, 228, 237, 243, 249, and 25 4 ) and a VL CDR1 (SEQ ID NOS: 4, 10, 16, 21, 30, 36, 42, 47, 56, 62, 68, 73, 82, 88, 94, 99, 108, 114, 120, 125 , 134, 140, 146, 151, 160, 166, 172, 177, 186, 192, 198, 203, 212, 218, 224, 229, 238, 244, 250, and 255), a VH CDR2 (SEQ ID NOS: 2, 8, 14, 19, 24, 28, 34, 40, 45, 50, 54, 60, 66, 71, 76, 80, 86, 92, 97, 102, 106, 112, 118, 123, 128, 132, 138, 144, 149, 154, 158, 164, 170, 175, 180, 184, 190, 196, 201, 206, 210, 216, 222, 227, 232, 236, 242, 248, 3, and 258), a VH CDR3 (SEQ ID NOS: 3, 9, 15, 20, 29, 35, 41, 46, 55, 61, 67, 72, 81, 87, 93, 98, 107, 113, 119 , 124, 133, 139, 145, 150, 159, 165, 171, 176, 185, 191, 197, 202, 211, 217, 223, 228, 237, 243, 249, and 254) and a CDR2 of VL ( SEQ ID NOS: 5, 11, 22, 31, 37, 48, 57, 63, 74, 83, 89, 100, 109, 115, 126, 135, 141, 152, 161, 167, 178, 187, 193, 204, 213, 219, 230, 239, 245, and 256); a VH CDR2 (SEQ ID NOS: 2, 8, 14, 19, 24, 28, 34, 40, 45, 50, 54, 60, 66, 71, 76, 80, 86, 92, 97, 102, 106 , 112, 118, 123, 128, 132, 138, 144, 149, 154, 158, 164, 170, 175, 180, 184, 190, 196, 201, 206, 210, 216, 222, 227, 232, 236 , 242, 248, 253, and 258), a VH CDR3 (SEQ ID NOS: 3, 9, 15, 20, 29, 35, 41, 46, 55, 61, 67, 72, 81, 87, 93, 98, 107, 113, 119, 124, 133, 139, 145, 150, 159, 165, 171, 176, 185, 191, 197, 202, 211, 217, 223, 228, 237, 243, 249, and 25 4 ) and a VL CDR3 (SEQ ID NOS: 6, 17, 23, 32, 43, 49, 58, 69, 75, 84, 95, 101, 110, 121, 127, 136, 147, 153, 162, 173 , 179, 188, 199, 205, 214, 225, 231, 240, 251, and 257); a VH CDR1 (SEQ ID NOS: 1, 7, 12, 13, 18, 27, 33, 38, 39, 44, 53, 59, 64, 65, 70, 79, 85, 90, 91, 96, 105 , 111, 116, 117, 122, 131, 137, 142, 143, 148, 157, 163, 168, 169, 174, 183, 189, 194, 195, 200, 209, 215, 220, 221, 226, 235 , 241, 246, 247, and 252.), a VL CDR1 (SEQ ID NOS: 4, 10, 16, 21, 30, 36, 42, 47, 56, 62, 68, 73, 82, 88, 94 , 99, 108, 114, 120, 125, 134, 140, 146, 151, 160, 166, 172, 177, 186, 192, 198, 203, 212, 218, 224, 229, 238, 244, 250, and 255) and a CDR2 of VL (SEQ ID NOS: 5, 11, 22, 31, 37, 48, 57, 63, 74, 83, 89, 100, 109, 115, 126, 135, 141, 152, 161, 167, 178, 187, 193, 204, 213, 219, 230, 239, 245, and 256); a VH CDR1 (SEQ ID NOS: 1, 7, 12, 13, 18, 27, 33, 38, 39, 44, 53, 59, 64, 65, 70, 79, 85, 90, 91, 96, 105 , 111, 116, 117, 122, 131, 137, 142, 143, 148, 157, 163, 168, 169, 174, 183, 189, 194, 195, 200, 209, 215, 220, 221, 226, 235 , 241, 246, 247, and 252), a VL CDR1 (SEQ ID NOS: 4, 10, 16, 21, 30, 36, 42, 47, 56, 62, 68, 73, 82, 88, 94, 99, 108, 114, 120, 125, 134, 140, 146, 151, 160, 166, 172, 177, 186, 192, 198, 203, 212, 218, 224, 229, 238, 244, 250, and 25 5 ) and a VL CDR3 (SEQ ID NOS: 6, 17, 23, 32, 43, 49, 58, 69, 75, 84, 95, 101, 110, 121, 127, 136, 147, 153, 162, 173 , 179, 188, 199, 205, 214, 225, 231, 240, 251, and 257); a VH CDR1 (SEQ ID NOS: 1, 7, 12, 13, 18, 27, 33, 38, 39, 44, 53, 59, 64, 65, 70, 79, 85, 90, 91, 96, 105 , 111, 116, 117, 122, 131, 137, 142, 143, 148, 157, 163, 168, 169, 174, 183, 189, 194, 195, 200, 209, 215, 220, 221, 226, 235 , 241, 246, 247, and 252), a VL CDR2 (SEQ ID NOS: 5, 11, 22, 31, 37, 48, 57, 63, 74, 83, 89, 100, 109, 115, 126, 135, 141, 152, 161, 167, 178, 187, 193, 204, 213, 219, 230, 239, 245, and 256) and a CDR3 of VL (SEQ ID NOS: 6, 17, 23, 32, 43 , 49, 58, 69, 75, 84, 95, 101, 110, 121, 127, 136, 147, 153, 162, 173, 179, 188, 199, 205, 214, 225, 231, 240, 251, and 257); a VH CDR2 (SEQ ID NOS: 2, 8, 14, 19, 24, 28, 34, 40, 45, 50, 54, 60, 66, 71, 76, 80, 86, 92, 97, 102, 106 , 112, 118, 123, 128, 132, 138, 144, 149, 154, 158, 164, 170, 175, 180, 184, 190, 196, 201, 206, 210, 216, 222, 227, 232, 236 , 242, 248, 253, and 258), a VL CDR1 (SEQ ID NOS: 4, 10, 16, 21, 30, 36, 42, 47, 56, 62, 68, 73, 82, 88, 94, 99, 108, 114, 120, 125, 134, 140, 146, 151, 160, 166, 172, 177, 186, 192, 198, 203, 212, 218, 224, 229, 238, 244, 250, and 25 5 ) and a VL CDR2 (SEQ ID NOS: 5, 11, 22, 31, 37, 48, 57, 63, 74, 83, 89, 100, 109, 115, 126, 135, 141, 152, 161, 167 , 178, 187, 193, 204, 213, 219, 230, 239, 245, and 256); a VH CDR2 (SEQ ID NOS: 2, 8, 14, 19, 24, 28, 34, 40, 45, 50, 54, 60, 66, 71, 76, 80, 86, 92, 97, 102, 106 , 112, 118, 123, 128, 132, 138, 144, 149, 154, 158, 164, 170, 175, 180, 184, 190, 196, 201, 206, 210, 216, 222, 227, 232, 236 , 242, 248, 253, and 258), a VL CDR1 (SEQ ID NOS: 4, 10, 16, 21, 30, 36, 42, 47, 56, 62, 68, 73, 82, 88, 94, 99, 108, 114, 120, 125, 134, 140, 146, 151, 160, 166, 172, 177, 186, 192, 198, 203, 212, 218, 224, 229, 238, 244, 250, and 25 5 ) and a VL CDR3 (SEQ ID NOS: 6, 17, 23, 32, 43, 49, 58, 69, 75, 84, 95, 101, 110, 121, 127, 136, 147, 153, 162, 173 , 179, 188, 199, 205, 214, 225, 231, 240, 251, and 257); a VH CDR2 (SEQ ID NOS: 2, 8, 14, 19, 24, 28, 34, 40, 45, 50, 54, 60, 66, 71, 76, 80, 86, 92, 97, 102, 106 , 112, 118, 123, 128, 132, 138, 144, 149, 154, 158, 164, 170, 175, 180, 184, 190, 196, 201, 206, 210, 216, 222, 227, 232, 236 , 242, 248, 253, and 258), a VL CDR2 (SEQ ID NOS: 5, 11, 22, 31, 37, 48, 57, 63, 74, 83, 89, 100, 109, 115, 126, 135, 141, 152, 161, 167, 178, 187, 193, 204, 213, 219, 230, 239, 245, and 256) and a CDR3 of VL (SEQ ID NOS: 6, 17, 23, 32, 43 , 49, 58, 69, 75, 84, 95, 101, 110, 121, 127, 136, 147, 153, 162, 173, 179, 188, 199, 205, 214, 225, 231, 240, 251, and 257); a VH CDR3 (SEQ ID NOS: 3, 9, 15, 20, 29, 35, 41, 46, 55, 61, 67, 72, 81, 87, 93, 98, 107, 113, 119, 124, 133 , 139, 145, 150, 159, 165, 171, 176, 185, 191, 197, 202, 211, 217, 223, 228, 237, 243, 249, and 254), a VL CDR1 (SEQ ID NOS: 4, 10, 16, 21, 30, 36, 42, 47, 56, 62, 68, 73, 82, 88, 94, 99, 108, 114, 120, 125, 134, 140, 146, 151, 160, 166, 172, 177, 186, 192, 198, 203, 212, 218, 224, 229, 238, 244, 250, and 255) and a CDR2 of VL (SEQ ID NOS: 5, 11, 22, 31, 37 , 48, 57, 63, 74, 83, 89, 100, 109, 115, 126, 135, 141, 152, 161, 167, 178, 187, 193, 204, 213, 219, 230, 239, 245, and 256); a VH CDR3 (SEQ ID NOS: 3, 9, 15, 20, 29, 35, 41, 46, 55, 61, 67, 72, 81, 87, 93, 98, 107, 113, 119, 124, 133 , 139, 145, 150, 159, 165, 171, 176, 185, 191, 197, 202, 211, 217, 223, 228, 237, 243, 249, and 254), a VL CDR1 (SEQ ID NOS: 4, 10, 16, 21, 30, 36, 42, 47, 56, 62, 68, 73, 82, 88, 94, 99, 108, 114, 120, 125, 134, 140, 146, 151, 160, 166, 172, 177, 186, 192, 198, 203, 212, 218, 224, 229, 238, 244, 250, and 255) and a CDR3 of VL (SEQ ID NOS: 6, 17, 23, 32, 43 , 49, 58, 69, 75, 84, 95, 101, 110, 121, 127, 136, 147, 153, 162, 173, 179, 188, 199, 205, 214, 225, 231, 240, 251, and 257); a VH CDR3 (SEQ ID NOS: 3, 9, 15, 20, 29, 35, 41, 46, 55, 61, 67, 72, 81, 87, 93, 98, 107, 113, 119, 124, 133 , 139, 145, 150, 159, 165, 171, 176, 185, 191, 197, 202, 211, 217, 223, 228, 237, 243, 249, and 254), a VL CDR2 (SEQ ID NOS: 5, 11, 22, 31, 37, 48, 57, 63, 74, 83, 89, 100, 109, 115, 126, 135, 141, 152, 161, 167, 178, 187, 193, 204, 213, 219, 230, 239, 245, and 256) and a VL CDR3 (SEQ ID NOS: 6, 17, 23, 32, 43, 49, 58, 69, 75, 84, 95, 101, 110, 121, 127 , 136, 147, 153, 162, 173, 179, 188, 199, 205, 214, 225, 231, 240, 251, and 257); a VH CDR1 (SEQ ID NOS: 1, 7, 12, 13, 18, 27, 33, 38, 39, 44, 53, 59, 64, 65, 70, 79, 85, 90, 91, 96, 105 , 111, 116, 117, 122, 131, 137, 142, 143, 148, 157, 163, 168, 169, 174, 183, 189, 194, 195, 200, 209, 215, 220, 221, 226, 235 , 241, 246, 247, and 252), a VH CDR2 (SEQ ID NOS: 2, 8, 14, 19, 24, 28, 34, 40, 45, 50, 54, 60, 66, 71, 76, 80, 86, 92, 97, 102, 106, 112, 118, 123, 128, 132, 138, 144, 149, 154, 158, 164, 170, 175, 180, 184, 190, 196, 201, 206, 210, 216, 222, 227, 232, 236, 242, 248, 253, and 258), a VH CDR3 (SEQ ID NOS: 3, 9, 15, 20, 29, 35, 41, 46, 55, 61 , 67, 72, 81, 87, 93, 98, 107, 113, 119, 124, 133, 139, 145, 150, 159, 165, 171, 176, 185, 191, 197, 202, 211, 217, 223 , 228, 237, 243, 249, and 254) and a VL CDR1 (SEQ ID NOS: 4, 10, 16, 21, 30, 36, 42, 47, 56, 62, 68, 73, 82, 88, 94, 99, 108, 114, 120, 125, 134, 140, 146, 151, 160, 166, 172, 177, 186, 192, 198, 203, 212, 218, 224, 229, 238, 244, 250, and 255); a VH CDR1 (SEQ ID NOS: 1, 7, 12, 13, 18, 27, 33, 38, 39, 44, 53, 59, 64, 65, 70, 79, 85, 90, 91, 96, 105 , 111, 116, 117, 122, 131, 137, 142, 143, 148, 157, 163, 168, 169, 174, 183, 189, 194, 195, 200, 209, 215, 220, 221, 226, 235 , 241, 246, 247, and 252), a VH CDR2 (SEQ ID NOS: 2, 8, 14, 19, 24, 28, 34, 40, 45, 50, 54, 60, 66, 71, 76, 80, 86, 92, 97, 102, 106, 112, 118, 123, 128, 132, 138, 144, 149, 154, 158, 164, 170, 175, 180, 184, 190, 196, 201, 206, 210, 216, 222, 227, 232, 236, 242, 248, 253, and 258), a VH CDR3 (SEQ ID NOS: 3, 9, 15, 20, 29, 35, 41, 46, 55, 61 , 67, 72, 81, 87, 93, 98, 107, 113, 119, 124, 133, 139, 145, 150, 159, 165, 171, 176, 185, 191, 197, 202, 211, 217, 223 , 228, 237, 243, 249, and 254) and a VL CDR2 (SEQ ID NOS: 5, 11, 22, 31, 37, 48, 57, 63, 74, 83, 89, 100, 109, 115, 126, 135, 141, 152, 161, 167, 178, 187, 193, 204, 213, 219, 230, 239, 245, and 256); a VH CDR1 (SEQ ID NOS: 1, 7, 12, 13, 18, 27, 33, 38, 39, 44, 53, 59, 64, 65, 70, 79, 85, 90, 91, 96, 105 , 111, 116, 117, 122, 131, 137, 142, 143, 148, 157, 163, 168, 169, 174, 183, 189, 194, 195, 200, 209, 215, 220, 221, 226, 235 , 241, 246, 247, and 252), a VH CDR2 (SEQ ID NOS: 2, 8, 14, 19, 24, 28, 34, 40, 45, 50, 54, 60, 66, 71, 76, 80, 86, 92, 97, 102, 106, 112, 118, 123, 128, 132, 138, 144, 149, 154, 158, 164, 170, 175, 180, 184, 190, 196, 201, 206, 210, 216, 222, 227, 232, 236, 242, 248, 253, and 258), a VH CDR3 (SEQ ID NOS: 3, 9, 15, 20, 29, 35, 41, 46, 55, 61 , 67, 72, 81, 87, 93, 98, 107, 113, 119, 124, 133, 139, 145, 150, 159, 165, 171, 176, 185, 191, 197, 202, 211, 217, 223 , 228, 237, 243, 249, and 254), and a VL CDR3 (SEQ ID NOS: 6, 17, 23, 32, 43, 49, 58, 69, 75, 84, 95, 101, 110, 121 , 127, 136, 147, 153, 162, 173, 179, 188, 199, 205, 214, 225, 231, 240, 251, and 257); a VH CDR1 (SEQ ID NOS: 1, 7, 12, 13, 18, 27, 33, 38, 39, 44, 53, 59, 64, 65, 70, 79, 85, 90, 91, 96, 105 , 111, 116, 117, 122, 131, 137, 142, 143, 148, 157, 163, 168, 169, 174, 183, 189, 194, 195, 200, 209, 215, 220, 221, 226, 235 , 241, 246, 247, and 252), a VH CDR2 (SEQ ID NOS: 2, 8, 14, 19, 24, 28, 34, 40, 45, 50, 54, 60, 66, 71, 76, 80, 86, 92, 97, 102, 106, 112, 118, 123, 128, 132, 138, 144, 149, 154, 158, 164, 170, 175, 180, 184, 190, 196, 201, 206, 210, 216, 222, 227, 232, 236, 242, 248, 253, and 258), a CDR1 of VL (SEQ ID NOS: 4, 10, 16, 21, 30, 36, 42, 47, 56, 62 , 68, 73, 82, 88, 94, 99, 108, 114, 120, 125, 134, 140, 146, 151, 160, 166, 172, 177, 186, 192, 198, 203, 212, 218, 224 , 229, 238, 244, 250, and 255) and a VL CDR2 (SEQ ID NOS: 5, 11, 22, 31, 37, 48, 57, 63, 74, 83, 89, 100, 109, 115, 126, 135, 141, 152, 161, 167, 178, 187, 193, 204, 213, 219, 230, 239, 245, and 256); a VH CDR1 (SEQ ID NOS: 1, 7, 12, 13, 18, 27, 33, 38, 39, 44, 53, 59, 64, 65, 70, 79, 85, 90, 91, 96, 105 , 111, 116, 117, 122, 131, 137, 142, 143, 148, 157, 163, 168, 169, 174, 183, 189, 194, 195, 200, 209, 215, 220, 221, 226, 235 , 241, 246, 247, and 252), a VH CDR2 (SEQ ID NOS: 2, 8, 14, 19, 24, 28, 34, 40, 45, 50, 54, 60, 66, 71, 76, 80, 86, 92, 97, 102, 106, 112, 118, 123, 128, 132, 138, 144, 149, 154, 158, 164, 170, 175, 180, 184, 190, 196, 201, 206, 210, 216, 222, 227, 232, 236, 242, 248, 253, and 258), a CDR1 of VL (SEQ ID NOS: 4, 10, 16, 21, 30, 36, 42, 47, 56, 62 , 68, 73, 82, 88, 94, 99, 108, 114, 120, 125, 134, 140, 146, 151, 160, 166, 172, 177, 186, 192, 198, 203, 212, 218, 224 , 229, 238, 244, 250, and 255) and a VL CDR3 (SEQ ID NOS: 6, 17, 23, 32, 43, 49, 58, 69, 75, 84, 95, 101, 110, 121, 127, 136, 147, 153, 162, 173, 179, 188, 199, 205, 214, 225, 231, 240, 251, and 257); a VH CDR1 (SEQ ID NOS: 1, 7, 12, 13, 18, 27, 33, 38, 39, 44, 53, 59, 64, 65, 70, 79, 85, 90, 91, 96, 105 , 111, 116, 117, 122, 131, 137, 142, 143, 148, 157, 163, 168, 169, 174, 183, 189, 194, 195, 200, 209, 215, 220, 221, 226, 235 , 241, 246, 247, and 252), a VH CDR2 (SEQ ID NOS: 2, 8, 14, 19, 24, 28, 34, 40, 45, 50, 54, 60, 66, 71, 76, 80, 86, 92, 97, 102, 106, 112, 118, 123, 128, 132, 138, 144, 149, 154, 158, 164, 170, 175, 180, 184, 190, 196, 201, 206, 210, 216, 222, 227, 232, 236, 242, 248, 253, and 258), a VL CDR2 (SEQ ID NOS: 5, 11, 22, 31, 37, 48, 57, 63, 74, 83 , 89, 100, 109, 115, 126, 135, 141, 152, 161, 167, 178, 187, 193, 204, 213, 219, 230, 239, 245, and 256) and a VL CDR3 (SEQ ID NOS:6, 17, 23, 32, 43, 49, 58, 69, 75, 84, 95, 101, 110, 121, 127, 136, 147, 153, 162, 173, 179, 188, 199, 205, 214, 225, 231, 240, 251, and 257); a VH CDR1 (SEQ ID NOS: 1, 7, 12, 13, 18, 27, 33, 38, 39, 44, 53, 59, 64, 65, 70, 79, 85, 90, 91, 96, 105 , 111, 116, 117, 122, 131, 137, 142, 143, 148, 157, 163, 168, 169, 174, 183, 189, 194, 195, 200, 209, 215, 220, 221, 226, 235 , 241, 246, 247, and 252), a VH CDR3 (SEQ ID NOS: 3, 9, 15, 20, 29, 35, 41, 46, 55, 61, 67, 72, 81, 87, 93, 98, 107, 113, 119, 124, 133, 139, 145, 150, 159, 165, 171, 176, 185, 191, 197, 202, 211, 217, 223, 228, 237, 243, 249, and 25 4 ), a CDR1 of VL (SEQ ID NOS:4, 10, 16, 21, 30, 36, 42, 47, 56, 62, 68, 73, 82, 88, 94, 99, 108, 114, 120, 125 , 134, 140, 146, 151, 160, 166, 172, 177, 186, 192, 198, 203, 212, 218, 224, 229, 238, 244, 250, and 255) and a VL CDR2 (SEQ ID NOS:5, 11, 22, 31, 37, 48, 57, 63, 74, 83, 89, 100, 109, 115, 126, 135, 141, 152, 161, 167, 178, 187, 193, 204, 213, 219, 230, 239, 245, and 256); a VH CDR1 (SEQ ID NOS: 1, 7, 12, 13, 18, 27, 33, 38, 39, 44, 53, 59, 64, 65, 70, 79, 85, 90, 91, 96, 105 , 111, 116, 117, 122, 131, 137, 142, 143, 148, 157, 163, 168, 169, 174, 183, 189, 194, 195, 200, 209, 215, 220, 221, 226, 235 , 241, 246, 247, and 252), a VH CDR3 (SEQ ID NOS: 3, 9, 15, 20, 29, 35, 41, 46, 55, 61, 67, 72, 81, 87, 93, 98, 107, 113, 119, 124, 133, 139, 145, 150, 159, 165, 171, 176, 185, 191, 197, 202, 211, 217, 223, 228, 237, 243, 249, and 25 4 ), a CDR1 of VL (SEQ ID NOS:4, 10, 16, 21, 30, 36, 42, 47, 56, 62, 68, 73, 82, 88, 94, 99, 108, 114, 120, 125 , 134, 140, 146, 151, 160, 166, 172, 177, 186, 192, 198, 203, 212, 218, 224, 229, 238, 244, 250, and 255) and a VL CDR3 (SEQ ID NOS: 6, 17, 23, 32, 43, 49, 58, 69, 75, 84, 95, 101, 110, 121, 127, 136, 147, 153, 162, 173, 179, 188, 199, 205, 214, 225, 231, 240, 251, and 257); a VH CDR1 (SEQ ID NOS: 1, 7, 12, 13, 18, 27, 33, 38, 39, 44, 53, 59, 64, 65, 70, 79, 85, 90, 91, 96, 105 , 111, 116, 117, 122, 131, 137, 142, 143, 148, 157, 163, 168, 169, 174, 183, 189, 194, 195, 200, 209, 215, 220, 221, 226, 235 , 241, 246, 247, and 252), a VH CDR3 (SEQ ID NOS: 3, 9, 15, 20, 29, 35, 41, 46, 55, 61, 67, 72, 81, 87, 93, 98, 107, 113, 119, 124, 133, 139, 145, 150, 159, 165, 171, 176, 185, 191, 197, 202, 211, 217, 223, 228, 237, 243, 249, and 25 4 ), a VL CDR2 (SEQ ID NOS:5, 11, 22, 31, 37, 48, 57, 63, 74, 83, 89, 100, 109, 115, 126, 135, 141, 152, 161, 167 , 178, 187, 193, 204, 213, 219, 230, 239, 245, and 256) and a VL CDR3 (SEQ ID NOS: 6, 17, 23, 32, 43, 49, 58, 69, 75, 84, 95, 101, 110, 121, 127, 136, 147, 153, 162, 173, 179, 188, 199, 205, 214, 225, 231, 240, 251, and 257); a VH CDR2 (SEQ ID NOS:2, 8, 14, 19, 24, 28, 34, 40, 45, 50, 54, 60, 66, 71, 76, 80, 86, 92, 97, 102, 106 , 112, 118, 123, 128, 132, 138, 144, 149, 154, 158, 164, 170, 175, 180, 184, 190, 196, 201, 206, 210, 216, 222, 227, 232, 236 , 242, 248, 253, and 258), a VH CDR3 (SEQ ID NOS: 3, 9, 15, 20, 29, 35, 41, 46, 55, 61, 67, 72, 81, 87, 93, 98, 107, 113, 119, 124, 133, 139, 145, 150, 159, 165, 171, 176, 185, 191, 197, 202, 211, 217, 223, 228, 237, 243, 249, and 25 4 ), a CDR1 of VL (SEQ ID NOS:4, 10, 16, 21, 30, 36, 42, 47, 56, 62, 68, 73, 82, 88, 94, 99, 108, 114, 120, 125 , 134, 140, 146, 151, 160, 166, 172, 177, 186, 192, 198, 203, 212, 218, 224, 229, 238, 244, 250, and 255) and a VL CDR2 (SEQ ID NOS:5, 11, 22, 31, 37, 48, 57, 63, 74, 83, 89, 100, 109, 115, 126, 135, 141, 152, 161, 167, 178, 187, 193, 204, 213, 219, 230, 239, 245, and 256); a VH CDR2 (SEQ ID NOS:2, 8, 14, 19, 24, 28, 34, 40, 45, 50, 54, 60, 66, 71, 76, 80, 86, 92, 97, 102, 106 , 112, 118, 123, 128, 132, 138, 144, 149, 154, 158, 164, 170, 175, 180, 184, 190, 196, 201, 206, 210, 216, 222, 227, 232, 236 , 242, 248, 253, and 258), a VH CDR3 (SEQ ID NOS: 3, 9, 15, 20, 29, 35, 41, 46, 55, 61, 67, 72, 81, 87, 93, 98, 107, 113, 119, 124, 133, 139, 145, 150, 159, 165, 171, 176, 185, 191, 197, 202, 211, 217, 223, 228, 237, 243, 249, and 25 4 ), a CDR1 of VL (SEQ ID NOS:4, 10, 16, 21, 30, 36, 42, 47, 56, 62, 68, 73, 82, 88, 94, 99, 108, 114, 120, 125 , 134, 140, 146, 151, 160, 166, 172, 177, 186, 192, 198, 203, 212, 218, 224, 229, 238, 244, 250, and 255); a VH CDR2 (SEQ ID NOS:2, 8, 14, 19, 24, 28, 34, 40, 45, 50, 54, 60, 66, 71, 76, 80, 86, 92, 97, 102, 106 , 112, 118, 123, 128, 132, 138, 144, 149, 154, 158, 164, 170, 175, 180, 184, 190, 196, 201, 206, 210, 216, 222, 227, 232, 236 , 242, 248, 253, and 258), a VH CDR3 (SEQ ID NOS: 3, 9, 15, 20, 29, 35, 41, 46, 55, 61, 67, 72, 81, 87, 93, 98, 107, 113, 119, 124, 133, 139, 145, 150, 159, 165, 171, 176, 185, 191, 197, 202, 211, 217, 223, 228, 237, 243, 249, and 25 4 ), a VL CDR2 (SEQ ID NOS:5, 11, 22, 31, 37, 48, 57, 63, 74, 83, 89, 100, 109, 115, 126, 135, 141, 152, 161, 167 , 178, 187, 193, 204, 213, 219, 230, 239, 245, and 256) and a VL CDR3 (SEQ ID NOS: 6, 17, 23, 32, 43, 49, 58, 69, 75, 84, 95, 101, 110, 121, 127, 136, 147, 153, 162, 173, 179, 188, 199, 205, 214, 225, 231, 240, 251, and 257); a VH CDR1 (SEQ ID NOS: 1, 7, 12, 13, 18, 27, 33, 38, 39, 44, 53, 59, 64, 65, 70, 79, 85, 90, 91, 96, 105 , 111, 116, 117, 122, 131, 137, 142, 143, 148, 157, 163, 168, 169, 174, 183, 189, 194, 195, 200, 209, 215, 220, 221, 226, 235 , 241, 246, 247, and 252), a VH CDR2 (SEQ ID NOS: 2, 8, 14, 19, 24, 28, 34, 40, 45, 50, 54, 60, 66, 71, 76, 80, 86, 92, 97, 102, 106, 112, 118, 123, 128, 132, 138, 144, 149, 154, 158, 164, 170, 175, 180, 184, 190, 196, 201, 206, 210, 216, 222, 227, 232, 236, 242, 248, 253, and 258), a VH CDR3 (SEQ ID NOS: 3, 9, 15, 20, 29, 35, 41, 46, 55, 61 , 67, 72, 81, 87, 93, 98, 107, 113, 119, 124, 133, 139, 145, 150, 159, 165, 171, 176, 185, 191, 197, 202, 211, 217, 223 , 228, 237, 243, 249, and 254), a VL CDR1 (SEQ ID NOS: 4, 10, 16, 21, 30, 36, 42, 47, 56, 62, 68, 73, 82, 88, 94, 99, 108, 114, 120, 125, 134, 140, 146, 151, 160, 166, 172, 177, 186, 192, 198, 203, 212, 218, 224, 229, 238, 244, 250, and 255) and a CDR2 of VL (SEQ ID NOS:5, 11, 22, 31, 37, 48, 57, 63, 74, 83, 89, 100, 109, 115, 126, 135, 141, 152, 161 , 167, 178, 187, 193, 204, 213, 219, 230, 239, 245, and 256); a VH CDR1 (SEQ ID NOS: 1, 7, 12, 13, 18, 27, 33, 38, 39, 44, 53, 59, 64, 65, 70, 79, 85, 90, 91, 96, 105 , 111, 116, 117, 122, 131, 137, 142, 143, 148, 157, 163, 168, 169, 174, 183, 189, 194, 195, 200, 209, 215, 220, 221, 226, 235 , 241, 246, 247, and 252), a VH CDR2 (SEQ ID NOS: 2, 8, 14, 19, 24, 28, 34, 40, 45, 50, 54, 60, 66, 71, 76, 80, 86, 92, 97, 102, 106, 112, 118, 123, 128, 132, 138, 144, 149, 154, 158, 164, 170, 175, 180, 184, 190, 196, 201, 206, 210, 216, 222, 227, 232, 236, 242, 248, 253, and 258), a VH CDR3 (SEQ ID NOS: 3, 9, 15, 20, 29, 35, 41, 46, 55, 61 , 67, 72, 81, 87, 93, 98, 107, 113, 119, 124, 133, 139, 145, 150, 159, 165, 171, 176, 185, 191, 197, 202, 211, 217, 223 , 228, 237, 243, 249, and 254), a VL CDR1 (SEQ ID NOS: 4, 10, 16, 21, 30, 36, 42, 47, 56, 62, 68, 73, 82, 88, 94, 99, 108, 114, 120, 125, 134, 140, 146, 151, 160, 166, 172, 177, 186, 192, 198, 203, 212, 218, 224, 229, 238, 244, 250, and 255) and a CDR3 of VL (SEQ ID NOS: 6, 17, 23, 32, 43, 49, 58, 69, 75, 84, 95, 101, 110, 121, 127, 136, 147, 153, 162 , 173, 179, 188, 199, 205, 214, 225, 231, 240, 251, and 257); a VH CDR1 (SEQ ID NOS: 1, 7, 12, 13, 18, 27, 33, 38, 39, 44, 53, 59, 64, 65, 70, 79, 85, 90, 91, 96, 105 , 111, 116, 117, 122, 131, 137, 142, 143, 148, 157, 163, 168, 169, 174, 183, 189, 194, 195, 200, 209, 215, 220, 221, 226, 235 , 241, 246, 247, and 252), a VH CDR2 (SEQ ID NOS: 2, 8, 14, 19, 24, 28, 34, 40, 45, 50, 54, 60, 66, 71, 76, 80, 86, 92, 97, 102, 106, 112, 118, 123, 128, 132, 138, 144, 149, 154, 158, 164, 170, 175, 180, 184, 190, 196, 201, 206, 210, 216, 222, 227, 232, 236, 242, 248, 253, and 258), a VH CDR3 (SEQ ID NOS: 3, 9, 15, 20, 29, 35, 41, 46, 55, 61 , 67, 72, 81, 87, 93, 98, 107, 113, 119, 124, 133, 139, 145, 150, 159, 165, 171, 176, 185, 191, 197, 202, 211, 217, 223 , 228, 237, 243, 249, and 254), a VL CDR2 (SEQ ID NOS: 5, 11, 22, 31, 37, 48, 57, 63, 74, 83, 89, 100, 109, 115, 126, 135, 141, 152, 161, 167, 178, 187, 193, 204, 213, 219, 230, 239, 245, and 256) and a VL CDR3 (SEQ ID NOS:6, 17, 23, 32 , 43, 49, 58, 69, 75, 84, 95, 101, 110, 121, 127, 136, 147, 153, 162, 173, 179, 188, 199, 205, 214, 225, 231, 240, 251 , and 257); a VH CDR1 (SEQ ID NOS: 1, 7, 12, 13, 18, 27, 33, 38, 39, 44, 53, 59, 64, 65, 70, 79, 85, 90, 91, 96, 105 , 111, 116, 117, 122, 131, 137, 142, 143, 148, 157, 163, 168, 169, 174, 183, 189, 194, 195, 200, 209, 215, 220, 221, 226, 235 , 241, 246, 247, and 252), a VH CDR2 (SEQ ID NOS: 2, 8, 14, 19, 24, 28, 34, 40, 45, 50, 54, 60, 66, 71, 76, 80, 86, 92, 97, 102, 106, 112, 118, 123, 128, 132, 138, 144, 149, 154, 158, 164, 170, 175, 180, 184, 190, 196, 201, 206, 210, 216, 222, 227, 232, 236, 242, 248, 253, and 258), a CDR1 of VL (SEQ ID NOS: 4, 10, 16, 21, 30, 36, 42, 47, 56, 62 , 68, 73, 82, 88, 94, 99, 108, 114, 120, 125, 134, 140, 146, 151, 160, 166, 172, 177, 186, 192, 198, 203, 212, 218, 224 , 229, 238, 244, 250, and 255), a VL CDR2 (SEQ ID NOS: 5, 11, 22, 31, 37, 48, 57, 63, 74, 83, 89, 100, 109, 115, 126, 135, 141, 152, 161, 167, 178, 187, 193, 204, 213, 219, 230, 239, 245, and 256), and a CDR3 of VL (SEQ ID NOS:6, 17, 23, 32, 43, 49, 58, 69, 75, 84, 95, 101, 110, 121, 127, 136, 147, 153, 162, 173, 179, 188, 199, 205, 214, 225, 231, 240, 251, and 257); a VH CDR1 (SEQ ID NOS: 1, 7, 12, 13, 18, 27, 33, 38, 39, 44, 53, 59, 64, 65, 70, 79, 85, 90, 91, 96, 105 , 111, 116, 117, 122, 131, 137, 142, 143, 148, 157, 163, 168, 169, 174, 183, 189, 194, 195, 200, 209, 215, 220, 221, 226, 235 , 241, 246, 247, and 252), a VH CDR3 (SEQ ID NOS: 3, 9, 15, 20, 29, 35, 41, 46, 55, 61, 67, 72, 81, 87, 93, 98, 107, 113, 119, 124, 133, 139, 145, 150, 159, 165, 171, 176, 185, 191, 197, 202, 211, 217, 223, 228, 237, 243, 249, and 25 4 ), a VL CDR1 (SEQ ID NOS:4, 10, 16, 21, 30, 36, 42, 47, 56, 62, 68, 73, 82, 88, 94, 99, 108, 114, 120, 125 , 134, 140, 146, 151, 160, 166, 172, 177, 186, 192, 198, 203, 212, 218, 224, 229, 238, 244, 250, and 255), a CDR2 of VL (SEQ ID NOS:5, 11, 22, 31, 37, 48, 57, 63, 74, 83, 89, 100, 109, 115, 126, 135, 141, 152, 193, 204, 213, 219, 230, 239, 245, and 256), and a CDR3 of VL (SEQ ID NOS: 6, 17, 23, 32, 43, 49, 58, 69, 75, 84, 95, 101, 110, 121, 127, 136, 147, 153, 162, 173, 179, 188, 199, 205, 214, 225, 231, 240, 251, and 257); a VH CDR2 (SEQ ID NOS:2, 8, 14, 19, 24, 28, 34, 40, 45, 50, 54, 60, 66, 71, 76, 80, 86, 92, 97, 102, 106 , 112, 118, 123, 128, 132, 138, 144, 149, 154, 158, 164, 170, 175, 180, 184, 190, 196, 201, 206, 210, 216, 222, 227, 232, 236 , 242, 248, 253, and 258), a VH CDR3 (SEQ ID NOS: 3, 9, 15, 20, 29, 35, 41, 46, 55, 61, 67, 72, 81, 87, 93, 98, 107, 113, 119, 124, 133, 139, 145, 150, 159, 165, 171, 176, 185, 191, 197, 202, 211, 217, 223, 228, 237, 243, 249, and 25 4 ), a CDR1 of VL (SEQ ID NOS:4, 10, 16, 21, 30, 36, 42, 47, 56, 62, 68, 73, 82, 88, 94, 99, 108, 114, 120, 125 , 134, 140, 146, 151, 160, 166, 172, 177, 186, 192, 198, 203, 212, 218, 224, 229, 238, 244, 250, and 255), a CDR2 of VL (SEQ ID NOS:5, 11, 22, 31, 37, 48, 57, 63, 74, 83, 89, 100, 109, 115, 126, 135, 141, 152, 161, 167, 178, 187, 193, 204, 213, 219, 230, 239, 245, and 256) or any combination thereof of the VH CDRs and VL CDRs listed in Tables 1-10.

