Binding proteins and methods of use thereof

Abstract

Binding proteins, eg, antibodies, that bind to GDNF family receptor alpha-like (GFRAL) proteins, including human GFRAL proteins, and methods for their use are provided. [Solution] An antibody or fragment thereof that binds to GDNF family receptor alpha-like (GFRAL) protein, wherein the antibody or fragment thereof comprises a heavy chain variable (VH) region and a light chain variable (VL) region, and (i) the VH region comprises complementarity-determining regions (CDRs) 1, CDR2, and CDR3 that contain specific amino acid sequences; and (ii) the VL region comprises CDR1, CDR2, and CDR3 that contain specific amino acid sequences.

Description

[Technical Field]

[0001] (Field) This application is incorporated by reference in its entirety. The benefit of provisional application No. 62 / 316,516 is claimed.

[0002] The present disclosure generally relates to GDNF family receptor alpha-like (GFRAL) proteins, including human GFRAL proteins. The present invention relates to binding proteins, e.g., antibodies, that bind to GFRAL (globulin receptor agonist) proteins and methods of using the same. [Background technology]

[0003] (background) Growth differentiation factor 15 (GDF15) belongs to the transforming growth factor beta (TGF-β) superfamily. GDF15 is a protein that acts as a regulator of TGF-PL, MIC-1, PDF, PLAB, NAG-1, and PTGFB. It is also known that GDF15 mRNA is most abundant in the liver, Its expression in the liver is higher than that in the liver, kidney, heart, and It can be significantly upregulated upon injury to organs such as the lungs.

[0004] GDF15 is involved in the regulation of inflammatory and apoptotic pathways in injured tissues and during disease processes. GDF15 has been reported to play a role in mediating cachexia in various diseases. However, cachexia is a complex and incompletely understood condition. Furthermore, at least some tumors overexpress and secrete GDF15, Elevated serum GDF15 levels have been associated with various cancers. GDF-15 inhibits TNF-α, IL-1, and IL- 2 and MCS-F release, thereby suppressing local inflammatory signal transduction similar to the effects of TGF-β. It is described as a negative regulator of macrophage activation by inhibiting the positive feedback of cell proliferation. Monoclonal antibodies against GDF15 have shown potential for the treatment of cachexia and cancer. The receptor for GDF15 is unknown.

[0005] Wasting diseases such as cachexia or sarcopenia and inflammation such as systemic inflammation or acute inflammatory responses for therapeutic agents effective in treating weight loss associated with several diseases and conditions, including the There is significant unmet need: cancer, chronic kidney disease, chronic obstructive pulmonary disease, AIDS, tuberculosis Involved in chronic inflammatory diseases, systemic inflammation, and involuntary weight loss and / or muscle loss There is a met need for effective therapeutic agents to treat chronic diseases, including muscle wasting diseases, including those There are also significant needs that are not addressed. Summary of the Invention

[0006] (overview) The present disclosure provides methods for the treatment of GDNF family members, including binding proteins such as antibodies that bind to GFRAL proteins. Proteins that bind to GFRAL protein, including antibodies, are provided. Such binding proteins may be GFRAL polypeptides, GFRAL fragments, and / or GFRAL epitopes. Such binding proteins, including antibodies, are capable of binding to the antagonists. (e.g., inhibits the binding of GDF15 protein to GFRAL protein, inhibits the binding of GDF15 protein to GFRAL protein, It inhibits the binding of RET protein to proteins and blocks GDF15-induced signal transduction. and / or inhibit the formation of GDF15 / GFRAL or GDF15 / GFRAL / RET receptor complexes This can be done.

[0007] The present disclosure provides a method for detecting a GDF15 protein that (i) binds to a GFRAL protein and (ii) binds to a GFRAL protein. and / or (iii) inhibiting the binding of the RET protein to the GFRAL protein. Also provided are binding proteins, including antibodies (e.g., monoclonal antibodies) or fragments thereof, that bind to the do.

[0008] In some embodiments, the anti-GFRAL antibody is a GFRAL polypeptide, a GFRAL fragment, or a GFRAL polypeptide. In one embodiment, the anti-GFRAL antibody is a humanized antibody that binds to the GFRAL epitope. 1C1, 3P10, 12A3, 5F12, 5A20, 8D8, 17J16, 25M22, 2B8, 22N5, 2 I23, 6N16, 1B3, 19K19, 2B3, 8C10, 2A9, 24G2, 6G9, 2B11, 1A3, P1B6, P1H8, or are the VH CDR1, VH CDR2, VH CDR3 of the monoclonal antibody designated P8G4 or its humanized variants. In certain embodiments, the anti-GFRAL antibody comprises a VL CDR1, a VL CDR2, and / or a VL CDR3. VH FR1, VH FR2, VH FR3, VH FR4 of a human immunoglobulin amino acid sequence or a variant thereof , VL FR1, VL FR2, VL FR3, and / or VL FR4.

[0009] In some embodiments, the binding protein (e.g., an anti-GFRAL antibody) comprises six CDRs or In some embodiments, the binding protein (e.g., anti-GFRAL) comprises fewer than six CDRs. antibody) is selected from VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2, and / or VL CDR3 In some embodiments, the recombinant polypeptide comprises one, two, three, four, five, or six CDRs. The conjugate protein (e.g., anti-GFRAL antibody) can be any of the antibodies 1C1, 3P10, 12A3, 5F12, 5F16, 5F18, 5F19, 5F20, 5F21, 5F22, 5F23, 5F24, 5F25, 5F26, 5F27, 5F28, 5F29, 5 5A20, 8D8, 17J16, 25M22, 2B8, 22N5, 2I23, 6N16, 1B3, 19K19, 2B3, 8C10, 2A9, 24G2 , 6G9, 2B11, 1A3, P1B6, P1H8, or P8G4 or a monoclonal antibody thereof From the VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2, and / or VL CDR3 of the humanized variant In some embodiments, the CDRs comprise one, two, three, four, five, or six selected CDRs. Thus, the binding protein (e.g., anti-GFRAL antibody) may contain a human immunoglobulin amino acid sequence or sequences thereof. VH FR1, VH FR2, VH FR3, VH FR4, VL FR1, VL FR2, VL FR3, and / or VL FR 4.

[0010] In some embodiments, the antibody is a humanized antibody, a monoclonal antibody, or a recombinant antibody. , an antigen-binding fragment, or any combination thereof. The present invention relates to a GFRAL polypeptide (e.g., cell surface-expressed or soluble GFRAL), a GFRAL fragment, or a GFRAL polypeptide. It is a humanized monoclonal antibody or antigen-binding fragment thereof that binds to the FRAL epitope.

[0011] The present disclosure provides: (i) a method for detecting an antibody against a GFRAL polypeptide (e.g., a cell surface antigen) that binds to a GFRAL polypeptide; surface-expressed or soluble GFRAL), a GFRAL fragment, or a GFRAL epitope (e.g., and / or (ii) competitively blocks the GFRAL signaling pathway by the anti-GFRAL antibodies provided herein. Also included are binding proteins, e.g., anti-GFRAL antibodies, that bind to the GFRAL epitope bound by In other embodiments, the binding protein, e.g., an anti-GFRAL antibody, is a 2. The monoclonal antibody 25M22, 3P10, 8D8, or 5F12 described in 2. above, or a humanized variant thereof. The body may express a GFRAL polypeptide (e.g., a cell surface-expressed or soluble GFRAL protein), GFRAL The fragment or fragments competitively block (eg, in a dose-dependent manner) binding to the GFRAL epitope. In other embodiments, the binding protein, e.g., the anti-GFRAL antibody, is a and monoclonal antibodies 25M22, 3P10, 8D8, or 5F12, or humanized variants thereof. Binds to (e.g., recognizes) the GFRAL epitope.

[0012] The present disclosure provides (i) a heavy chain having the amino acid sequence of SEQ ID NO: 3, 7, 11, or 15, respectively; a light chain variable region having the amino acid sequence of SEQ ID NO: 4, 8, 12, or 16; (ii) binds to an epitope of the GFRAL protein that is recognized by the antibody; or Binding to the protein was assessed by the use of antibodies having the amino acid sequences of SEQ ID NOs: 3, 7, 11, or 15, respectively. a heavy chain variable region having the amino acid sequence of SEQ ID NO: 4, 8, 12, or 16; and a light chain variable region having the amino acid sequence of SEQ ID NO: 4, 8, 12, or 16. Also provided are binding proteins, including antibodies or fragments thereof, that compete with antibodies comprising In one embodiment, one or more amino acids of a GFRAL protein (e.g., a human GFRAL protein) are Binding proteins, including antibodies or fragments thereof, that bind to regions of the acid, including epitopes. In some embodiments, the binding tag comprises an antibody or a fragment thereof. The protein is located in a region of the GFRAL protein (e.g., one or more amino acid residues in the extracellular domain). (i) domain 1 of the GFRAL protein (e.g., the amino acid sequence of SEQ ID NO: 1797) amino acid residues Q20 to S130; (ii) domain 2 of the GFRAL protein (e.g., amino acid residues Q20 to S130 of SEQ ID NO: 1797); (iii) domain 3 of the GFRAL protein (e.g., amino acid residues C131 to C210); residues C220 to C316); or (iv) the extracellular domain of a GFRAL protein (e.g., the amino acid sequence of SEQ ID NO: 1797). These include those that bind to amino acid residues Q20 to E351.

[0013] In some embodiments, a specific epitope of a GFRAL protein (e.g., one or more Binding proteins, including antibodies or fragments thereof, that bind to the amino acid residues of These include, for example: (i) amino acid residues SER156, GLN of the GFRAL protein (SEQ ID NO: 1797) 147, LEU148, ALA149, SER150, TYR151, LEU152, LYS153, ALA154, CYS155, PHE174, TYR 175, GLU136, ALA137, CYS138, VAL139, GLY140, ASP141, VAL142, VAL143, CYS144, ASN 145, ALA146, LEU186, CYS189, CYS191, ALA192, GLN193, SER194, ASP195, ILE196, PRO 197, CYS198, GLN199, GLN200, SER201, LYS202, GLU203, ALA204, LEU205, HIS206, SER 207, SER130, CYS131, LEU132, GLU133, VAL134, or at least one of ALA135 (ii) an epitope of a GFRAL protein comprising (e.g., one or more) the amino acid sequence of SEQ ID NO: 1797; Amino acid residues LEU132, GLU133, VAL134, ALA135, GLU136, ALA137, CYS138, VAL139, GLY140 , ASP141, VAL142, VAL143, CYS144, ASN145, ALA146, GLN147, LEU148, ALA149, SER150 , TYR151, PHE174, TYR175, ALA169, ALA170, ILE171, ARG172, PHE173, GLN176, ASN177 , ILE178, PRO179, PHE180, ASN181, ILE182, ALA183, GLN184, MET185, LEU186, ALA187 , PHE188, or CYS189. (iii) an epitope of the L protein; (iii) amino acid residues LEU164, LYS208, VAL212, A of SEQ ID NO: 1797 SN213, MET214, VAL215, PRO216, PRO217, PRO218, THR219, CYS220, LEU221, VAL223, T RP245, LEU267, CYS269, GLN28, VAL289, GLN290, CYS291, THR292, CYS293, ARG294, TH R295, ILE296, THR297, GLN298, SER299, GLU300, GLU301, SER302, LEU303, CYS304, LY S305, ILE306, PHE307, GLN308, HIS309, MET310, LEU311, HIS312, ARG313, LYS314, SE containing at least one (e.g., one or more) of R315, CYS316, or PHE317 an epitope of the GFRAL protein; (iv) amino acid residues CYS233, ARG234, and ARG235 of SEQ ID NO: 1797; , HIS236, TYR237, ARG238, THR239, PHE240, GLN241, SER242, LYS243, CYS244, TRP245 , GLN246, ARG247, VAL248, THR249, ARG250, LYS251, CYS252, HIS253, GLU254, ASP255 , GLU256, ASN257, CYS258, ILE259, SER260, THR261, LEU262, SER263, LYS264, ASP266 , LEU267, THR268, SER272, ASP274, CYS275, ALA278, CYS269, SER270, SER302, LEU303 , ILE306, HIS309, LEU311, MET310, SER315, or at least one of CYS316 (e.g., (v) an epitope of the GFRAL protein comprising, for example, one or more of the amino acids of SEQ ID NO: 1797; Acid residues GLY140, LEU148, ALA149, ALA146, VAL142, ASN145, VAL139, ALA135, GLU136, LE U152, LEU132, SER201, ALA204, LEU205, LYS153, ILE196, PRO197, or GLN200 (vi) an epitope of a GFRAL protein comprising at least one (e.g., one or more) of the following: Amino acid residues GLU136, ALA137, VAL139, GLY140, ASP141, VAL142, and VAL143 of SEQ ID NO: 1797 , CYS144, ASN145, ALA146, GLN147, PHE173, ASN177, ILE178, PRO179, ASN181, ILE182 or a GFRAL protein containing at least one (e.g., one or more) of MET185. (vii) an epitope of at least one of amino acid residues GLN298 or GLU301 of SEQ ID NO: 1797; or (viii) a sequence comprising at least one (e.g., one or more) epitopes of a GFRAL protein; Amino acid residues ARG234, ARG238, GLN241, SER242, LYS243, TRP245, and GLN246 in column 1797 THR249, ARG250, LYS251, CYS252, HIS253, ASP255, ASN257, CYS258, SER260, THR261, or LEU262 of a GFRAL protein containing at least one (e.g., one or more) of Such antibodies provided above include those that bind to the epitope In one embodiment, for example, GDF15-induced signaling in cells expressing a GFRAL protein. Neural transduction and / or signal transduction of the GFRAL / GDF15 or RET / GFRAL / GDF15 receptor complexes. Furthermore, in some embodiments, the antibody is , monoclonal antibodies, for example, humanized, human, or chimeric antibodies.

[0014] In some embodiments, the binding proteins provided herein, e.g., anti-GFRA The antibody may be coupled to a diagnostic agent, a detectable agent (e.g., a radioisotope, an enzyme, a fluorescent compound, a bioluminescent conjugated or recombinantly linked to a fluorescent compound (e.g., a fluorescent compound, or a chemiluminescent compound) In some embodiments, the binding proteins provided herein, e.g., The anti-GFRAL antibody is used (e.g., administered) in conjunction with a therapeutic agent. In this case, the therapeutic agent is an inhibitor of activin-A, an inhibitor of ActRIIB, an inhibitor of IL-6, or an inhibitor of IL- 6R inhibitors, ghrelin, ghrelin mimetics, or GHS-RIa agonists, SARMs, TNFα inhibitors agents, IL-Ia inhibitors, myostatin inhibitors, beta-blockers, melanocortin peptide inhibitors The drug may include one or more drugs such as a melanocortin receptor inhibitor, a melanocortin receptor inhibitor, or an anti-cancer drug.

[0015] In certain embodiments, the binding proteins described herein, e.g., anti-GFRAL antibodies, Also provided herein are compositions comprising the conjugates described herein. A pharmaceutical composition comprising a protein, for example, an anti-GFRAL antibody.

[0016] The present disclosure provides methods for detecting GFRAL polypeptides, GFRAL polypeptide fragments, or GFRAL epitopes (e.g., , one or more amino acids of the GFRAL protein, including the extracellular domain of the GFRAL protein) Immunoglobulin heavy chains and immunoglobulin light chains of binding proteins (e.g., anti-GFRAL antibodies) , VH region, VL region, VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2, and / or VL CDR3 Also provided is an isolated nucleic acid molecule encoding the nucleic acid molecule are the same as those described herein: 1C1, 3P10, 12A3, 5F12, 5A20, 8D8, 17J16, 25M22, 2B8, 22N 5, 2I23, 6N16, 1B3, 19K19, 2B3, 8C10, 2A9, 24G2, 6G9, 2B11, 1A3, P1B6, P1H8, also or the VH region, VL region, and VH region of the monoclonal antibody designated P8G4 or its humanized variant. Each of the VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2, and / or VL CDR3 encodes a VH CDR1, VL CDR2, and / or VL CDR3. In some embodiments, the nucleic acid molecule comprises a human immunoglobulin amino acid sequence or a variant thereof. and / or VL FR4 of The scaffold or framework regions further encode the polypeptides provided herein. Provided are vectors containing nucleic acid molecules encoding binding proteins, e.g., anti-GFRAL antibodies. The host cells provided herein can be used in combination with a binding protein, e.g., By culturing under conditions that promote the production of anti-GFRAL antibodies, binding proteins, e.g. A method for producing anti-GFRAL antibodies.

[0017] The present disclosure provides a method for treating GDF15-mediated diseases, disorders, or conditions (e.g., one or more symptoms), including GDF15-mediated diseases, disorders, or conditions. A method for treating, preventing, or ameliorating a medical condition, disorder, or disease in a person in need thereof.

[0023] Administering a binding protein provided herein, e.g., an anti-GFRAL antibody, to a subject, including a subject having and administering a therapeutically effective amount of the compound of formula (I) to treat, prevent, or alleviate the disease, disorder, or condition. In some embodiments, the disease, disorder, or condition is The present invention relates to a method for detecting a GDF15 protein (e.g., a human GDF15 protein) and / or a GFRAL protein (e.g., a human GDF15 protein). These may be caused by or otherwise associated with a protein (e.g., the human GFRAL protein). Related to this, for example, is GDF15-induced signaling in a subject. In certain embodiments, the disease, disorder, or condition is cachexia, sarcopenia, or By reducing the incidence, frequency, or severity of muscle wasting, bone wasting, or involuntary weight loss In certain embodiments, the disease, disorder, or condition is cachexia. In certain embodiments, the disease, disorder, or condition is cancer. The disease, disorder, or condition is a cardiovascular disease. In certain embodiments, the disease, disorder, Or the disease is a chronic inflammatory disease (e.g., chronic kidney disease, chronic obstructive pulmonary disease). In embodiments, the disease, disorder, or condition is induced by or is caused by a GDF15 protein. The effects of chemotherapy (e.g., anti-tumor agents) associated with the GDF15 protein, including elevated levels of GDF15, In cancers with reduced sensitivity (e.g., resistance) to anti-cancer antibodies (e.g., trastuzumab), be.

[0018] In some embodiments, the disease, disorder, or condition is cachexia, sarcopenia, muscle atrophy, or muscular atrophy. Wasting or muscle loss, bone wasting, involuntary weight loss (e.g., associated with illness, disorder, or disease) weight loss associated with or resulting from these. In this embodiment, the disease, disorder, or condition is selected from the group consisting of, but not limited to, cancer, chronic kidney disease, chronic pancreatic cancer, and ovarian cancer. obstructive pulmonary disease, AIDS, tuberculosis, chronic inflammatory diseases, sepsis and other forms of systemic inflammation, muscle Wasting, including muscular dystrophy, as well as the eating disorder known as anorexia nervosa Selected from the group of underlying diseases associated with cachexia.

[0019] In some embodiments, the methods of treating, preventing, or ameliorating include improving weight gain. or reduce weight loss, or improve muscle mass gain or reduce muscle mass loss. In some embodiments, the methods of treating, preventing, or ameliorating include methods of treating, preventing, or ameliorating The occurrence, frequency, or severity of fluid loss, sarcopenia or muscle wasting, bone wasting, or involuntary weight loss Improved methods of treating cancer by preventing, minimizing, or reducing its severity.

[0020] The present disclosure provides a method for detecting GFRAL in a sample, the method comprising: , comprising contacting the antibody with a binding protein described herein, e.g., an anti-GFRAL antibody. In one embodiment, the sample contains cells expressing GFRAL on their surface. include.

[0021] The present disclosure provides a method for producing a GFRAL polypeptide, a GFRAL fragment, or a GFRAL epitope described herein. Kits containing binding proteins that bind to the polypeptide, e.g., anti-GFRAL antibodies, are also provided. [Brief explanation of the drawings]

[0022] BRIEF DESCRIPTION OF THE DRAWINGS [Figure 1] Figure 1 shows a sequence alignment between various GFRAL proteins, with SEQ ID NOs listed in bold text within brackets.

[0023] [Figure 2] Figure 2 shows a sequence alignment between various GDF15 proteins, with SEQ ID NOs listed in bold text within brackets.

[0024] [Figure 3] FIG. 3 shows the results of experiments demonstrating the binding affinity of exemplary anti-GFRAL antibodies 1C1 and 3P10 to GFRAL protein.

[0025] [Figure 4] 4A-4B show alignments of the VH and VL sequences of exemplary anti-GFRAL antibodies, with SEQ ID NOs listed in bold text within brackets.

[0026] [Figure 5]Figures 5A-5F show alignments of the VH and VL sequences of exemplary anti-GFRAL antibodies that bind to Domain 1 (Figures 5A-5B), Domain 2 (Figures 5C-5D), or Domain 3 (Figures 5E-5F) of GFRAL. SEQ ID NOs are listed in bold text within parentheses.

[0027] [Figure 6] 6A-6B show the alignment of the VH and VL sequences of the humanized 1C1 antibody, with SEQ ID NOs listed in bold text within brackets.

[0028] [Figure 7] 7A-7B show the alignment of the VH and VL sequences of the humanized 25M22 antibody, with SEQ ID NOs listed in bold text within brackets.

[0029] [Figure 8] 8A-8B show the alignment of the VH and VL sequences of the humanized 17J16 antibody, with SEQ ID NOs listed in bold text within brackets.