[0210] In yet another aspect, the CDRs disclosed herein include related derived consensus sequences (see, for example, Tables 1-10). As described herein, a "consensus sequence" refers to amino acid sequences having conserved amino acids common among a series of sequences and variable amino acids that vary within a given amino acid sequence. The CDR consensus sequences provided include CDRs corresponding to CDRH1, CDRH2, CDRH3, CDRL1, CDRL2 and/or CDRL3. The CDR consensus sequences of anti-Klotho beta antibodies are shown in Figure 2.

[0211] In certain embodiments, an antibody of a fragment thereof described herein comprises a VH region comprising: (1) a VH FR1 having an amino acid sequence selected from the group consisting of SEQ ID NOS: 278, 279 , 280 and 378; (2) a VH FR2 having an amino acid sequence selected from the group consisting of SEQ ID NOS: 281, 282, and 283; (3) a VH FR3 having an amino acid sequence selected from the group consisting of SEQ ID NOS: 284, 285, 286, 287 and 379-381; and/or (4) a VH FR4 having an amino acid of SEQ ID NO: 288. Accordingly, in some aspects, the humanized antibody comprises a VH region that includes a VH FR1 having an amino acid sequence selected from the group consisting of SEQ ID NOS: 278, 279, 280 and 378. In some aspects, the humanized antibody comprises a VH region that includes a VH FR2 having an amino acid sequence selected from the group consisting of SEQ ID NOS: 281, 282, and 283. In some aspects, the humanized antibody comprises a VH region that includes a VH FR3 having an amino acid sequence selected from the group consisting of SEQ ID NOS: 284, 285, 286, 287 and 379-381. In some aspects, the humanized antibody comprises a VH region that includes a VH FR4 having an amino acid of SEQ ID NO: 288.

[0212] In certain embodiments, an antibody of a fragment thereof described herein comprises a VL region comprising: (1) a VL FR1 having an amino acid sequence selected from the group consisting of SEQ ID NOS: 289, 290 and 382-384; (2) a VL FR2 having an amino acid sequence selected from the group consisting of SEQ ID NOS: 291, 292 and 385-392; (3) a VL FR3 having an amino acid sequence selected from the group consisting of SEQ ID NOS: 293, 294, 295 and 393-404; and/or (4) a VL FR4 having an amino acid of SEQ ID NO: 296 and 405-407. Thus, in some aspects, the humanized antibody comprises a VL region that includes a VL FR1 having an amino acid sequence selected from the group consisting of SEQ ID NOS: 289, 290 and 382-384. In some aspects, the humanized antibody comprises a VL region that includes a VL FR2 having an amino acid sequence selected from the group consisting of SEQ ID NOS: 291, 292 and 385-392. In some aspects, the humanized antibody comprises a VL region that includes a VL FR3 having an amino acid sequence selected from the group consisting of SEQ ID NOS: 293, 294, 295 and 393-404. In some aspects, the humanized antibody comprises a VL region that includes a VL FR4 having an amino acid of SEQ ID NO: 296 and 405-407.

[0213] In certain embodiments, an antibody of a fragment thereof described herein comprises a VH region and a VL region, wherein the VH region further comprises: (1) a VH FR1 having an amino acid sequence selected from of the group consisting of SEQ ID NOS: 278, 279, 280 and 378; (2) a VH FR2 having an amino acid sequence selected from the group consisting of SEQ ID NOS: 281, 282, and 283; (3) a VH FR3 having an amino acid sequence selected from the group consisting of SEQ ID NOS: 284, 285, 286, 287 and 379-381; and/or (4) a VH FR4 having an amino acid sequence of SEQ ID NO: 288; and wherein the VL region further comprises: (1) a VL FR1 having an amino acid sequence selected from the group consisting of SEQ ID NOS: 289, 290 and 382-384; (2) a VL FR2 having an amino acid sequence selected from the group consisting of SEQ ID NOS: 291, 292 and 385-392; (3) a VL FR3 having an amino acid sequence selected from the group consisting of SEQ ID NOS: 293, 294, 295 and 393-404; and/or (4) a VL FR4 having an amino acid of SEQ ID NO: 296 and 405-407.