[0030] [Figure 9] 9A-9B show the alignment of the VH and VL sequences of the humanized 5F12 antibody, with SEQ ID NOs listed in bold text within brackets.

[0031] [Figure 10] Figures 10A-10B show the alignment of the VH and VL sequences of the humanized 3P10 antibody, with SEQ ID NOs listed in bold text within brackets.

[0032] [Figure 11] FIG. 11 shows the results of a receptor antagonist assay using humanized GFRAL antibodies.

[0033] [Figure 12] FIG. 12 shows the results of an Elk1 reporter assay showing the humanized antibodies.

[0034] [Figure 13] 13A-13C show the results of experiments demonstrating the specificity of exemplary humanized anti-GFRAL antibodies.

[0035] [Figure 14] Figures 14A-14B show the results of experiments demonstrating anti-GFRAL antibody inhibition of GDF15-induced weight loss in DIO mice. For Figure 14A, from left to right, the treatments administered were PBS, 3P10, 1C1, 17J16, 5A20, 25M22, 5F12, and 1MO3 (on days d1, d2, and d3, respectively). For Figure 14B, from left to right, the treatments administered were PBS, 8D8, and 12A3 (on days d1, d2, and d3, respectively).

[0036] [Figure 15] FIG. 15 shows the results of experiments demonstrating anti-GFRAL antibodies in a model of GDF15-induced weight loss.

[0037] [Figure 16] FIG. 16 shows the results of experiments demonstrating the effect of anti-GFRAL antibodies on food intake.

[0038] [Figure 17] FIG. 17 shows the results of an experiment demonstrating the effect of anti-GFRAL antibodies on body weight in DIO mice.

[0039] [Figure 18] FIG. 18 shows the results of experiments demonstrating the effect of anti-GFRAL antibodies on food intake in DIO mice.

[0040] [Figure 19] FIG. 19 shows the results of experiments demonstrating the effect of anti-GFRAL antibodies on fat mass.

[0041] [Figure 20] FIG. 20 shows the results of experiments demonstrating the effect of anti-GFRAL antibodies on GDF15-induced weight loss and fat and lean mass loss in DIO mice.

[0042] [Figure 21] FIG. 21 shows the results of experiments demonstrating the effect of anti-GFRAL antibodies on GDF15-induced increased energy expenditure and decreased food intake in DIO mice.

[0043] [Figure 22] FIG. 22 shows the results of experiments demonstrating the effect of anti-GFRAL antibodies on GDF15-induced weight loss and fat and lean mass loss in lean mice.

[0044] [Figure 23] FIG. 23 shows the results of experiments demonstrating the effect of anti-GFRAL antibodies on GDF15-induced RER changes and GDF15-induced reduction in food intake in lean mice.

[0045] [Figure 24] FIG. 24 shows the results of an experiment demonstrating the effect of anti-GFRAL antibody (3P10) on body weight in lean mice.

[0046] [Figure 25] FIG. 25 shows the results of an experiment demonstrating the effect of administration of anti-GFRAL antibodies on food intake in lean mice.

[0047] [Figure 26] 26A-26C show the results of experiments demonstrating the effects of dietary adenine.

[0048] [Figure 27] FIG. 27A shows the results of an experiment demonstrating the effect of dietary adenine.

[0049] Figure 27B shows the effect of an exemplary anti-GFRAL antibody (3P10) on mice with chronic kidney injury. The results of the experiment are shown.

[0050] [Figure 28] FIG. 28 shows the results of an experiment demonstrating the effect of an exemplary anti-GFRAL antibody (3P10) on mice with chronic kidney injury.

[0051] [Figure 29] FIG. 29 shows the results of experiments demonstrating the effect of an exemplary humanized anti-GFRAL antibody (h3P10) on GDF15-induced weight loss.

[0052] [Figure 30] FIG. 30 shows an exemplary crystal of a complex of GFRAL and GDF15 proteins.

[0053] [Figure 31] FIG. 31 shows an exemplary GFRAL electron density map.

[0054] [Figure 32] Figure 32 shows an exemplary ribbon diagram of the GFRAL / GDF15 complex formed in the asymmetric GFRAL / GDF15 crystal unit, with GFRAL protein domains D2 and D3 designated as GFRAL D2 and GFRAL D3.

[0055] [Figure 33] Figure 33 shows an exemplary ribbon diagram of a dimer of two GFRAL / GDF15 complexes, with GFRAL protein domains D2 and D3 shown as GFRAL D2 and GFRAL D3.

[0056] [Figure 34] Figures 34A-34B show different surface representations of the dimers of the two GFRAL / GDF15 complexes.

[0057] [Figure 35] FIG. 35 shows the GFRAL amino acid residues interacting with GDF15 amino acid residues.

[0058] [Figure 36] Figures 36A-36D show the GFRAL / GDF15 interface. GFRAL protein domains D2 and D3 are designated GFRAL D2 and GFRAL D3.

[0059] [Figure 37] 37A-37B show different aspects of the superposition of GFRAL protein and GFRα1 shown as ribbon diagrams.

[0060] [Figure 38] Figures 38A to 38D show different aspects of the interaction between GFRAL and RET proteins in the RET / GFRAL / GDF15 model.

[0061] [Figure 39] Figures 39A-39B show the amino acid residues at the RET protein interface of the GFRAL protein.

[0062] [Figure 40] FIG. 40 shows exemplary crystals of a complex with GFRAL protein, 3P10 Fab, and 25M22 Fab produced under crystallization conditions B11, D11, and H8.

[0063] [Figure 41] FIG. 41 shows exemplary crystals of a complex with GFRAL protein, 8D8 Fab, and 5F12 Fab produced under crystallization conditions C6, E11, and C2.

[0064] [Figure 42]FIG. 42 shows an exemplary electron density map of the 3P10 Fab and 25M22 Fab CDR regions in the crystal structure of the GFRAL / 3P10 / 25M22 Fab complex.

[0065] [Figure 43] FIG. 43 shows an exemplary electron density of the GFRAL / 8D8 / 5F12 Fab complex.

[0066] [Figure 44] FIG. 44 shows an exemplary ribbon diagram of the GFRAL / 3P10 / 25M22 Fab complex formed in an asymmetric GFRAL / 3P10 / 25M22 Fab complex crystal unit.

[0067] [Figure 45] Figure 45 shows an alignment of the 3P10 Fab and 25M22 Fab CDR sequences (top row) with the GFRAL amino acid residues involved in 3P10 Fab and 25M22 Fab binding (bottom row). Residues involved in the GFRAL-Fab interaction are boxed. For 3P10 Fab, amino acid residues Q1-S120 of SEQ ID NO: 1824 are shown for Hc, and amino acid residues D1-F120 of SEQ ID NO: 1825 are shown for Lc. For 25M22 Fab, amino acid residues Q1-S120 of SEQ ID NO: 1826 are shown for Hc.

[0068] [Figure 46] Figure 46 shows a ribbon diagram illustrating the interaction of the GFRAL 3P10 Fab epitope with the 3P10 Fab heavy chain CDR regions.

[0069] [Figure 47] Figure 47 shows a ribbon diagram illustrating the interaction of the GFRAL 3P10 Fab epitope with the 3P10 light chain CDR regions.

[0070] [Figure 48]Figure 48 shows a ribbon diagram illustrating the interaction of the GFRAL 25M22 epitope with the 25M22 Fab heavy chain CDR regions.

[0071] [Figure 49] Figure 49 shows the amino acid sequences of the GFRAL protein (residues S130-N318 of SEQ ID NO: 1797) and 25M22 Fab heavy chain (HC) residues Q1-P138 and G146-P225 of SEQ ID NO: 1826 and light chain (LC) residues D1-E218 of SEQ ID NO: 1827. Residues with a gray background and white text indicate core interaction interface amino acids of both the GFRAL protein and the 25M22 Fab. Residues on the Fab HC or LC with a gray background and black or white lettering indicate exemplary CDR sequences of the 25M22 Fab.

[0072] [Figure 50] Figures 50A-50B show the core interacting interface amino acid residues on the GFRAL protein and on 25M22 Fab that are involved in the GFRAL / 25M22 Fab interaction.

[0073] [Figure 51] Figure 51 shows the boundary interaction interface amino acid residues on the GFRAL protein involved in the GFRAL / 25M22 Fab interaction.

[0074] [Figure 52] FIG. 52 shows an exemplary side view of a ribbon diagram showing the overlapping 25M22 Fab and GDF15 epitopes (core interacting interface amino acids) on the GFRAL protein as a space-filling surface model.

[0075] [Figure 53] FIG. 53 shows a top view of a ribbon diagram showing the overlapping 25M22 Fab and GDF15 epitopes (core interacting interface amino acids) on the GFRAL protein as a space-filling surface model.

[0076] [Figure 54] Figure 54 shows the amino acid sequences of the GFRAL protein (residues S130-N318 of SEQ ID NO: 1797) and the 3P10 Fab heavy chain (HC) residues Q1-A130 and G138-C221 of SEQ ID NO: 1824 and the light chain (LC) residues D1-C218 of SEQ ID NO: 1825. Residues with a gray background and white text indicate core interacting interface amino acids of both the GFRAL protein and the 3P10 Fab. Residues on the Fab HC or LC with a gray background and black or white lettering indicate exemplary CDR sequences of the 3P10 Fab.

[0077] [Figure 55] Figures 55A-55B show ribbon diagrams depicting the crystal structure of the GFRAL / 3P10 Fab complex, with interacting interface residues shown as stick models (Figure 55A) or space-filling surface models (Figure 55B).

[0078] [Figure 56] Figures 56A-56B show the interface residues of the GFRAL 3P10 Fab epitope as a space-filling surface model in a ribbon diagram of GFRAL, with Figure 56A showing the core interacting interface residues and Figure 56B showing the boundary interacting interface residues.

[0079] [Figure 57] Figure 57 shows the residues of the GFRAL protein that bind to the overlapping 3P10 Fab and the residues of the GFRAL protein that bind to the RET protein as a space-filling surface model in a ribbon diagram of the GFRAL protein.

[0080] [Figure 58] Figure 58 shows the combined coverage of the boundary interaction interface residues of GFRAL protein bound to 3P10 Fab and 25M22 Fab.

[0081] [Figure 59]FIG. 59 shows an exemplary ribbon diagram of the GFRAL / 8D8 / 5F12 Fab complex formed in an asymmetric GFRAL / 8D8 / 5F12 Fab complex crystal unit.

[0082] [Figure 60] Figure 60 shows an exemplary ribbon diagram of the 8D8 Fab and 5F12 Fab binding sites on the GFRAL protein. Residues on the GFRAL protein important for Fab binding are shown as stick models.

[0083] [Figure 61] Figure 61 shows the amino acid sequences of the GFRAL protein (residues S130-N318 of SEQ ID NO: 1797) and the 8D8 Fab heavy chain (HC) residues Q1-K217 of SEQ ID NO: 1828 and light chain (LC) residues D1-R211 of SEQ ID NO: 1829. Residues with a gray background and white text indicate core interaction interface amino acids of both the GFRAL protein and the 8D8 Fab. Residues on the Fab HC or LC with a gray background and black or white lettering indicate exemplary CDR sequences of the 8D8 Fab.

[0084] [Figure 62] Figures 62A, 62B, 62C, and 62D show the core and boundary amino acid residues in the GFRAL / 8D8 Fab interaction interface.

[0085] [Figure 63] Figures 63A, 63B, 63C, and 63D show the core and boundary amino acid residues in the GFRAL / 5F12 Fab interaction interface.

[0086] [Figure 64]Figure 64 shows the amino acid sequences of GFRAL (residues S130-N318 of SEQ ID NO: 1797) and 5F12 Fab heavy chain (HC) residues Q1-K223 of SEQ ID NO: 1830 and light chain (LC) residues N1-E217 of SEQ ID NO: 1831. Residues with a gray background and white text indicate core interaction interface amino acids of both the GFRAL protein and the 5F12 Fab. Residues on the Fab HC or LC with a gray background and black or white lettering indicate exemplary CDR sequences of a 5F12 Fab. DETAILED DESCRIPTION OF THE INVENTION

[0087] (Detailed explanation) Binding proteins that bind to GFRAL proteins, including human GFRAL proteins, e.g. Antibodies are provided herein. Such binding proteins, including the antibodies disclosed herein, A unique property of proteins is their antagonistic properties, which include blocking the effects of GDF15 protein. and / or inhibit the binding of GDF15 protein to GFRAL protein. or the ability to inhibit the binding of the RET protein to the GFRAL protein, including These include cases where inhibition of binding reduces (e.g., blocks) GDF15 signaling. Notably and specifically, binding proteins to the GFRAL protein disclosed herein, e.g. For example, the antibody (i) binds to the GFRAL protein and (ii) binds to the GDF15 protein. and / or (iii) inhibiting the binding of the RET protein to the GFRAL protein. This can be measured, for example, by several in vitro cell-based assays. GDF15 / GFRAL protein complex or GDF15 / GFRAL / RET protein complex, including Such assays include blocking the formation of steroid hormones or GDF15 signaling. (1) ELK1-luciferase reporter assay (see, e.g., Example 3); and / or (2) U2 Examples include ERK phosphorylation assay in OS cells (see, for example, Example 4). Thus, the binding proteins described herein, e.g., anti-GFRAL antibodies, can be used to inhibit in vivo ( For example, it is thought to inhibit GDF15 activity (which is associated with the signaling function of GDF15). Due to their nature, the disclosed binding proteins, including anti-GFRAL antibodies, bind to GDF15 proteins (e.g., For example, human GDF15 protein) and / or GFRAL protein (for example, human GFRAL protein) Diseases, disorders, or illnesses caused by or otherwise associated with said protein , for example, for the treatment of conditions associated with GDF15-induced signaling in a subject. It can be a useful therapeutic agent.

[0088] The binding proteins provided herein, e.g., antibodies that bind to GFRAL proteins, can be i) binding of GDF15 protein to GFRAL protein and / or (ii) binding of GDF15 protein to GFRAL protein The present invention relates to a method for treating RET protein binding, and the method for treating RET protein binding is also provided herein. The anti-GFRAL antibodies include 1C1, 3P10, 1C2, 1C3, 1C4, 1C5, 1C6, 1C7, 1C8, 1C9, 1C10, 1C11, 1C12, 1C13, 1C14, 1C15, 1C16, 1C17, 1C18, 1C19, 1C20, 1 2A3, 5F12, 5A20, 8D8, 17J16, 25M22, 2B8, 22N5, 2I23, 6N16, 1B3, 19K19, 2B3, 8C10 , 2A9, 24G2, 6G9, 2B11, 1A3, P1B6, P1H8, and / or P8G4. The present invention also includes humanized anti-GFRAL antibodies, including humanized anti-GFRAL antibodies based on the present invention. Such anti-GFRAL antibodies bind to specific domains of the GFRAL protein, including: For example: (i) domain 1 of the GFRAL protein (e.g., amino acid residues Q20 to S130 of SEQ ID NO: 1797) (ii) domain 2 of the GFRAL protein (e.g., amino acid residues C131 to C210 of SEQ ID NO: 1797); ( iii) domain 3 of the GFRAL protein (e.g., amino acid residues C220 to C316 of SEQ ID NO: 1797); or (iv) the extracellular domain of the GFRAL protein (e.g., amino acid residues Q20 to E35 of SEQ ID NO: 1797) 1) is included.

[0089] In some embodiments of the present disclosure, the binding protein, e.g., the anti-GFRAL antibody, is selected from the group consisting of antibodies listed in Table 1. 24. An immunoglobulin variable region containing one or more complementarity-determining regions (CDRs) as set forth in In such a binding protein (e.g., an anti-GFRAL antibody), the CDRs may comprise one or more The scaffold or framework regions may be connected to the CDRs. The CDRs are oriented to achieve the appropriate antigen binding characteristics of the anti-GFR Such binding proteins, including AL antibodies, include: (i) binding proteins for GDF15 protein and GFRAL protein; interaction and / or (ii) inhibiting the interaction of the RET protein with the GFRAL protein (e.g., Such binding proteins, including the anti-GFRAL antibodies described herein, can be used to block the binding of GFRAL to GFRAL. The protein can inhibit (e.g., block) GDF15 signaling.

[0090] In some embodiments of the present disclosure, the binding protein, e.g., an anti-GFRAL antibody, comprises: (i) G Amino acid residues SER156, GLN147, LEU148, ALA149, and SER150 of the FRAL protein (SEQ ID NO: 1797) , TYR151, LEU152, LYS153, ALA154, CYS155, PHE174, TYR175, GLU136, ALA137, CYS138 , VAL139, GLY140, ASP141, VAL142, VAL143, CYS144, ASN145, ALA146, LEU186, CYS189 , CYS191, ALA192, GLN193, SER194, ASP195, ILE196, PRO197, CYS198, GLN199, GLN200 , SER201, LYS202, GLU203, ALA204, LEU205, HIS206, SER207, SER130, CYS131, LEU132 , GLU133, VAL134, or ALA135 (e.g., one or more) (ii) an epitope of the GFRAL protein comprising amino acid residues LEU132, GLU133, and VA of SEQ ID NO: 1797; L134, ALA135, GLU136, ALA137, CYS138, VAL139, GLY140, ASP141, VAL142, VAL143, CY S144, ASN145, ALA146, GLN147, LEU148, ALA149, SER150, TYR151, PHE174, TYR175, AL A169, ALA170, ILE171, ARG172, PHE173, GLN176, ASN177, ILE178, PRO179, PHE180, AS N181, ILE182, ALA183, GLN184, MET185, LEU186, ALA187, PHE188, or CYS189 an epitope of the GFRAL protein comprising at least one (e.g., one or more) of the following; ) Amino acid residues LEU164, LYS208, VAL212, ASN213, MET214, VAL215, PRO21 of SEQ ID NO: 1797 6, PRO217, PRO218, THR219, CYS220, LEU221, VAL223, TRP245, LEU267, CYS269, GLN28 , VAL289, GLN290, CYS291, THR292, CYS293, ARG294, THR295, ILE296, THR297, GLN298 , SER299, GLU300, GLU301, SER302, LEU303, CYS304, LYS305, ILE306, PHE307, GLN308 , HIS309, MET310, LEU311, HIS312, ARG313, LYS314, SER315, CYS316, or PHE317 an epitope of a GFRAL protein comprising at least one (e.g., one or more) of: (iv) amino acid residues CYS233, ARG234, ARG235, HIS236, TYR237, ARG238, TH of SEQ ID NO: 1797 R239, PHE240, GLN241, SER242, LYS243, CYS244, TRP245, GLN246, ARG247, VAL248, TH R249, ARG250, LYS251, CYS252, HIS253, GLU254, ASP255, GLU256, ASN257, CYS258, IL E259, SER260, THR261, LEU262, SER263, LYS264, ASP266, LEU267, THR268, SER272, AS P274, CYS275, ALA278, CYS269, SER270, SER302, LEU303, ILE306, HIS309, LEU311, ME containing at least one (e.g., one or more) of T310, SER315, or CYS316 (v) an epitope of the GFRAL protein; (v) amino acid residues GLY140, LEU148, and ALA149 of SEQ ID NO: 1797 , ALA146, VAL142, ASN145, VAL139, ALA135, GLU136, LEU152, LEU132, SER201, ALA204 , LEU205, LYS153, ILE196, PRO197, or GLN200 (e.g., 1 (vi) an epitope of the GFRAL protein comprising one or more amino acid residues of SEQ ID NO: 1797; LU136, ALA137, VAL139, GLY140, ASP141, VAL142, VAL143, CYS144, ASN145, ALA146, G At least one of LN147, PHE173, ASN177, ILE178, PRO179, ASN181, ILE182, or MET185 (vii) an epitope of a GFRAL protein comprising at least one (e.g., one or more) of the sequence At least one of amino acid residues GLN298 or GLU301 (e.g., one or more of GLN298 and GLU301) at position 1797 or (viii) an epitope of a GFRAL protein comprising amino acid residue A of SEQ ID NO: 1797; or RG234, ARG238, GLN241, SER242, LYS243, TRP245, GLN246, THR249, ARG250, LYS251, C At least one of YS252, HIS253, ASP255, ASN257, CYS258, SER260, THR261, or LEU262 binds to at least one (e.g., one or more) epitopes of the GFRAL protein Such antibodies as provided above may, in some embodiments, be used in combination with, for example, GFRAL. GDF15-induced signaling and / or GFRAL / GDF15 signaling in cells expressing the protein or inhibiting signal transduction or activation of the RET / GFRAL / GDF15 receptor complex. Furthermore, in some embodiments of the present disclosure, the antibody is a monoclonal antibody, e.g. For example, a humanized antibody.

[0091] (General technique) The techniques and procedures described or referred to herein are generally well understood by those skilled in the art. and / or conventional methodologies, e.g., Sambrook et al., Molecular Cloning: Experimental Methods. Molecular Cloning: A Laboratory Manual, 3rd Edition (2001) Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY; Current Protocols in Molecular Biology Protocols in Molecular Biology (eds. FM Ausubel et al. (2003)); Therapeutic monoclonal antibodies :Therapeutic Monoclonal Antibodies: From Bench to Clinic, Z. An, ed., Wiley, Hoboken NJ (2009); Monoclonal Antibodies: Methods and Protocols l Antibodies: Methods and Protocols), edited by M. Albitar, Humana Press, Totawa, NJ (2 010); and Antibody Engineering, 2nd Edition, Vol. 1 and 2, Kont A widely used method is described in "Theoretical and Applied Chemistry of Biomolecules," edited by Ermann and Dubel, Springer-Verlag, Heidelberg, 2010. This includes commonly used methodologies.