[0214] Also provided herein are antibodies comprising one or more (e.g., one, two, three or four) VH FRs and one or more VL FRs listed in Table 19. In particular, provided herein is an antibody comprising an FR1 of VH (SEQ ID NOS:278, 279, 280 and 378) and an FR1 of VL (SEQ ID NOS:289, 290 and 382-384); an FR1 of VH (SEQ ID NOS:278, 279, 280 and 378) and an FR2 of VL (SEQ ID NOS:291, 292 and 385-392); an FR1 of VH (SEQ ID NOS: 278, 279, 280 and 378) and an FR3 of VL (SEQ ID NOS: 293, 294, 295 and 393-404); an FR1 of VH (SEQ ID NOS:278, 279, 280 and 378) and an FR4 of VL (SEQ ID NO:296 and 405-407); an FR2 of VH (SEQ ID NOS:281, 282, and 283) and an FR1 of VL (SEQ ID NOS:289, 290 and 382-384); an FR2 of VH (SEQ ID NOS:281, 282, and 283) and an FR2 of VL (SEQ ID NOS:291, 292 and 385-392); an FR2 of VH (SEQ ID NOS:281, 282, and 283) and an FR3 of VL (SEQ ID NOS:293, 294, 295 and 393-404); an FR2 of VH (SEQ ID NOS:281, 282, and 283) and an FR4 of VL (SEQ ID NO:296 and 405-407); a VH FR3 (SEQ ID NOS:284, 285, 286, 287 and 379-381) and a VH FR1 (SEQ ID NOS:278, 279, 280 and 378); an FR3 of VH (SEQ ID NOS:284, 285, 286, 287 and 379-381) and an FR2 of VL (SEQ ID NOS:291, 292 and 385-392); an FR3 of VH (SEQ ID NOS:284, 285, 286, 287 and 379-381) and an FR3 of VL (SEQ ID NOS:293, 294, 295 and 393-404); an FR3 of VH (SEQ ID NOS:284, 285, 286, 287 and 379-381) and an FR4 of VL (SEQ ID NO:296 and 405-407); an FR1 of VH (SEQ ID NOS:278, 279, 280 and 378), an FR2 of VH (SEQ ID NOS:281, 282, and 283) and an FR1 of VL (SEQ ID NOS:289, 290 and 382384) ; an FR1 of VH (SEQ ID NOS:278, 279, 280 and 378), an FR2 of VH (SEQ ID NOS:281, 282, and 283) and an FR2 of VL (SEQ ID NOS:291, 292 and 385- 392); an FR1 of VH (SEQ ID NOS:278, 279, 280 and 378), an FR2 of VH (SEQ ID NOS:281, 282, and 283) and an FR3 of VL (SEQ ID NOS:293, 294, 295 and 393-404); a VH FR1 (SEQ ID NOS:278, 279, 280 and 378), a VH FR2 (SEQ ID NOS:281, 282, and 283) and a VL FR4 (SEQ ID NO:296 and 405-407); a FR2 of VH (SEQ ID NOS:281, 282, and 283), an FR3 of VH (SEQ ID NOS:284, 285, 286, 287 and 379381) and an FR1 of VL (SEQ ID NOS:289, 290 and 382-384), a VH FR2 (SEQ ID NOS:281, 282, and 283), a VH FR3 (SEQ ID NOS:284, 285, 286, 287 and 379-381) and an FR2 of VL (SEQ ID NOS:291, 292 and 385-392); an FR2 of VH (SEQ ID NOS:281, 282, and 283), an FR3 of VH (SEQ ID NOS:284, 285, 286, 287 and 379-381) and an FR3 of VL (SEQ ID NOS:293, 294, 295 and 393-404); an FR2 of VH (SEQ ID NOS:281, 282, and 283), an FR3 of VH (SEQ ID NOS:284, 285, 286, 287 and 379,381) and an FR4 of VL (SEQ ID NO:296 and 405 -407); an FR1 of VH (SEQ ID NOS:278, 279, 280 and 378), an FR1 of VL (SEQ ID NOS:289, 290 and 382-384) and an FR2 of VL (SEQ ID NOS:291, 292 and 385 -392); an FR1 of VH (SEQ ID NOS:278, 279, 280 and 378), an FR1 of VL (SEQ ID NOS:289, 290 and 382-384) and an FR3 of VL (SEQ ID NOS:293, 294, 295 and 393-404); an FR1 of VH (SEQ ID NOS:278, 279, 280 and 378), an FR1 of VL (SEQ ID NOS:289, 290 and 382-384) and an FR4 of VL (SEQ ID NO:296 and 405-407 ); an FR1 of VH (SEQ ID NOS:278, 279, 280 and 378), an FR2 of VL (SEQ ID NOS:291, 292 and 385-392) and an FR3 of VL (SEQ ID NOS:293, 294, 295 and 393-404); an FR1 of VH (SEQ ID NOS:278, 279, 280 and 378), an FR2 of VL (SEQ ID NOS:291, 292 and 385-392) and an FR4 of VL (SEQ ID NO:296 and 405-407 ); an FR2 of VH (SEQ ID NOS:281, 282, and 283), an FR1 of VL (SEQ ID NOS:289, 290 and 382384) and an FR2 of VL (SEQ ID NOS:291, 292 and 385-392) ; an FR2 of VH (SEQ ID NOS:281, 282, and 283), an FR1 of VL (SEQ ID NOS:289, 290 and 382-384) and an FR3 of VL (SEQ ID NOS:293, 294, 295 and 393404); an FR2 of VH (SEQ ID NOS:281, 282, and 283), an FR1 of VL (SEQ ID NOS:289, 290 and 382-384) and an FR4 of VL (SEQ ID NO:296 and 405-407) ; an FR2 of VH (SEQ ID NOS:281, 282, and 283), an FR2 of VL (SEQ ID NOS:291, 292 and 385-392) and an FR3 of VL (SEQ ID NOS:293, 294, 295 and 393-404); an FR2 of VH (SEQ ID NOS:281, 282, and 283), an FR2 of VL (SEQ ID NOS:291, 292 and 385-392) and an FR4 of VL (SEQ ID NO:296 and 405-407) ; an FR3 of VH (SEQ ID NOS:284, 285, 286, 287 and 379-381), an FR1 of VL (SEQ ID NOS:289, 290 and 382-384) and an FR2 of VL (SEQ ID NOS:291 , 292 and 385-392); an FR3 of VH (SEQ ID NOS:284, 285, 286, 287 and 379-381), an FR1 of VL (SEQ ID NOS:289, 290 and 382-384) and an FR3 of VL (SEQ ID NOS:293 , 294, 295 and 393-404); an FR3 of VH (SEQ ID NOS:284, 285, 286, 287 and 379381), an FR1 of VL (SEQ ID NOS:289, 290 and 382-384) and an FR4 of VL (SEQ ID NO:296 and 405 -407); an FR3 of VH (SEQ ID NOS:284, 285, 286, 287 and 379-381), an FR2 of VL (SEQ ID NOS:291, 292 and 385-392) and an FR3 of VL (SEQ ID NOS:293 , 294, 295 and 393-404); an FR3 of VH (SEQ ID NOS:284, 285, 286, 287 and 379-381), an FR2 of VL (SEQ ID NOS:291, 292 and 385-392) and an FR4 of VL (SEQ ID NO:296 and 405-407); an FR4 of VH (SEQ ID NO:288), an FR1 of VL (SEQ ID NOS:289, 290 and 382-384) and an FR2 of VL (SEQ ID NOS:291, 292 and 385-392); an FR4 of VH (SEQ ID NO:288), an FR1 of VL (SEQ ID NOS:289, 290 and 382-384) and an FR3 of VL (SEQ ID NOS:293, 294, 295 and 393-404); an FR4 of VH (SEQ ID NO:288), an FR1 of VL (SEQ ID NOS:289, 290 and 382-384) and an FR4 of VL (SEQ ID NO:296 and 405-407); an FR4 of VH (SEQ ID NO:288), an FR2 of VL (SEQ ID NOS:291, 292 and 385-392) and an FR3 of VL (SEQ ID NOS:293, 294, 295 and 393404); an FR4 of VH (SEQ ID NO:288), an FR2 of VL (SEQ ID NOS:291, 292 and 385-392) and an FR4 of VL (SEQ ID NO:296 and 405-407); a VH FR1 (SEQ ID NOS:278, 279, 280 and 378), a VH FR2 (SEQ ID NOS:281, 282, and 283), a VH FR3 (SEQ ID NOS:284, 285, 286, 287 and 379-381) and an FR1 of VL (SEQ ID NOS:289, 290 and 382-384); a VH FR1 (SEQ ID NOS:278, 279, 280 and 378), a VH FR2 (SEQ ID NOS:281, 282, and 283), a VH FR3 (SEQ ID NOS:284, 285, 286, 287 and 379-381) and an FR2 of VL (SEQ ID NOS:291, 292 and 385-392); a VH FR1 (SEQ ID NOS:278, 279, 280 and 378), a VH FR2 (SEQ ID NOS:281, 282, and 283), a VH FR3 (SEQ ID NOS:284, 285, 286, 287 and 379-381) and an FR3 of VL (SEQ ID NOS:293, 294, 295 and 393404); a VH FR1 (SEQ ID NOS:278, 279, 280 and 378), a VH FR2 (SEQ ID NOS:281, 282, and 283), a VH FR3 (SEQ ID NOS:284, 285, 286, 287 and 379-381) and an FR4 of VL (SEQ ID NO:296 and 405-407); an FR1 of VH (SEQ ID NOS:278, 279, 280 and 378), an FR2 of VH (SEQ ID NOS:281, 282, and 283), an FR4 of VH (SEQ ID NO:288) and an FR1 of VL (SEQ ID NOS:289, 290 and 382-384); an FR1 of VH (SEQ ID NOS:278, 279, 280 and 378), an FR2 of VH (SEQ ID NOS:281, 282, and 283), an FR4 of VH (SEQ ID NO:288) and an FR2 of VL (SEQ ID NOS:291, 292 and 385-392); an FR1 of VH (SEQ ID NOS:278, 279, 280 and 378), an FR2 of VH (SEQ ID NOS:281, 282, and 283), an FR4 of VH (SEQ ID NO:288) and an FR3 of VL (SEQ ID NOS:293, 294, 295 and 393404); an FR1 of VH (SEQ ID NOS:278, 279, 280 and 378), an FR2 of VH (SEQ ID NOS:281, 282, and 283), an FR4 of VH (SEQ ID NO:288) and an FR4 of VL (SEQ ID NO:296 and 405-407); an FR1 of VH (SEQ ID NOS:278, 279, 280 and 378), an FR3 of VH (SEQ ID NOS:284, 285, 286, 287 and 379-381), an FR4 of VH (SEQ ID NO:288 ) and an FR1 of VL (SEQ ID NOS:289, 290 and 382-384); an FR1 of VH (SEQ ID NOS:278, 279, 280 and 378), an FR3 of VH (SEQ ID NOS:284, 285, 286, 287 and 379-381), an FR4 of VH (SEQ ID NO:288 ) and an FR2 of VL (SEQ ID NOS:291, 292 and 385-392); an FR1 of VH (SEQ ID NOS:278, 279, 280 and 378), an FR3 of VH (SEQ ID NOS:284, 285, 286, 287 and 379-381), an FR4 of VH (SEQ ID NO:288 ) and an FR3 of VL (SEQ ID NOS:293, 294, 295 and 393-404); an FR1 of VH (SEQ ID NOS:278, 279, 280 and 378), an FR3 of VH (SEQ ID NOS:284, 285, 286, 287 and 379-381), an FR4 of VH (SEQ ID NO:288 ) and an FR4 of VL (SEQ ID NO:296 and 405-407); an FR2 of VH (SEQ ID NOS:281, 282, and 283), an FR3 of VH (SEQ ID NOS:284, 285, 286, 287 and 379-381), an FR4 of VH (SEQ ID NO:288) and an FR1 of VL (SEQ ID NOS:289, 290 and 382-384); an FR2 of VH (SEQ ID NOS:281, 282, and 283), an FR3 of VH (SEQ ID NOS:284, 285, 286, 287 and 379-381), an FR4 of VH (SEQ ID NO:288) and an FR2 of VL (SEQ ID NOS:291, 292 and 385-392); an FR2 of VH (SEQ ID NOS:281, 282, and 283), an FR3 of VH (SEQ ID NOS:284, 285, 286, 287 and 379-381), an FR4 of VH (SEQ ID NO:288) and an FR3 of VL (SEQ ID NOS:293, 294, 295 and 393-404); an FR2 of VH (SEQ ID NOS:281, 282, and 283), an FR3 of VH (SEQ ID NOS:284, 285, 286, 287 and 379,381), an FR4 of VH (SEQ ID NO:288) and an FR4 of VL (SEQ ID NO:296 and 405-407); an FR1 of VH (SEQ ID NOS:278, 279, 280 and 378), an FR2 of VH (SEQ ID NOS:281, 282, and 283), an FR1 of VL (SEQ ID NOS:289, 290 and 382- 384) and an FR2 of VL (SEQ ID NOS:291, 292 and 385-392); an FR1 of VH (SEQ ID NOS:278, 279, 280 and 378), an FR2 of VH (SEQ ID NOS:281, 282, and 283), an FR1 of VL (SEQ ID NOS:289, 290 and 382- 384) and an FR3 of VL (SEQ ID NOS:293, 294, 295 and 393-404); an FR1 of VH (SEQ ID NOS:278, 279, 280 and 378), an FR2 of VH (SEQ ID NOS:281, 282, and 283), an FR1 of VL (SEQ ID NOS:289, 290 and 382- 384) and an FR4 of VL (SEQ ID NO:296 and 405-407); an FR1 of VH (SEQ ID NOS:278, 279, 280 and 378), an FR3 of VH (SEQ ID NOS:284, 285, 286, 287 and 379-381), an FR1 of VL (SEQ ID NOS:289 , 290 and 382-384) and an FR2 of VL (SEQ ID NOS:291, 292 and 385-392); an FR1 of VH (SEQ ID NOS:278, 279, 280 and 378), an FR3 of VH (SEQ ID NOS:284, 285, 286, 287 and 379-381), an FR1 of VL (SEQ ID NOS:289 , 290 and 382-384) and a FR3 of VL (SEQ ID NOS:293, 294, 295 and 393-404); an FR1 of VH (SEQ ID NOS:278, 279, 280 and 378), an FR3 of VH (SEQ ID NOS:284, 285, 286, 287 and 379-381), an FR1 of VL (SEQ ID NOS:289 , 290 and 382-384) and an FR4 of VL (SEQ ID NO:296 and 405-407); an FR1 of VH (SEQ ID NOS:278, 279, 280 and 378), an FR4 of VH (SEQ ID NO:288), an FR1 of VL (SEQ ID NOS:289, 290 and 382-384) and an FR2 of VL (SEQ ID NOS:291, 292 and 385-392); an FR1 of VH (SEQ ID NOS:278, 279, 280 and 378), an FR4 of VH (SEQ ID NO:288), an FR1 of VL (SEQ ID NOS:289, 290 and 382-384) and an FR3 of VL (SEQ ID NOS:293, 294, 295 and 393-404); an FR1 of VH (SEQ ID NOS:278, 279, 280 and 378), an FR4 of VH (SEQ ID NO:288), an FR1 of VL (SEQ ID NOS:289, 290 and 382-384) and an FR4 of VL (SEQ ID NO:296 and 405-407); an FR2 of VH (SEQ ID NOS:281, 282, and 283), an FR3 of VH (SEQ ID NOS:284, 285, 286, 287 and 379-381), an FR1 of VL (SEQ ID NOS:289, 290 and 382-384) and an FR2 of VL (SEQ ID NOS:291, 292 and 385-392); an FR2 of VH (SEQ ID NOS:281, 282, and 283), an FR3 of VH (SEQ ID NOS:284, 285, 286, 287 and 379-381), an FR1 of VL (SEQ ID NOS:289, 290 and 382-384) and an FR3 of VL (SEQ ID NOS:293, 294, 295 and 393-404); an FR2 of VH (SEQ ID NOS:281, 282, and 283), an FR3 of VH (SEQ ID NOS:284, 285, 286, 287 and 379-381), an FR1 of VL (SEQ ID NOS:289, 290 and 382-384) and an FR4 of VL (SEQ ID NO:296 and 405-407); an FR2 of VH (SEQ ID NOS:281, 282, and 283), an FR4 of VH (SEQ ID NO:288), an FR1 of VL (SEQ ID NOS:289, 290 and 382-384) and an FR2 of VL (SEQ ID NOS:291, 292 and 385-392); an FR2 of VH (SEQ ID NOS:281, 282, and 283), an FR4 of VH (SEQ ID NO:288), an FR1 of VL (SEQ ID NOS:289, 290 and 382-384) and an FR3 of VL (SEQ ID NOS:293, 294, 295 and 393-404); an FR2 of VH (SEQ ID NOS:281, 282, and 283), an FR4 of VH (SEQ ID NO:288), an FR1 of VL (SEQ ID NOS:289, 290 and 382-384) and an FR4 of VL (SEQ ID NO:296 and 405-407); an FR3 of VH (SEQ ID NOS:284, 285, 286, 287 and 379-381), an FR4 of VH (SEQ ID NO:288), an FR1 of VL (SEQ ID NOS:289, 290 and 382-384 ) and an FR2 of VL (SEQ ID NOS:291, 292 and 385-392); an FR3 of VH (SEQ ID NOS:284, 285, 286, 287 and 379-381), an FR4 of VH (SEQ ID NO:288), an FR1 of VL (SEQ ID NOS:289, 290 and 382-384 ) and an FR3 of VL (SEQ ID NOS:293, 294, 295 and 393-404); an FR3 of VH (SEQ ID NOS:284, 285, 286, 287 and 379-381), an FR4 of VH (SEQ ID NO:288), an FR1 of VL (SEQ ID NOS:289, 290 and 382-384 ) and an FR4 of VL (SEQ ID NO:296 and 405-407); an FR1 of VH (SEQ ID NOS:278, 279, 280 and 378), an FR2 of VH (SEQ ID NOS:281, 282, and 283), an FR2 of VL (SEQ ID NOS:291, 292 and 385- 392) and an FR3 of VL (SEQ ID NOS:293, 294, 295 and 393-404); an FR1 of VH (SEQ ID NOS:278, 279, 280 and 378), an FR2 of VH (SEQ ID NOS:281, 282, and 283), an FR2 of VL (SEQ ID NOS:291, 292 and 385- 392) and an FR4 of VL (SEQ ID NO:296 and 405-407); an FR1 of VH (SEQ ID NOS:278, 279, 280 and 378), an FR3 of VH (SEQ ID NOS:284, 285, 286, 287 and 379-381), an FR2 of VL (SEQ ID NOS:291 , 292 and 385-392) and a FR3 of VL (SEQ ID NOS:293, 294, 295 and 393-404); an FR1 of VH (SEQ ID NOS:278, 279, 280 and 378), an FR3 of VH (SEQ ID NOS:284, 285, 286, 287 and 379-381), an FR2 of VL (SEQ ID NOS:291 , 292 and 385-392) and an FR4 of VL (SEQ ID NO:296 and 405-407); an FR1 of VH (SEQ ID NOS:278, 279, 280 and 378), an FR4 of VH (SEQ ID NO:288), an FR2 of VL (SEQ ID NOS:291, 292 and 385-392) and an FR3 of VL (SEQ ID NOS:293, 294, 295 and 393-404); an FR1 of VH (SEQ ID NOS:278, 279, 280 and 378), an FR4 of VH (SEQ ID NO:288), an FR2 of VL (SEQ ID NOS:291, 292 and 385-392) and an FR4 of VL (SEQ ID NO:296 and 405-407); an FR2 of VH (SEQ ID NOS:281, 282, and 283), an FR3 of VH (SEQ ID NOS:284, 285, 286, 287 and 379381), an FR2 of VL (SEQ ID NOS:291, 292 and 385-392) and an FR3 of VL (SEQ ID NOS:293, 294, 295 and 393-404); an FR2 of VH (SEQ ID NOS:281, 282, and 283), an FR3 of VH (SEQ ID NOS:284, 285, 286, 287 and 379-381), an FR2 of VL (SEQ ID NOS:291, 292 and 385-392) and an FR4 of VL (SEQ ID NO:296 and 405-407); an FR2 of VH (SEQ ID NOS:281, 282, and 283), an FR4 of VH (SEQ ID NO:288), an FR2 of VL (SEQ ID NOS:291, 292 and 385-392) and an FR3 of VL (SEQ ID NOS:293, 294, 295 and 393-404); an FR2 of VH (SEQ ID NOS:281, 282, and 283), an FR4 of VH (SEQ ID NO:288), an FR2 of VL (SEQ ID NOS:291, 292 and 385-392) and an FR4 of VL (SEQ ID NO:296 and 405-407); an FR3 of VH (SEQ ID NOS:284, 285, 286, 287 and 379-381), an FR4 of VH (SEQ ID NO:288), an FR2 of VL (SEQ ID NOS:291, 292 and 385-392 ) and an FR3 of VL (SEQ ID NOS:293, 294, 295 and 393-404); an FR3 of VH (SEQ ID NOS:284, 285, 286, 287 and 379-381), an FR4 of VH (SEQ ID NO:288), an FR2 of VL (SEQ ID NOS:291, 292 and 385-392 ) and an FR4 of VL (SEQ ID NO:296 and 405-407); an FR1 of VH (SEQ ID NOS:278, 279, 280 and 378), an FR2 of VH (SEQ ID NOS:281, 282, and 283), an FR3 of VL (SEQ ID NOS:293, 294, 295 and 393-404) and an FR4 of VL (SEQ ID NO:296 and 405-407); an FR1 of VH (SEQ ID NOS:278, 279, 280 and 378), an FR3 of VH (SEQ ID NOS:284, 285, 286, 287 and 379-381), an FR3 of VL (SEQ ID NOS:293 , 294, 295 and 393-404) and an FR4 of VL (SEQ ID NO:296 and 405-407); an FR1 of VH (SEQ ID NOS:278, 279, 280 and 378), an FR4 of VH (SEQ ID NO:288), an FR3 of VL (SEQ ID NOS:293, 294, 295 and 393-404) and an FR4 of VL (SEQ ID NO:296 and 405-407); an FR2 of VH (SEQ ID NOS:281, 282, and 283), an FR3 of VH (SEQ ID NOS:284, 285, 286, 287 and 379-381), an FR3 of VL (SEQ ID NOS:293, 294, 295 and 393-404) and an FR4 of VL (SEQ ID NO:296 and 405-407); an FR2 of VH (SEQ ID NOS:281, 282, and 283), an FR4 of VH (SEQ ID NO:288), an FR3 of VL (SEQ ID NOS:293, 294, 295 and 393-404) and a FR4 of VL (SEQ ID NO:296 and 405-407); an FR3 of VH (SEQ ID NOS:284, 285, 286, 287 and 379-381), an FR4 of VH (SEQ ID NO:288), an FR3 of VL (SEQ ID NOS:293, 294, 295 and 393 -404) and an FR4 of VL (SEQ ID NO:296 and 405-407); an FR1 of VH (SEQ ID NOS:278, 279, 280 and 378), an FR1 of VL (SEQ ID NOS:289, 290 and 382-384), an FR2 of VL (SEQ ID NOS:291, 292 and 385 -392) and an FR3 of VL (SEQ ID NOS:293, 294, 295 and 393-404); an FR1 of VH (SEQ ID NOS:278, 279, 280 and 378), an FR1 of VL (SEQ ID NOS:289, 290 and 382-384), an FR2 of VL (SEQ ID NOS:291, 292 and 385 -392) and an FR4 of VL (SEQ ID NO:296 and 405-407); an FR1 of VH (SEQ ID NOS:278, 279, 280 and 378), an FR1 of VL (SEQ ID NOS:289, 290 and 382-384), an FR3 of VL (SEQ ID NOS:293, 294, 295 and 393-404) and an FR4 of VL (SEQ ID NO:296 and 405-407); an FR1 of VH (SEQ ID NOS:278, 279, 280 and 378), an FR2 of VL (SEQ ID NOS:291, 292 and 385-392), an FR3 of VL (SEQ ID NOS:293, 294, 295 and 393-404) and an FR4 of VL (SEQ ID NO:296 and 405-407); an FR2 of VH (SEQ ID NOS:281, 282, and 283), an FR1 of VL (SEQ ID NOS:289, 290 and 382-384), an FR2 of VL (SEQ ID NOS:291, 292 and 385- 392) and an FR3 of VL (SEQ ID NOS:293, 294, 295 and 393404); an FR2 of VH (SEQ ID NOS:281, 282, and 283), an FR1 of VL (SEQ ID NOS:289, 290 and 382-384), an FR2 of VL (SEQ ID NOS:291, 292 and 385- 392) and an FR4 of VL (SEQ ID NO:296 and 405-407); an FR2 of VH (SEQ ID NOS:281, 282, and 283), an FR1 of VL (SEQ ID NOS:289, 290 and 382-384), an FR3 of VL (SEQ ID NOS:293, 294, 295 and 393-404) and an FR4 of VL (SEQ ID NO:296 and 405-407); an FR2 of VH (SEQ ID NOS:281, 282, and 283), an FR2 of VL (SEQ ID NOS:291, 292, and 385-392), an FR3 of VL (SEQ ID NOS:293, 294, 295 and 393-404) and an FR4 of VL (SEQ ID NO:296 and 405-407); an FR3 of VH (SEQ ID NOS:284, 285, 286, 287 and 379-381), an FR1 of VL (SEQ ID NOS:289, 290 and 382-384), an FR2 of VL (SEQ ID NOS:291 , 292 and 385-392) and a FR3 of VL (SEQ ID NOS:293, 294, 295 and 393-404); an FR3 of VH (SEQ ID NOS:284, 285, 286, 287 and 379-381), an FR1 of VL (SEQ ID NOS:289, 290 and 382-384), an FR2 of VL (SEQ ID NOS:291 , 292 and 385-392) and an FR4 of VL (SEQ ID NO:296 and 405-407); an FR3 of VH (SEQ ID NOS:284, 285, 286, 287 and 379-381), an FR1 of VL (SEQ ID NOS:289, 290 and 382-384), an FR3 of VL (SEQ ID NOS:293 , 294, 295 and 393-404) and an FR4 of VL (SEQ ID NO:296 and 405-407); an FR3 of VH (SEQ ID NOS:284, 285, 286, 287 and 379-381), an FR2 of VL (SEQ ID NOS:291, 292 and 385-392), an FR3 of VL (SEQ ID NOS:293 , 294, 295 and 393-404) and an FR4 of VL (SEQ ID NO:296 and 405-407); an FR4 of VH (SEQ ID NO:288), an FR1 of VL (SEQ ID NOS:289, 290 and 382-384), an FR2 of VL (SEQ ID NOS:291, 292 and 385-392) and an FR3 of VL (SEQ ID NOS:293, 294, 295 and 393-404); an FR4 of VH (SEQ ID NO:288), an FR1 of VL (SEQ ID NOS:289, 290 and 382-384), an FR2 of VL (SEQ ID NOS:291, 292 and 385-392) and an FR4 of VL (SEQ ID NO:296 and 405-407); an FR4 of VH (SEQ ID NO:288), an FR1 of VL (SEQ ID NOS:289, 290 and 382-384), an FR3 of VL (SEQ ID NOS:293, 294, 295 and 393-404) and an FR4 of VL (SEQ ID NO:296 and 405-407); an FR4 of VH (SEQ ID NO:288), an FR2 of VL (SEQ ID NOS:291, 292 and 385-392), an FR3 of VL (SEQ ID NOS:293, 294, 295 and 393-404) and an FR4 of VL (SEQ ID NO:296 and 405-407); a VH FR1 (SEQ ID NOS:278, 279, 280 and 378), a VH FR2 (SEQ ID NOS:281, 282, and 283), a VH FR3 (SEQ ID NOS:284, 285, 286, 287 and 379-381), an FR4 of VH (SEQ ID NO:288) and an FR1 of VL (SEQ ID NOS:289, 290 and 382-384); a VH FR1 (SEQ ID NOS:278, 279, 280 and 378), a VH FR2 (SEQ ID NOS:281, 282, and 283), a VH FR3 (SEQ ID NOS:284, 285, 286, 287 and 379-381), an FR4 of VH (SEQ ID NO:288) and an FR2 of VL (SEQ ID NOS:291, 292 and 385-392); a VH FR1 (SEQ ID NOS:278, 279, 280 and 378), a VH FR2 (SEQ ID NOS:281, 282, and 283), a VH FR3 (SEQ ID NOS:284, 285, 286, 287 and 379-381), an FR4 of VH (SEQ ID NO:288) and an FR3 of VL (SEQ ID NOS:293, 294, 295 and 393-404); a VH FR1 (SEQ ID NOS:278, 279, 280 and 378), a VH FR2 (SEQ ID NOS:281, 282, and 283), a VH FR3 (SEQ ID NOS:284, 285, 286, 287 and 379-381), an FR4 of VH (SEQ ID NO:288) and an FR4 of VL (SEQ ID NO:296 and 405-407); a VH FR1 (SEQ ID NOS:278, 279, 280 and 378), a VH FR2 (SEQ ID NOS:281, 282, and 283), a VH FR3 (SEQ ID NOS:284, 285, 286, 287 and 379-381), an FR1 of VL (SEQ ID NOS:289, 290 and 382-384) and an FR2 of VL (SEQ ID NOS:291, 292 and 385-392); a VH FR1 (SEQ ID NOS:278, 279, 280 and 378), a VH FR2 (SEQ ID NOS:281, 282, and 283), a VH FR3 (SEQ ID NOS:284, 285, 286, 287 and 379-381), an FR1 of VL (SEQ ID NOS:289, 290 and 382-384) and an FR3 of VL (SEQ ID NOS:293, 294, 295 and 393-404); a VH FR1 (SEQ ID NOS:278, 279, 280 and 378), a VH FR2 (SEQ ID NOS:281, 282, and 283), a VH FR3 (SEQ ID NOS:284, 285, 286, 287 and 379-381), an FR1 of VL (SEQ ID NOS:289, 290 and 382-384) and an FR4 of VL (SEQ ID NO:296 and 405-407); a VH FR1 (SEQ ID NOS:278, 279, 280 and 378), a VH FR2 (SEQ ID NOS:281, 282, and 283), a VH FR3 (SEQ ID NOS:284, 285, 286, 287 and 379-381), an FR2 of VL (SEQ ID NOS:291, 292 and 385-392) and an FR3 of VL (SEQ ID NOS:293, 294, 295 and 393-404); a VH FR1 (SEQ ID NOS:278, 279, 280 and 378), a VH FR2 (SEQ ID NOS:281, 282, and 283), a VH FR3 (SEQ ID NOS:284, 285, 286, 287 and 379-381), an FR2 of VL (SEQ ID NOS:291, 292 and 385-392) and an FR4 of VL (SEQ ID NO:296 and 405-407); a VH FR1 (SEQ ID NOS:278, 279, 280 and 378), a VH FR2 (SEQ ID NOS:281, 282, and 283), a VH FR3 (SEQ ID NOS:284, 285, 286, 287 and 379-381), an FR3 of VL (SEQ ID NOS:293, 294, 295 and 393-404) and an FR4 of VL (SEQ ID NO:296 and 405-407); an FR1 of VH (SEQ ID NOS:278, 279, 280 and 378), an FR2 of VH (SEQ ID NOS:281, 282, and 283), an FR4 of VH (SEQ ID NO:288), an FR1 of VL (SEQ ID NOS:289, 290 and 382-384) and an FR2 of VL (SEQ ID NOS:291, 292 and 385-392); an FR1 of VH (SEQ ID NOS:278, 279, 280 and 378), an FR2 of VH (SEQ ID NOS:281, 282, and 283), an FR4 of VH (SEQ ID NO:288), an FR1 of VL (SEQ ID NOS:289, 290 and 382-384) and an FR3 of VL (SEQ ID NOS:293, 294, 295 and 393-404); an FR1 of VH (SEQ ID NOS:278, 279, 280 and 378), an FR2 of VH (SEQ ID NOS:281, 282, and 283), an FR4 of VH (SEQ ID NO:288), an FR1 of VL (SEQ ID NOS:289, 290 and 382-384) and an FR4 of VL (SEQ ID NO:296 and 405-407); an FR1 of VH (SEQ ID NOS:278, 279, 280 and 378), an FR2 of VH (SEQ ID NOS:281, 282, and 283), an FR4 of VH (SEQ ID NO:288), an FR2 of VL (SEQ ID NOS:291, 292 and 385-392) and an FR3 of VL (SEQ ID NOS:293, 294, 295 and 393-404); an FR1 of VH (SEQ ID NOS:278, 279, 280 and 378), an FR2 of VH (SEQ ID NOS:281, 282, and 283), an FR4 of VH (SEQ ID NO:288), an FR2 of VL (SEQ ID NOS:291, 292 and 385-392) and an FR4 of VL (SEQ ID NO:296 and 405-407); an FR1 of VH (SEQ ID NOS:278, 279, 280 and 378), an FR2 of VH (SEQ ID NOS:281, 282, and 283), an FR4 of VH (SEQ ID NO:288), an FR3 of VL (SEQ ID NOS:293, 294, 295 and 393-404) and an FR4 of VL (SEQ ID NO:296 and 405-407); an FR2 of VH (SEQ ID NOS:281, 282, and 283), an FR3 of VH (SEQ ID NOS:284, 285, 286, 287 and 379-381), an FR4 of VH (SEQ ID NO:288) , an FR1 of VL (SEQ ID NOS:289, 290 and 382-384) and an FR2 of VL (SEQ ID NOS:291, 292 and 385-392); an FR2 of VH (SEQ ID NOS:281, 282, and 283), an FR3 of VH (SEQ ID NOS:284, 285, 286, 287 and 379-381), an FR4 of VH (SEQ ID NO:288) , an FR1 of VL (SEQ ID NOS:289, 290 and 382-384) and an FR3 of VL (SEQ ID NOS:293, 294, 295 and 393-404); an FR2 of VH (SEQ ID NOS:281, 282, and 283), an FR3 of VH (SEQ ID NOS:284, 285, 286, 287 and 379-381), an FR4 of VH (SEQ ID NO:288) , an FR1 of VL (SEQ ID NOS:289, 290 and 382-384) and an FR4 of VL (SEQ ID NO:296 and 405-407); an FR2 of VH (SEQ ID NOS:281, 282, and 283), an FR3 of VH (SEQ ID NOS:284, 285, 286, 287 and 379-381), an FR4 of VH (SEQ ID NO:288) , an FR2 of VL (SEQ ID NOS:291, 292 and 385-392) and an FR3 of VL (SEQ ID NOS:293, 294, 295 and 393-404); an FR2 of VH (SEQ ID NOS:281, 282, and 283), an FR3 of VH (SEQ ID NOS:284, 285, 286, 287 and 379-381), an FR4 of VH (SEQ ID NO:288) , an FR2 of VL (SEQ ID NOS:291, 292 and 385-392) and an FR4 of VL (SEQ ID NO:296 and 405-407); an FR2 of VH (SEQ ID NOS:281, 282, and 283), an FR3 of VH (SEQ ID NOS:284, 285, 286, 287 and 379-381), an FR4 of VH (SEQ ID NO:288) , an FR3 of VL (SEQ ID NOS:293, 294, 295 and 393-404) and an FR4 of VL (SEQ ID NO:296 and 405-407); an FR1 of VH (SEQ ID NOS:278, 279, 280 and 378), an FR2 of VH (SEQ ID NOS:281, 282, and 283), an FR1 of VL (SEQ ID NOS:289, 290 and 382- 384), an FR2 of VL (SEQ ID NOS:291, 292 and 385-392), and an FR3 of VL (SEQ ID NOS:293, 294, 295 and 393-404); an FR1 of VH (SEQ ID NOS:278, 279, 280 and 378), an FR2 of VH (SEQ ID NOS:281, 282, and 283), an FR1 of VL (SEQ ID NOS:289, 290 and 382- 384), an FR2 of VL (SEQ ID NOS:291, 292 and 385-392), and an FR4 of VL (SEQ ID NO:296 and 405-407); an FR1 of VH (SEQ ID NOS:278, 279, 280 and 378), an FR2 of VH (SEQ ID NOS:281, 282, and 283), an FR1 of VL (SEQ ID NOS:289, 290 and 382- 384), an FR3 of VL (SEQ ID NOS:293, 294, 295 and 393-404), and an FR4 of VL (SEQ ID NO:296 and 405-407); an FR1 of VH (SEQ ID NOS:278, 279, 280 and 378), an FR2 of VH (SEQ ID NOS:281, 282, and 283), an FR2 of VL (SEQ ID NOS:291, 292 and 385- 392), an FR3 of VL (SEQ ID NOS:293, 294, 295 and 393-404), and an FR4 of VL (SEQ ID NO:296 and 405-407); an FR1 of VH (SEQ ID NOS:278, 279, 280 and 378), an FR3 of VH (SEQ ID NOS:284, 285, 286, 287 and 379-381), an FR1 of VL (SEQ ID NOS:289 , 290 and 382-384), an FR2 of VL (SEQ ID NOS:291, 292 and 385-392), and an FR3 of VL (SEQ ID NOS:293, 294, 295 and 393-404); an FR1 of VH (SEQ ID NOS:278, 279, 280 and 378), an FR3 of VH (SEQ ID NOS:284, 285, 286, 287 and 379-381), an FR1 of VL (SEQ ID NOS:289 , 290 and 382-384), an FR2 of VL (SEQ ID NOS:291, 292 and 385-392), and an FR4 of VL (SEQ ID NO:296 and 405-407); an FR1 of VH (SEQ ID NOS:278, 279, 280 and 378), an FR3 of VH (SEQ ID NOS:284, 285, 286, 287 and 379381), an FR1 of VL (SEQ ID NOS:289, 290 and 382-384), an FR3 of VL (SEQ ID NOS:293, 294, 295 and 393-404), and an FR4 of VL (SEQ ID NO:296 and 405-407); an FR1 of VH (SEQ ID NOS:278, 279, 280 and 378), an FR3 of VH (SEQ ID NOS:284, 285, 286, 287 and 379-381), an FR2 of VL (SEQ ID NOS:291 , 292 and 385-392), an FR3 of VL (SEQ ID NOS:293, 294, 295 and 393-404), and an FR4 of VL (SEQ ID NO:296 and 405-407); an FR1 of VH (SEQ ID NOS:278, 279, 280 and 378), an FR4 of VH (SEQ ID NO:288), an FR1 of VL (SEQ ID NOS:289, 290 and 382384), an FR2 of VL (SEQ ID NOS:291, 292 and 385-392), and an FR3 of VL (SEQ ID NOS:293, 294, 295 and 393-404); an FR1 of VH (SEQ ID NOS:278, 279, 280 and 378), an FR4 of VH (SEQ ID NO:288), an FR1 of VL (SEQ ID NOS:289, 290 and 382-384), an FR2 of VL (SEQ ID NOS:291, 292 and 385-392), and an FR4 of VL (SEQ ID NO:296 and 405-407); an FR1 of VH (SEQ ID NOS:278, 279, 280 and 378), an FR4 of VH (SEQ ID NO:288), an FR1 of VL (SEQ ID NOS:289, 290 and 382-384), an FR3 of VL (SEQ ID NOS:293, 294, 295 and 393-404), and an FR4 of VL (SEQ ID NO:296 and 405-407); an FR1 of VH (SEQ ID NOS:278, 279, 280 and 378), an FR4 of VH (SEQ ID NO:288), an FR2 of VL (SEQ ID NOS:291, 292 and 385-392), an FR3 of VL (SEQ ID NOS:293, 294, 295 and 393-404), and an FR4 of VL (SEQ ID NO:296 and 405-407); an FR2 of VH (SEQ ID NOS:281, 282, and 283), an FR3 of VH (SEQ ID NOS:284, 285, 286, 287 and 379-381), an FR1 of VL (SEQ ID NOS:289, 290 and 382-384), an FR2 of VL (SEQ ID NOS:291, 292 and 385-392), and an FR3 of VL (SEQ ID NOS:293, 294, 295 and 393-404); an FR2 of VH (SEQ ID NOS:281, 282, and 283), an FR3 of VH (SEQ ID NOS:284, 285, 286, 287 and 379-381), an FR1 of VL (SEQ ID NOS:289, 290 and 382-384), an FR2 of VL (SEQ ID NOS:291, 292 and 385-392), and an FR4 of VL (SEQ ID NO:296 and 405-407); an FR2 of VH (SEQ ID NOS:281, 282, and 283), an FR3 of VH (SEQ ID NOS:284, 285, 286, 287 and 379-381), an FR1 of VL (SEQ ID NOS:289, 290 and 382-384), an FR3 of VL (SEQ ID NOS:293, 294, 295 and 393-404), and an FR4 of VL (SEQ ID NO:296 and 405-407); an FR2 of VH (SEQ ID NOS:281, 282, and 283), an FR3 of VH (SEQ ID NOS:284, 285, 286, 287 and 379-381), an FR2 of VL (SEQ ID NOS:291, 292 and 385-392), an FR3 of VL (SEQ ID NOS:293, 294, 295 and 393-404), and an FR4 of VL (SEQ ID NO:296 and 405-407); an FR2 of VH (SEQ ID NOS:281, 282, and 283), an FR4 of VH (SEQ ID NO:288), an FR1 of VL (SEQ ID NOS:289, 290 and 382-384), an FR2 of VL (SEQ ID NOS:291, 292 and 385-392), and an FR3 of VL (SEQ ID NOS:293, 294, 295 and 393-404); an FR2 of VH (SEQ ID NOS:281, 282, and 283), an FR4 of VH (SEQ ID NO:288), an FR1 of VL (SEQ ID NOS:289, 290 and 382-384), an FR2 of VL (SEQ ID NOS:291, 292 and 385-392), and an FR4 of VL (SEQ ID NO:296 and 405-407); an FR2 of VH (SEQ ID NOS:281, 282, and 283), an FR4 of VH (SEQ ID NO:288), an FR1 of VL (SEQ ID NOS:289, 290 and 382-384), an FR3 of VL (SEQ ID NOS:293, 294, 295 and 393-404), and an FR4 of VL (SEQ ID NO:296 and 405-407); an FR2 of VH (SEQ ID NOS:281, 282, and 283), an FR4 of VH (SEQ ID NO:288), an FR2 of VL (SEQ ID NOS:291, 292 and 385-392), an FR3 of VL (SEQ ID NOS:293, 294, 295 and 393-404), and an FR4 of VL (SEQ ID NO:296 and 405-407); an FR3 of VH (SEQ ID NOS:284, 285, 286, 287 and 379381), an FR4 of VH (SEQ ID NO:288), an FR1 of VL (SEQ ID NOS:289, 290 and 382-384), an FR2 of VL (SEQ ID NOS:291, 292 and 385-392), and an FR3 of VL (SEQ ID NOS:293, 294, 295 and 393-404); an FR3 of VH (SEQ ID NOS:284, 285, 286, 287 and 379-381), an FR4 of VH (SEQ ID NO:288), an FR1 of VL (SEQ ID NOS:289, 290 and 382-384 ), an FR2 of VL (SEQ ID NOS:291, 292 and 385-392), and an FR4 of VL (SEQ ID NO:296 and 405-407); an FR3 of VH (SEQ ID NOS:284, 285, 286, 287 and 379 381), an FR4 of VH (SEQ ID NO:288), an FR1 of VL (SEQ ID NOS:289, 290 and 382-384) , an FR3 of VL (SEQ ID NOS:293, 294, 295 and 393-404), and an FR4 of VL (SEQ ID NO:296 and 405-407); an FR3 of VH (SEQ ID NOS:284, 285, 286, 287 and 379-381), an FR4 of VH (SEQ ID NO:288), an FR2 of VL (SEQ ID NOS:291, 292 and 385-392 ), an FR3 of VL (SEQ ID NOS:293, 294, 295 and 393-404), and an FR4 of VL (SEQ ID NO:296 and 405-407); an FR1 of VH (SEQ ID NOS:278, 279, 280 and 378), an FR1 of VL (SEQ ID NOS:289, 290 and 382-384), an FR2 of VL (SEQ ID NOS:291, 292 and 385 -392), an FR3 of VL (SEQ ID NOS:293, 294, 295 and 393-404), and an FR4 of VL (SEQ ID NO:296 and 405-407); an FR2 of VH (SEQ ID NOS:281, 282, and 283), an FR1 of VL (SEQ ID NOS:289, 290 and 382-384), an FR2 of VL (SEQ ID NOS:291, 292 and 385- 392), an FR3 of VL (SEQ ID NOS:293, 294, 295 and 393-404), and an FR4 of VL (SEQ ID NO:296 and 405-407); an FR3 of VH (SEQ ID NOS:284, 285, 286, 287 and 379-381), an FR1 of VL (SEQ ID NOS:289, 290 and 382-384), an FR2 of VL (SEQ ID NOS:291 , 292 and 385-392), an FR3 of VL (SEQ ID NOS:293, 294, 295 and 393-404), and an FR4 of VL (SEQ ID NO:296 and 405-407); an FR4 of VH (SEQ ID NO:288), an FR1 of VL (SEQ ID NOS:289, 290 and 382-384), an FR2 of VL (SEQ ID NOS:291, 292 and 385-392), an FR3 of VL (SEQ ID NOS:293, 294, 295 and 393 404), and an FR4 of VL (SEQ ID NO:296 and 405-407); a VH FR1 (SEQ ID NOS:278, 279, 280 and 378), a VH FR2 (SEQ ID NOS:281, 282, and 283), a VH FR3 (SEQ ID NOS:284, 285, 286, 287 and 379 381), an FR4 of VH (SEQ ID NO:288), FR1 of VL (SEQ ID NOS:289, 290 and 382-384) and an FR2 of VL (SEQ ID NOS:291, 292 and 385- 392); a VH FR1 (SEQ ID NOS:278, 279, 280 and 378), a VH FR2 (SEQ ID NOS:281, 282, and 283), a VH FR3 (SEQ ID NOS:284, 285, 286, 287 and 379-381), an FR4 of VH (SEQ ID NO:288), FR1 of VL (SEQ ID NOS:289, 290 and 382-384) and an FR3 of VL (SEQ ID NOS:293, 294, 295 and 393-404); a VH FR1 (SEQ ID NOS:278, 279, 280 and 378), a VH FR2 (SEQ ID NOS:281, 282, and 283), a VH FR3 (SEQ ID NOS:284, 285, 286, 287 and 379-381), an FR4 of VH (SEQ ID NO:288), FR1 of VL (SEQ ID NOS:289, 290 and 382-384) and an FR4 of VL (SEQ ID NO:296 and 405-407 ); a VH FR1 (SEQ ID NOS:278, 279, 280 and 378), a VH FR2 (SEQ ID NOS:281, 282, and 283), a VH FR3 (SEQ ID NOS:284, 285, 286, 287 and 379-381), an FR4 of VH (SEQ ID NO:288), FR2 of VL (SEQ ID NOS:291, 292 and 385-392) and an FR3 of VL (SEQ ID NOS:293, 294, 295 and 393-404); a VH FR1 (SEQ ID NOS:278, 279, 280 and 378), a VH FR2 (SEQ ID NOS:281, 282, and 283), a VH FR3 (SEQ ID NOS:284, 285, 286, 287 and 379-381), an FR4 of VH (SEQ ID NO:288), FR2 of VL (SEQ ID NOS:291, 292 and 385-392) and an FR4 of VL (SEQ ID NO:296 and 405-407 ); a VH FR1 (SEQ ID NOS:278, 279, 280 and 378), a VH FR2 (SEQ ID NOS:281, 282, and 283), a VH FR3 (SEQ ID NOS:284, 285, 286, 287 and 379-381), an FR4 of VH (SEQ ID NO:288), FR3 of VL (SEQ ID NOS:293, 294, 295 and 393 404) and an FR4 of VL (SEQ ID NO:296 and 405- 407); a VH FR1 (SEQ ID NOS:278, 279, 280 and 378), a VH FR2 (SEQ ID NOS:281, 282, and 283), a VH FR3 (SEQ ID NOS:284, 285, 286, 287 and 379-381), an FR1 of VL (SEQ ID NOS:289, 290 and 382-384), an FR2 of VL (SEQ ID NOS:291, 292 and 385-392) and an FR3 of VL (SEQ ID NOS :293, 294, 295 and 393-404); a VH FR1 (SEQ ID NOS:278, 279, 280 and 378), a VH FR2 (SEQ ID NOS:281, 282, and 283), a VH FR3 (SEQ ID NOS:284, 285, 286, 287 and 379-381), an FR1 of VL (SEQ ID NOS:289, 290 and 382-384), an FR2 of VL (SEQ ID NOS:291, 292 and 385-392) and an FR4 of VL (SEQ ID NO :296 and 405-407); a VH FR1 (SEQ ID NOS:278, 279, 280 and 378), a VH FR2 (SEQ ID NOS:281, 282, and 283), a VH FR3 (SEQ ID NOS:284, 285, 286, 287 and 379-381), an FR1 of VL (SEQ ID NOS:289, 290 and 382-384), an FR3 of VL (SEQ ID NOS:293, 294, 295 and 393-404) and an FR4 of VL (SEQ ID NO:296 and 405-407); a VH FR1 (SEQ ID NOS:278, 279, 280 and 378), a VH FR2 (SEQ ID NOS:281, 282, and 283), a VH FR3 (SEQ ID NOS:284, 285, 286, 287 and 379-381), an FR2 of VL (SEQ ID NOS:291, 292 and 385-392), an FR3 of VL (SEQ ID NOS:293, 294, 295 and 393-404) and an FR4 of VL (SEQ ID NO:296 and 405-407); a VH FR1 (SEQ ID NOS:278, 279, 280 and 378), a VH FR2 (SEQ ID NOS:281, 282, and 283), a VH FR3 (SEQ ID NOS:284, 285, 286, 287 and 379-381), an FR1 of VL (SEQ ID NOS:289, 290 and 382-384), an FR2 of VL (SEQ ID NOS:291, 292 and 385-392) and an FR3 of VL (SEQ ID NOS :293, 294, 295 and 393-404); an FR1 of VH (SEQ ID NOS:278, 279, 280 and 378), an FR2 of VH (SEQ ID NOS:281, 282, and 283), an FR4 of VH (SEQ ID NO:288), an FR1 of VL (SEQ ID NOS:289, 290 and 382-384), FR2 of VL (SEQ ID NOS:291, 292 and 385-392) and an FR4 of VL (SEQ ID NO:296 and 405-407); an FR1 of VH (SEQ ID NOS:278, 279, 280 and 378), an FR2 of VH (SEQ ID NOS:281, 282, and 283), an FR4 of VH (SEQ ID NO:288), an FR1 of VL (SEQ ID NOS:289, 290 and 382-384), FR3 of VL (SEQ ID NOS:293, 294, 295 and 393-404) and an FR4 of VL (SEQ ID NO:296 and 405-407); an FR1 of VH (SEQ ID NOS:278, 279, 280 and 378), an FR2 of VH (SEQ ID NOS:281, 282, and 283), an FR4 of VH (SEQ ID NO:288), an FR2 of VL (SEQ ID NOS:291, 292 and 385-392), FR3 of VL (SEQ ID NOS:293, 294, 295 and 393-404) and an FR4 of VL (SEQ ID NO:296 and 405-407); an FR1 of VH (SEQ ID NOS:278, 279, 280 and 378), an FR3 of VH (SEQ ID NOS:284, 285, 286, 287 and 379-381), an FR4 of VH (SEQ ID NO:288 ), an FR1 of VL (SEQ ID NOS:289, 290 and 382-384), an FR2 of VL (SEQ ID NOS:291, 292 and 385-392) and an FR3 of VL (SEQ ID NOS:293, 294, 295 and 393-404); an FR1 of VH (SEQ ID NOS:278, 279, 280 and 378), an FR3 of VH (SEQ ID NOS:284, 285, 286, 287 and 379-381), an FR4 of VH (SEQ ID NO:288 ), an FR1 of VL (SEQ ID NOS:289, 290 and 382-384), an FR2 of VL (SEQ ID NOS:291, 292 and 385-392) and an FR4 of VL (SEQ ID NO:296 and 405- 407); an FR1 of VH (SEQ ID NOS:278, 279, 280 and 378), an FR3 of VH (SEQ ID NOS:284, 285, 286, 287 and 379-381), an FR4 of VH (SEQ ID NO:288 ), an FR1 of VL (SEQ ID NOS:289, 290 and 382-384), an FR3 of VL (SEQ ID NOS:293, 294, 295 and 393-404) and an FR4 of VL (SEQ ID NO:296 and 405-407); an FR1 of VH (SEQ ID NOS:278, 279, 280 and 378), an FR3 of VH (SEQ ID NOS:284, 285, 286, 287 and 379 381), an FR4 of VH (SEQ ID NO:288) , an FR2 of VL (SEQ ID NOS:291, 292 and 385-392), an FR3 of VL (SEQ ID NOS:293, 294, 295 and 393-404) and an FR4 of VL (SEQ ID NO:296 and 405 -407); an FR2 of VH (SEQ ID NOS:281, 282, and 283), an FR3 of VH (SEQ ID NOS:284, 285, 286, 287 and 379-381), an FR4 of VH (SEQ ID NO:288) , an FR1 of VL (SEQ ID NOS:289, 290 and 382-384), an FR2 of VL (SEQ ID NOS:291, 292 and 385-392) and an FR3 of VL (SEQ ID NOS:293, 294, 295 and 393-404); an FR2 of VH (SEQ ID NOS:281, 282, and 283), an FR3 of VH (SEQ ID NOS:284, 285, 286, 287 and 379-381), an FR4 of VH (SEQ ID NO:288) , an FR1 of VL (SEQ ID NOS:289, 290 and 382-384), an FR2 of VL (SEQ ID NOS:291, 292 and 385-392) and an FR4 of VL (SEQ ID NO:296 and 405-407 ); an FR2 of VH (SEQ ID NOS:281, 282, and 283), an FR3 of VH (SEQ ID NOS:284, 285, 286, 287 and 379-381), an FR4 of VH (SEQ ID NO:288) , an FR1 of VL (SEQ ID NOS:289, 290 and 382-384), an FR3 of VL (SEQ ID NOS:293, 294, 295 and 393-404) and an FR4 of VL (SEQ ID NO:296 and 405 -407); an FR2 of VH (SEQ ID NOS:281, 282, and 283), an FR3 of VH (SEQ ID NOS:284, 285, 286, 287 and 379-381), an FR4 of VH (SEQ ID NO:288) , an FR2 of VL (SEQ ID NOS:291, 292 and 385-392), an FR3 of VL (SEQ ID NOS:293, 294, 295 and 393-404) and an FR4 of VL (SEQ ID NO:296 and 405 -407); an FR1 of VH (SEQ ID NOS:278, 279, 280 and 378), an FR2 of VH (SEQ ID NOS:281, 282, and 283), an FR1 of VL (SEQ ID NOS:289, 290 and 382- 384), an FR2 of VL (SEQ ID NOS:291, 292 and 385-392), an FR3 of VL (SEQ ID NOS:293, 294, 295 and 393-404) and an FR4 of VL (SEQ ID NO:296 and 405-407); an FR1 of VH (SEQ ID NOS:278, 279, 280 and 378), an FR3 of VH (SEQ ID NOS:284, 285, 286, 287 and 379-381), an FR1 of VL (SEQ ID NOS:289 , 290 and 382-384), an FR2 of VL (SEQ ID NOS:291, 292 and 385-392), an FR3 of VL (SEQ ID NOS:293, 294, 295 and 393-404) and an FR4 of VL ( SEQ ID NO:296 and 405-407); an FR1 of VH (SEQ ID NOS:278, 279, 280 and 378), an FR4 of VH (SEQ ID NO:288), an FR1 of VL (SEQ ID NOS:289, 290 and 382-384), an FR2 of VL (SEQ ID NOS:291, 292 and 385-392), FR3 of VL (SEQ ID NOS:293, 294, 295 and 393-404) and an FR4 of VL (SEQ ID NO:296 and 405-407) ; an FR2 of VH (SEQ ID NOS:281, 282, and 283), an FR3 of VH (SEQ ID NOS:284, 285, 286, 287 and 379-381), an FR1 of VL (SEQ ID NOS:289, 290 and 382-384), an FR2 of VL (SEQ ID NOS:291, 292 and 385-392), an FR3 of VL (SEQ ID NOS:293, 294, 295 and 393-404) and an FR4 of VL (SEQ ID NO:296 and 405-407); an FR2 of VH (SEQ ID NOS:281, 282, and 283), an FR4 of VH (SEQ ID NO:288), an FR1 of VL (SEQ ID NOS:289, 290 and 382-384), an FR2 of VL (SEQ ID NOS:291, 292 and 385-392), FR3 of VL (SEQ ID NOS:293, 294, 295 and 393-404) and an FR4 of VL (SEQ ID NO:296 and 405-407); an FR3 of VH (SEQ ID NOS:284, 285, 286, 287 and 379-381), an FR4 of VH (SEQ ID NO:288), an FR1 of VL (SEQ ID NOS:289, 290 and 382-384 ), an FR2 of VL (SEQ ID NOS:291, 292 and 385-392), an FR3 of VL (SEQ ID NOS:293, 294, 295 and 393-404) and an FR4 of VL (SEQ ID NO:296 and 405-407); a VH FR1 (SEQ ID NOS:278, 279, 280 and 378), a VH FR2 (SEQ ID NOS:281, 282, and 283), a VH FR3 (SEQ ID NOS:284, 285, 286, 287 and 379 381), an FR4 of VH (SEQ ID NO:288), FR1 of VL (SEQ ID NOS:289, 290 and 382-384), FR2 of VL (SEQ ID NOS:291, 292 and 385-392 ) and an FR3 of VL (SEQ ID NOS:293, 294, 295 and 393-404); a VH FR1 (SEQ ID NOS:278, 279, 280 and 378), a VH FR2 (SEQ ID NOS:281, 282, and 283), a VH FR3 (SEQ ID NOS:284, 285, 286, 287 and 379 381), an FR4 of VH (SEQ ID NO:288), FR1 of VL (SEQ ID NOS:289, 290 and 382-384), FR2 of VL (SEQ ID NOS:291, 292 and 385-392 ) and an FR4 of VL (SEQ ID NO:296 and 405-407); a VH FR1 (SEQ ID NOS:278, 279, 280 and 378), a VH FR2 (SEQ ID NOS:281, 282, and 283), a VH FR3 (SEQ ID NOS:284, 285, 286, 287 and 379-381), an FR4 of VH (SEQ ID NO:288), FR2 of VL (SEQ ID NOS:291, 292 and 385-392), FR3 of VL (SEQ ID NOS:293, 294, 295 and 393-404) and an FR4 of VL (SEQ ID NO:296 and 405-407); a VH FR1 (SEQ ID NOS:278, 279, 280 and 378), a VH FR2 (SEQ ID NOS:281, 282, and 283), a VH FR3 (SEQ ID NOS:284, 285, 286, 287 and 379-381), an FR1 of VL (SEQ ID NOS:289, 290 and 382-384), an FR2 of VL (SEQ ID NOS:291, 292 and 385-392), FR3 of VL (SEQ ID NOS: 293, 294, 295 and 393-404) and an FR4 of VL (SEQ ID NO:296 and 405-407); an FR1 of VH (SEQ ID NOS:278, 279, 280 and 378), an FR2 of VH (SEQ ID NOS:281, 282, and 283), an FR4 of VH (SEQ ID NO:288), an FR1 of VL (SEQ ID NOS:289, 290 and 382-384), FR2 of VL (SEQ ID NOS:291, 292 and 385-392), FR3 of VL (SEQ ID NOS:293, 294, 295 and 393-404) and an FR4 of VL (SEQ ID NO:296 and 405-407); an FR2 of VH (SEQ ID NOS:281, 282, and 283), an FR3 of VH (SEQ ID NOS:284, 285, 286, 287 and 379-381), an FR4 of VH (SEQ ID NO:288) , an FR1 of VL (SEQ ID NOS:289, 290 and 382-384), FR2 of VL (SEQ ID NOS:291, 292 and 385-392), FR3 of VL (SEQ ID NOS:293, 294, 295 and 393-404) and an FR4 of VL (SEQ ID NO:296 and 405-407); or any combination thereof of VH FRs (SEQ ID NOS: 278, 279, 280, 378, 281, 282, 283, 284, 285, 286, 287, 379-381 and 288) and VL FRs (SEQ ID NOS : 289, 290, 382-384, 291, 292, 385-392, 293, 294, 295, 393-404, 296, 405-407) listed in Table 19.