[0092] (technical term) Unless otherwise defined, all technical and scientific terms used herein are understood by those skilled in the art. For the purposes of interpreting this specification, the following terms shall have the same meaning as commonly understood: Wherever appropriate, explanations of words apply and terms used in the singular include the plural and vice versa. All patents, applications, published applications, and other publications are hereby fully incorporated by reference. Any explanation of a described term is incorporated herein by reference. In the event of any conflict with the terms in this document, the following explanations of terms shall prevail.

[0093] "GDNF family receptor alpha-like," "growth differentiation factor 15 receptor," "GFRAL," or The term "GFRAL protein" and similar terms refer to polypeptides (e.g., PEG-1, PEG-2, PEG-3, PEG-4, PEG-5, PEG-6, PEG-7, PEG-8, PEG-9, PEG-10, PEG-11, PEG-12, PEG-13, PEG-14, PEG-15, PEG-16, PEG-17, PEG-18, PEG-19, "Polypeptide" and "protein" are used interchangeably herein) or mammalian , for example, primates (e.g., humans, cynomolgus monkeys (cyno)), dogs, and rodents (e.g., refers to any native GFRAL from any vertebrate source, including mice and rats, and In certain embodiments, this includes related GFRAL polypeptides, including SNP variants thereof. GFRAL is a gene encoding "C6orf144," "chromosome 6 open reading frame 144," Also known in the art as "BA360D14.1", "IVFI9356", and "UNQ9356".

[0094] The amino acid sequence of the full-length precursor human GFRAL is provided below, which includes the signal peptide The peptide sequence (underlined lowercase residues) includes: [ka]

[0095] The amino acid sequence of the mature human GFRAL polypeptide is provided below: [ka]

[0096] In some embodiments, GFRAL has the amino acid sequence of mature human GFRAL (SEQ ID NO: 1798). A binding protein refers to a protein that is at least 55% identical to a The anti-GFRAL antibodies shown bind to GFRAL and / or bind to the signaling domain as described herein. In some embodiments, the antibodies described herein can modulate signal transduction. , binds to human GFRAL.

[0097] Human GFRAL contains an extracellular domain (e.g., residues 20-351 of SEQ ID NO: 1797), a transmembrane domain (e.g., residues 352-371 of SEQ ID NO: 1797), and the cytoplasmic domain (e.g., It has residues 372 to 394).

[0098] The nucleic acid sequence encoding the precursor GFRAL polypeptide is provided below: [ka] TIFF0007610541000004.tif112170.

[0099] As used herein, "GFRAL" refers to human GFRAL and, without limitation, to mouse GFRAL. RAL, rat GFRAL, cynomolgus monkey GFRAL, and its orthologues, including its variants. GFRAL is a member of the TGFβRII family (NCBI reference sequences: NM_001024847.2 (GI:133908632); NM_003242. 5 (GI:133908633)) or its ortholog. GFRAL is not related to TGFβRI (NCBI reference sequence: NP_ 001124388.1 (GI:195963412); NP_004603.1 (GI:4759226)) or its orthologs. In some embodiments, GFRAL is at least 55%, 60%, 65%, 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100%, 101%, 102%, 103%, 104%, 105%, 10 5%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or more identical Such exemplary GFRAL proteins include proteins having an amino acid sequence similar to that of the GFRAL protein. As shown in Figure 1, the results were obtained from chimpanzees (99%), cynomolgus monkeys (92%), and giant pandas. Dad (82%), Dog (81%), Cat (80%), Pig (77%), Cow (75%), Mouse (70%), Rat (70%), Chinese hamster (65%), and platypus (59%).

[0100] GFR from cynomolgus monkeys (cyno), scientific name Macaca fascicularis The amino acid sequence of the AL protein is provided below, and includes the signal peptide sequence (underlined residues) include: [ka] .

[0101] The coding nucleic acid sequence for the cynomolgus monkey GFRAL protein is provided below: [ka]

[0102] Amino acid sequence of the GFRAL protein from mouse, Mus musculus is provided below, including the signal peptide sequence (underlined residues). Can be: [ka]

[0103] The coding nucleic acid sequence for the mouse GFRAL protein is provided below: [ka] TIFF0007610541000009.tif249170TIFF0007610541000010.tif19170

[0104] The GFRAL protein from rats (Rattus norvegicus) The amino acid sequence is provided below, including the signal peptide sequence (underlined residues) include: [ka]

[0105] The coding nucleic acid sequence for the rat GFRAL protein is provided below: [ka]

[0106] The GFRAL protein or GFRAL may contain (e.g., variants and / or species) of one or more amino acid residues. have modifications (at positions not conserved beyond the conserved region) but retain the conserved domain, and It also refers to proteins that have biological activity similar to that of naturally occurring GFRAL. A DNA sequence that differs by one or more bases from an existing DNA sequence, but which nevertheless does not exist in nature due to the degeneracy of the genetic code. It may be encoded by a nucleic acid sequence that is translated into an amino acid sequence corresponding to a protein present in the The GFRAL protein may contain one or more conservative substitutions and / or substitutions with the naturally occurring sequence of GFRAL. It also refers to proteins that differ by the amount of the amino acid sequence and / or the conjugate.

[0107] The term "GFRAL" or "GFRAL protein" refers to the "full-length," unprocessed GFRAL protein. GF includes unmodified GFRAL and any form of GFRAL produced by intracellular processing. The term RAL or "GFRAL protein" includes: allelic variants (e.g., SNP variants); isoforms; fragments; derivatives; substitution, deletion, and insertion variants; fusion polypeptides and interspecies homologs, preferably those that retain GFRAL activity and / or elicit an anti-GFRAL immune response. As will be appreciated by those skilled in the art, the methods provided herein include those that are sufficient to generate The provided anti-GFRAL antibodies may be directed against GFRAL polypeptide fragments, GFRAL antigens, and / or GFRAL epitopes. The epitope can bind to the GFRAL protein, including the GFRAL antigen. The GFRAL antigen may be part of a larger GFRAL polypeptide fragment. and the GFRAL polypeptide fragment is a portion of a larger GFRAL protein. The GFRAL protein may be present in its native or denatured form. The GFRAL proteins described herein can be obtained from a variety of sources, for example, human tissue. isolated from the original form or from another source, or prepared by recombinant or synthetic methods The GFRAL protein has the same amino acid sequence as the corresponding naturally occurring GFRAL polypeptide. The GFRAL protein may be cleaved or secreted. and modified forms of the GFRAL polypeptide (e.g., extracellular domain sequence), mutations of the polypeptide. The present invention encompasses multiple forms (e.g., alternatively spliced ​​forms) and allelic variants. The GFRAL polypeptides described herein (e.g., human GFRAL) can be obtained from a variety of sources, e.g., human isolated from the target tissue type or from another source, or prepared by recombinant or synthetic methods. It is possible.

[0108] The GFRAL protein has at least 5 amino acids compared to the naturally occurring full-length GFRAL polypeptide. , at least 10, up to at least 50, or more amino acids. For example, a GFRAL protein can be prepared by cleaving a GFRAL polypeptide with the amino acid sequence of a naturally occurring GFRAL polypeptide. The GFRAL protein may also contain a signal sequence based on the naturally occurring G It may contain the same or similar post-translational modifications as the FRAL polypeptide, or may be free of post-translational modifications. For example, the protein may not contain a GFRAL polypeptide identical to a naturally occurring GFRAL polypeptide. It may have the same or a similar glycosylation pattern, or may contain no glycosylation at all. In other embodiments, the GFRL protein is a naturally occurring GFRL protein. Compared to the protein sequence, glycosyl groups are present at positions not present in the naturally occurring GFRAL protein. It contains a mutation that introduces a transcription site.

[0109] In one embodiment, the GFRAL protein is a gene encoding a GFRAL protein that expresses the GFRAL protein on its cell surface. The isolated GDF1 can be expressed by recombinant cells that have been genetically modified to express the GDF1. In some cases, the cells may be in a composition comprising a RET protein. For example, the cells may be genetically engineered to contain an exogenous sequence encoding a RET protein. The RET protein may be endogenously expressed without genetic modification. The cells may not express detectable levels of RET protein and may not express the exogenous sequence The RET protein may be genetically modified to express the RET protein derived from the recombinant human ovarian tumor.

[0110] Also disclosed herein are fragments of the GFRAL protein, e.g., native GFRAL. The intracellular domain present in the protein, or the native GFRAL protein, e.g. It lacks the intracellular and transmembrane domains present in native GFRAL, as shown in Figure 1. As noted above, fragments of the GFRAL protein also contain fragments of the native GFRAL. and may or may not contain a heterologous signal sequence. The fragment may lack the intracellular domain present in the native GFRAL protein. but contains a transmembrane domain.

[0111] The term "GFRAL-extracellular domain" ("GFRAL-ECD") includes the full-length GFRAL ECD, GFRAL EC As used herein, "GFRAL ECD" includes the GFRAL ECD fragment, the GFRAL ECD variant, and the GFRAL ECD variant. The term refers to a gene that lacks an intracellular and / or transmembrane domain, has or has a signal peptide. In some embodiments, the GFRAL ECD refers to a GFRAL polypeptide that does not The amino acid sequence of the amino acid sequence of the human full-length GFRAL ECD is at least 70% identical. It refers to a protein having the amino acid sequence: [ka]

[0112] As used herein, the term "full-length GFRAL ECD" refers to the last amino acid of the extracellular domain. GFRAL ECD, which may or may not include an N-terminal signal peptide, extending to the 5'-amino acid However, the term "full-length GFRAL ECD" refers to a sequence that includes the amino acid residues of the transmembrane domain. GFRAL-ECD extended at the C-terminus by 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 amino acids Note that the present invention also encompasses the following (provided that the polypeptide is soluble): Such a GFRAL ECD is long enough to prevent it from being anchored to the plasma membrane. The term "full-length GFRAL ECD" refers to a GFRAL ECD that lacks the transmembrane domain of the signal peptide. extended by 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 amino acids at the N-terminus to include amino acid residues In one embodiment, the GFRAL ECD is a GFRAL ECD as shown in FIG. Contiguous amino acid sequence and at least 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or more identical and having at least one C-terminal amino acid sequence identical to the GFRAL sequence shown in FIG. Lacking at least 30, 33, 35, 40, 45, 50, or 55 amino acids, or more; It refers to a continuous amino acid sequence.

[0113] The GFRAL ECD is the ECD of TGFβRII (accession numbers: NM_001024847.2; NM_003242.5). GFRAL ECD is not a member of TGFβRI (accession number: NP_00112438 8.1; NP_004603.1) or its orthologs. D is a compound having a nucleotide sequence identical to SEQ ID NO: 1806 by at least 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or more. %, 99% or more identical amino acid sequence to the protein of interest.