[0215] In yet another aspect, antibodies are provided to compete with one of the exemplified antibodies or functional fragments for binding to (i) Klotho beta or (ii) a complex comprising Klotho beta and one of FGFR1c, FGFR2c, FGFR3c, and FGFR4. Such antibodies may also bind to the same epitope as one of the antibodies exemplified herein, or an overlapping epitope. Antibodies and fragments that compete with or bind to the same epitope as the exemplified antibodies are expected to show similar functional properties. Exemplified antigen-binding proteins and fragments include those with the VH and VL regions, and CDRs provided herein, including those in Tables 1-10. Thus, as a specific example, antibodies that are provided include those that compete with an antibody that comprises: (a) 1, 2, 3, 4, 5 or all 6 CDRs listed for an antibody listed in Tables 1-10; (b) a VH and a VL selected from VH and VL regions listed for an antibody listed in Tables 1-10, such as for antibody 5H23 (Table 1), or (c) two light chains and two heavy chains comprising a VH and a VL as specified for an antibody listed in Tables 1-10.

[0216] In yet another aspect, antibodies are provided herein that bind to a region, including an epitope, of human Klotho beta or Cyno Klotho beta. For example, in some embodiments, an antibody provided herein binds to a KLB2 domain of human Klotho beta comprising amino acid residues 509-1044 of SEQ ID NO: 297. As another example, in some embodiments, an antibody provided herein binds to a glycosyl hydrolase 1 region of a KLB2 domain of human Klotho beta comprising amino acid residues 517967 of SEQ ID NO: 297. As yet another example, in some embodiments, an antibody provided herein binds to a region of human Klotho beta comprising amino acid residues 657-703 of SEQ ID NO: 297. As yet another example, in some embodiments, an antibody provided herein binds to a Klotho beta region of cyno comprising amino acid residues 657-703 of SEQ ID NO: 299.

[0217] In another aspect, antibodies are provided here that bind to a specific epitope of human Klotho beta. For example, in some embodiments, an antibody provided herein binds to a human Klotho beta epitope comprising at least one of amino acid residues 657, 701 and/or 703 of human Klotho beta (SEQ ID NO: 297). Therefore, in some embodiments, an antibody provided herein binds to a human Klotho beta epitope, wherein the human Klotho beta epitope comprises at least amino acid residue 657 of SEQ ID NO: 297. In some embodiments, an antibody provided herein binds to a human Klotho beta epitope, wherein the human Klotho beta epitope comprises at least amino acid residue 701 of SEQ ID NO: 297. In some embodiments, an antibody provided herein binds to a Klotho epitope. human beta, wherein the human Klotho beta epitope comprises at least amino acid residue 703 of SEQ ID NO: 297. In some embodiments, an antibody provided herein binds to a human Klotho beta epitope, wherein the Klotho epitope human beta comprises at least amino acid residues 657 and 701 of SEQ ID NO: 297. In some embodiments, an antibody provided herein binds to a human Klotho beta epitope, wherein the human Klotho beta epitope comprises at least the residues amino acid residues 657 and 703 of SEQ ID NO: 297. In some embodiments, an antibody provided herein binds to a human Klotho beta epitope, wherein the human Klotho beta epitope comprises at least amino acid residues 701 and 703 of SEQ ID NO: 297. In some embodiments, an antibody provided herein binds to a human Klotho beta epitope, wherein the human Klotho beta epitope comprises at least amino acid residues 657, 701 and 703 of SEQ ID NO: 297. Such antibodies provided above can, in some embodiments, induce FGF19-like signaling and/or FGF21-like signaling in a cell expressing human Klotho beta and an FGF receptor. Furthermore, in some embodiments, the antibody is a humanized, human or chimeric antibody.

1. Polyclonal Antibodies

[0218] The antibodies of the present invention may comprise polyclonal antibodies. Methods of preparing polyclonal antibodies are known to those skilled in the art. Polyclonal antibodies can be raised in a mammal, for example, by one or more injections of an immunizing agent and, if desired, an adjuvant. Typically, the immunizing agent and/or adjuvant will be injected into the mammal through multiple subcutaneous or intraperitoneal injections. The immunizing agent may include a Klotho beta polypeptide or a fusion protein thereof. It may be useful to conjugate the immunizing agent with a protein that is known to be immunogenic in the mammal to be immunized or to immunize the mammal with the protein and one or more adjuvants. Examples of such immunogenic proteins include, but are not limited to, keyhole limpet hemocyanin, serum albumin, bovine thyroglobulin and soybean trypsin inhibitor. Examples of adjuvants that can be used include Ribi, CpG, poly 1C, complete Freund's adjuvant and MPL-TDM adjuvant (monophosphoryl-Lipid A, synthetic trehalose dicorynomycolate). The immunization protocol can be selected by one skilled in the art without undue experimentation. The mammal can then be bled, and the serum assayed for Klotho beta antibody titer. If desired, the mammal can be boosted until the antibody titer increases or reaches a plateau. Additionally or alternatively, lymphocytes can be obtained from the immunized animal for fusion and preparation of monoclonal antibodies from the hybridoma as described below.

2. Monoclonal Antibodies

[0219] The antibodies of the present invention may, alternatively, be monoclonal antibodies. Monoclonal antibodies can be produced using the hybridoma method first described by Kohler et al, Nature, 256:495 (1975), or can be produced by recombinant DNA methods (see, for example, US Patent No. 4,816,567).

[0220] In the hybridoma method, a mouse or other appropriate host animal, such as a hamster, is immunized as described above to elicit lymphocytes that produce or are capable of producing antibodies that will specifically bind to the protein used for immunization. Alternatively, lymphocytes can be immunized in vitro. After immunization, lymphocytes are isolated and then fused with a myeloma cell line using a suitable fusing agent, such as polyethylene glycol, to form a hybridoma cell (Goding, Monoclonal Antibodies: Principles and Practice, pp.59-103 (Goding, Monoclonal Antibodies: Principles and Practice, pp.59-103). Academic Press, 1986)).

[0221] The hybridoma cells thus prepared are seeded and grown in a suitable culture medium that preferably contains one or more substances that inhibit the growth or survival of unfused parental myeloma cells (also referred to as fusion partners). For example, if the progenitor myeloma cells lack the enzyme hypoxanthine guanine phosphoribosyl transferase (HGPRT or HPRT), the selective culture medium for hybridomas will typically include hypoxanthine, aminopterin, and thymidine (HAT medium), the substances of which prevent the growth of HGPRT-deficient cells.

[0222] Preferred fusion partner myeloma cells are those that fuse efficiently, support stable high-level antibody production by the selected antibody-producing cells, and are sensitive to a selective medium that selects against the unfused parental cells. Preferred myeloma cell lines are murine myeloma lines such as SP-2 and derivatives, e.g., X63-Ag8-653 cells available from the American Type Culture Collection, Manassas, Virginia, USA, and those derived from tumors of Mouse MOPC-21 and MPC-11 available from the Salk Institute Cell Distribution Center, San Diego, California, USA. Human myeloma and mouse-human heteromyeloma cell lines have also been described for the production of human monoclonal antibodies (Kozbor, J., Immunol., 133:3001 (1984); and Brodeur et al., Monoclonal Antibody Production Techniques and Applications , pp. 51-63 (Marcel Dekker, Inc., New York, 1987)).

[0223] The culture medium in which the hybridoma cells are growing is tested for the production of monoclonal antibodies directed against the antigen. The binding specificity of monoclonal antibodies produced by hybridoma cells is determined by immunoprecipitation or by an in vitro binding assay, such as radioimmunoassay (RIA) or an enzyme-linked immunosorbent assay (ELISA). The binding affinity of the monoclonal antibody can, for example, be determined by the Scatchard analysis described in Munson et al., Anal. Biochem., 107:220 (1980).

[0224] Once hybridoma cells that produce antibodies with the desired specificity, affinity and/or activity are identified, clones can be subcloned by limiting dilution procedures and grown by standard methods (Goding, Monoclonal Antibodies: Principles and Practice , pp.59-103 (Academic Press, 1986)). Suitable culture media for this purpose include, for example, D-MEM medium or RPMI-1640 medium. Furthermore, hybridoma cells can be grown in vivo as ascites tumors in an animal, for example, by i.p. injection. of cells in mice.

[0225] The monoclonal antibodies secreted by the subclones are suitably separated from the culture medium, ascites fluid, or serum by conventional antibody purification procedures, such as, for example, affinity chromatography (e.g., using protein A or protein G -Sepharose) or ion exchange chromatography, hydroxylapatite chromatography, gel electrophoresis, dialysis, etc.

[0226] DNA encoding monoclonal antibodies is readily isolated and sequenced using conventional procedures (for example, using oligonucleotide probes that are capable of specifically binding to genes encoding the heavy and light chains of murine antibodies). Hybridoma cells serve as a preferred source of such DNA. Once isolated, the DNA can be placed into expression vectors, which are then transfected into host cells such as E. coli cells, simian COS cells, Chinese Hamster Ovary (CHO) cells, or myeloma cells that otherwise do not produce antibody protein, to obtain the synthesis of monoclonal antibodies in recombinant host cells. Review articles on recombinant expression in bacteria of antibody-encoding DNA include Skerra et al., Curr. Opinion in Immunol., 5:256-262 (1993) and Plückthun, Immunol. Revs. 130:151-188 (1992).

[0227] In some embodiments, an antibody that binds to a Klotho beta epitope comprises an amino acid sequence of a VH domain and/or an amino acid sequence of a VL domain encoded by a nucleotide sequence that hybridizes to ( 1) the complement of a nucleotide sequence encoding any of VH and/or VL described herein under stringent conditions (e.g., hybridization to filter-bound DNA in sodium chloride/sodium citrate (SSC) 6x at about 45 °C, followed by one or more washes in 0.2xSSC/0.1% SDS at about 50-65°C) under highly stringent conditions (e.g., filter-bound nucleic acid hybridization in 6xSSC at about of 45°C, followed by one or more washes in 0.1xSSC/0.2% SDS at about 68°C), or under other stringent hybridization conditions that are known to those skilled in the art (see, e.g., Ausubel , F.M. et al., eds., 1989, Current Protocols in Molecular Biology, Vol. I, Green Publishing Associates, Inc. and John Wiley & Sons, Inc., New York on pages 6.3.1-6.3.6 and 2.10. 3).

[0228] In some embodiments, an antibody that binds to a Klotho beta epitope comprises an amino acid sequence of a VH CDR or an amino acid sequence of a VL CDR encoded by a nucleotide sequence that hybridizes to the complement of a nucleotide sequence encoding any of the VH CDR and/or VL CDR represented in Tables 1-10 under stringent conditions (e.g., hybridization of filter-bound DNA in 6X SSC at about 45°C, followed by one or plus washes in 0.2X SSC/0.1% SDS at about 50-65°C), under highly stringent conditions (e.g., filter-bound nucleic acid hybridization in 6X SSC at about 45°C, followed by one or more washes in 0.1X SSC/0.2% SDS at about 68°C), or under other stringent hybridization conditions that are known to those skilled in the art (see, for example, Ausubel, F.M. et al., eds., 1989, Current Protocols in Molecular Biology, Vol. I, Green Publishing Associates, Inc. and John Wiley & Sons, Inc., New York at pages 6.3.1-6.3.6 and 2.10.3)

[0229] In another embodiment, monoclonal antibodies or antibody fragments can be isolated from antibody phage libraries generated using the techniques described in, for example, Antibody Phage Display: Methods and Protocols, P.M. O'Brien and R. Aitken, eds, Humana Press, Totawa N.J., 2002. In principle, synthetic antibody clones are selected by screening phage libraries containing phage that displays various antibody variable region (Fv) fragments fused to the phage coat protein . Such phage libraries are screened against the desired antigen. Clones expressing Fv fragments capable of binding the desired antigen are adsorbed to the antigen and then separated from the non-binding clones in the library. Binding clones are then eluted from the antigen, and can be further enriched by additional cycles of antigen adsorption/elution.

[0230] The variable domains can be functionally displayed on the phage, either as single-chain Fv fragments (scFv), in which VH and VL are covalently linked via a short, flexible peptide, or as Fab fragments, in which they are, each, fused to a constant domain and interact non-covalently, as described, for example, in Winter et al., Ann. Rev. Immunol., 12: 433-455 (1994).

[0231] The VH and VL gene repertoires can be cloned separately by polymerase chain reaction (PCR) and randomly recombined into phage libraries, which can then be screened for antigen-binding clones as described in Winter et al. al., supra. Immunized source libraries provide high affinity antibodies to the immunogen without the requirement to construct hybridomas. Alternatively, the naive repertoire can be cloned to provide a single source of human antibodies to a wide range of non-self and also self-antigens without any immunization as described by Griffiths et al, EMBO J., 12:725-734 ( 1993). Finally, naïve libraries can also be produced synthetically by cloning the non-rearranged V gene segments from stem cells, and using PCR primers containing the random sequence to encode the highly variable CDR3 regions and to carry out the rearrangement in vitro as described. , for example, by Hoogenboom and Winter, J. Mol. Biol., 227: 381-388 (1992).

[0232] Library screening can be carried out by various techniques known in the art. For example, Klotho beta, (e.g., a Klotho beta polypeptide, fragment, or epitope) can be used to coat the wells of adsorption plates, expressed in host cells attached to adsorption plates or used in cell separation, or conjugated with biotin for capture with streptavidin-coated beads, or used in any other method to mine display libraries. Selection of antibodies with slow dissociation kinetics (e.g., good binding affinities) can be promoted by the use of long washes and monovalent phage display as described in Bass et al., Proteins, 8: 309-314 (1990) and in WO 92/09690, and a low coating density of antigen as described in Marks et al, Biotechnol., 10:779-783 (1992).

[0233] Anti-Klotho beta antibodies can be obtained by devising a suitable antigen screening procedure to select the phage clone of interest followed by construction of a full-length anti-Klotho beta antibody clone using VH sequences and/or VL (e.g., Fv sequences), or various CDR sequences from VH and VL sequences, from the phage clone of interest and the appropriate constant region sequences (e.g., Fc) described in Kabat et al. al., Sequences of Proteins of Immunological Interest, Fifth Edition, publication NIH 91-3242, Bethesda MD (1991), vols. 1-3.

3. Antibody Fragments

[0234] The present invention provides antibodies and antibody fragments that bind to Klotho beta. In certain circumstances, there are advantages to using antibody fragments rather than whole antibodies. The smaller size of the fragments allows for rapid clearance and may lead to better access to cells, tissues or organs. For a review of certain antibody fragments, see Hudson et al. (2003) Nat Med 9:129-134.

[0235] Several techniques have been developed for the production of antibody fragments. Traditionally, these fragments were derived via proteolytic digestion of intact antibodies (see, for example, Morimoto et al., Journal of Biochemical and Biophysical Methods 24:107-117 (1992); and Brennan et al., Science, 229:81 (1985 )). However, these fragments can now be produced directly by recombinant host cells. The Fab, Fv, and ScFv antibody fragments can all be expressed in, and secreted by, E. coli or yeast cells, thus allowing the facile production of large quantities of these fragments. Antibody fragments can be isolated from the antibody phage libraries discussed above. Alternatively, Fab'-SH fragments can be directly recovered from E. coli and chemically coupled to form F(ab')2 fragments (Cárter et al, Bio/Technology 10:163-167 (1992)). According to another approach, F(ab')2 fragments can be isolated directly from recombinant host cell cultures. Fab and F(ab')2 fragments with increased in vivo half-life comprising salvage receptor-binding epitope residues are described, for example, in the US Patent. No. 5,869,046. Other techniques for producing antibody fragments will be apparent to one skilled in the art. In certain embodiments, an antibody is a single-chain Fv fragment (scFv) (see, for example, WO 93/16185; US Patent Nos. 5,571,894; and 5,587,458). Fv and scFv have intact combining sites that are devoid of constant regions; thus, they may be suitable for reducing nonspecific binding during in vivo use. scFv fusion proteins can be constructed in order to obtain the fusion of an effector protein at either the amino or carboxy terminus of a scFv. (See, for example, Antibody Engineering, ed. Borrebaeck, supra). The antibody fragment may also be a "linear antibody", for example, as described, for example, in the references cited above. Such linear antibodies may be monospecific or multispecific, such as bispecific.

[0236] Smaller antibody-derived binding structures are separate variable domains (V domains) also called single variable domain antibodies (SdAbs). Certain types of organisms, the camelids and cartilaginous fish, have unique high-affinity V-like domains assembled on an equivalent Fc domain structure as part of their immune system. (Woolven et al, Immunogenetics 50:98-101, 1999; Streltsov et al, Proc Natl Acad Sci USA 101:12444-12449, 2004.). V-like domains (called VhH in camelids and V-NAR in sharks) often exhibit long surface loops, which allow penetration of target antigen cavities. They also stabilize isolated VH domains by masking hydrophobic surface patches.

[0237] These VhH and V-NAR domains were used to design sdAbs. Human V domain variants have been engineered using phage library selection and other approaches that have resulted in stable, high-VL and high-binding VH-derived domains.

[0238] Antibodies that bind to Klotho beta as provided herein include, but are not limited to, synthetic antibodies, monoclonal antibodies, recombinantly produced antibodies, multispecific antibodies (including bispecific antibodies), human antibodies, humanized antibodies, camelized antibodies, chimeric antibodies, intrabodies, anti-idiotypic antibodies (anti-Id), and functional fragments, (e.g., Klotho beta-binding fragments) of any of the above. Non-limiting examples of functional fragments (e.g., fragments that bind to Klotho beta) include single-chain Fvs (scFv) (e.g., including monospecific, bispecific, etc.), Fab fragments, F(ab') fragments, F (ab)2, F(ab')2 fragments, disulfide-linked Fvs (sdFv), Fd fragments, Fv fragments, diabody, triabody, tetrabody and minibody.

[0239] Antibodies provided herein include, but are not limited to, immunoglobulin molecules and immunologically active portions of immunoglobulin molecules, for example, molecules containing an antigen binding site that bind to a Klotho beta epitope. The immunoglobulin molecules provided herein may be of any type (e.g., IgG, IgE, IgM, IgD, IgA and IgY), class (e.g., IgG1, IgG2, IgG3, IgG4, IgA1 and IgA2) or subclass of immunoglobulin molecule. immunoglobulin.

[0240] Antibody variants and derivatives include functional antibody fragments that retain the ability to bind to a Klotho beta epitope. Exemplary functional fragments include Fab fragments (e.g., an antibody fragment that contains the antigen-binding domain and comprises a light chain and part of a heavy chain linked by a disulfide bond); Fab' (e.g., an antibody fragment containing a single anti-binding domain comprising a Fab and an additional portion of the heavy chain through the hinge region); F(ab')2 (for example, two Fab' molecules joined by interchain disulfide bonds in the hinge regions of the heavy chains; Fab' molecules can be directed to the same or different epitopes); a bispecific Fab (e.g., a Fab molecule with two antigen-binding domains, each of which can be targeted to a different epitope); a single-chain Fab chain comprising a variable region, also known as, an sFv (e.g., the variable antigen-binding determining region of a single light chain and heavy chain of an antibody linked together by a 10-chain 25 amino acids); a disulfide-linked Fv, or dsFv (e.g., the variable antigen-binding determining region of a single light chain and heavy chain of an antibody linked together by a disulfide bridge); a camelized VH (e.g., the variable antigen-binding determining region of a single antibody heavy chain in which some amino acids at the VH interface are those found in the naturally occurring camel antibody heavy chain); a bispecific sFv (e.g., an sFv or dsFv molecule having two antigen-binding domains, each of which can be directed to a different epitope); a diabody (e.g., a dimerized sFv formed when the VH domain of a first sFv assembles with the VL domain of a second sFv and the VL domain of the first sFv assembles with the VH domain of the second sFv; the two antigen-binding regions of the body can be directed to the same or different epitopes); and a triabody (e.g., a trimerized sFv, formed in a manner similar to a diabody, but in which three antigen-binding domains are created in a single complex; the three antigen-binding domains may be directed to the same or different epitopes). Antibody derivatives also include one or more CDR sequences of an antibody combining site. CDR sequences can be linked together in a scaffold when two or more CDR sequences are present. In certain embodiments, the antibody comprises a single Fv chain ("scFv"). scFvs are antibody fragments that comprise the VH and VL domains of an antibody, where these domains are present in a single polypeptide chain. The scFv polypeptide may further comprise a polypeptide linker between the VH and VL domains that allows the scFv to form the desired structure for binding to the antigen. For a review of scFv, see Pluckthun in The Pharmacology of Monoclonal Antibodies, vol. 113, Rosenburg and Moore eds. Springer-Verlag, New York, pp. 269-315 (1994).

4. Humanized Antibodies

[0241] The present invention provides humanized antibodies that bind Klotho beta, including human and/or Cyno beta Klotho. The humanized antibodies of the present invention may comprise one or more CDRs as set forth in Tables 110. Various methods for humanized non-human antibodies are known in the art. For example, a humanized antibody may have one or more amino acid residues introduced into it from a source that is non-human. These non-human amino acid residues are often referred to as "import" residues, which are typically taken from an "import" variable domain. Humanization can be carried out, for example, following the method of Winter and colleagues (Jones et al. (1986) Nature 321:522-525; Riechmann et al. (1988) Nature 332:323-327; Verhoeyen et al. ( 1988) Science 239:1534-1536), by replacing hypervariable region sequences with corresponding sequences from a human antibody.

[0242] In some cases, humanized antibodies are constructed by CDR grafting, in which the amino acid sequences of the six complementarity-determining regions (CDRs) of the original non-human (e.g., rodent) antibody are grafted onto an antibody structure human. For example, Padlan et al. (FASEB J. 9:133-139, 1995) determined that only about a third of the residues in the CDRs actually come into contact with the antigen, and termed these the "specificity determining residues", or SDRs. In the SDR grafting technique, only the SDR residues are grafted onto the human antibody structure (see, for example, Kashmiri et al., Methods 36: 25-34, 2005).

[0243] The choice of human variable domains, both light and heavy, to be used in the preparation of humanized antibodies can be important to reduce antigenicity. For example, according to the so-called "best fit" method, the variable domain sequence of a non-human (e.g., rodent) antibody is searched against the entire library of known human variable domain sequences. The human sequence that is closest to the rodent can be selected as the human framework for the humanized antibody (Sims et al. (1993) J. Immunol. 151:2296; Chothia et al. (1987) J. Mol. Biol. 196:901. Another method uses a particular structure derived from the consensus sequence of all human antibodies of a particular subgroup of light or heavy chains. The same structure can be used for several different humanized antibodies (Carter et al. (1992) Proc. Natl. Acad. Sci. USA, 89:4285; Presta et al. (1993) J. Immunol., 151:2623. subgroup I (VL6I) and VH subgroup III (VHIII). In another method, human germline genes are used in the source of the framework regions.

[0244] In an alternative paradigm based on the comparison of CDRs, called superhumanization, FR homology is irrelevant. The method consists of comparing the non-human sequence with the repertoire of functional human germline genes. Those genes encoding the same canonical structures or closely related to the murine sequences are then selected. Then, within the genes that share canonical structures with the non-human antibody, those with the highest homology within the CDRs are chosen as FR donors. Finally, non-human CDRs are grafted onto these FRs (see, for example, Tan et al, J. Immunol. 169:1119-1125, 2002).

[0245] It is still generally desirable for antibodies to be humanized with retention of their affinity for the antigen and other favorable biological properties. To achieve this objective, according to one method, humanized antibodies are prepared by a process of analyzing the parental sequences and various conceptual humanized products using three-dimensional models of the parental and humanized sequences. Three-dimensional immunoglobulin models are commonly available and are familiar to those skilled in the art. Computer programs are available which illustrate and present likely three-dimensional conformational structures of selected candidate immunoglobulin sequences. These include, for example, WAM (Whitelegg and Rees, Protein Eng. 13: 819-824, 2000), Modeller (Sall and Blundell, J. Mol Biol 234: 779815, 1993), and Swiss PDB Viewer (Guex and Peitsch, Electrophoresis 18:2714-2713, 1997). Inspection of these displays allows analysis of the likely role of residues in the functioning of the candidate immunoglobulin sequence, for example, analysis of residues that influence the ability of the candidate immunoglobulin to bind its antigen. In this way, FR residues can be selected and combined from the receptor sequences and imported so that the desired antibody characteristic, such as greater affinity to the target antigen(s), is achieved. In general, residues in the hypervariable region are directly and very substantially involved in influencing antigen binding.

[0246] Another method for humanizing antibodies is based on an antibody humanity metric called Human Chain Content (HSC). This method compares the mouse sequence to the human germline gene repertoire and the differences are marked as HSC. The target sequence is then humanized, maximizing its HSC rather than using a global identity measure to generate multiple diverse humanized variants. (Lazar et al, Mol Immunol 44: 1986-1998, 2007).

[0247] In addition to the methods described above, empirical methods can be used to generate and select humanized antibodies. These methods include those that are based on generating large libraries of humanized variants and selecting the best clones using enrichment technologies or high-throughput screening techniques. Antibody variants can be isolated from phage libraries, ribosome display libraries, and yeast, as well as by screening bacterial colonies (see, e.g., Hoogenboom, Nat. Biotechnol. 23:1105-1116, 2005; Dufner et al., Trends Biotechnol. 24: 523-529, 2006; 2004).