[0114] As used herein, the term "GFRAL ECD fragment" refers to a fragment of GFRAL ECD that is missing one or more residues from the full-length ECD. GFRAL EC, which is deleted from the N- and / or C-terminus of GFRAL and retains the ability to bind to GDF15. In some instances, the GFRAL ECD fragment may include an N-terminal signal peptide. In some examples, the GFRAL ECD fragment may be present at the N-terminus of the following sequence: a human GFRAL ECD fragment lacking 1, 5, 10, 15, 16, 17, 18, or 19 residues present in: [ka]

[0115] Another exemplary GFRAL ECD fragment corresponds to Q20 to C316 of the full-length human precursor GFRAL protein. It contains the following amino acid sequence: [ka]

[0116] Yet another exemplary GFRAL ECD fragment is from W115 to E351 of the full-length human precursor GFRAL protein. It contains the corresponding amino acid sequence: [ka] The above exemplary GFRAL ECD fragments may be used in combination with the GFRAL protein and the GDF15 protein or the GFRAL protein. Examples include those for producing crystals of complexes comprising proteins and exemplary anti-GFRAL antibodies. were used in a variety of ways as described in

[0117] Within the GFRAL protein or GFRAL ECD, there are three domains: domain 1 (D1), domain 2 ( In some embodiments, the D1 domain includes domain 2 (D2), and domain 3 (D3). The amino acid sequence is residues Q20 to S130 of SEQ ID NO: 1797. An exemplary amino acid sequence of the D2 domain is residues C131 to C210 of SEQ ID NO: 1797. In embodiments, the amino acid sequence of an exemplary D3 domain is residues C220 to C3 of SEQ ID NO: 1797. 16. Certain properties of GFRAL proteins are related to the activity of these domains, including those within the ECD. and / or binding. For example, as described herein, The amino acid residues in this region are the core interaction for GFRAL protein binding to GDF15 protein. The amino acids identified are interface amino acids and / or boundary interaction interface amino acids. As described herein, the amino acid residues in D3 are GFRAL targets for the RET protein. Core and / or boundary interaction interface amino acids for protein binding It has been identified that there is.

[0118] The term "core interaction interface amino acid" or its grammatical equivalents refers to at least one Atoms of a given protein are within 4.5 Å or less of the interacting protein. acid residues (e.g., GDF15 protein or GFRAL protein interacting with RET protein) The distance of 4.5 Å is the van der Waals radius plus the possible water The atoms within the hydrogen bond allow the molecule to form a bond with the interacting protein.

[0119] The term "boundary interacting interface amino acid" or its grammatical equivalents refers to at least one Atoms of a given protein are within 5 Å or less of a core interface amino acid on a given protein. The amino acid residues of proteins (e.g., GFRAL, which interact with GDF15 protein or RET protein) (amino acids in the GFRAL protein that are within 5 Å of the core interaction interface amino acids of the protein) A distance of 5 Å or less refers to a distance less than 5 Å from a core interaction interface amino acid on a given protein. Protein binding to residues far apart is a function of the van der Waals interaction of the interacting proteins. This allows the device to be within the radius of the grid.

[0120] As used herein, the term "GFRAL ECD variant" refers to a variant of a GFRAL ECD that has an amino acid addition, deletion, or deletion. "GFRAL ECD" refers to a GFRAL ECD that contains a deletion or substitution and is still capable of binding to GDF15. Such variants should have at least 80%, 85%, 90%, 92%, 95%, 97%, or 99% similarity to the parent GFRAL ECD. It can be 8% or 99% identical.

[0121] In the art, MIC-1 (macrophage inhibitory cytokine-1), PDF (prostatic differentiation factor (child), PLAB (placental bone morphogenetic protein), NAG-1 (nonsteroidal anti-inflammatory drug (NSAID)-activated gene Growth differentiation factor 15 (GDF15), also known as TGF-PL, TGF-PL, and PTGFB, is a GDF15 is a member of the transforming growth factor β (TGF-β) superfamily. It is synthesized as a 62 kDa intracellular precursor protein that is subsequently cleaved by ATPase to form a 25 kDa It is secreted as a disulfide-bonded protein (see, e.g., Fairlie et al., J. Leukoc. Biol 65:2-5 (1999). GDF15 mRNA is expressed in tissues including the liver, kidney, pancreas, colon, and placenta. GDF15 expression in the liver is found in several tissues, and GDF15 expression in the liver is associated with other organs such as the liver, kidney, heart, and lung. It can be significantly upregulated upon injury to the vessels.

[0122] The GDF15 precursor contains a 29 amino acid signal peptide, a 167 amino acid pro-domain, and A 112 amino acid mature domain is excised from the pro-domain by a furin-like protease. It is a 308 amino acid polypeptide (NCBI reference sequence NP_004855.2; GI:153792495) containing be.

[0123] The amino acid sequence of the precursor human GDF15 polypeptide is provided below: [ka] Such a 308-amino acid GDF15 polypeptide is referred to as a "full-length" GDF15 polypeptide; The 112-amino acid GDF15 polypeptide (amino acids 197-308 of "full-length" GDF15) is the "mature" GDF1 5 polypeptide.

[0124] As used herein, "GDF15" refers to a polypeptide that includes the amino acid sequence of the mature human GDF15 polypeptide and a small amount of GDF15. The mature human GDF15 polypeptide includes proteins having an amino acid sequence that is at least 65% identical to the mature human GDF15 polypeptide. The amino acid sequence of the polypeptide is provided below: [ka]

[0125] The above exemplary mature human GDF15 is referred to as the GFRAL protein and the GDF15 protein or GFRAL protein. Various methods described in the Examples, including those for producing crystals of complexes containing proteins, are also described. was used in the method.

[0126] Unless otherwise indicated, the term "GDF15" refers to the 112 amino acid mature human sequence (e.g., Additionally, numerical references to specific GDF15 residues refer to the 112 amino acid sequence. The mature sequence is referred to (eg, residue 1 is Ala (A) and residue 112 is Ile (I) in SEQ ID NO: 1811). For example, from the GDF15 precursor amino acid sequence, three putative forms of “mature” human GDF15 (e.g. Three excision sites are predicted, resulting in 112 amino acids (110, 112, and 115 amino acids), but the 112 amino acid product The mature sequence is accepted as correct.

[0127] In the context of the present disclosure, "GDF15" or "GDF15 protein" refers to GDF15 derived from mouse (NP_035949 ; GI:170784848), chimpanzee (XP_009433302.1; GI:694973734), orangutan (XP_ 009251261.1 GI:686757768), Rhesus macaque (EHH29815; GI:355703324), Giant bread Da (XP_002912774; GI:301753921), gibbon (XP_004089328.1; GI:441627981), guinea pig rats (XP_003465238; GI:348558868), ferrets (AER98997; GI:355689945), and cattle (NP_00 1193227; GI:329664989), pig (NP_001167527; GI:291291599), dog (XP_541938; GI:571 01740), and the platypus ( Ornithorhynchus anatinus ); GDF15 orthologs from other mammalian species and their modifications, including AFV61279; GI:410111209 Such exemplary GDF15 proteins are shown in FIG. , which includes an alignment of various exemplary GDF15 proteins. The mature form of human GDF15 shares approximately 67% amino acid identity with the mouse orthologue.

[0128] In the art, there are known genes for the Ret proto-oncogene, cadherin-related family member 16, trans Rearranged During Transfection, RET Receptor Tyrosine Kinase enzyme, cadherin family member 12, proto-oncogene C-Ret, EC 2.7.10.1, CDHF12, CDHR 16, RET51, PTC, hydroxyaryl-protein kinase, RET transforming sequence, and receptor RET, also known as a cytoplasmic tyrosine kinase, transduces signals for cell growth and differentiation. RET is a receptor tyrosine kinase that acts as a co-receptor on the cell surface. , glial cell line-derived GDNF receptors when bound to members of the GDNF receptor alpha (GFRα) co-receptor GDNF (GDNF) ligands (in humans, GDNF, artemin, neurturin, and parsley) RET proteins (e.g., RET-ECD) are known as the primary signaling receptors for the thymocyte endothelial cells (TECs). ) consists of four consecutive cadherin-like domains (CLD1 to CLD4) followed by a membrane-juxtamembrane cysteine. As disclosed herein, the RET protein comprises a GF-receptor-enriched domain (CRD). It is a co-receptor with the RAL protein and GDF15 protein (e.g., a co-receptor with the RET protein). The receptor complexes described herein include GFRAL proteins, e.g., Examples include the RET / GFRAL complex, the GFRAL / GDF15 complex, and the RET / GFRAL / GDF15 complex.

[0129] As used herein, "Ret" or "RET" refers to a gene having the amino acid sequence of SEQ ID NO: 1813 or a sequence similar to that of SEQ ID NO: 1813. at least 75% identical, e.g., 77%, 79%, 80%, 85%, 90%, 95%, 96%, 97%, RE refers to proteins having amino acid sequences that are 98%, 99%, or more identical. T is distinct from TGFβRI and TGFβRII. SEQ ID NO: 1812 is the mature human TGFβRII lacking the signal peptide. The RET9 sequence is: [ka]

[0130] The amino acid sequence of the full-length precursor human RET protein is provided below, which includes: The signal peptide sequence (underlined lowercase residues) includes: [ka]

[0131] Therefore, as used herein, "RET" or "RET protein" refers to human RET and its related proteins. and variants thereof, including orthologues thereof, such as mouse RET, cynomolgus monkey RET, etc. In one embodiment, RET is selected from the group consisting of SEQ ID NO: 1813, SEQ ID NO: 1814, SEQ ID NO: 1815, SEQ ID NO: 1816, SEQ ID NO: 1817, SEQ ID NO: 1818, SEQ ID NO: 1819, SEQ ID NO: 1820, SEQ ID NO: 1821, SEQ ID NO: 1822, SEQ ID NO and at least 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or more The amino acid sequence may be identical to that of the target protein.

[0132] In some embodiments, an isolated RET extracellular domain (RET-ECD) polypeptide is provided. RET-ECD, when present in the isolated protein complex of the present disclosure, binds to the GFRAL protein. The "RET-extracellular domain" ("RET-ECD") may be bound to a ligand such as a protein. The term "RET ECD" includes full-length RET ECD, RET ECD fragments, and RET ECD variants. As used, the term "RET ECD" refers to a gene that lacks the intracellular and transmembrane domains. , refers to a RET polypeptide with or without a signal peptide. In the present invention, the RET ECD has the amino acid sequence of at least one amino acid sequence similar to that of the full-length human RET ECD having the amino acid sequence shown below. refers to proteins having an amino acid sequence that is at least 75% identical to: [ka]

[0133] In another exemplary embodiment, the RET ECD is a human full-length RET having the amino acid sequence: Refers to a protein having an amino acid sequence that is at least 75% identical to the amino acid sequence of the ECD : [ka]

[0134] As used herein, the term "full-length RET ECD" refers to the last amino acid sequence of the extracellular domain. refers to the RET ECD, which may or may not include an N-terminal signal peptide, and extends to the ribosomal amino acid sequence. However, the "full-length RET ECD" is a C-terminal fragment containing the amino acid residues of the transmembrane domain. Also included are RET-ECDs extended at the termini by 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 amino acids. Note that the polypeptide is soluble. D lacks a transmembrane domain of sufficient length so that it is not anchored to the cell membrane. The phrase "full length RET ECD" refers to the N-terminal sequence of the RET ECD, which is a sequence that includes the amino acid residues of the signal peptide. - Also includes RET-ECDs extended at the termini by 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 amino acids In certain embodiments, the RET fragment comprises consecutive amino acids of RET as described herein. sequence and at least 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or more than identical to and at least 30 at the C-terminus of the RET sequence described herein , 33, 35, 40, 45, 50, or 55 amino acids or more, It refers to the amino acid sequence.

[0135] As used herein, the term "RET ECD fragment" refers to a fragment in which one or more residues are N-terminal to the N-terminal of the full-length ECD. and / or R deleted from the C-terminus and retaining the ability to bind to the GFRAL protein. In some instances, a RET ECD fragment includes an N-terminal signal peptide. In some examples, the RET ECD fragment may or may not include the N-terminus of the following sequence: A human RET ECD fragment lacking 1, 5, 10, 15, 16, 17, 18, or 19 residues present in: [ka]

[0136] The above exemplary RET ECD fragments are used in the RET protein, the GFRAL protein, and the GDF15 protein. Various methods described in the Examples, including those for creating models of complexes containing proteins, are also described. used in law.

[0137] In an alternative embodiment of the RET ECD, the RET-ECD is the R of the human full-length RET ECD of SEQ ID NO: 1814. The ET contains a C64R, N75Q, N166Q, or C183S mutation in the ECD sequence.

[0138] The phrase "modulates activity and / or signaling" refers to a binding protein, e.g., When applied to antibodies that bind to GFRAL of the present disclosure, the binding protein (e.g., antibody) is (ii) GDF15 protein and GFRAL protein; or (iii) GDF15 protein In vitro or in vivo induced by binding of the GFRAL protein and the RET protein The term "anti-GFRAL antibody" refers to a compound that mimics or modulates the biological action of a substance. In assessing, for example, an antibody or The fragments are designed to enhance the biological response of wild-type GFRAL proteins (e.g., mature human GFRAL proteins). 95% or less of the activity of the original form, preferably 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45% %, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, or 95% or less The antibody or fragment thereof that binds to GFRAL (e.g., human GFRAL) is considered to induce the response. A fragment is also believed to induce a biological response when it possesses one or more of the following properties: performed: (1) an ELK1-luciferase reporter assay (see, e.g., Example 3); or (2) In an ERK-phosphorylation assay in U2OS cells (see, e.g., Example 4), For example, IC of 90nM, 80nM, 70nM, 60nM, 50nM, 40nM, 30nM, 20nM, 10nM, 1nM, 0.1nM, 0.01nM 50, indicating an efficacy level of 95% or less of the GFRAL standard.

[0139] The term "binding protein" refers to a protein that binds to a GFRAL protein, including the human GFRAL protein. a binding portion (e.g., one or more binding regions, e.g., CDRs), and optionally, the binding portion steric structures that promote binding of the conjugate protein to a GFRAL polypeptide, fragment, or epitope A scaffold or framework portion (e.g., one or more structural members) that allows the structure to be taken up. The term refers to proteins containing such binding proteins (scaffold or framework regions). Examples of proteins include antibodies, e.g., human antibodies, humanized antibodies; chimeric antibodies; recombinant antibodies; monoclonal antibodies; Monoclonal antibody; diabody; triabody; tetrabody; Fab fragment; F(ab')2 fragment; IgD antibody; Ig Examples of antibodies include E antibodies, IgM antibodies, IgG1 antibodies, IgG2 antibodies, IgG3 antibodies, and IgG4 antibodies, as well as fragments thereof. Binding proteins can be, for example, alternative proteins into which CDRs or CDR derivatives have been grafted. Such scaffolds may include solid or artificial scaffolds. The fold may include, for example, a protrusion introduced to stabilize the three-dimensional structure of the binding protein. antibody-derived scaffolds containing natural mutations and fully synthetic, e.g., biocompatible polymers Scaffolds include, but are not limited to, those described in, for example, Korndorfer et al. 2003, Proteins: Structure, Function, and Bioinformatics (2003); Roque et al., Biotechnol. Pr og. 20:639-654 (2004). Additionally, peptide antibody mimetics ("PAMs") and Antibody mimetic-based scaffolds utilizing fibronectin components as scaffolds In the context of the present disclosure, a binding protein can be used, e.g. , dissociation constant (K D ) is ≦10 -8 When M, it specifically or selectively binds to GFRAL. Binding proteins (e.g., antibodies) are known to bind to K D is ≦10 -9 M or K D is ≦10 -10 M In some embodiments, the antibody can specifically bind to GFRAL with high affinity. In the present invention, the binding protein (e.g., antibody) binds to GFRAL, e.g., about 10 -7 M ~ about 10 -12 K of M D in In other embodiments, the binding protein (e.g., antibody) is 1-2x 10 -9 K of M D can be combined with

[0140] The terms "antibody" and "immunoglobulin" or "Ig" are used interchangeably herein. It is used in the broadest sense, specifically, for example, the following individual anti GFRAL monoclonal antibodies (agonist, antagonist, neutralizing antibody, full-length or intact monoclonal antibody) Anti-GFRAL antibody combinations with multi-epitope or single-epitope specificity (including clonal antibodies) A composition, polyclonal or monovalent antibody, a multivalent antibody, formed from at least two intact antibodies Multispecific antibodies (e.g., bispecific antibodies), single chain antibodies, as long as they exhibit the desired biological activity. Antibodies include anti-GFRAL antibodies and fragments of anti-GFRAL antibodies. Antibodies include human antibodies, humanized antibodies, chimeric antibodies, and the like. and / or affinity matured antibodies, as well as antibodies derived from other species, e.g., mouse, rabbit, etc. The term "antibody" refers to a molecule capable of binding to a specific antigen. and polypeptides of the immunoglobulin class composed of two identical pairs of polypeptide chains. It is intended to include polypeptide products of B cells within a range of pairs, where each pair is one It has one heavy chain (approximately 50-70 kDa) and one light chain (approximately 25 kDa), and the amino terminal portion of each chain is Each chain contains a variable region of about 100 to about 130 or more amino acids, The hydroxy-terminal portion contains the constant region (Borrebaeck (ed.) (1995), Antibody Engineering dy Engineering, 2nd ed., Oxford University Press.; Kuby (1997), Immunology (See, for example, "Theory," 3rd ed., W.H. Freeman and Company, New York). Specific molecular antigens that can be bound by the antibodies provided herein include GF Antibodies include RAL polypeptides, GFRAL fragments, or GFRAL epitopes. Antibodies include synthetic antibodies, Monoclonal antibodies, recombinantly produced antibodies, multispecific antibodies (including bispecific antibodies), human antibodies Antibodies, humanized antibodies, camelized antibodies, chimeric antibodies, intrabodies, anti-idiotypes (anti-Ids) Antibodies, and functional fragments of any of the above (e.g., antigen-binding fragments, e.g., GFRAL binding fragments) a fragment of an antibody heavy chain or fragments thereof which retain some or all of the binding activity of the antibody from which the fragment is derived refers to a portion of a light chain polypeptide). For example, non-limiting examples of antigen-binding fragments (e.g., GFRAL-binding fragments) include single-chain Fvs (scFvs) ( For example, monospecific, bispecific, etc.), Fab fragments, F(ab') fragments, F(ab)2 fragments, F(ab ')2 fragments, disulfide-linked Fv (sdFv), Fd fragments, Fv fragments, diabodies, triabodies, In particular, the antibodies provided herein include immunoglobulins, tetrabodies, and minibodies. Immunoglobulin molecules and immunologically active portions of immunoglobulin molecules, e.g., antigen-binding domains or an antigen-binding site that binds to a GFRAL antigen (e.g., one or more complementarity-determining regions of an anti-GFRAL antibody). Such antibody fragments include molecules containing the CDRs (chimeric antibody fragments) of the antibody family. Reference, Antibodies: A Laboratory Manual, Cold Spring Harbor Laboratory Aboratory, New York (1989); Myers (ed.), Molecular Biology and Biotechnology: A Comprehensive Desk Reference (Mo lec. Biology and Biotechnology: A Comprehensive Desk Reference), New York: VCH P Hublisher, Inc.; Huston et al., Cell Biophysics, 22:189-224 (1993); Pluckthun and Sker ra, Meth. Enzymol., 178:497-515 (1989), and Day, ED, Advanced Immunochemistry (A Advanced Immunochemistry, 2nd ed., Wiley-Liss, New York, NY (1990). The antibodies provided herein can be found to be any of the immunoglobulin molecules. Type (e.g., IgG, IgE, IgM, IgD, IgA, and IgY), any class (e.g., IgG1, IgG IgG1, IgG2, IgG3, IgG4, IgA1, and IgA2), or any subclass (e.g., IgG2a and IgG2b) The anti-GFRAL antibody can be an agonistic or antagonistic antibody. The antibodies provided herein include antagonistic antibodies against GFRAL, e.g., antibodies that inhibit GFRAL signaling. Exemplary anti-GFRAL antibodies include those having the CDRs shown in Tables 1-24. Antibodies that

[0141] The term "about" or "approximately" means within 20% of a given value or range. , within 15%, within 10%, within 9%, within 8%, within 7%, within 6%, within 5%, within 4%, within 3% This means within, within 2%, or within 1%, or less.

[0142] An "antigen" is a predetermined antigen to which an antibody can selectively bind. peptides, carbohydrates, nucleic acids, lipids, haptens, or other natural or synthetic compounds. In some embodiments, the target antigen is a polypeptide.

[0143] The terms "antigen-binding fragment," "antigen-binding domain," "antigen-binding region," and similar terms The term refers to a binding agent that interacts with an antigen and transfers its specificity and affinity for the antigen to a binding substance. It refers to the portion of an antibody (eg, the complementarity determining region (CDR)) that contains the amino acid residues to be conferred.