[0248] In the FR library approach, a collection of residue variants are introduced at specific positions in the FR followed by library selection to select the FR that best supports the grafted CDR. Residues to be replaced may include some or all of the "Vernier" residues identified as potentially contributing to the structure of the CDR (see, e.g., Foote and Winter, J. Mol Biol 224:487-499, 1992), or the from the more limited set of target residues identified by Baca et al. (J. Biol Chem 272: 10678-10684, 1997).

[0249] In FR shuffling, entire FRs are combined with non-human CDRs, rather than creating combinatorial libraries of selected residue variants (see, for example, Dall'Acqua et al, Methods 36: 43-60, 2005) . Libraries can be screened for linkage in a two-step selection process, first humanizing VL, followed by VH. Alternatively, a one-step FR shuffling process can be used. Such a process has been demonstrated to be more effective than two-step screening, as the resulting antibodies exhibited improved biochemical and physicochemical properties, including increased expression, increased affinity, and thermal stability (see, for example, Damschroder et al. , Mol. Immunol. 44: 3049-60, 2007).

[0250] The "humaneering" method is based on the experimental identification of minimal essential specificity determinants (MSDs) and is based on the sequential replacement of non-human fragments in human FR libraries and assessment of binding. It begins with the CDR3 regions of non-human VH and VL chains and progressively replaces other non-human antibody regions in human FRs, including the CDR1 and CDR2 of both VL and VH. This methodology typically results in epitope retention and identification of antibodies from multiple subclasses with distinct human V segment CDRs. Humaneering allows the isolation of antibodies that are 91-96% homologous to human germline gene antibodies. (See, for example, Alfenito, Cambridge Healthtech Institute’s Third Annual PEGS, The Protein Engineering Summit, 2007).

[0251] The "human engineering" method involves altering a non-human antibody or antibody fragment, such as a mouse or chimeric antibody or antibody fragment, by making specific changes to the amino acid sequence of the antibody so as to produce a modified antibody with reduced immunogenicity in a human, however, which retains the desirable binding properties of the original non-human antibodies. Generally, the technique involves classifying amino acid residues of a non-human (e.g., mouse) antibody as "low risk", "moderate risk", or "high risk" residues. The ranking is performed using an overall risk/reward calculation that evaluates the anticipated advantages of making the particular substitution (e.g., for immunogenicity in humans) against the risk that the substitution will affect the folding of the resulting antibody and/or be replaced by human waste. The particular human amino acid residue to be substituted at a given position (e.g., low or moderate risk) of a non-human (e.g., mouse) antibody sequence can be selected by aligning a variable region amino acid sequence. of the non-human antibody with the corresponding region of a specific or consensus human antibody sequence. Amino acid residues at low or moderate risk positions in the non-human sequence can be replaced with corresponding residues from the human antibody sequence according to the alignment. Techniques for producing proteins by human engineering are described in greater detail in Studnicka et al, Protein Engineering, 7:805-814 (1994), US Patents 5,766,886, 5,770,196, 5,821,123, and 5,869,619, and PCT WO patent application 93/11794.

5. Human Antibodies

[0252] Human anti-Klotho beta antibodies can be constructed by combining Fv clone variable domain sequence(s) selected from phage display libraries of human origin with known human constant domain sequence(s). Alternatively, the anti-Klotho-beta human monoclonal antibodies of the present invention can be made by the hybridoma method. Human myeloma and mouse-human heteromyeloma cell lines for the production of human monoclonal antibodies have been described, for example, by Kozbor J. Immunol., 133: 3001 (1984); Brodeur et al., Monoclonal Antibody Production Techniques and Applications, pp. 51-63 (Marcel Dekker, Inc., New York, 1987); and Boerner et al., J. Immunol., 147: 86 (1991).

[0253] It is also possible to produce transgenic animals (e.g., mice) that are capable, after immunization, of producing a complete repertoire of human antibodies in the absence of endogenous immunoglobulin production. Transgenic mice expressing human antibody repertoires have been used to generate high-affinity human sequence monoclonal antibodies against a wide variety of potential drug targets (see, e.g., Jakobovits, A., Curr. Opin. Biotechnol. 1995, 6(5):561-6; 1117-1125, 2005).

[0254] Alternatively, the human antibody may be prepared by immortalizing human B lymphocytes producing an antibody directed against a target antigen (e.g., such B lymphocytes may be recovered from an individual or may have been immunized in vitro ) (see, for example, Cole et al, Monoclonal Antibodies and Cancer Therapy, Alan R. Liss, p 77 (1985); Boerner et al, J. Immunol, 147 (1):86-95 (1991); and Patent No. 5,750,373).

[0255] Gene shuffling can also be used to derive human antibodies from non-human, e.g., rodent, antibodies, wherein the human antibody has similar affinities and specificities to the starting non-human antibody. According to this method, which is also called "epitope printing" or "guided selection", either the heavy or light chain variable region of a non-human antibody fragment obtained by phage display techniques as described herein is replaced with a repertoire of human V domain genes, creating a population of non-human chain/human chain scFv or Fab chimeras. Selection with antigen results in the isolation of a non-human chain/chimeric human chain scFv or Fab in that the human chain restores the antigen-binding site destroyed upon removal of the corresponding non-human chain in the primary phage display clone, (e.g., the epitope guides (prints) the choice of human chain partner). When the process is repeated to replace the remaining non-human chain, a human antibody is obtained (see, for example, PCT WO 93/06213; and Osbourn et al., Methods., 36, 61-68, 2005). . Unlike traditional humanization of non-human antibodies by CDR grafting, this technique provides completely human antibodies, which have no FR or CDR residues of non-human origin. Examples of guided selection to humanize mouse antibodies against cell surface antigens include the folate-binding protein present on ovarian cancer cells (see, for example, Figini et al., Cancer Res., 58, 991-996, 1998) and CD147, which is highly expressed in hepatocellular carcinoma (see, for example, Bao et al., Cancer Biol. Ther., 4, 1374-1380, 2005).

[0256] A potential disadvantage of the guided selection approach is that shuffling one antibody chain while keeping the other constant can result in epitope drift. In order to maintain the epitope recognized by the non-human antibody, CDR retention can be applied (see, for example, Klimka et al., Br. J. Cancer., 83, 252-260, 2000; VH CDR2 Beiboer et al., J. Mol. Biol., 296, 833-49, 2000). In this method, a non-human VH CDR3 is normally retained, as this CDR may be at the center of the antigen-binding site and may prove to be the most important region of the antibody for antigen recognition. In some cases, however, VH CDR3 and VL CDR3, as well as VH CDR3, VL CDR3, and VL CFR1, of the non-human antibody may be retained.

6. Bispecific Antibodies

[0257] Bispecific antibodies are monoclonal antibodies that have binding specificities for at least two different antigens. In certain embodiments, the bispecific antibodies are human or humanized antibodies. In certain embodiments, one of the binding specificities is for Klotho beta and the other is for any other antigen. In some embodiments, one of the binding specificities is for Klotho beta, and the other is for another surface antigen expressed on cells expressing Klotho beta and an FGF receptor (e.g., FGFR1c, FGFR2c, FGFR3c, FGFR4). In certain embodiments, the bispecific antibodies can bind to two different Klotho beta epitopes. Bispecific antibodies can be prepared as full-length antibodies or antibody fragments (e.g., F(ab') 2 bispecific antibodies).

[0258] Methods for preparing bispecific antibodies are known in the art, such as, for example, by co-expression of two immunoglobulin heavy chain-light chain pairs, wherein the two heavy chains have different specificities (see, e.g. example, Milstein and Cuello, Nature, 305:537 (1983)). For more details on the generation of bispecific antibodies see, for example, Bispecific Antibodies, Kontermann, ed., Springer-Verlag, Hiedelberg (2011).

7. Multivalent Antibodies

[0259] A multivalent antibody can be internalized (and/or catabolized) more quickly than a bivalent antibody by a cell that expresses an antigen to which the antibodies bind. The antibodies of the present invention can be multivalent antibodies (which are other than those of the IgM class) with three or more antigen-binding sites (e.g., tetravalent antibodies), which can be readily produced by recombinant expression of nucleic acid encoding the antibody polypeptide chains. The multivalent antibody may comprise a dimerization domain and three or more antigen-binding sites. In certain embodiments, the dimerization domain comprises (or consists of) an Fc region or a hinge region. In this scenario, the antibody will comprise an Fc region and three or more antigen-binding sites amino-terminal to the Fc region. In certain embodiments, a multivalent antibody comprises (or consists of) three to about eight antigen binding sites. In such an embodiment, a multivalent antibody comprises (or consists of) four antigen-binding sites. The multivalent antibody comprises at least one polypeptide chain (e.g., two polypeptide chains), wherein the polypeptide chain(s) comprises two or more variable domains. For example, the polypeptide chain(s) may comprise VD1-(X1)n-VD2-(X2)n-Fc, wherein VD1 is a first variable domain, VD2 a second variable domain, Fc is a polypeptide chain of an Fc region, X1 and X2 represent an amino acid or a polypeptide, and n is 0 or 1. For example, the polypeptide chain(s) may comprise: VH-CH1-flexible linker-VH-CH1-Fc chain region; or VH-CH1-VH-CH1-Fc chain region. The multivalent antibody described herein may further comprise at least two (e.g., four) light chain variable domain polypeptides. The multivalent antibody described herein may, for example, comprise from about two to about eight light chain variable domain polypeptides. The light chain variable domain polypeptides contemplated herein comprise a light chain variable domain and, optionally, further comprise a CL domain.

8. FC Engineering

[0260] It may be desirable to modify an antibody to Klotho beta through Fc engineering, including, with respect to effector function, for example, so as to decrease or eliminate antigen-dependent cell-mediated cytotoxicity (ADCC) and/or or complement-dependent cytotoxicity (CDC) of the antibody. This can be accomplished by introducing one or more amino acid substitutions into an Fc region of the antibody. For example, substitutions in human IgG1 using Ig2 residues such as positions 233-236 and IgG4 residues at positions 327, 330, and 331 have been shown to greatly reduce ADCC and CDC (see, e.g., Armor et al., Eur. J . Immunol. 29:(8):2613-24 (1999);

[0261] To increase the serum half-life of the antibody, a salvage receptor binding epitope can be incorporated into the antibody (especially an antibody fragment), for example, as described in US Patent 5,739,277. The term "salvage receptor binding epitope" refers to an epitope in the Fc region of an IgG molecule (e.g., IgG1, IgG2, IgG3, or IgG4) that is responsible for the in vivo increase in half-life in IgG molecule serum.

9. Alternative Liaison Agents

[0262] The present invention encompasses non-immunoglobulin binding agents that specifically bind to the same epitope as an anti-Klotho beta antibody disclosed herein. In some embodiments, a non-immunoglobulin binding agent is identified as an agent that displaces or is displaced by an anti-Klotho beta antibody of the present disclosure in a competitive binding assay. These alternative binding agents may include, for example, any of the engineered protein scaffolds known in the art. Such frameworks may comprise one or more CDRs as presented in Tables 1-10. Such scaffolds include, for example, anticalins, which are based on the lipocalin scaffold, a protein structure characterized by a rigid beta-barrel that supports four hypervariable loops that form the ligand-binding site. New binding specificities can be manipulated by targeted random mutagenesis in the loop regions, in combination with functional display and guided selection (see, for example, Skerra (2008) FEBS J. 275:2677-2683). Other suitable scaffolds may include, for example, adnectins, or monobodies, based on the tenth extracellular domain of human fibronectin III (see, for example, Koide and Koide (2007) Methods Mol Biol 352:95-109); affibodies, based on the Z domain of staphylococcal protein A (see, for example, Nygren et al (2008) FEBS J. 275:2668-2676)); DARPins, based on ankyrin repeat proteins (see, for example, Stumpp et al. (2008) Drug. Discov. Today 13: 695-701); finamers, based on the SH3 domain of the human Fyn protein kinase (Grabulovski et al. (2007) J. Biol. Chem. 282:3196-3204); afilins, based on Sac7d from Sulfolobus acidolarius (see, for example, Krehenbrink et al. (2008) J. Mol. Biol. 383: 1058 1068); afilins, based on human γ-B-crystallin (see, for example, Ebersbach et al (2007) J. Mol Biol 372:172-185); avimers, based on the A domains of membrane receptor proteins (see, for example, Silverman et al (2005) Biotechnol. 23:1556-1561); cysteine-rich knottin peptides (see, for example, Kolmar (2008) FEBS J. 275: 2684-2690); and engineered Kunitz-type inhibitors (see, for example, Nixon and Wood (2006) Curr. Opin. Drug. Discov. Dev. 9: 261-268). For a review, see, for example, Gebauer and Skerra (2009) Curr. Opinion. Chem. Biol. 13: 245-255.

Antibody Variants

[0263] In some embodiments, amino acid sequence modification(s) of the antibodies that bind to Klotho beta or described herein are contemplated. For example, it may be desirable to improve the binding affinity and/or other biological properties of the antibody, including, but not limited to, specificity, thermostability, expression level, effector functions, glycosylation, solubility, or reduced immunogenicity. This, in addition to the anti-Klotho beta antibodies described herein, is contemplated that anti-Klotho beta antibody variants can be prepared. For example, anti-Klotho beta antibody variants can be prepared by introducing appropriate nucleotide changes into the coding DNA, and/or by synthesizing the desired polypeptide or antibody. Those skilled in the art should note that amino acid changes may alter post-translational processes of the anti-Klotho beta antibody, such as altering the number or position of glycosylation sites or altering membrane anchoring characteristics.

[0264] In some embodiments, the antibodies provided here are chemically modified, for example, by covalently attaching any type of molecule to the antibody. Antibody derivatives may include antibodies that have been chemically modified, for example, by glycosylation, acetylation, pegylation, phosphorylation, amidation, derivatization by known protecting/blocking groups, proteolytic cleavage, binding to a cellular ligand or other protein, etc. Any of numerous chemical modifications can be carried out by known techniques, including, but not limited to, specific chemical cleavage, acetylation, formulation, metabolic synthesis of tunicamycin, etc. Furthermore, the antibody may contain one or more non-classical amino acids.

[0265] Variations may be a substitution, deletion or insertion of one or more codons encoding the antibody or polypeptide that results in a change in the amino acid sequence compared to the native sequence antibody or polypeptide. Amino acid substitutions may be the result of replacing an amino acid with another amino acid having similar structural and/or chemical properties, such as replacing a leucine with a serine, for example, conservative amino acid substitutions. The insertions or deletions may optionally be in the range of about 1 to 5 amino acids. In certain embodiments, the substitution, deletion or insertion includes fewer than 25 amino acid substitutions, fewer than 20 amino acid substitutions, fewer than 15 amino acid substitutions, fewer than 10 amino acid substitutions, fewer than 5 amino acid substitutions , less than 4 amino acid substitutions, less than 3 amino acid substitutions, or less than 2 amino acid substitutions relative to the original molecule. In a specific embodiment, the substitution is a conservative amino acid substitution made at one or more predicted non-essential amino acid residues. The variation allowed can be determined by systematically making insertions, deletions, or substitutions of amino acids in the sequence and testing the resulting variants against the activity exhibited by the mature or full-length native sequence.

[0266] Insertions in the amino acid sequence include amino and/or carboxyl-terminal fusions ranging in length from one residue to polypeptides containing one hundred or more residues, as well as intra-sequence insertions of single or multiple amino acid residues. Examples of terminal insertions include an antibody with an N-terminal methionyl residue. Other insertion variants of the antibody molecule include fusion at the N- or C-terminus of the antibody to an enzyme (e.g., for antibody-directed enzyme prodrug therapy) or a polypeptide that increases the serum half-life of the antibody. .

[0267] Substantial modifications to the biological properties of the antibody are achieved by selecting substitutions that differ significantly in their effect on maintaining (a) the backbone structure of the polypeptide in the area of the substitution, for example, as a sheet or helical conformation, (b) the charge or hydrophobicity of the molecule at the target site, or (c) the volume of the side chain. Alternatively, conservative substitutions (e.g., within a group of amino acids with similar properties and/or side chains) can be made, so as to maintain or not significantly alter the properties. Amino acids can be grouped according to similarities in the properties of their side chains (see, for example, AL Lehninger, in Biochemistry, 2nd Ed, pp 73-75, Worth Publishers, New York (1975)): (1) non- polar: Ala (A), Val (v), Leu (L), Ile (I), Pro (P), Phe (F), Trp (W), Met (M); (2) polar uncharged: Gly (G), Ser (S), Thr (T), Cys (C), Tyr (Y), Asn (N), Gln (Q); (3) acids: Asp (D), Glu (E); and (4) basic: Lys (K), Arg (R), His (H).

[0268] Alternatively, naturally occurring residues can be divided into groups based on common side chain properties: (1) hydrophobic: Norleucine, Met, Ala, Val, Leu, Ile; (2) neutral hydrophilic: Cys, Ser, Thr, Asn, Gln; (3) acids: Asp, Glu; (4) basic: his, lys, Arg; (5) residues that influence chain orientation: Gly, Pro; and (6) aromatic: Trp, Tyr, Phe.

[0269] Non-conservative substitutions will imply exchanging a member of one of these classes for another class. Such substituted residues can also be introduced at conservative substitution sites or at remaining (non-conservative) sites. Therefore, in one embodiment, an antibody or fragment thereof that binds to a Klotho beta epitope comprises an amino acid sequence that is at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identical to amino acid sequence of a murine monoclonal antibody described herein. In one embodiment, an antibody or fragment thereof that binds to a Klotho beta epitope comprises an amino acid sequence that is at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identical to a sequence of amino acids represented in Tables 1-10. In yet another embodiment, an antibody or fragment thereof that binds to a Klotho beta epitope comprises a VH CDR and/or a VL CDR amino acid sequence that is at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identical to a VH CDR amino acid sequence depicted in Tables 1-10 and/or a VL CDR amino acid sequence depicted in Tables 1-10. Variations can be made using methods known in the art such as oligonucleotide-mediated (site-directed) mutagenesis, alanine scanning, and PCR mutagenesis. Site-directed mutagenesis (see, e.g., Carter et al, Nucl Acids Res., 13:4331 (1986); Zoller et al, Nucl Acids Res., 10:6487 (1987)), cassette mutagenesis (see, e.g., example, Wells et al, Gene, 34:315 (1985)), restriction selection mutagenesis (see, e.g., Wells et al, Philos Trans R. Soc. London SerA, 317:415 (1986)) or other techniques Known techniques can be performed on the cloned DNA to produce the anti-Klotho beta antibody variant DNA.

[0270] Any cysteine residue not involved in maintaining the proper conformation of the Anti-Klotho beta antibody can also be replaced, for example, with another amino acid such as alanine or serine, to improve the oxidative stability of the molecule and prevent aberrant cross-linking. On the other hand, cysteine linkage(s) can be added to the anti-Klotho beta antibody to improve its stability (for example, where the antibody is an antibody fragment such as an Fv fragment).

[0271] In some embodiments, an anti-Klotho beta antibody molecule of the present disclosure is a "de-immunized" antibody. A "de-immunized" anti-Klotho beta antibody is an antibody derived from a humanized or chimeric Anti-Klotho beta antibody, which has one or more changes in its amino acid sequence, resulting in a reduction in the immunogenicity of the antibody compared to its counterpart. original non-de-immunized antibody. One of the processes for generating such antibody mutants involves identifying and eliminating T cell epitopes from the antibody molecule. In a first step, the immunogenicity of the antibody molecule can be determined by various methods, for example, by in vitro determination of T cell epitopes or in silico prediction of such epitopes, as is known in the art. Once residues critical for T cell epitope function have been identified, mutations can be made to remove immunogenicity and retain antibody activity. For a review, see, for example, Jones et al, Methods in Molecular Biology 525:405-423, 2009.

1. In vitro affinity maturation

[0272] In some embodiments, antibody variants having an improved property, such as affinity, stability or expression level as compared to a parental antibody, can be prepared by in vitro affinity maturation. Like the natural prototype, in vitro affinity maturation is based on the principles of mutation and selection. Antibody libraries are displayed as Fab, scFv, or V-domain fragments either on the surface of an organism (e.g., phage, bacteria, yeast, or mammalian cells) or in association (e.g., covalently or noncovalently) with the mRNA or coding DNA. Affinity selection of the antibodies displayed allows the isolation of organisms that are complex or carry the genetic information coding for the antibodies. Two or three rounds of mutation and selection using display methods such as phage display usually results in antibody fragments with affinities in the low nanomolar range. Preferred affinity-matured antibodies will have nanomolar or even picomolar affinities for the target antigen.

[0273] Phage display is a widespread method for displaying and selecting antibodies. Antibodies are displayed on the surface of Fd or M13 bacteriophages as fusions to the bacteriophage coat protein. Selection involves exposure to antigen to allow phage-displayed antibodies to bind to their targets, a process called "panning." The antigen-bound phages are recovered and infected into bacteria to produce phages for further rounds of selection. For review, see, for example, Hoogenboom, Methods. Mol. Biol. 178: 1-37, 2002; Bradbury and Marks, J. Immuno. Methods 290: 29-49, 2004).

[0274] In a yeast display system (see, for example, Boder et al., Nat. Biotech. 15:553-57, 1997; Chao et al., Nat. Protocols 1:755-768, 2006), The antibody can be displayed in the form of single-chain variable fusions (scFv) in which the heavy and light chains are linked by a flexible linker. The scFv is fused to the adhesion subunit of the yeast Aga2p agglutinin protein, which binds to the yeast cell wall via disulfide bonds to Aga1p. Display of a protein through Aga2p projects the protein away from the cell surface, minimizing possible interactions with other molecules in the yeast cell wall. Magnetic separation and flow cytometry are used to screen the library to select antibodies with improved affinity or stability. Binding to a soluble antigen of interest is determined by labeling yeast with biotinylated antigen and a secondary reagent such as streptavidin conjugated to a fluorophore. Variations in antibody surface expression can be measured by immunofluorescence labeling of either hemagglutinin or epitope tagging of c-Myc flanking the scFv. Expression has been shown to correlate with the stability of the displayed protein and thus antibodies can be selected for better stability as well as affinity (see, e.g., Shusta et al., J. Mol. Biol. 292: 949 -956, 1999). An additional advantage of yeast display is that the displayed proteins are folded in the endoplasmic reticulum of eukaryotic yeast cells, taking advantage of the endoplasmic reticulum chaperones and quality control machinery. Once maturation is complete, antibody affinity can be conveniently 'titrated' while displayed on the yeast surface, eliminating the need for expression and purification of each clone. A theoretical limitation of yeast surface display is the potentially smaller functional library size than that of other display methods; however, a recent approach utilizes the yeast cell docking system to create combinatorial diversity estimated at 1014 in size (see, for example, US Patent Publication 2003/0186,374; Blaise et al, Gene 342:211 -218, 2004).

[0275] In ribosome display, antibody-ribosome-mRNA (ARM) complexes are generated for selection in a cell-free system. The DNA library coding for a given antibody library is genetically fused to a spacer sequence that lacks a stop codon. This spacer sequence, when translated, is still bound to the peptidyl-tRNA and occupies the ribosomal tunnel, and thus allows the protein of interest to protrude out of the ribosome and fold. The resulting complex of mRNA, ribosome, and protein can bind to the surface-bound ligand, allowing simultaneous isolation of the antibody and its encoding mRNA through ligand affinity capture. The ribosome-bound mRNA is then reverse transcribed back into cDNA, which can then be subjected to mutagenesis and used in the next round of selection (see, for example, Fukuda et al., Nucleic Acids Res. 34, E127 , 2006). In mRNA display, a covalent bond between the antibody and the mRNA is established using puromycin as an adapter molecule (Wilson et al., Proc. Natl. Acad. Sci. USA 98, 3750-3755, 2001).

[0276] As these methods are carried out entirely in vitro, they provide two main advantages over other selection technologies. Firstly, library diversity is not limited by the transformation efficiency of bacterial cells, but only by the number of ribosomes and different mRNA molecules present in the test tube. Second, random mutations can easily be introduced after each selection cycle, for example, by uncorrecting polymerases, as no library should be transformed after any diversification step.

[0277] Diversity can be introduced into the CDRs or entire V genes of antibody libraries in a selective manner or through random introduction. The first approach includes sequentially targeting all CDRs of an antibody through a high or low level of mutagenesis or by targeting isolated hot spots of somatic hypermutations (see, e.g., Ho, et al., J. Biol. Chem. 280: 607 -617, 2005) or residues suspected of affecting affinity on experimental basis or structural reasons. Random mutations can be introduced throughout the entire V gene using mutant strains of E. coli, error-prone replication with DNA polymerases (see, e.g., Hawkins et al, J. Mol Biol 226:889-896, 1992 ) or RNA replicases. Diversity can also be introduced by replacing regions that are naturally diverse through DNA rearrangement or similar techniques (see, e.g., Lu et al, J. Biol Chem 278:43496-43507, 2003; U.S. Patent No. 5,565,332; U.S. Pat. No. 6,989,250). Alternative techniques target hypervariable loops that extend into framework region residues (see, e.g., Bond et al, J. Mol Biol 348:699-709, 2005), employ deletions and insertions in CDRs, or use hybridization-based diversification ( see, for example, US Patent Publication No. 2004/0005709). Additional methods for generating diversity in CDRs are described, for example, in U.S. Pat. US No. 7,985,840.

[0278] Library screening can be carried out by various techniques known in the art. For example, Klotho beta can be immobilized on solid supports, columns, pins or cellulose/poly(vinylidene fluoride)/other filter membranes, expressed in host cells attached to adsorption plates or used in cell separation, or conjugated with biotin for capture with streptavidin-coated beads, or used in any other method to mine display libraries.

[0279] For review of in vitro affinity maturation methods, see, for example, Hoogenboom, Nature Biotechnology 23:1105-1116, 2005 and Quiroz and Sinclair, Revista Ingeneria Biomedia 4: 39-51, 2010 and references therein.

2. Anti-Klotho Beta Antibody Modifications

[0280] Covalent modifications of anti-Klotho beta antibodies are included within the scope of the present disclosure. Covalent modifications include reacting target amino acid residues of an anti-Klotho beta antibody with an organic derivatizing agent that is capable of reacting with selected side chains or the N- or C-terminal residues of the anti-Klotho beta antibody. Other modifications include deamidation of glutaminyl and asparaginyl residues to the corresponding glutamyl and aspartyl residues, respectively, hydroxylation of proline and lysine, phosphorylation of hydroxyl groups of seryl or threonyl residues, methylation of the α-amino groups of the side chains of lysine, arginine, and histidine ( see, for example, T.E. Creighton, Proteins: Structure and Molecular Properties, W.H. Freeman & Co., San Francisco, pp. 79-86 (1983)), acetylation of the N-terminal amine, and amidation of any C-terminal carboxyl group .

[0281] Other types of covalent modifications of the Anti-Klotho beta antibody included within the scope of the present invention include altering the native glycosylation pattern of the antibody or polypeptide (see, for example, Beck et al., Curr. Pharm. Biotechnol. 9: 482-501, 2008; Walsh, Drug Discov. Today 15: 773-780, 2010), and binding the antibody to one of a variety of non-proteinaceous polymers, for example, polyethylene glycol (PEG), polypropylene glycol or polyoxyalkylenes, in the manner set forth, for example, in US Patent Nos. 4,640,835; 4,496,689; 4,301,144; 4,670,417; 4,791,192 or 4,179,337.

[0282] An Anti-Klotho beta antibody of the present disclosure can also be modified so as to form chimeric molecules comprising an Anti-Klotho beta antibody fused to another heterologous polypeptide or amino acid sequence, for example, an epitope tag (see, e.g. example, Terpe, Appl. Microbiol. Biotechnol. 60: 523-533, 2003) or the Fc region of an IgG molecule (see, for example, Aruffo, “Immunoglobulin fusion proteins” in Antibody Fusion Proteins, S.M. Chamow and A. Ashkenazi, eds., Wiley-Liss, New York, 1999, pp. 221-242).

[0283] Also provided herein are fusion proteins comprising an antibody provided herein that binds to a Klotho beta antigen and a heterologous polypeptide. In some embodiments, the heterologous polypeptide to which the antibody is fused is useful for targeting the antibody to cells having Klotho beta expressed on the cell surface.

[0284] Furthermore, panels of antibodies that bind to a Klotho beta antigen are provided here. In specific embodiments, the antibody panels have different association rate constants, different dissociation rate constants, different affinities to the Klotho beta antigen, and/or different specificities for a Klotho beta antigen. In some embodiments, the panels comprise or consist of about 10, about 25, about 50, about 75, about 100, about 125, about 150, about 175, about 200, about 250, about 300, about 350, about 400, about 450, about 500, about 550, about 600, about 650, about 700, about 750, about 800, about 850, about 900, about 950, or about 1000 antibodies or more. Antibody panels can be used, for example, in 96-well or 384-well plates, such as for assays such as ELISA assays.

Preparation of Anti-Klotho beta Antibodies

[0285] Anti-Klotho beta antibodies can be produced by cells transformed or transfected with a vector containing nucleic acids encoding the anti-Klotho beta antibody. The polynucleotide sequences encoding the antibody polypeptide components of the present specification can be obtained using conventional recombinant techniques. Desired polynucleotide sequences can be isolated and sequenced from antibody-producing cells, such as hybridoma cells. Alternatively, polynucleotides can be synthesized using nucleotide synthesizers or PCR techniques. Once obtained, the sequences that encode the polypeptides are inserted into a recombinant vector capable of replicating and expressing heterologous polynucleotides in host cells. Many vectors that are available and known in the art can be used for the purposes of the present specification. The selection of an appropriate vector will depend primarily on the size of the nucleic acids to be inserted into the vector and the particular host cell to be transformed with the vector. Suitable host cells for expression of antibodies of the present invention include prokaryotic cells, such as archaebacteria and eubacteria, including Gram-negative or Gram-positive organisms, eukaryotic microbes such as filamentous fungi or yeast, invertebrate cells, such as insect cells or plant, and vertebrate cells such as mammalian host cell lines. Host cells are transformed with the expression vectors described above and grown in conventional nutrient media modified as appropriate to induce promoters, select transformants, or amplify genes encoding the desired sequences. Antibodies produced by host cells are purified using standard protein purification methods as known in the art.

[0286] Methods for producing antibodies including vector construction, expression and purification are further described in Plückthun et al., (1996) in Antibody Engineering: Producing antibodies in Escherichia coli: From PCR to fermentation (McCafferty, J. , Hoogenboom, H. R., and Chiswell, D. J., eds), 1 Ed., pp. 203-252, IRL Press, Oxford; Kwong, K. & Rader, C., E. coli expression and purification of Fab antibody fragments, Current protocols in protein science editorial board John E Coligan et al., Chapter 6, Unit 6.10 (2009); Tachibana and Takekoshi, “Production of Antibody Fab Fragments in Escherischia coli,” in Antibody Expression and Production, M. Al-Rubeai, Ed., Springer, New York, 2011; Therapeutic Monoclonal Antibodies: From Bench to Clinic (ed Z. An), John Wiley & Sons, Inc., Hoboken, NJ, USA.

[0287] It is, of course, contemplated that alternative methods, which are well known in the art, may be employed to prepare anti-Klotho beta antibodies. For example, the appropriate amino acid sequence, or portions thereof, can be produced through direct peptide synthesis using solid phase techniques (see, e.g., Stewart et al., SolidPhase Peptide Synthesis, W.H. Freeman Co., San Francisco, CA (1969); Merrifield, J. Am. Soc., 85:2149-2154 (1963)). In vitro protein synthesis can be carried out using manual techniques or by automation. Various portions of the anti-Klotho beta antibody can be chemically synthesized separately and combined using chemical or enzymatic methods to produce the desired anti-Klotho beta antibody. Alternatively, antibodies can be purified from cells or body fluids, such as milk, of a transgenic animal engineered to express the antibody, as disclosed, for example, in U.S. Pat. U.S. Pat. No. 5,545,807 and U.S. Pat. US No. 5,827,690.

Immunoconjugates

[0288] The present invention also provides conjugates comprising any of the anti Klotho-beta antibodies of the present invention covalently linked by a synthetic linker to one or more non-antibody agents.

[0289] A variety of radioactive isotopes are available for the production of radioconjugated antibodies. Examples include At211, I4, I4, Y4, Re4, Re4, SM4, BI4, P4, Pb4, and radioactive isotopes of Lu. When the conjugate is used for detection, it may comprise a radioactive atom for scintigraphic studies, for example tc4 or I4, or a spin label for nuclear magnetic resonance (NMR) imaging (also known as magnetic resonance imaging, MRI ), such as iodine-123 again, iodine-131, indium-111, fluorine-19, carbon-13, nitrogen-15, oxygen-17, gadolinium, manganese or iron. Radioisotopes can be incorporated into the conjugate in known ways as described, for example, in Reilly, “The radiochemistry of monoclonal antibodies and peptides,” in Monoclonal Antibody and Peptide-Targeted Radiotherapy of Cancer, R.M. Reilly, ed., Wiley, Hoboken N.J. , 2010.

[0290] In some embodiments, the antibodies provided herein are conjugated or recombinantly fused to a diagnostic, detectable or therapeutic agent or any other molecule. Conjugated or recombinantly fused antibodies may be useful, for example, for monitoring or prognosing the onset, development, progression and/or severity of a Klotho beta-mediated disease as part of a clinical testing procedure, such as determining efficacy. of a particular therapy.

[0291] Such diagnosis and detection can be carried out, for example, by coupling the antibody to detectable substances, including, but not limited to, various enzymes, such as, but not limited to, horseradish peroxidase, alkaline phosphatase, beta- galactosidase, or acetylcholinesterase; prosthetic groups, such as, but not limited to, streptavidin/biotin and avidin/biotin; fluorescent materials, such as, but not limited to, umbelliferone, fluorescein, fluorescein isothiocyanate, rhodamine, dichlorotriazinylamine fluorescein, dansyl chloride or phycoerythrin; luminescent materials, such as, but not limited to, luminol; bioluminescent materials such as, but not limited to, luciferase, luciferin and aequorin; chemiluminescent material, such as, but not limited to, an acridinium-based compound or a HaloTag; radioactive materials, such as, but not limited to, iodine (131I, 125I, 123I, and 121I), carbon (14C), sulfur (35S), tritium (3H), indium (115In, 113In, 112In, and 111In, ), technetium (99Tc), thallium (201Ti), gallium (68Ga, 67Ga), palladium (103Pd), molybdenum (99Mo), xenon (133Xe), fluorine (18F), 153Sm, 177Lu, 159Gd, 149Pm, 140La, 175Yb , 166Ho, 90Y, 47Sc, 186Re, 188Re, 142Pr, 105Rh, 97Ru, 68Ge, 57Co, 65Zn, 85Sr, 32P, 153Gd, 169Yb, 51Cr, 54Mn, 75Se, 113Sn, and ; and positron-emitting metals using various positron emission tomography and non-radioactive paramagnetic metal ions.

[0292] Furthermore, antibodies that are conjugated or recombinantly fused with a therapeutic moiety (or one or more therapeutic moieties), as well as their uses, are provided herein. The antibody may be recombinantly conjugated or fused to a therapeutic moiety, including a cytotoxin, such as a cytostatic or cytocidal agent, a therapeutic agent, or a radioactive metal ion, such as alpha-emitters. A cytotoxin or cytotoxic agent includes any agent that is harmful to cells.

[0293] Furthermore, an antibody provided herein can be conjugated or recombinantly fused with a therapeutic moiety or drug moiety that modifies a given biological response. Therapeutic moieties or drug moieties should not be interpreted as limited to classical chemical therapeutic agents. For example, the drug moiety may be a protein, peptide, or polypeptide having a desired biological activity. Such proteins may include, for example, a toxin such as abren, ricin A, pseudomonas exotoxin, cholera toxin, or diphtheria toxin; a protein such as tumor necrosis factor, Y-interferon, α-interferon, nerve growth factor, platelet-derived growth factor, tissue plasminogen activator, apoptotic agent, e.g., TNF-Y, TNF-Y, AIM I (see, for example, International publication WO No. 97/33899), AIM II (see, for example, International publication WO No. 97/34911), Fas ligand (see, for example, Takahashi et al. 1994, J. Immunol., 6:1567 1574), and VEGF (see, for example, International publication WO No. 99/23105), an anti-angiogenic agent, including, for example, angiostatin, endostatin or a component of the coagulation (e.g. tissue factor); or, a biological response modifier such as, for example, a lymphokine (e.g., interferon gamma, interleukin-1 ("IL-1"), interleukin-2 ("IL-2"), interleukin-5 ("IL -5"), interleukin-6 ("IL-6"), interleukin-7 ("IL-7"), interleukin-9 ("IL-9"), interleukin-10 ("IL-10"), interleukin -12 ("IL-12"), interleukin-15 ("IL-15"), interleukin-23 ("IL-23"), granulocyte macrophage colony-stimulating factor ("GM-CSF"), and granulocyte colony-stimulating agent ("L-CSF")), or a growth factor (e.g., growth hormone ("GH")), or a clotting agent (e.g., calcium, vitamin K, tissues, such as, but not limited to, Hageman factor (factor XII), high molecular weight kininogen (HMWK), prekallikrein (PK), protein clotting factors II (prothrombin), factor V, Xlla, VIII , XIIIa, XI, XIa, IX, IXa, X, phospholipid, and fibrin monomer).

[0294] Furthermore, antibodies are provided herein that are recombinantly fused or chemically conjugated (covalent or non-covalent conjugations) to a heterologous protein or polypeptide (or fragment thereof, for example, to a polypeptide of about 10, about 20 , about 30, about 40, about 50, about 60, about 70, about 80, about 90 or about 100 amino acids) to generate fusion proteins, as well as their uses. In particular, provided herein are fusion proteins comprising an antigen-binding fragment of an antibody provided herein (e.g., a Fab fragment, Fd fragment, Fv fragment, F(ab)2 fragment, a VH domain, a VH CDR , a VL domain or a VL CDR) and a heterologous protein, polypeptide or peptide. In one embodiment, the heterologous protein, polypeptide, peptide or antibody to which it is fused is useful for targeting the antibody to a particular type of cell, such as a cell expressing Klotho beta or a Klotho beta receptor. For example, an antibody that binds to a cell surface receptor expressed by a particular cell type (e.g., an immune cell) can be fused or conjugated to a modified antibody provided herein.

[0295] In addition, an antibody provided herein can be conjugated with therapeutic moieties, such as a radioactive metal ion, such as alpha-emitters, such as 213Bi or macrocyclic chelators useful for the conjugation of radiometallic ions, including, but not limited to a, 131In, 131LU, 131Y, 131Ho, 131Sm, a polypeptides. In certain embodiments, the macrocyclic chelating agent is 1,4,7,10-tetraazacyclododecane-N,N',N'',N'''-tetraacetic acid (DOTA), which can be linked to the antibody through a binding molecule. Such binding molecules are commonly known in the art and described, for example, in Denardo et al., 1998, Clin Cancer Res. 4(10):2483-90; Peterson et al., 1999, Bioconjug. Chem. 10(4):553-7; and Zimmerman et al., 1999, Nucl. Med. Biol. 26(8):943-50.

[0296] Furthermore, the antibodies provided herein can be fused to marker "tag" sequences, such as a peptide to facilitate purification. In specific embodiments, the amino acid marker or tag sequence is a hexahistidine peptide, such as the tag provided in a pQE vector (see, for example, Qiagen, Inc.), among others, many of which are commercially available. For example, as described in Gentz et al., 1989, Proc. Natl. Academic. Sci. USA 86:821-824, hexahistidine provides convenient purification of the fusion protein. Other peptide signals useful for purification include, but are not limited to, the hemagglutinin ("HA") tag, which corresponds to an epitope derived from the influenza hemagglutinin protein (Wilson et al, 1984, Cell. 37:767), and the "FLAG" marking.

[0297] Methods for fusing or conjugating therapeutic moieties (including polypeptides) to antibodies are well known, (see, for example, Arnon et al., “Monoclonal Antibodies For Immunotargeting Of Drugs In Cancer Therapy”, in Monoclonal Antibodies And Cancer Therapy, Reisfeld et al. (eds.), pp. 243-56 (Alan R. Liss, Inc. 1985); Hellstrom et al., “Antibodies For Drug Delivery,” in Controlled Drug Delivery (2nd Ed.), Robinson et al. . (eds.), pp. 623-53 (Marcel Dekker, Inc. 1987); Thorpe, “Antibody Carriers Of Cytotoxic Agents In Cancer Therapy: A Review,” in Monoclonal Antibodies 84: Biological And Clinical Applications, Pinchera et al. (eds.), pp. 475-506 (1985); “Analysis, Results, And Future Prospective Of The Therapeutic Use Of Radiolabeled Antibody In Cancer Therapy”, in Monoclonal Antibodies For Cancer Detection And Therapy, Baldwin et al. ), pp. 303-16 (Academic Press 1985), Thorpe et al., 1982, Immunol. Rev. 62:119-58; 5, 5,783,181, 5,908,626 , 5,844,095, and 5,112,946; EP 307,434; EP 367,166; EP 394,827; PCT Publications WO 91/06570, WO 96/04388, WO 96/22024, WO 97/34631, and WO 99/04813; Ashkenazi et al., Proc. Natl. Academic. Sci. USA, 88: 10535-10539, 1991; Traunecker et al., Nature, 331:84-86, 1988; Zheng et al., J. Immunol., 154:5590-5600, 1995; Vil et al., Proc. Natl. Academic. Sci. USA, 89:11337-11341, 1992).

[0298] Fusion proteins can be generated, for example, through the techniques of gene shuffling, motif shuffling, exon shuffling, and/or codon shuffling (collectively referred to as "DNA shuffling"). DNA shuffling can be employed to alter the activities of anti-Klotho beta antibodies as provided herein, including, for example, antibodies with higher affinities and lower dissociation rates (see, e.g., U.S. Patent Nos. 5,605,793 , 5,811,238, 5,830,721, 5,834,252, and 5,837,458; 99 , J. Mol. Biol. 287:265-76; and Lorenzo and Blasco, 1998, Biotechniques 24(2):308-313). Antibodies, or encoded antibodies, can be altered by being subjected to random mutagenesis by error-prone PCR, random nucleotide insertion, or other methods prior to recombination. A polynucleotide encoding an antibody provided herein can be recombined with one or more components, motifs, sections, parts, domains, fragments, etc., of one or more heterologous molecules.

[0299] An antibody provided herein can also be conjugated with a second antibody to form an antibody heteroconjugate, as described, for example, in US Patent No. 4,676,980.

[0300] The therapeutic group or drug conjugated or recombinantly fused to an antibody provided herein that binds to Klotho beta (e.g., a Klotho beta polypeptide, fragment, epitope) must be chosen to obtain the effect(s). s) desired prophylactic or therapeutic. In certain embodiments, the antibody is a modified antibody. A physician or other medical personnel may consider, for example, the following when deciding which therapeutic group or drug to conjugate or recombinantly fuse to an antibody provided herein: the nature of the disease, the severity of the disease, and the condition of the individual.

[0301] Antibodies that bind to Klotho beta as provided herein can also be bound to solid supports, which are particularly useful for immunoassays or purification of the target antigen. Such solid supports include, but are not limited to, glass, cellulose, polyacrylamide, nylon, polystyrene, polyvinyl chloride or polypropylene.

[0302] The linker may be a "cleavable linker" facilitating the release of the conjugating agent into the cell, but non-cleavable linkers are also contemplated here. Linkers for use in the conjugates of the present invention include, without limitation, acid labile linkers (e.g., hydrazone linkers), disulfide-containing linkers, peptidase-sensitive linkers (e.g., peptide linkers comprising amino acids, e.g., valine and /or citrulline, such as citrulline-valine or phenylalanine-lysine), photolabile linkers, dimethyl linkers (see, for example, Chari et al, Cancer Research 52: 127-131 (1992); US Patent No. 5,208,020), thioether ligands or hydrophilic ligands designed to evade transporter-mediated multidrug resistance (see, for example, Kovtun et al., Cancer Res. 70: 2528-2537, 2010).

[0303] Antibody and agent conjugates can be prepared using a variety of bifunctional protein coupling agents, such as BMPS, EMCS, GMBS, HBVS, LC-SMCC, MBS, MPBH, SBAP, SIA, SIAB, SMCC, SMPB , SMPH, sulfo-EMCS, sulfo-GMBS, sulfo-KMUS, sulfo-MBS, sulfo-SIAB, sulfo-SMCC, and sulfo-SMPB, and SVSB (succinimidyl-(4-vinylsulfone)benzoate). The present disclosure further contemplates that conjugates of antibodies and agents may be prepared using any suitable methods as disclosed in the art, (see, for example, in Bioconjugate Techniques, 2nd Ed., G.T. Hermanson, ed., Elsevier, San Francisco, 2008).

[0304] Conventional conjugation strategies for antibodies and agents have been based on random conjugation chemistries involving the ε-amino group of Lys residues or the thiol group of Cys residues, which results in heterogeneous conjugates. Recently developed techniques allow site-specific conjugation to antibodies, resulting in homogeneous loading and avoiding conjugated subpopulations with antigen binding or altered pharmacokinetics. These include engineering "TioMab" comprising cysteine substitutions at positions in the heavy and light chains that provide reactive thiol groups and do not disrupt immunoglobulin folding and assembly or alter antigen binding (see, e.g., Junutula et al., J . Immunol. Meth. 332: 41-52 (2008); In another method, selenocysteine is co-translationally inserted into an antibody sequence by recoding the UGA stop codon from the end to the selenocysteine insertion, allowing site-specific covalent conjugation to the nucleophilic selenol group of selenocysteine in the presence of the other amino acids. natural (see, for example, Hofer et al., Proc. Natl. Acad. Sci. USA 105: 12451-12456 (2008); Hofer et al., Biochemistry 48(50): 12047-12057, 2009).

Pharmaceutical Formulations

[0305] Anti-Klotho beta antibodies of the present invention can be administered by any route appropriate to the condition being treated. The antibody will typically be administered parenterally, for example, by infusion, subcutaneously, intramuscularly, intravenously, intradermally, intrathecally and epidurally. The dose of antibodies may vary, including depending on the nature and/or severity of the disease, as well as the individual's condition, may include doses between 1 mg and 100 mg. Doses may also include those between 1 mg/kg and 15 mg/kg. In some embodiments, the dose is between about 5 mg/kg and about 7.5 mg/kg. In some embodiments, the dose is about 5 mg/kg. In some embodiments, the dose is about 7.5 mg/kg. Fixed doses selected from the group consisting of: (a) 375-400 mg every two weeks and (b) 550-600 mg every three weeks. In some embodiments, the fixed dose is 375-400 mg every two weeks. In some embodiments, the fixed dose is 550-600 mg every three weeks. In some embodiments, the fixed dose is 400 mg every two weeks. In some embodiments, the fixed dose is 600 mg every three weeks. In some sequential dosing embodiments, a first dose and a second dose are each between 1 mg/kg and 15 mg/kg, with the second dose following the first by between 1 and 4 weeks. In some embodiments, the first dose and the second dose are each between 5 mg/kg and 7.5 mg/kg and the second dose follows the first dose by between 2 and 3 weeks. In some embodiments, the first dose and the second dose are each 5 mg/kg and the second dose follows the first dose by 2 weeks. In some embodiments, the first dose and the second dose are each 7.5 mg/kg and the second dose follows the first dose by 3 weeks.

[0306] For the treatment of diseases, disorders and conditions, the antibody in some embodiments is administered via intravenous infusion. The dose administered via infusion is in the range of about 1 μg/m2 and about 10,000 μg/m2 per dose, generally one dose per week for a total of one, two, three, or four doses. Alternatively, the dosage range is about 1 μg/m2 to about 1000 μg/m2, about 1 μg/m2 to about 800 μg/m2, about 1 μg/m2 to about 600 μg/m2, about 1 μg/m2 to about 400 μg/m2; Alternatively, about 10 μg/m2 to about 500 μg/m2, about 10 μg/m2 to about 300 μg/m2, about 10 μg/m2 to about 200 μg/m2 and about 1 μg/m2 at about 200 μg/m2. The dose may be administered once a day, once a week, several times a week but less than once a day, several times a month but less than once a day, several times a month but less than than once a week, once a month or intermittently to alleviate or lessen the symptoms of the disease, disorder or condition. Administration may continue at any of the intervals described until improvement of the disease, disorder or condition, or improvement of the symptoms of the disease, disorder or condition being treated. Administration may continue after remission or relief of symptoms is achieved when such remission or relief is prolonged by said continued administration.

[0307] In one aspect, the present invention further provides pharmaceutical formulations comprising at least one anti-Klotho beta antibody of the present disclosure. In some embodiments, a pharmaceutical formulation comprises 1) an anti-Klotho beta antibody, and 2) a pharmaceutically acceptable carrier. In some embodiments, a pharmaceutical formulation comprises 1) an anti-Klotho beta antibody and/or an immunoconjugate thereof and, optionally, 2) at least one additional therapeutic agent.

[0308] Pharmaceutical formulations comprising an antibody are prepared for storage by mixing the antibody having the desired degree of purity with optional physiologically acceptable carriers, excipients or stabilizers, (see, for example, Remington's Pharmaceutical Sciences 16th edition, Osol, A. Ed. (1980)) in the form of aqueous solutions or freeze-dried or other dry formulations. The formulations of this invention may also contain more than one active compound as necessary for the particular indication being treated, preferably those with complementary activities that do not adversely affect each other. For example, in addition to an Anti-Klotho beta antibody, it may be desirable to include in the formulation, an additional antibody, e.g., a second Anti-Klotho beta antibody that binds to a different epitope on the Klotho beta polypeptide, or an antibody to another target. Alternatively, or additionally, the composition may further comprise another agent, including, for example, a chemotherapeutic agent, a cytotoxic agent, cytokine, growth inhibitory agent, antihormonal and/or cardioprotective agent. In some embodiments, the formulation includes an alkylating agent (e.g., chlorambucil, bendamustine hydrochloride, or cyclophosphamide), a nucleoside analog (e.g., fludurabine, pentostatin, cladribine, or cytarabine) a corticosteroid (e.g., prednisone, prednisolone, or methylprednisolone). ), an immunomodulatory agent (e.g., lenalidomide), an antibiotic (e.g., doxorubicin, daunorubicin, idarubicin, or mitoxentron), a synthetic flavon (e.g., flavopiridol), a Bcl2 antagonist, (e.g., oblimersen or ABT-263 ), a hypomethylating agent (e.g., azacitidine or decitabine), a FLT3 inhibitor (e.g., midostaurin, sorafenib, and AC220). Such molecules are suitably present in combination in amounts that are effective for the intended purpose.

[0309] The antibodies of the present invention can be formulated in any form suitable for delivery to a target cell/tissue, for example, as microcapsules or macroemulsions (Remington's Pharmaceutical Sciences, 16th edition, Osol, A. Ed. (1980); Park et al., Molecules 10: 146161 (2005); Malik et al., Curr. as sustained release formulations (Putney and Burke, Nature Biotechnol. 16: 153-157, (1998)) or in liposomes (Maclean et al., Int. J. Oncol. 11: 235-332 (1997); Kontermann, Curr Opin. Mol. Ther. 8: 39-45 (2006)).

[0310] An antibody provided herein can also be entrapped in a microcapsule prepared, for example, by coacervation techniques or by interfacial polymerization, for example, hydroxymethylcellulose or gelatin microcapsule and poly(methylmethacrylate) microcapsule, respectively, in delivery systems of colloidal drugs (e.g. liposomes, albumin microspheres, microemulsions, nanoparticles and nanocapsules) or in macroemulsions. Such techniques are described, for example, in Remington's Pharmaceutical Sciences (1990) Mack Publishing Co., Easton, PA.

[0311] Various delivery systems are known and can be used to administer a prophylactic or therapeutic agent (e.g., an antibody that binds to Klotho beta as described herein), including, but not limited to, encapsulation in liposomes, microparticles , microcapsules, recombinant cells capable of expressing the antibody, receptor-mediated endocytosis (see, e.g., Wu and Wu, J. Biol. Chem. 262:4429-4432 (1987)), construction of a nucleic acid as part of a retroviral vector or other, etc. In another embodiment, a prophylactic or therapeutic agent, or a composition provided herein may be delivered in a controlled-release or sustained-release system. In one embodiment, a pump may be used to achieve controlled or sustained release (see, e.g., Langer, supra; Sefton, 1987, CRC Crit. Ref. Biomed. Eng. 14:20; Buchwald et al., 1980, Surgery 88:507; Saudek et al., 1989, N. J. Med. 321:574). In another embodiment, polymeric materials can be used to achieve controlled or sustained release of a prophylactic or therapeutic agent (e.g., an antibody that binds to Klotho beta as described herein) or a composition of the invention (see, e.g., Medical Applications of Controlled Release, Langer and Wise (eds.), CRC Pres., Boca Raton, Florida (1974); Controlled Drug Bioavailability, Drug Product Design and Performance, Smolen and Ball (eds.), Wiley, New York (1984) ;Ranger and Peppas, 1983, J., Sci. Rev. Macromol., 1989, Ann. :351; Howard et al., 1989, J. Neurosurg 7 1:105); US Patent No. 5,679,377; US Patent No. 5,916,597; US Patent No. 5,912,015; US Patent No. 5,989,463; US Patent No. 5,128,326; PCT Publication No. WO 99/15154; and PCT Publication No. WO 99/20253). Examples of polymers used in sustained release formulations include, but are not limited to, poly(2-hydroxy ethyl methacrylate), poly(methyl methacrylate), poly(acrylic acid), poly(ethylene-co-vinyl acetate), poly (methacrylic acid), polyglycolides (PLG), polyanhydrides, poly(N-vinyl pyrrolidone), poly(vinyl alcohol), polyacrylamide, poly(ethylene glycol), polylactides (PLA), poly(lactide-co-glycolide) (PLGA ), and polyorthoesters. In one embodiment, the polymer used in a sustained release formulation is inert, free of leachable impurities, storage stable, sterile, and biodegradable.

[0312] In yet another embodiment, a controlled or sustained release system can be placed in proximity to the therapeutic target, e.g., the nasal passages or lungs, thus requiring only a fraction of the systemic dose (see, e.g., Goodson, at Medical Applications of Controlled Release, supra, vol. 2, pp. 115-138 (1984)). Controlled release systems are discussed, for example, by Langer (1990, Science 249:1527-1533). Any technique known to one skilled in the art can be used to produce sustained release formulations comprising one or more antibodies that bind to Klotho beta as described herein. (See, for example, U.S. Patent No. 4,526,938, PCT Publication WO 91/05548, PCT Publication WO 96/20698, Ning et al., 1996, “Intratumoral Radioimmunotherapy of a Human Colon Cancer Xenograft Using a Sustained-Release Gel, ” Radiotherapy & Oncology 39:179- 189, Song et al., 1995, “Antibody Mediated Lung Targeting of Long-Circulating Emulsions,” PDA Journal of Pharmaceutical Science & Technology 50:372-397, Cleek et al., 1997, “ Biodegradable Polymeric Carriers for a bFGF Antibody for Cardiovascular Application,” Pro. Int'l. Rel. Bioact. Local Delivery,” Int’l Symp. Bioact.

Therapeutic methods

[0313] An antibody of the present invention can be used in, for example, in vitro, ex vivo and in vivo therapeutic methods. In one aspect, the present invention provides methods for treating or preventing a disease, disorder or condition, whether in vivo or in vitro, the method comprising exposing a cell to an anti-Klotho beta antibody.

[0314] In one aspect, an antibody of the present disclosure is used to treat or prevent a disease, disorder or condition, including, for example, type 2 diabetes, obesity, dyslipidemia, NASH, cardiovascular disease, metabolic syndrome or broadly any disease, disorder or condition in which it is desirable to mimic or enhance the in vivo effects of FGF19 and/or FGF21.

[0315] In one aspect, methods are provided for treating a disease, disorder or condition comprising administering to an individual an effective amount of an anti-Klotho beta antibody or a fragment thereof. In certain embodiments, a method for treating a disease, disorder or condition comprises administering to an individual an effective amount of a pharmaceutical formulation comprising an Anti-Klotho beta antibody and, optionally, at least one additional therapeutic agent, such as those described here.

[0316] An anti-Klotho beta antibody or a fragment thereof can be administered to a human for therapeutic purposes. Additionally, an anti-Klotho beta antibody or fragment thereof can be administered to a non-human Klotho beta-expressing mammal with which the antibody cross-reacts (e.g., a primate, pig, rat, or mouse) for purposes veterinarians, or as an animal model of human disease. With respect to the latter, these animal models may be useful for evaluating the therapeutic efficacy of the antibodies of the present invention (e.g., dose testing and administration time courses).

[0317] The antibodies of the present disclosure can be used alone or in combination with other compositions in a therapy. For example, an anti-Klotho beta antibody of the present invention can be co-administered with at least one additional therapeutic agent and/or adjuvant. In some embodiments, the additional compound is a therapeutic antibody other than an anti-Klotho beta antibody.

[0318] Such combination therapies mentioned above encompass combined administration (in which two or more therapeutic agents are included in the same or separate formulations), and separate administration, in which case, the administration of an Anti-Klotho beta antibody or a fragment thereof of the present disclosure may occur before, simultaneously, and/or after, the administration of the agent and/or an additional therapeutic adjuvant. The antibodies of the present disclosure may also be used in combination with additional therapeutic regimens, including, without limitation, those described herein.

[0319] An antibody of the present invention (and any additional therapeutic agent or an adjuvant) can be administered by any suitable means, including parenteral, subcutaneous, intraperitoneal, intrapulmonary and intranasal administration, and, if desired for local treatment, intralesional. Parenteral infusions include intramuscular, intravenous, intra-arterial, intraperitoneal, or subcutaneous administration. Furthermore, the antibody or conjugate is suitably administered by pulse infusion, in particular with decreasing doses of the antibody or fragment thereof. Dosage may be by any suitable route, for example, by injections, such as intravenous or subcutaneous injections, depending in part on whether administration is brief or chronic.

[0320] The antibodies of the present disclosure could be formulated, dosed, and administered in a manner consistent with good medical practice. Factors for consideration in this context include the particular disorder being treated, the particular mammal being treated, the clinical condition of the individual patient, the cause of the disorder, the site of delivery of the agent, the method of administration, the schedule of administration, and other factors known to medical practitioners. The anti-Klotho beta antibody need not be, but is optionally formulated with, one or more agents currently used to prevent or treat the disorder in question. The effective amount of such other agents depends on the amount of antibody or immunoconjugate present in the formulation, the type of disorder or treatment, and other factors discussed above. These are generally used in the same dosages and with routes of administration as described herein, or about 1 to 99% of the doses described herein or in any dose and by any route that is empirically/clinically determined to be suitable.

[0321] For the prevention or treatment of a disease, disorder or condition, the appropriate dosage of an Anti-Klotho beta antibody of the present invention (when used alone or in combination with one or more other additional therapeutic agents, such as agents described herein ) will depend on the type of disease, disorder or condition being treated, the type of antibody, the severity and course of the disease, disorder or condition, whether the antibody is administered for preventive or therapeutic purposes, prior therapy, the patient's clinical history and the antibody response, and the discretion of the attending physician. The anti-Klotho beta antibody is appropriately administered to the patient all at once or over a series of treatments. Depending on the type and severity of the disease, about 1 μg/kg to 100 mg/kg (e.g. 0.1 mg/kg - 20 mg/kg, 1 mg/kg - 15 mg/kg, etc.) of antibody may be an initial dosage candidate for administration to the patient, whether, for example, by one or more separate administrations, or by continuous infusion. A typical daily dosage can range from about 1 μg/kg to 100 mg/kg or more, depending on the factors mentioned above. For repeated administrations over several days or more, depending on the condition, treatment would generally be continued until a desired suppression of disease symptoms occurs. Exemplary antibody dosages may range from about 0.05 mg/kg to about 10.0 mg/kg. Thus, one or more doses of about 0.5 mg/kg, 1.0 mg/kg, 2.0 mg/kg, 3.0 mg/kg, 4.0 mg/kg, 5.0 mg/kg , 6.0 mg/kg, 7.0 mg/kg, 8.0 mg/kg, 9.0 mg/kg, or 10.0 mg/kg (or any combination thereof) of antibody may be administered to the patient . Such doses may be administered intermittently, for example, every week or every three weeks (e.g., such that the patient receives from about two to about twenty, or, for example, about six doses of the antibody ). An initial higher loading dose, followed by one or more low doses, may be administered. An exemplary dosage regimen comprises administering an initial loading dose followed by a maintenance dose (e.g., weekly) of the antibody. The initial loading dose may be higher than the maintenance dose. However, other dosage regimens may be useful. The progress of this therapy is easily monitored by conventional techniques and assays.

Diagnostic methods and detection methods

[0322] In one aspect, the anti-Klotho beta antibodies and fragments thereof of the present invention are useful for detecting the presence of Klotho beta in a biological sample. Such anti-Klotho beta antibodies may include those that bind human and/or Cyno Klotho beta, but do not induce FGF19-like signaling activity and/or FGF21-like signaling activity. The term "detection" as used herein encompasses qualitative or quantitative detection. In certain embodiments, a biological sample comprises a cell or tissue.

[0323] In one aspect, the present invention provides a method of detecting the presence of Klotho beta in a biological sample. In certain embodiments, the method comprises contacting the biological sample with an Anti-Klotho beta antibody under conditions permissive for binding of the Anti-Klotho beta antibody to Klotho beta, and detecting whether a complex is formed between the Anti-Klotho beta antibody and Klotho beta.

[0324] In one aspect, the present invention provides a method of diagnosing a disorder associated with the expression of Klotho beta. In certain embodiments, the method comprises contacting a test cell with an anti-Klotho beta antibody; determining the level of expression (either quantitatively or qualitatively) of Klotho beta by the test cell by detecting binding of the anti-Klotho beta antibody to Klotho beta; and comparing the level of Klotho beta expression by the test cell with the level of Klotho beta expression by a control cell (e.g., a normal cell from the same tissue of origin as the test cell, or a cell expressing Klotho beta at levels comparable to such a normal cell), wherein a higher level of Klotho beta expression by the test cell compared to the control cell indicates the presence of a disease associated with increased Klotho beta expression. In certain embodiments, the test cell is obtained from an individual suspected of having a disease, disorder or condition associated with Klotho beta expression and/or a disease, disorder or condition in which it is desirable to mimic or enhance the effects of Klotho beta. live of FGF19 and/or FGF21. In certain embodiments, the disease, disorder or condition is, for example, type 2 diabetes, obesity, dyslipidemia, NASH, cardiovascular disease or metabolic syndrome. Such exemplary diseases, disorders or conditions can be diagnosed using an anti-Klotho beta antibody of the present disclosure.

[0325] In certain embodiments, a diagnostic or detection method, such as those described above, comprises detecting the binding of an anti-Klotho beta antibody to Klotho beta expressed on the surface of a cell or in a membrane preparation obtained from of a cell that expresses Klotho beta on its surface. In certain embodiments, the method comprises contacting a cell with an anti-Klotho beta antibody under conditions permissive for binding of the anti-Klotho beta antibody to Klotho beta, and detecting whether a complex is formed between the anti-Klotho beta antibody and Klotho beta on the cell surface. An exemplary assay for detecting the binding of an anti-Klotho beta antibody to Klotho beta expressed on the surface of a cell is a "FACS" assay.

[0326] Certain other methods can be used to detect the binding of anti-Klotho beta antibodies to Klotho beta. Such methods include, but are not limited to, antigen binding assays that are well known in the art, such as western blots, radioimmunoassays, ELISA (enzyme-linked immunosorbent assay), sandwich immunoassays, immunoprecipitation assays, fluorescent stains, protein A immunoassays, and immunohistochemistry (IHC).

[0327] In certain embodiments, anti-Klotho beta antibodies are labeled. Tags include, but are not limited to, tags or groups that are detected directly (e.g., fluorescent, chromophoric, electron-dense, chemiluminescent, and radioactive tags), as well as groups, such as enzymes or ligands, that are detected indirectly. , for example, through an enzymatic reaction or molecular interaction. Exemplary labels include, but are not limited to, radioisotopes 32P, 14C, 125I, 3H, and 131I, fluorophores such as rare earth chelates or fluorescein and its derivatives, rhodamine and its derivatives, dansyl, umbelliferone, luceriferases, e.g. firefly and bacterial luciferase (see, e.g., U.S. Pat. No. 4,737,456), luciferin, 2,3-dihydrophthalazinediones, horseradish peroxidase (HRP), alkaline phosphatase, β-galactosidase, glucoamylase, lysozyme, saccharide oxidases, e.g. , glucose oxidase, galactose oxidase, and glucose-6-phosphate dehydrogenase, heterocyclic oxidases such as uricase and xanthine oxidase, coupled with an enzyme that uses hydrogen peroxide to oxidize a dye precursor such as HRP, lactoperoxidase, or microperoxidase, biotin /avidin, spin tags, bacteriophage tags, stable free radicals, and the like.

[0328] In certain embodiments, anti-Klotho beta antibodies are immobilized in an insoluble matrix. Immobilization involves separating the anti-Klotho beta antibody from any Klotho beta that remains free in solution. This is conventionally achieved either by insolubilization of the Anti-Klotho beta antibody prior to the assay procedure, by adsorption to a water-insoluble matrix or surface (see, e.g., Bennich et al., US 3,720,760), or by covalent coupling (e.g. example, using cross-linking with glutaraldehyde), or by insolubilization of the anti-Klotho beta antibody after the formation of a complex between the anti-Klotho beta antibody and Klotho beta, for example, by immunoprecipitation.

[0329] Any of the above diagnostic or detection embodiments can be performed using an immunoconjugate of the present disclosure, in place of, or in addition to, an anti-Klotho beta antibody.

Essay

[0330] The anti-Klotho beta antibodies of the present invention can be characterized by their physical/chemical properties and/or biological activities by various assays known in the art.

1. Activity trials

[0331] In one aspect, assays are provided to identify anti-Klotho beta antibodies thereof having biological activity. Biological activity may include, for example, assays that measure effects on glucose and/or lipid metabolism. For example, a blood glucose assay may be used. Blood glucose (e.g., in a rat tail clipping or a human blood sample) can be measured using ACCU-CHEK Active Assay Strips read by an Active Accu-Chek Meter (Roche Diagnostics, Indianapolis, IN), following the manufacturer's instructions. Additionally, for example, a lipid profile assay may be used. Whole blood (e.g., from rat tail clippings or from a human blood sample) can be collected in simple capillary tubes (BD Clay Adams SurePrep, Becton Dickenson and Co. Sparks, MD). Serum and blood cells can be separated by centrifuging the tubes in an Autocrit Ultra 3 (Becton Dickinson and Co. Sparks, MD). Serum samples can be analyzed for lipid profile (triglycerides, total cholesterol, HDL, and non-HDL) using the Integra 400 Clinical Analyzer (Roche Diagnostics, Indianapolis, IN) following the manufacturer's instructions.

2. Binding assays and other assays

[0332] In one aspect, an anti-Klotho beta antibody is tested for its antigen binding activity. For example, in certain embodiments, an anti-Klotho beta antibody is tested for its ability to bind exogenous or endogenous Klotho beta expressed on the surface of a cell. A FACS assay can be used for such tests.

[0333] A panel of monoclonal antibodies raised against Klotho beta can be grouped based on the epitopes they recognize, a process known as epitope binning. Epitope binning is typically performed using competition assays, which evaluate the ability of an antibody to bind to an antigen in the presence of another antibody. In an exemplary competition assay, immobilized Klotho beta is incubated in a solution comprising a first labeled antibody that binds to Klotho beta and a second unlabeled antibody that is being tested for its ability to compete with the first antibody to bind to Klotho beta. . The second antibody may be present in a hybridoma supernatant. As a control, the immobilized Klotho beta is incubated in a solution comprising the labeled first antibody but not the unlabeled second antibody. After incubation under conditions permissive for binding of the first antibody to Klotho beta, excess unbound antibody is removed, and the amount of label associated with the immobilized Klotho beta is measured. If the amount of label associated with immobilized Klotho beta is substantially reduced in the test sample relative to the control sample, then this indicates that the second antibody competes with the first antibody to bind to Klotho beta. In certain embodiments, the immobilized Klotho beta is present on the surface of a cell or in a membrane preparation obtained from a cell that expresses Klotho beta on its surface.

[0334] High-throughput epitope binning methods are also known in the art (see, for example, Jia et al, J. Immunol Methods 2004, 288 (1-2):91-98, describing a binning method of competitive multiplexing antibody for the characterization of monoclonal antibodies; and Miller et al, J. Immunol Methods 2011, 365 (1-2):118-25, describing epitope binning of murine monoclonal antibodies by a multiplexed matching assay) .

3. Epitope Mapping

[0335] Epitope mapping is the process of identifying the binding sites, or epitopes of an antibody, on its target protein antigen. Antibody epitopes can be linear epitopes or conformational epitopes. Linear epitopes are formed by a continuous sequence of amino acids in a protein. Conformational epitopes are formed from amino acids that are discontinuous in the protein sequence, but are brought together by folding the protein into its three-dimensional structure.

[0336] A variety of methods are known in the art for mapping antibody epitopes onto target protein antigens. These include mutagenesis methods, peptide tracking methods, display methods, methods involving mass spectroscopy, and structural determination.

[0337] The site-directed mutagenesis method involves target site-directed mutagenesis where critical amino acids are identified by systematically introducing substitutions along the protein sequence and then determining the effects of each substitution on antibody binding . This can be done by "alanine scanning mutagenesis", as described, for example, by Cunningham and Wells (1989) Science 244: 1081-1085, or some other form of point mutagenesis of amino acid residues of human Klotho beta. Mutagenesis studies, however, may also reveal amino acid residues that are crucial to the overall three-dimensional structure of Klotho beta but which are not directly involved in antibody-antigen contacts, and therefore other methods may be needed to confirm a functional epitope determined using this method.

[0338] Shotgun mutagenesis mapping utilizes a complete plasmid-mutation library for the target gene, with each of the clones in the library having a single amino acid mutation and the entire library covering every amino acid in the target protein. The clones that constitute the mutation library are individually arranged in microplates, expressed within living mammalian cells, and tested for immunoreactivity with the antibodies of interest. Critical amino acids for antibody epitopes are identified by a loss of reactivity and are then mapped to a protein structure to visualize epitopes. By automating the analysis, new epitope maps can be derived within days or weeks. Because it uses the native structure of proteins within mammalian cells, the technique allows both conformational and linear epitope structures to be mapped onto complex proteins. (See, for example, Paes et al., J. Am. Chem. Soc. 131(20): 6952-6954 (2009); Banik and Doranz, Genetic Engineering and Biotechnology News 3(2): 25-28 (2010 ).

[0339] The epitope bound by an anti-Klotho beta antibody can also be determined using peptide tracking methods. In peptide screening, libraries of short peptide sequences from overlapping segments of the target protein, Klotho beta, are tested for their ability to bind antibodies of interest. Peptides are synthesized and screened for binding, for example, by means of ELISA or BIAcore, or on a chip, by any of several methods for solid phase screening (see, for example, Reineke et al, Curr. Opin. Biotechnol. 12: 59-64, 2001) as in the "pepscan" method (see, for example, WO 84/03564; WO 93/09872). Such peptide screening methods may not be able to detect some functional epitopes, that is, functional discontinuous epitopes involving amino acid residues that are not contiguous along the primary sequence of the Klotho beta polypeptide chain.

[0340] A recently developed technology called CLIPS (chemical binding of peptides to scaffolds) can be used to map conformational epitopes. The loose ends of the peptides are affixed onto synthetic scaffolds, so that the "scaffolded" peptide may be able to adopt the same spatial structure as the corresponding sequence in the intact protein. CLIPS technology is used to fix linear peptides into cyclic structures ("single loop" shape), and to bring together different parts of a protein binding site ("double loop", "triple loop" shape, etc.), as well as to create conformational epitopes that can be assayed for antibody binding (see, e.g., U.S. Pat. No. 7,972,993).

[0341] The antibody-bound epitopes of the present disclosure can also be mapped using display techniques, including, for example, phage display, microbial display, and ribosome/mRNA display as described above. In these methods, libraries of peptide fragments are displayed on the surface of the phage or cells. The epitopes are then mapped by screening mAbs against these fragments using selective binding assays. A number of computational tools have been developed that allow the prediction of conformational epitopes based on linear affinity-selected peptides obtained using phage display (see, for example, Mayrose et al, Bioinformatics 23:32443246, 2007). Methods are also available for the detection of conformational epitopes by phage display. Microbial visualization systems can also be used to express correctly folded antigenic fragments on the cell surface for the identification of conformational epitopes (see, for example, Cochran et al., J. Immunol. Meth. 287: 147-158, 2004; Rockberg et al., Nature Methods 5: 1039-1045, 2008).

[0342] Methods involving proteolysis and mass spectroscopy can also be used to determine antibody epitopes (see, for example, Baerga-Ortiz et al, Protein. Sci. 2002 June; 11 (6): 1300-1308). In limited proteolysis, the antigen is cleaved by different proteases in the presence and absence of the antibody, and the fragments are identified by mass spectrometry. The epitope is the region of the antigen that is protected against proteolysis upon antibody binding (see, for example, Suckau et al., Proc. Natl. Acad. Sci. USA 87:9848-9852, 1990). Additional methods based on proteolysis include, for example, selective chemical modification (see, e.g., Fiedler et al, Bioconjugate Chemistry 1998, 9(2):236-234, 1998), epitope excision (see, e.g., van de Water et al, Clin Immunol Immunopathol 1997, 85 (3):229-235, 1997), and the recently developed hydrogen-deuterium (H/D) exchange method (see, e.g., Flanagan, N., Genetic Engineering and Biotechnology News 3(2): 25-28, 2010).

[0343] The antibody-bound epitope of the present disclosure can also be determined by structural methods, such as X-ray crystal structure determination (see, for example, WO 2005/044853), molecular modeling and nuclear magnetic resonance spectroscopy (NMR), including NMR determination of the H-D exchange rates of labile amide hydrogens when free and when bound in a complex with an antibody of interest (see, e.g., Zinn-Justin et al. (1992) Biochemistry 31:11335 -11347; Zinn-Justin et al. (1993) Biochemistry 32:6884-6891).

[0344] Additional antibodies that bind to the same epitope as an antibody of the present invention can be obtained, for example, by screening antibodies raised against Klotho beta for binding to the epitope, by immunizing an animal with a peptide comprising a fragment of human Klotho beta comprising the epitope sequence, or by selection of antibodies using phage display for binding to the epitope sequence. Antibodies that bind to the same functional epitope can be expected to exhibit similar biological activities, such as blocking a Klotho beta biological activity, and these activities can be confirmed by functional assays of the antibodies.

Additional Activity Trials

[0345] In one embodiment, an anti-Klotho beta antibody of the present disclosure is an antagonist antibody that inhibits a biological activity of Klotho beta. The anti-Klotho beta antibodies of the present disclosure can be tested to determine whether they inhibit the biological activity of Klotho beta.

[0346] In one aspect, purified Anti-Klotho beta antibodies can be further characterized by a series of assays including, but not limited to, N-terminal sequencing, amino acid analysis, high pressure size exclusion liquid chromatography and -denaturing (HPLC), mass spectrometry, ion exchange chromatography and papain digestion.

[0347] In one embodiment, the present invention contemplates an altered antibody that has some, but not all, effector functions, which makes it a desirable candidate for many applications in which the half-life of the antibody in vivo is important, even though certain functions effectors (such as complement and ADCC) are unnecessary or deleterious. In certain embodiments, the Fc activities of the antibody are measured to ensure that only the desired properties are maintained. In vitro and/or in vivo cytotoxicity assays can be performed to confirm the reduction/depletion of CDC and/or ADCC activities. For example, Fc receptor (FcR) binding assays can be performed to ensure that the antibody lacks FcyR binding (thus likely lacking ADCC activity) but retains FcRn binding capacity. An in vitro assay for evaluating the ADCC activity of a molecule of interest is described, for example, in US Patent No. 5,500,362 or 5,821,337. Useful effector cells for such assays include peripheral blood mononuclear cells (PBMC) and natural killer (NK) cells. Alternatively, or additionally, the ADCC activity of the molecule of interest can be evaluated in vivo, for example, in an animal model such as that disclosed in Clynes et al. PNAS (USA) 95:652-656 (1998). C1q binding assays can also be performed to confirm that the antibody is capable of binding to C1q and therefore lacks CDC activity. To assess complement activation, a CDC assay, for example, as described in Gazzano-Santoro et al., J. Immunol. Methods 202: 163 (1996), can be accomplished. FcRn binding and in vivo clearance/half-life determinations can also be performed using methods known in the art.

[0348] Although the present foregoing description has been described in some detail by way of illustration and example for purposes of clarity of understanding, the descriptions and examples should not be construed as limiting the scope of the present disclosure. Disclosures of all patent and scientific literature cited herein are expressly incorporated in their entirety by reference.

EXAMPLES

[0349] The following are examples of methods and compositions of the present disclosure.

EXAMPLE 1: GENERATION OF ANTIBODIES TO KLOTHO BETA

[0350] Antibodies to Klotho beta were generated, for example, by immunizations of mice (i) with cells expressing human Klotho beta (HuKLB) and FGF receptor 1c (FGFRIc or RIc) and (ii) with HuKLB and Klotho protein beta from cynomologists (KLB from cyno).

[0351] For example, cells expressing Klotho beta were prepared as follows. 293EXPI cells (Invitrogen) were transiently co-transfected with nucleic acid sequences encoding a variant of FGFR1c with a mutation at amino acid position 623 (see, for example, SEQ ID NO: 308, but with a D623N mutation) and HuKLB ( SEQ ID NO: 297). Cells were analyzed for expression of R1c and HuKLB by respective specific antibodies by FACS. Cells were washed twice in PBS, pelleted by centrifugation and frozen in individual vials at 6 x 107 cells for immunization. 129/B6 animals were immunized with 1 x 107 cells with adjuvants (Ribi, CpG, and PolyIC). Animals were boosted every 2 weeks for the duration necessary to induce an adequate titer. Animals were boosted with HuKLB and CyKLB protein after 4 boosts with 293EXPI cells overexpressing R1C and HuKLB. Titers were determined by ELISA and FACS. Single cell suspensions of lymphocytes were obtained from the spleen and draining lymph nodes of animals with appropriate titers. The cells were fused with SP2/0 myeloma cells at a ratio of 1:2 by electrofusion. The fused cells were plated at 2.5 x 106 cells per plate in 70 μL in twenty-four 384-well plates in the presence of HAT selection. After 7 days, 50 μL of supernatant was removed and replaced with fresh medium containing HAT. After 10-14 days of culture, supernatants were collected and subjected to screening by FACS using 293EXPI cells overexpressing R1c and HuKLB or by Biacore using HuKLB protein to confirm binding. Positive clones were further selected and subjected to subcloning.

[0352] In a first campaign of immunizations and fusions, at least 25-30 384-well plates were screened for binding to HuKLB (e.g., HuKLB proteins and/or HuKLB-expressing cells). In a second campaign for immunizations and mergers, a similar number of plates were tracked as described for the first campaign. Thousands of clones were screened and hundreds of clones were selected for further study, including in binding, affinity, and epitope specificity assays, as described in Examples 2 and 3. Hundreds of hybridoma supernatants were also tested in functional assays, such as described in examples 4 and 5, including FGF receptor-like agonist activity, FGF19 ligand and/or FGF21 (e.g., FGF19-like and/or FGF21-like signaling activity).

EXAMPLE 2: SCREENING AND SELECTION OF ANTIBODIES FOR KLOTHO BETA

[0353] Antibodies to Klotho beta were generated from hybridomas, for example, as described in Example 1. Hybridoma supernatants were screened for binding to Klotho beta (e.g., human and/or Cyno Klotho beta). in FACS and/or Biacore-based assays.

[0354] For example, after 2 weeks of culture, hybridoma supernatants were screened for monoclonal antibodies that bind to human Klotho beta by a FACS-based binding scan. Briefly, hybridoma supernatants were co-incubated with cells overexpressing human Klotho beta for 30 minutes at 4°C. After washing with PBS/1% BSA/0.1% azide, cells overexpressing human Klotho beta were co-incubated with labeled mouse anti-Fc (Jackson Immunoresearch) for 30 minutes at 4°C. After washing with PBS/1% BSA/0.1% azide, cells were acquired on flow cytometer (FACS Calibur) and analyzed by cytometry analysis software (FlowJo). A binding antibody is one that shows displacement of cells incubated with anti-mouse Fc antibodies only.

[0355] For example, after 2 weeks of culture, hybridoma supernatants were screened for monoclonal antibodies that bind to human Klotho beta by a Biacore-based binding screen. Briefly, mouse anti-Fc antibody (Sigma-Aldrich, St. Louis, MO) was immobilized on all four flow cells of a CM5 chip using amine coupling reagents (GE Healthcare Life Sciences, Piscataway, NJ). Hybridoma supernatants were diluted three times with PBS-P buffer (PBS containing 0.005% P20) and injected for 30 seconds into flow cells 2, 3, and 4 to capture antibody (flow cell 1 was used as a reference ). This was followed by a short injection of human Klotho beta (25 nM, R&D Systems, Minneapolis, MN) for 60 seconds at a flow rate of 30 μL/min to test for binding to the antibody captured on each flow cell.

[0356] From two immunization and fusion campaigns as described in Example 1, fifty to sixty 384-well plates of hybridoma supernatants were assayed for binding by FACS and/or Biacore. From these assays, approximately 250 antibodies were identified as ligands for human Klotho beta. These antibodies were purified and subsequently tested for their binding affinity for human Klotho beta and cyno Klotho beta by Biacore and for their functional activity by reporter assays as described in Example 3.

[0357] In additional Biacore/screening-based binding assays, the binding affinity of antibodies to human Klotho beta and Cyno was measured. For example, antibodies were ranked based on their binding affinity to human Klotho beta and cyno Klotho beta by low-resolution KD measurement by Biacore. Briefly, mouse anti-Fc antibody (Sigma-Aldrich, St. Louis, MO) was immobilized on all four flow cells of a CM5 chip using amine coupling reagents (GE Healthcare Life Sciences, Piscataway, NJ). Purified antibodies were captured (~100 RUs) in flow cells 2, 3, and 4, using flow cell 1 as a reference. This was followed by injection of human Klotho beta or Cyno (25 nM in PBS-P buffer) at a flow rate of 70 μL/min and monitoring binding kinetics at 25°C.

[0358] Binding affinity measurements were also made in additional Biacore-based assays. For example, dissociation equilibrium constant (KD) measurements were performed with purified antibodies to assess their binding to human Klotho beta and Cyno Klotho beta. As mentioned above, anti-mouse Fc antibody (Sigma-Aldrich, St. Louis, MO) was immobilized on all four flow cells of a CM5 chip using amine coupling reagents (GE Healthcare Life Sciences, Piscataway, NJ ). Purified antibodies were captured (~100 RUs) in flow cells 2, 3, and 4, using flow cell 1 as a reference. This was followed by injection of different concentrations of human Klotho beta or Cyno (1.56 nM to 25 nM, two-fold dilutions in PBS-P buffer) at a flow rate of 70 μL/min and the binding kinetics were evaluated at 25° C.

[0359] Representative results are presented as KD (nM) values as shown in Table 11 below. Table 11

EXAMPLE 3: SCREENING AND SELECTION OF ANTIBODIES FOR KLOTHO BETA

[0360] Antibodies that were Klotho beta, for example, as evaluated in epitope binning binding assays.

[0361] For example, for competition binding assays by FACS analysis, antibody standards were prepared that were conjugated to a fluorochrome using an A488 or A647 antibody labeling kit (Invitrogen) following the manufacturer's instructions. Dose titration of the conjugated antibody standard was evaluated using cells overexpressing HuKLB. The plateau of maximum antibody binding signal is EC = 100 and the background signal is EC = 0. FACS competition against the fluorochrome-labeled antibody was performed by preincubating cells overexpressing HuKLB with hybridoma supernatants for 15 minutes at room temperature. Without washing, an EC=10 concentration of labeled antibody standard A488 or A647 was added. EC=10 for an individual antibody was determined by 10% signal using the maximum signal as (100%) and the background signal as (0%). After 30 minutes at 4°C, the cells are washed and analyzed by FACS. In these assays, a competing antibody is one that exhibits a signal comparable to competition for 5H23. A non-competing antibody is one that shows the same signal as the labeled antibody alone. A partial competing antibody is a sample that exhibits signal between the labeled antibody alone and the background. Antibodies that show complete competition against the same standard antibody are considered to be in the same position.

[0362] In exemplary FACS competition binding experiments, the 5H23 or 3I13 antibody was used as an antibody standard for a positive control (competition antibody) or a negative control (non-competing antibody), respectively. Representative results are shown in Table 12 below presented as mean fluorescence intensity (MFI). For these experiments, the signal comparable to the labeled antibody alone is a non-competing antibody, while the signal comparable to competition for 5H23 is a competing antibody. Table 12

[0363] To further evaluate antibody binding sites on human Klotho beta, competition experiments were also created on Biacore. For example, two antibodies were immobilized on two flow cells of a CM5 chip. Human beta antibody-Klotho complexes were prepared with different antibodies (antibody concentration was titrated 0.1-50 nM, keeping the Klotho beta concentration constant at 5 nM) in a 96-well microplate and injected onto the antibody surfaces. The measured signal (Response Unit, RU) was plotted against the antibody concentration in the solution [nM]. If the antibody in solution recognized the same epitope as the antibody immobilized on the chip surface, then a decrease in RU was observed with increasing concentration of antibody in solution (demonstrating competition for the binding site on Klotho beta). However, if the antibody in solution recognized a distinct epitope relative to the immobilized antibody, an increase in RU was observed. In the latter case, the antibody-Klotho complex could bind to the surface of immobilized antibodies, leading to the observed increase in signal.

[0364] In exemplary competition binding experiments by Biacore, the 5H23 antibody competed with itself for binding to HuKLB and additional antibodies 1C17, 1D19, 2L12, 3L3, 3N20, 4P5, 5C23, 5F7 and 1G19 competed with 5H23. These antibodies were designated as members of the 5H23 epitope bin. The sequences for these epitope-related antibodies are aligned and shown in Figures 1 and 2. Figure 2 also shows the conserved amino acid sequences for the CDRs of these related antibodies.

EXAMPLE 4: FUNCTIONAL TESTS

[0365] Antibodies to Klotho beta generated, for example, as described in Example 1, were tested for their functional activity in cell-based reporter assays.

[0366] For example, ELK1-luciferase reporter assays, which measure FGFRIc/Klotho beta signaling, were performed using transiently transfected HEK293, HEK293T, or L6 cells (ATCC). The transfection plasmids consisted of two reporter plasmids Gal4-Elk1 and 5*UAS-Luc (Agilent Technologies PathDetect Elk1 trans-reporting system Cat# 219005) and the plasmids encoding human Klotho beta (GeneCopoeia Cat# EX-E1104-MO2) or Klotho beta from cynomolgus monkey (Klotho beta de cyno) and human FGFR1c (GeneCopoeia Cat# EX-A0260-MO2). In these assays, activation of the FGFR1c/Klotho beta receptor complex recombinantly expressed in cells induces intracellular signaling transduction, which subsequently leads to the phosphorylation of ERK and Elk1. Once Gal4-Elkl is phosphorylated, Gal4-Elk1 binds to the 5*UAS promoter region and turns the transcription of the luciferase reporter gene. Luciferase activity is then measured in luciferase enzyme assays.

[0367] For these experiments, the four aforementioned plasmids (e.g., 2 reporter plasmids, Klotho beta, R1c) were transfected into freshly collected cells in suspension using FuGene6 or Fugene HD transfection reagent (Promega). Cell density and amount of transfection reagent were optimized for each cell type and each batch of Fugene. The ratio of Klotho beta to FGFR1c DNA in transfection was optimized for each cell line and varied between 6:1 to 27:1. Transfected cells were seeded in 96-well plates (30000 cells/100 μl/well), or 384-well plates (7500 cells/25μl/well) in normal growth medium. After overnight incubation at 37°C, a variety of antibodies to Klotho beta were added. After 6 hours of 37°C incubation with the antibodies, an equal volume of Bright-Glo reagent (Promega) was added and the luminescence signal was read using the Enspire reader (Perkin Elmer).

[0368] Representative results using human Klotho beta and Cyno beta Klotho, transfected into HEK 293 cells, are reported as EC50 values as shown in Table 13 and Table 14, respectively, below. Table 13 *control mAB comprises SEQ ID NO: 358 and SEQ ID NO: 360 Table 14 * Control mAB comprises SEQ ID NO: 358 and SEQ ID NO: 360

[0369] Representative results, using human Klotho beta, transfected into L6 cells, are reported as EC50 values as shown in Table 15 below. Table 15

[0370] L6 cells lack endogenous receptors and are often used to investigate antibody specificity to various subtypes of transfected FGF receptors. Receptor activation through FGFR1c/Klotho beta signaling in the absence of ligand (e.g., FGF19 (e.g., SEQ ID NO: 304) or FGF21 (e.g., SEQ ID NO: 429)) by anti-Klotho beta antibodies exemplary of the present disclosure was observed with L6 cells transfected with FGFR1c (R1c), but not with L6 cells transfected with FGFR2c (R2c), FGFR3c (R3c), or FGFR4 (R4), whereas activation by controlling FGF19 was observed with L6 cells transfected with R1c, R2c, R3c and R4.

EXAMPLE 5: ADDITIONAL FUNCTIONAL TESTS

[0371] Antibodies to Klotho beta generated, for example, as described in Example 1, were tested for their functional activity in a cell-based assay, such as an adipocyte assay, which measures FGFR1/Klotho beta signaling. endogenous. FGF19 or FGF21 stimulate ERK phosphorylation, increase glucose uptake and lipolysis in cultured adipocytes. Adipocytes are considered physiologically relevant to demonstrate the functional activity of receptor ligands or agonist antibodies that mimic the function of ligands (e.g., receptor signaling by ligands).

[0372] For example, frozen human preadipocytes (Lonza Cat# PT-5005) were thawed on day 1, differentiated on day 3, and maintained in differentiation medium for about two weeks before the experiment (e.g., then starving on the 17th, and rehearsed on the 18th). The seeding medium was 1:1 DMEM/F12K + 10% FBS. Seeding cell density was 25,000 cells/100 μL/well in 96-well plates. On day 3, the medium was replaced with human adipocyte differentiation medium (Cell Applications Inc). Thereafter, fresh differentiation medium was added to the cells every 2-3 days. On day 17 (the day before the assay), cells were washed twice and left in DMEM/0.1% BSA (Sigma cat# A3803 essential fatty acid-free BSA) overnight. The next day, fresh DMEM/0.1% BSA medium was added for 1 hour before cells were treated with test anti-Klotho beta antibodies for 15 minutes at 37°C. The Cis-Bio Cellul'erk Assay Kit (Cat# 64ERKPEH) was used to assay the level of ERK phosphorylation following the manufacturer's protocol.

[0373] Representative results using human adipocytes are reported as EC50 values as shown in Table 16 below: Table 16

EXAMPLE 6: LINK COMPETITION

[0374] Ligand competition assays (FGF19 or FGF21) were performed to evaluate whether the antibody-human Klotho beta interaction influences the binding of Klotho beta to its natural ligand, FGF19 or FGF21.

[0375] For example, Biacore-based competition assays were created in which FGF19 (e.g., SEQ ID NO: 304) or FGF21 (e.g., SEQ ID NO: 429) was immobilized in a flow cell (Fc2) of a CM5 chip (using Fc1 as a reference surface). Human Klotho beta-antibody complexes were prepared with exemplary antibodies of the present disclosure, such as 5H23 (e.g., VH of SEQ ID NO: 25 and VL of SEQ ID NO: 26) or a humanized 5H23 (e.g., VH of SEQ ID NO: 271 and VL of SEQ ID NO: 276)). For example, concentrations of 5H23 and a control antibody were titrated from 0.1-67 nM, keeping the concentration of Klotho beta constant at 5 nM in a 96-well microplate and injected onto the surface of FGF19. For another example, the concentrations of a humanized 5H23 (e.g., VH of SEQ ID NO: 271 and VL of SEQ ID NO: 276) were titrated to 0.00167 nM, keeping the concentration of Klotho beta constant at 2.5 nM in a 96-well microplate and injected onto the surface of FGF21. The measured signal (Response Unit, RU) was plotted against the antibody concentration in solution [nM]. If the antibody in solution recognized the same epitope as the FGF19 ligand or FGF21 ligand immobilized on the chip surface, then a decrease in RU was observed with increasing concentration of the antibody in solution, which demonstrates competition with the FGF19 ligand or ligand. of FGF21 to the binding site on Klotho beta. However, if the antibody in solution recognized a distinct epitope relative to the FGF19 ligand or immobilized FGF21 ligand, an increase in RU was observed. In the latter case, the antibody-Klotho complex could bind to the immobilized FGF19 surface ligand or immobilized FGF21 ligand surface leading to the observed increase in signal. In the exemplary data shown below in Table 17A, a control antibody partially competed with the FGF19 ligand resulting in a significant reduction in the RU signal, where 5H23 did not compete with the FGF19 ligand for binding to Klotho beta. In the exemplary data shown below in Table 17B, a control antibody competed with the FGF21 ligand, resulting in a significant reduction in RU signal, where a humanized 5H23 did not compete with the FGF21 ligand for binding to Klotho beta. Table 17A *Control antibody comprises SEQ ID NO:358 and SEQ ID NO:360 Table 17B *Control antibody comprises SEQ ID NO:358 and SEQ ID NO:360

[0376] Since 5H23 and a humanized 5H23 antibody bind to a different epitope of Klotho beta compared to endogenous ligands such as FGF19 and FGF21, experiments were conducted to test whether there were synergistic effects between FGF21 and 5H23 or a 5H23 antibody humanized. In a HEK293 reporter assay (see, e.g., Example 4), combinations of FGF21 and a humanized 5H23 antibody (e.g., VH of SEQ ID NO: 271 and VL of SEQ ID NO: 276) were tested at a molar ratio of 1:1 or fixing one and titrating the concentration of the other. No evidence of synergistic effects was observed; the maximum effect of FGF21 was not improved by the humanized 5H23 antibody, and vice versa.

EXAMPLE 7: HUMANIZATION

[0377] Humanized anti-Klotho beta antibodies were generated, including from selected antibodies as described in Examples 1-6.

[0378] A series of anti-Klotho beta antibodies were selected for sequencing and their VH and VL regions, including their CDRs, are shown in Tables 1-10 and Figures 1 and 2. An exemplary anti-Klotho beta antibody, 5H23 , was selected for humanization. Various methods for humanization were used. For some of the humanized antibodies, the method for humanization was empirical and based, in part, on structural information related to immunoglobulin variable regions including molecular models and structural stability requirements of the antibody (see, e.g., Ewert et al., 2004 , Methods 34:184-199; Honegger, 2008, Handb. Pharmacol 181:47-68; The method is also based, in part, on considerations of antigen contact residues and/or framework stability residues. For example, analysis of typical antigen contact residues depends on the size of the antigen in particular residues outside the CDRs that can contact the antigen, upper core, middle core and lower core divisions, VH:VL interface residues, Pro /Gly conserved (positive phi angles) and correspondence of correlated VH subtype residues (see, for example, Ewert et al., supra ; Honegger, supra ; Kügler et al., supra).

[0379] For example, human VH sequences homologous to the 5H23 VH framework sequences were sought and the human germline-encoded VH sequence IGHV1-3*01 (see, e.g., Ehrenmann et al., 2011, Cold Spring Harbor Protoc. G:737-749) was chosen as an acceptor for humanization. For some of the humanized antibodies, the 5H23 VH CDR sequences were first transferred to the corresponding positions of IGHV1-3*01. Next, a series of VH amino acid residues from 5H23 were replaced with the corresponding human residues individually or in combinations.

[0380] Furthermore, for example, human VL sequences homologous to the 5H23 VL framework sequences were sought and the human VK region encoded by IGKV4-1*01 (see, for example, Ehrennmann et al., supra) was chosen as an acceptor for humanization. For some of the humanized antibodies, the 5H23 VL CDR sequences were first transferred to the corresponding positions of IGKV4-1*01. Next, a series of VL amino acid residues from 5H23 were replaced with the corresponding human residues individually or in combinations.

[0381] For some of the humanized antibodies, the humanization method used an algorithm to construct a three-dimensional map of the mouse variable regions. This method also identified framework amino acids and residues important for the formation of the CDR structure or required for binding to Klotho beta. Furthermore, human VH and VL amino acid sequences with high homology to mouse sequences were selected for possible framework sequences for humanization. As described above, the 5H23 antibody CDR sequences can be transferred into such additional human framework sequences. A variety of human structural sequences, including germline sequences (e.g., IGHV1-3, IGHV1-46, IGHV1-69, IGKV4-1, IGKV1-39, or IGKV3-20) and individual mature sequences, may be suitable for the humanization method. Next, a series of amino acid residues from VH of 5H23 and/or VL of 5H23 can be replaced with the corresponding human residues individually or in combination.

[0382] For some of the humanized light chains, BLAST IG searches were used to identify human germline sequences that were closely matched in sequence to the VL of 5H23 and/or sequences that were commonly used, including, for example, IGKV1-39 and IGKV3-20. For some of the humanized light chains, the VL CDR sequences of 5H23 were first transferred to the corresponding positions of IGKV1-39 or IGKV3-20, and then certain amino acids were empirically selected for substation.

[0383] The amino acid sequences of resulting humanized VH (vH1-vH9) and VL (vL1 to vL5, v1-39a to v1-39p and v3-20a to v3-20j) sequences are shown with VH and VL sequences of 5H23 in Figure 3A-3D. For example, using the various humanization methods described in this example, a series of amino acid residues from VH and VL of 5H23 were substituted for the corresponding human residue to obtain humanized sequences, as shown in Figure 3A-3D.

[0384] Humanized Klotho beta antibodies can be prepared using any of the CDR sequences in Table 18, in combination with any of the framework sequences in Table 19. Table 18 CDR sequences for humanized Anti-Klotho beta antibodies Table 19 Framework Sequences for Humanized Anti-Klotho beta Antibodies

[0385] For example, a humanized anti-Klotho beta antibody may comprise a heavy chain variable region (VH) comprising: FR1 (e.g., SEQ ID NO: 278, 279, 280, or 378); CDR1 (e.g. SEQ ID NO: 1, 27, 53, 79, 105, 131, 157, 183, 209, 235); FR2 (e.g., SEQ ID NO: 281, 282, or 283); CDR2 (e.g., SEQ ID NO: 2, 28, 54, 80, 106, 132, 158, 184, 210, or 236); FR3 (e.g., SEQ ID NO: 284, 285, 286, 287, 379, 380, or 381); CDR3 (e.g., SEQ ID NO: 3, 29, 55, 81, 107, 133, 159, 185, 211, or 237); and/or FR4 (e.g. SEQ ID NO: 288); and/or a light chain variable region (VL) comprising: FR1 (e.g., SEQ ID NO: 289, 290, 382, 383, or 384); CDR1 (e.g., SEQ ID NO: 4, 30, 56, 82, 108, 134, 160, 186, 212, or 238); FR2 (e.g., SEQ ID NO: 291, 292, or 385-392); CDR2 (e.g., SEQ ID NO: 5, 31, 57, 83, 109, 135, 161, 187, 213, or 239); FR3 (e.g., SEQ ID NO: 293, 294, 295, or 393-404); CDR3 (e.g., SEQ ID NO: 6, 32, 58, 84, 110, 136, 162, 188, 214, 240); and/or FR4 (e.g., SEQ ID NO: 296, 405, 406, or 407).

[0386] As described in this example, humanized Anti-Klotho beta antibodies were empirically designed and expressed as Klotho beta binding proteins, including nine humanized variants of the VH region of the 5H23 antibody and thirty-one humanized variants of the VL region of the antibody 5H23 that were created. The sequences of these exemplary humanized 5H23 VH and VL regions are shown in Figure 3A-3D.

[0387] Humanized antibodies were prepared with humanized VH and humanized VL regions with sequences as shown in Figure 3A-3D. For example, eighteen (6 x 3) combinations of vH 1-6 and vL 1-3 were constructed using an IgG1 (Ala-Ala) constant region (SEQ ID NO: 316) and a kappa constant region (SEQ ID NO: 318): vH1-VL1, vH1-vL2, vH1-vL3, vH2-vL1, vH2-vL2, vH2-vL3, vH3-vL1, vH3-vL2, vH3-vL3, vH4-vL1, vH4-vL2, vH4-vL3 , vH5-vL1, vH5-vL2, vH5-vL3, vH6-vL1, vH6-vL2, vH6-vL3, with sequences as shown in Figure 3A-3D. Furthermore, humanized antibodies were constructed with one exemplary humanized VH region (e.g., vH3) and twenty-six humanized VL regions (v1-39a to v1-39p and v3-20a to v3-20j) with sequences as shown. in Figure 3A-3D.

[0388] The humanized antibodies were tested for their activity in a variety of assays, including, for example, as described in Examples 2-6. Expression of humanized antibodies with light chains comprising vL3 or v1-39c was low and these antibodies were not further tested. Exemplary results with a variety of humanized anti-Klotho beta antibodies are shown in Tables 20A and 20B below. Table 20A Table 20B Prep 1 = expressed preparation of humanized 5H23 antibody (e.g., VH of SEQ ID NO: 271 and VL of SEQ ID NO: 276) at the same time as the LC variants; Prep 2 = humanized 5H23 antibody preparation (e.g., VH of SEQ ID NO: 271 and VL of SEQ ID NO: 276) purified.

[0389] In additional assays, for example, reporter assays with HEK293T cells, as described in Example 4, in which the cells were transfected with plasmids encoding mouse Klotho beta (e.g., SEQ ID NO: 301), mouse Klotho beta rats (e.g., SEQ ID NO: 356), hamster beta Klotho (e.g., SEQ ID NO: 408), rabbit beta Klotho (e.g., SEQ ID NO: 410), or dog beta Klotho (e.g. , SEQ ID NO: 412) and were also transfected with plasmids encoding chimeric mouse FGFR1-βIIIc receptor (e.g., SEQ ID NO: 416), chimeric rat FGFR1-βIIIc receptor (e.g., SEQ ID NO: 419 ), chimeric hamster FGFR1-βIIIc receptor (e.g., SEQ ID NO: 417), chimeric rabbit FGFR1-βIIIc receptor (e.g., SEQ ID NO: 420), or chimeric dog FGFR1-βIIIc receptor (e.g., SEQ ID NO: 418), respectively, when treated with an Anti-Klotho beta antibody, such as a humanized 5H23 antibody (e.g., VH of SEQ ID NO: 271 and VL of SEQ ID NO: 276), did not activate the complex chimeric FGFR1c-Klotho beta receptor from mouse, rat, hamster, rabbit or dog, respectively. Anti-Klotho beta antibodies as described herein, including antibodies 5H23 and humanized 5H23, as well as antibodies that compete with 5H23 (e.g., 1C17, 1D19, 2L12, 3L3, 3N20, 4P5, 5C23, 5F7 and 1G19, as described in Example 3) with CDR sequences as shown in Tables 1-10, activate a human and Cyno FGF/Klotho beta receptor complex, but not mouse, rat, hamster, rabbit, or dog FGF/Klotho beta receptor complexes , as demonstrated by reporter assays described above. When a monovalent Fab of anti-Klotho beta antibody is prepared from a papain digestion of an Anti-Klotho beta antibody, such as a humanized 5H23 antibody (e.g., VH of SEQ ID NO: 271 and VL of SEQ ID NO: 276), was tested in a HEK293 reporter assay for its ability to activate the human FGFR1c/KLB receptor complex, the Fab did not show any antibody activity up to 67 nM, whereas the humanized 5H23 antibody showed activity at similar low nanomolar concentrations. to those shown in Table 20B.

EXAMPLE 8: STUDIES ON ANIMALS

[0390] The effects of anti-Klotho beta antibodies are evaluated in animal studies, including cynomolgus monkeys.

[0391] In studies of obese cynomolgus monkeys, an exemplary Anti-Klotho beta antibody that binds to human Klotho beta and cyno Klotho beta (e.g., 5H23 antibody or humanized variant thereof), as well as an antibody comprising one or more of the 5H23 CDRs as shown in Table 1, or alternatively, an antibody comprising one or more of the CDRs of an antibody or humanized variant thereof represented in Tables 2-10 that compete for binding of 5H23 to human Klotho beta, as described in Example 3, it is administered. Effects on a variety of metabolic parameters can be measured. Exemplary parameters include food intake, body weight, body mass index (BMI), waist circumference (AC), skinfold thickness (SFT), oral glucose tolerance test (OGTT), blood glucose levels (e.g. e.g. serum) fasting and/or fed (e.g. postprandial), insulin levels and/or triglyceride levels.

[0392] In a real study, twenty spontaneously obese cynomolgus monkeys with a body mass index of 40 or above are selected and randomized into vehicle (n = 10) and antibody treatment (n = 10) groups. Animals were injected subcutaneously with either vehicle or anti-Klotho beta antibody on days 1 and 14. Food intake for each meal is recorded and body weight is measured once a week. Blood samples are taken once a week for 7 weeks for measurements of glucose, insulin, lipids and plasma (alternatively serum) parameters of interest. On days 14, 28, and 49, an oral glucose tolerance test is conducted.

[0393] Exemplary treatment effects may include reduced food intake, decreased body weight, decreased BMI, AC and/or SFT, improved glucose tolerance, decreased insulin levels, decreased blood glucose levels ( alternatively, serum) fasted and/or fed (e.g., postprandial), reduced insulin levels and/or triglyceride levels. These effects indicate improvement in metabolic parameters with treatment with anti-Klotho beta antibodies.

[0394] For example, twenty male Cynomolgus monkeys were selected for treatment with a humanized 5H23 antibody (e.g., VH of SEQ ID NO: 271 and VL of SEQ ID NO: 276) or a vehicle control based on their BMI (> 40) and were trained in chair restraint, subcutaneous injection, blood collection, and oral probe. A routine feeding schedule was established.

[0395] Baseline values of several parameters of interest were measured before treatments. For example, on day -7, baseline body weight, BMI, waist circumference, and skinfold thickness were measured, and a dual-energy X-ray absorptiometry ("DEXA") scan was conducted for the cynomolgus monkeys under ketamine anesthesia to measure bone mineral density. Blood samples were taken on day -3, after an overnight fast. Baseline levels of serum glucose, insulin, total cholesterol, LDL, HDL, triglycerides, and a panel of hematology and clinical chemistry parameters were measured and analyzed. Immediately after the baseline samples, the animals underwent an oral glucose tolerance test (OGTT), receiving an esophageal tube of 4 g/kg glucose and were sampled at 5, 15, 30, 60, 120 and 180 minutes later. of the glucose challenge, and serum glucose and insulin were measured. Based on the baseline data, the animals were divided into two groups with 10 animals in each group (e.g., one antibody treatment group and the other group as a vehicle control group) to achieve similar baseline levels of the various parameters, for example, body weight, BMI, and serum glucose, insulin and triglyceride levels.

[0396] Starting on day 0, a group of animals (n = 10) received a subcutaneous injection dose of 10 mg/kg of an Anti-Klotho beta antibody, such as a humanized 5H23 antibody (e.g., VH of SEQ ID NO: 271 and VL of SEQ ID NO: 276) every two weeks (e.g., on days 0, 14, 28, and 42) for 4 doses. The control vehicle group received matching vehicles on the same days. Treatments were performed in the morning 30 minutes before the morning meal, and the dosage volume was 0.1 to 0.2 ml/kg.

[0397] Parameters of interest, for example, food intake, body weight, clinical chemistry, and OGTT, were monitored throughout the study. For example, food intake was measured daily. Body weight, BMI, waist circumference, and skinfold thickness were measured weekly, for example, on days 7, 14, 21, 28, 35, 42, 49, 56, 63, 70, 77, 84, 91, and 98 Blood samples were collected weekly, for example, on days 7, 14, 21, 28, 35, 42, 49, 56, 63 and 70, after an overnight fast, to measure glucose, insulin and lipids such as triglycerides. An additional blood sample was taken on day 98 after an overnight fast. OGTTs were performed after the start of the study, for example, on days 14, 28, and 56, where animals were gavaged with 4 g/kg glucose and sampled at 5, 15, 30, 60, 120 and 180 minutes after the glucose challenge and blood glucose and insulin were measured. A DEXA scan was performed on days 30 and 72. Additionally, a hematology and clinical chemistry panel was analyzed on days 28 and 70. Two animals from the vehicle group and two animals from the anti-Klotho beta antibody group were sacrificed and were necropsied. on day 50 for safety assessment. During the study, all animals were closely monitored for their health.

[0398] Examples of results from this study are presented in Tables 21 to 25 below. As shown in Table 21, the body weight of vehicle-treated animals remained constant (with a slight increase throughout the course); whereas the body weight of animals treated with the anti-Klotho beta antibody progressively decreased, and the body weight did not return to baseline for 8-14 weeks (e.g., recovery phase). Likewise, as shown in Table 22, animals treated with vehicle showed relatively stable BMI throughout the study, while animals treated with the anti-Klotho beta antibody showed a decrease in BMI level throughout the study. The BMI level also did not return to baseline values (e.g. during the recovery phase). These results suggest that treatment with anti-Klotho beta antibody resulted in a reduction in fat mass.

[0399] As shown in Table 23, serum insulin levels in vehicle-treated animals increased throughout the study; whereas serum insulin levels in animals treated with the anti-Klotho beta antibody significantly decreased. Serum glucose levels were also reduced in Anti-Klotho beta antibody-treated animals, as shown in Table 24. Similarly, as shown in Table 25, triglyceride levels in vehicle-treated animals increased throughout the study. ; while triglyceride levels in animals treated with the anti-Klotho beta antibody were significantly reduced.

[0400] Results of OGTTs demonstrated that there were significantly different differences between the vehicle and anti-Klotho beta antibody groups. In contrast, after treatment, there was a trend toward reduced glucose and insulin levels were reduced in animals treated with the anti-Klotho beta antibody compared to vehicle-treated animals.

[0401] In another exemplary study, forty spontaneously obese cynomolgus males were selected, trained and fed as described above.

[0402] Baseline values of several parameters were measured before treatment as discussed above. For example, baseline body weight, BMI, waist circumference, and skinfold thickness were measured on day -4, and baseline blood samples were taken for measurements of serum glucose, insulin, total cholesterol, LDL, HDL, and triglycerides. on day -3, following an overnight fast. Based on these baseline data, animals were divided into 5 groups (8 animals in each group) with 4 groups receiving various doses of an Anti-Klotho beta antibody such as a humanized 5H23 antibody (e.g., VH of SEQ ID NO : 271 and VL of SEQ ID NO: 276) and a group receiving a control vehicle.

[0403] On day 0, the first group of animals (n = 8) received a single dose of subcutaneous injection of 0.1 mg/kg of the anti-Klotho beta antibody; the second group of animals (n = 8) received a single dose of 1 mg/kg subcutaneous injection of the Anti-Klotho beta antibody, and the third group of animals (n = 8) received a single dose of 10 mg subcutaneous injection /kg of anti-Klotho beta antibody. Starting on day 0, the fourth group of animals (n = 8) received a subcutaneous injection dose of 0.1 mg/kg of anti-Klotho beta antibody once every 4 weeks over a period of 12 weeks. As a control, the fifth group of animals (n = 8) received a dose of vehicle once every 4 weeks for 12 weeks. Treatments were performed in the morning 30 minutes before the morning meal, and the dosage volume was 0.2 ml/kg.

[0404] Parameters of interest were monitored throughout the study. For example, food intake was measured for each meal. Body weight, BMI, waist circumference, and skinfold thickness were measured weekly. Samples of blood were collected at, for example, 3, 6, 12 and 24 hours and 3, 4, 7, 10, 14, 21, 28, 35, 42, 49, 56, 63, 70, 77, 84 and 112 days after dose(s), and parameters of interest, e.g., serum glucose, insulin, total cholesterol, LDL, HDL, and triglycerides, were measured. During the study, all animals were closely monitored for their health as described above.

[0405] Exemplary results from this dose-response study are shown in Tables 26-29. Table 26 shows the relative body weight changes in animals treated with the anti-Klotho beta antibody compared to the body weight changes in vehicle-treated animals. As shown, a single dose of subcutaneous injection of 0.1 mg/kg, 1 mg/kg, or 10 mg/kg of Anti-Klotho beta antibody, or four doses of subcutaneous injection of 1 mg/kg of Anti-Klotho antibody beta significantly reduced body weight. Furthermore, the reduction in body weight was maintained on day 112 for animals that received a single 10 mg/kg dose of Anti-Klotho beta antibody, or for animals that received four 1 mg/kg doses of Anti-Klotho beta antibody. Klotho beta compared to vehicle.

[0406] As shown in Table 27, a single dose of subcutaneous injection of 0.1 mg/kg, 1 mg/kg, or 10 mg/kg of the Anti-Klotho beta antibody, or four doses of subcutaneous injection of 1 mg/kg kg of anti-Klotho beta antibody reduced the serum insulin level compared to the vehicle control group. Furthermore, four doses of 1 mg/kg subcutaneous injection of the anti-Klotho beta antibody significantly reduced the serum glucose level, as shown in Table 28. Furthermore, serum triglyceride levels in animals treated with a single dose of 1 mg/kg subcutaneous injection, or 10 mg/kg of anti-Klotho beta antibody, or four doses of 1 mg/kg subcutaneous injection of anti-Klotho beta antibody, were reduced compared to vehicle-treated animals, as shown in Table 29. Table 26A Body Weight Table 26B Change in Body Weight (kg) Table 27 Insulin Table 28 Glucose Table 29 Triglycerides

[0407] The results of these animal studies demonstrate improvement in metabolic parameters with treatment with Anti-Klotho beta antibodies provided here, for example, such as a decrease in body weight, body mass index, waist circumference, skin folds, glucose ( e.g., serum glucose), insulin (e.g., serum insulin), and/or triglycerides (e.g., serum triglycerides).

EXAMPLE 9: EPITOPES AND DOMAIN MAPPING

[0408] Studies were carried out in order to localize the binding site on human KLB of anti-Klotho beta antibodies in the 5H23 epitope bin, including 5H23 as described in Example 3, with sequences shown in Tables 1-10 and Figures 1 -3, and human anti-Klotho beta antibodies in the 5H23 epitope bin, such as humanized 5H23 antibodies (e.g., VH of SEQ ID NO: 271 and VL of SEQ ID NO: 276). For example, FACS-based binding assays for domain mapping were performed on Expi293 cells (Life Technologies, A14635) that were transiently transfected with plasmids encoding KLB variants: human, mouse, cynomolgus, a chimeric version in which the sequence of the KL1 domain of mouse KLB (M1-F506) replaces the KL1 domain of human KLB (M1-F508) to create mouse-human KLB (SEQ ID NO: 376), and a second chimera in which the human KL1 sequence (M1-F508) replaces the KL1 domain of mouse KLB (M1-F506) to create human-mouse KLB (SEQ ID NO: 374). Furthermore, the pYD7 expression vector harboring no KLB sequence was transfected as a negative control.

[0409] In some studies, binding of a purified sample of a humanized 5H23 antibody (e.g., VH of SEQ ID NO: 271 and VL of SEQ ID NO: 276) to KLB variants was determined by FACS analysis. Two-day post-transfection cells were coincubated with purified antibodies: humanized 5H23 antibody (e.g., VH of SEQ ID NO: 271 and VL of SEQ ID NO: 276), a control antibody (e.g., VH of SEQ ID NO: 358 and VL of SEQ ID NO: 360), and a negative control antibody (e.g., keyhole limpet clam hemocyanin (KLH) antibody expressed from a construct comprising SEQ ID NO: 424 and 425 ) diluted to 1 μg/ml in PBS/1% BSA/0.1% azide for 30 minutes at 4°C. After washing with PBS/1% BSA/0.1% azide, the transfected cells were then co -incubated with labeled anti-human Fc (Jackson Immunoresearch) for 30 minutes at 4° C. After washing with PBS/1% BSA/0.1% azide, cells were acquired on the flow cytometer (FACS Calibur) and analyzed by software cytometry (FlowJo). To visualize the resulting data, graphs plotting cell number versus fluorescence intensity were generated, and the mean fluorescence intensity (MFI) was determined for each sample as shown in Table 30.Table 30 * Average fluorescence intensity calculated from FACS data using FlowJo analysis software; Negative Control. is an anti-KLH antibody.

[0410] An exemplary humanized 5H23 antibody (e.g., VH of SEQ ID NO: 271 and VL of SEQ ID NO: 276) bound to human KLB and cynomolgus KLB, as indicated by a large proportion of cells having high intensity of fluorescence compared to cells treated with the anti-KLH negative control antibody, but the exemplary humanized 5H23 antibody did not bind to mouse KLB. The exemplary humanized 5H23 antibody also bound to the mouse-human KLB chimeric protein, but not to the human-mouse KLB chimeric protein indicating that Anti-Klotho beta antibodies in the 5H23 epitope bin, including 5H23 as described in Example 3, with sequences shown in Tables 1-10 and Figures 1-3, and anti-human Klotho beta antibodies in the 5H23 epitope bin, such as humanized 5H23 antibodies (e.g., from VH of SEQ ID NO: 271 and VL of SEQ ID NO: 276) bind to the KL2 domain of human KLB. In contrast, the control antibody bound to the KL1 domain of human KLB as demonstrated by its binding to cells transfected with the human-mouse KLB chimeric protein, but not the mouse-human KLB chimeric protein.

[0411] In order to further identify specific binding residues within the KL2 domain of human Klotho beta, shotgun mutagenesis was used to separately mutate individual residues of the KL2 domain of human Klotho beta to an alanine (e.g. residues F508A-L1008A). The resulting Klotho beta mutant proteins were expressed within HEK-293T cells and assayed by fluorescence-activated cell sorting (FACS) for binding to Anti-Klotho beta antibodies in the 5H23 epitope bin, including 5H23 as described in Example 3 , with sequences shown in Tables 1-10 and Figures 1-3, and anti-human Klotho beta antibodies in the 5H23 epitope bin, such as a humanized 5H23 antibody (e.g., VH of SEQ ID NO: 271 and VL of SEQ ID NO: 276), or a monovalent Fab fragment of the humanized 5H23 antibody. For example, screening of Klotho beta mutant proteins was conducted at a concentration of 0.5 μg/ml for the humanized 5H23 antibody, 1.0 μg/ml for the Fab fragment, and 2.0 μg/ml for Antibodies. -Klotho beta polyclonal positive control.

[0412] The resulting mapping identified three specific binding residues, H657, Y701 and R703, which were negative for binding by the humanized 5H23 antibody, but were positive for control polyclonal anti-Klotho beta antibodies. These residues represented amino acids whose side changes made the greatest energetic contribution to the antibody-epitope interaction, as shown in Table 31. The locations of the three identified residues were modeled by showing them (dark spheres) in the equivalent positions in human cytosolic beta-glucosidase (PDB ID #2JFE; Tribolo et al, J. Mol Biol 370, 964-975 (2007)), identified by BLAST alignment of the two proteins, as shown in Figure 6. The structure shows the equivalent residues of Klotho beta 521- 963. The lower reactivity of the Y701A and R703A mutations with the humanized 5H23 antibody indicates that Y701 and R703 are the main energetic contributors to binding. Table 31

[0413] Therefore, the anti-Klotho beta antibodies provided herein, including 5H23 and antibodies in the 5H23 epitope bin recognize an epitope in the KLB2 domain comprising residues H657, Y701 and/or R703. These antibodies, as described in Example 3 and comprising CDR sequences in Tables 1-10 and Figures 1-3, are useful as agonist antibodies to induce FGF19-mediated and/or FGF21-mediated signaling, including, for example, to reduce body weight, food intake, BMI, insulin, glucose and/or triglycerides.

[0414] Furthermore, the anti-Klotho beta antibodies provided here share the common characteristic of competing with each other for Klotho beta binding (see, for example, Example 3 which describes antibodies in the 5H23 epitope bin). This competitive inhibition indicates that each antibody binds to the same region of Klotho beta (e.g., the same epitope), thus asserting similar effects. As further exemplified herein, Anti-Klotho beta antibodies include humanized anti-Klotho beta antibodies, including humanized Anti-Klotho beta antibodies derived from or based on 5H23, 1C17, 1D19, 2L12, 3L3, 3N20, 4P5, 5C23, 5F7 and /or 1G19 having the CDR sequence as described in Tables 1-10 or Figures 1-3, such as anti-Klotho beta antibodies, including humanized anti-Klotho beta antibodies, bind to a specific domain of human Klotho beta (e.g. , KL2 (residues S509-S1044) as described above). Furthermore, such binding can be attributed largely to the presence of particular amino acid residues within the KL2 region (e.g., H657, Y701, and R703, as described above), which comprise the epitope recognized by the anti-Klotho-beta antibodies described herein. . Taken together, these results demonstrate that the effects observed for the anti-Klotho beta antibody that is derived from or based on 5H23 or an antibody in the 5H23 epitope bin, including an antibody with one or more CDRs described in Tables 1-10 or Figures 1-3, can be extrapolated to other anti-Klotho beta antibodies described herein having the same or similar epitope specificity (e.g., the same or similar CDRs). For example, the in vitro activities of antibodies as shown in Examples 4-7 and above, as well as the in vivo effects demonstrated in Example 8 for an exemplary humanized anti-Klotho beta antibody, are representative of the activities and effects of anti-Klotho antibodies. - Klotho beta described here.

[0415] The embodiments of the present invention described above are intended to be exemplary only, and those skilled in the art will recognize, or be able to determine using no more than routine experimentation, numerous equivalents to the specific procedures described herein. All such equivalents are considered to be within the scope of the present description and are covered by the following claims. Furthermore, as used in the present specification and claims, the singular forms “a”, “an” and “the” and “a” include the plural forms unless the content clearly dictates otherwise. Thus, for example, reference to “an antibody” may include a mixture of two or more such antibodies, and the like. Furthermore, those skilled in the art will recognize that operational sequences must be established in some specific order for the purpose of explanation and claim, but the present invention contemplates various changes beyond such specific order.

[0416] The contents of all references described herein are incorporated herein by reference.

[0417] Other embodiments are within the following claims.

Claims (17)

1. Antibody characterized by the fact that it binds within the Klotho beta 2 (KLB2) domain comprising amino acid residues 657 to 703 of human Klotho beta (SEQ ID NO: 297), wherein the antibody comprises: (a) a region heavy chain variable comprising: a CDR1 comprising the amino acid sequence of SEQ ID NO: 1, SEQ ID NO: 12 or SEQ ID NO: 13; a CDR2 comprising the amino acid sequence of SEQ ID NO: 2 or SEQ ID NO: 14; and a CDR3 comprising the amino acid sequence of SEQ ID NO: 3 or SEQ ID NO: 15; and (b) a light chain variable region comprising: a CDR1 comprising the amino acid sequence of SEQ ID NO: 4 or SEQ ID NO: 16; a CDR2 comprising the amino acid sequence of SEQ ID NO: 5 or SEQ ID NO: 11; and a CDR3 comprising the amino acid sequence of SEQ ID NO: 6 or SEQ ID NO: 17. 2. Antibody according to claim 1, characterized in that the antibody comprises: (a) a heavy chain variable region comprising: a CDR1 comprising the amino acid sequence of SEQ ID NO: 1, a CDR2 comprising the sequence amino acid sequence of SEQ ID NO: 2, and a CDR3 comprising the amino acid sequence of SEQ ID NO: 3; and (b) a light chain variable region comprising: a CDR1 comprising the amino acid sequence of SEQ ID NO: 4, a CDR2 comprising the amino acid sequence of SEQ ID NO: 5, and a CDR3 comprising the amino acid sequence of SEQ ID NO: 6. 3. Antibody according to claim 1, characterized in that the antibody comprises: (a) a heavy chain variable region comprising a CDR1 comprising SEQ ID NO: 12, a CDR2 comprising SEQ ID NO: 2 and a CDR3 comprising SEQ ID NO: 3; and (b) a light chain variable region comprising a CDR1 comprising SEQ ID NO: 4, a CDR2 comprising SEQ ID NO: 5 and a CDR3 comprising SEQ ID NO: 6. 4. Antibody according to claim 1, characterized in that the antibody comprises: (a) a heavy chain variable region comprising a CDR1 comprising SEQ ID NO: 13, a CDR2 comprising SEQ ID NO: 14 and a CDR3 comprising SEQ ID NO: 15; and (b) a light chain variable region comprising a CDR1 comprising SEQ ID NO: 16, a CDR2 comprising SEQ ID NO: 11 and a CDR3 comprising SEQ ID NO: 17. 5. Antibody according to any one of claims 1 to 4, characterized by the fact that the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 25 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 26. 6. Antibody according to any one of claims 1 to 4, characterized by the fact that the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 271 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 276. 7. Antibody according to any one of claims 1 to 6, characterized in that the antibody binds to an epitope comprising at least one of amino acid residues 657, 701 and/or 703 of human Klotho beta (SEQ ID NO : 297). 8. Antibody according to any one of claims 1 to 7, characterized in that the antibody is a monoclonal antibody, a humanized antibody or a chimeric antibody. 9. Antibody according to any one of claims 1 to 7, characterized in that the antibody is a Fab, Fab', F(ab')2, Fv, scFv, (scFv)2, chain antibody molecule single, dual variable region antibody, linear antibody or a multispecific antibody formed from antibody fragments. 10. Antibody according to claim 2, characterized in that it comprises a heavy chain having amino acids 23-472 of SEQ ID NO: 317 and a light chain having amino acids 23-240 of SEQ ID NO: 319. 11. Antibody according to any one of claims 1 to 10 characterized by the fact that it is for use in a method for reducing blood glucose levels, reducing triglyceride levels, and/or reducing body weight in a human subject. 12. Host cell characterized by the fact that it comprises a polynucleotide or polynucleotides encoding the antibody as defined in any one of claims 1 to 10, wherein the host cell is a bacteria, filamentous fungus or yeast. 13. Pharmaceutical composition characterized by the fact that it comprises the antibody as defined in any one of claims 1 to 10 and a pharmaceutically acceptable carrier. 14. Use of the antibody as defined in any one of claims 1 to 10 characterized by the fact that it is in the preparation of a medicine for the treatment of reducing blood glucose levels, reducing triglyceride levels, and/or reducing body weight in a human individual. 15. Use according to claim 14, characterized by the fact that the human subject has Type 1 diabetes, Type 2 diabetes, obesity, dyslipidemia, (Non-alcoholic steatohepatitis) NASH, cardiovascular disease, metabolic syndrome, or liver disease non-alcoholic fatty acid (NAFLD). 16. Use according to claim 14 or claim 15, characterized in that the treatment comprises combination therapy with at least one additional therapeutic agent. 17. Use according to claim 16, characterized in that the at least one additional therapeutic agent is selected from the group consisting of biguanides, sulfonylureas, thiazolidinediones, GLP-1 analogues, PPAR gamma agonists, inhibitors of dipeptidyl peptidase-4 (DPP-4), bromocriptine, bile acid sequestrants, insulin, alpha-glucosidase inhibitors, metformin, SGLT-2 inhibitors, appetite suppressant agents and weight loss drugs.