[0144] The terms "binds" or "binding" refer to, for example, the formation of a complex. Complex refers to an interaction (e.g., covalent or non-covalent) between molecules, including the formation of a complex. A body is a group of two molecules held together by covalent or non-covalent bonds, interactions, or forces. The interactions may include, for example, hydrogen bonds, ionic bonds, loose bonds, non-covalent interactions, including aqueous and / or van der Waals interactions A single antigen-binding site on the antibody and a single target molecule, e.g., a single end of GFRAL, can be used. The strength of the overall non-covalent interactions between the epitopes determines the strength of the antibodies or is the affinity of the functional fragment. The ratio of antibody association (k1) to dissociation (k-1) for a monovalent antigen (k1 / k-1 ) is the association constant K, which is a measure of affinity. The value of K varies depending on the antibody-antigen complex. The association constant K of the antibodies provided herein varies depending on the The method can be determined using any of the methods provided in the document or any other method known to those skilled in the art. The affinity at one binding site does not always reflect the true strength of the interaction between antibody and antigen. Complex antigens containing multiple repeating antigenic determinants, e.g., multivalent GFRAL However, when exposed to an antibody containing multiple binding sites, the interaction of the antibody with the antigen at one site is The use of a multivalent antibody increases the likelihood of a reaction at a second site. The strength of such interactions is called avidity. The avidity of an antibody is determined by the strength of its individual binding sites. A high avidity can be a measure of the binding ability of a molecule that is superior to its affinity. For example, a high avidity can be This can compensate for the low affinity sometimes seen with pentameric IgM antibodies, and the pentameric IgM antibody It may have a lower affinity than G, but its multivalence results in I The high avidity of IgM allows it to bind to antigens efficiently.

[0145] "Antibodies that specifically bind to GFRAL" and "Antibodies that specifically bind to GFRAL epitopes" The terms and similar terms are also used interchangeably herein to refer to GFRAL polypeptides, e.g., GFRAL polypeptides. AL antigen, or fragment, or epitope (e.g., human GFRAL, e.g., human GFRAL polypeptide) GFRAL (e.g., human GFRAL) refers to an antibody that specifically binds to a specific target gene (e.g., a target gene, an antigen, or an epitope). The antibody that specifically binds to the extracellular domain of GFRAL or a peptide derived from the extracellular domain An antibody that specifically binds to a GFRAL antigen (e.g., human GFRAL) can bind to a peptide. can cross-react with related antigens (e.g., cynomolgus GFRAL). In the present study, antibodies that specifically bind to the GFRAL antigen do not cross-react with other antigens. Antibodies that specifically bind to the nucleotide sequence can be assayed using, for example, immunoassays, Biacore, or other assays known to those of skill in the art. Antibodies can be identified by techniques such as radioimmunoassay (RIA) and enzyme immunoassay. Any cross-reactivity as determined using laboratory techniques such as linked immunosorbent assay (ELISA) A compound that specifically binds to the GFRAL antigen is called a GFRAL antigen if it binds to the GFRAL antigen with higher affinity than the GFRAL antigen. Typically, a specific or selective reaction requires at least a small amount of background signal or noise. 2-fold and in some cases more than 10-fold above background. For a discussion, see Paul (ed.) (1989), Fundamental Immunology, 2nd ed., Raven P. See pages 332-336 of The Ress, New York. For example, an antibody that "binds" to a GFRAL (GFRAL) is one that targets cells or tissues that express the antigen. and which are useful as therapeutic agents in the treatment of cancer and do not significantly cross-react with other proteins. In such embodiments, "non-targeting" antibodies are antibodies that bind to the antigen with sufficient affinity. The extent of binding of the antibody to the protein can be determined, for example, by fluorescence activated cell sorting (FACS) analysis or radioactivity. The binding of an antibody to its specific target protein as determined by immunoprecipitation (RIA) With respect to the binding of an antibody to a target molecule, "specific binding" or specific polyclonal antibody binding is "specifically binds to" a polypeptide or epitope on a particular polypeptide target, or "Specific for" a particular polypeptide or epitope on a particular polypeptide target The term "specific binding" refers to binding that is measurably different from non-specific interactions. can be measured, for example, by determining binding of a molecule compared to binding of a control molecule. The control molecule is usually a molecule of similar structure that does not have binding activity. The differential 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 example, the binding of the labeled target to the probe can be determined by the excess of the unlabeled target. Specific binding is indicated if it is competitively inhibited by the labeled target. "Specific binding" or epitope binding on a particular polypeptide or a particular polypeptide target "specifically binds to" or on a particular polypeptide or a particular polypeptide target The term "specific for" an epitope means that a molecule is specific for, e.g., at least Also about 10 -4 M, or at least about 10 -5 M, or at least about 10 -6 M, or at least about 1 0 -7 M, or at least about 10 -8 M, or at least about 10 -9 M, or at least about 10 -10 M , or at least about 10 -11 M, or at least about 10 -12 M or higher Kd In one embodiment, the term "specific binding" can be expressed by The molecule does not substantially bind to any other polypeptide or polypeptide epitope. It refers to binding to a specific polypeptide or an epitope on a specific polypeptide. In some embodiments, the antibody that binds to GFRAL has a binding affinity of 10 nM, 5 nM, 4 nM, 3 nM, 2 nM, 1 nM, 0.9 nM, a dissociation constant (Kd) of less than M, 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 K D The lower the affinity of the anti-GFRAL antibody, the higher the affinity of the antibody. Anti-GFRAL antibodies can be used to differentiate GFRAL from different species (e.g., human GFRAL and cynomolgus monkey GFRAL). It binds to a conserved epitope of GFRAL (among

[0146] The term "compete" refers to binding to or to the same epitope or binding site on a target. Anti-GFRAL antibodies that compete for these (e.g., antagonistic antibodies that bind to GFRAL and antibodies that bind to GFRAL) When used in connection with a protein, the antibody (or binding fragment) under consideration However, a reference molecule (e.g., a reference ligand or is an assay that interferes with or inhibits the specific binding of a reference antigen-binding protein (e.g., a reference antibody). Many types of competitive binding assays can be used to test Determine whether the antibody competes with a reference antibody for binding to GFRAL (e.g., human GFRAL). Examples of assays that can be used include solid phase direct or indirect radioimmunoassays (RIAs). IA), solid-phase direct or indirect enzyme immunoassay (EIA), sandwich competitive assay (e.g., Sta (1983) Methods in Enzymology 9:242-253); solid-phase direct biotin-avidin Immunoassay (EIA) (e.g., Kirkland et al. (1986) J. Immunol. 137:3614-3619), solid-phase direct labeling assays, assay, solid-phase direct label sandwich assay (e.g., Harlow and Lane (1988), See Antibodies, A Laboratory Manual, Cold Spring Harbor Press. solid-phase direct labeling RIA using 1-125 label (see, e.g., Morel et al. (1988) Molec. Immunol 25:7-15); solid-phase direct biotin-avidin EIA (see, e.g., Cheung et al. (1990) Virol 176:546-552); and direct label RIA (Moldenhauer et al. (1990) Scand. J. Immunol. ol. 32:77-82). Typically, such assays involve the use of unlabeled test antigen-binding proteins. A labeled protein (e.g., a test anti-GFRAL antibody) or a labeled reference antigen-binding protein (e.g., a reference Purified antigen (e.g., GFRAL) bound to a solid surface or cells bearing either an anti-GFRAL antibody or an anti-GFRAL antibody. Competitive inhibition involves the use of a test antigen-binding protein (e.g., human GFRAL). This can be measured by determining the amount of label bound to the solid surface or cell. The test antigen-binding protein is usually present in excess. Antibodies identified by competitive assays ( Competing antibodies (competing antibodies) include antibodies that bind to the same epitope as the reference antibody and / or antibodies that bind to the same epitope as the reference antibody. Antibodies that bind to adjacent epitopes close enough to each other that the antibody sterically hindrances the epitope being bound. This includes methods for determining competitive binding and / or binding to the same epitope. Further details are provided herein. Typically, competing antibody proteins are present in excess. When a reference antibody is used, it reduces the specific binding of the reference antibody to a common antigen by at least 23%, e.g. , 40%, 45%, 50%, 55%, 60%, 65%, 70%, or 75% inhibition of binding. , at least 80%, 85%, 90%, 95%, 96%, or 97%, 98%, 99%, or more is greatly hindered.

[0147] The term "anti-GFRAL antibody" or "antibody that binds to GFRAL" refers to an antibody that targets GFRAL. and binds to GFRAL with sufficient affinity so as to be useful as a diagnostic and / or therapeutic agent when In some embodiments, the antibody can be used to identify unrelated, non-GFRAL proteins. The extent of binding of anti-GFRAL antibodies to the target protein can be determined, for example, by fluorescence-activated cell sorting (FACS) analysis or immunoassay. of antibodies to GFRAL as measured by an assay, e.g., a radioimmunoassay (RIA). Antibodies that "specifically bind to" or are "specific for" GFRAL have less than about 10% of the binding. In some embodiments, the GFRA described herein is used in combination with a GFRA antibody. The antibody binding to L was 10nM, 9nM, 8nM, 7nM, 6nM, 5nM, 4nM, 0.9nM, 0.8nM, 0.7nM, 0.6nM a dissociation constant (K In some embodiments, the anti-GFRAL antibody can be used to differentiate between GFRALs from different species (e.g., It binds to an epitope of GFRAL that is conserved between human GFRAL and cynomolgus monkey GFRAL.

[0148] As used herein, the terms "crystal" and "crystallized" refer to a substance that exists in crystalline form. A crystal is a form of a substance in its solid state, and This differs from other forms such as the amorphous solid state or the liquid crystal state. Crystals are formed by the separation of atoms, ions, molecules, and solids. molecules (e.g., proteins, e.g., antibodies), or molecular assemblies (e.g., ligand / receptor complexes). These three-dimensional arrays consist of regularly repeating three-dimensional arrays of antibodies (antigen / antibody complexes). are arranged according to specific mathematical relationships that are well understood in the art. The repeated basic unit, or building block, is called the asymmetric unit. The repetition of the asymmetric unit in an arrangement consistent with the crystallographic symmetry defined in Iteration of the unit cell by regular transformations in all three dimensions results in Giege, R. and Ducruix, A. Barrett, Nucleic Acid and Protein Crystals. Crystallization of Nucleic Acids and Proteins, a Practical Approach l Approach), 2nd edition, pp. 20 1-16, Oxford University Press, New York, NY, (1999) Please refer to.

[0149] An "isolated" antibody is free from cellular material or other contaminating proteins and other substances from a cell or tissue source. and / or are substantially free of other contaminants from which the antibody is derived, or are chemically synthesized. , chemical precursors, or other chemicals. "Substantially free of cellular material." The term "antibody" refers to a cellular component of the cell from which it is isolated or recombinantly produced. and preparations of antibodies that are isolated from the host and thus are substantially free of cellular material. contains less than about 30%, 25%, 20%, 15%, 10%, 5%, or 1% (by dry weight) foreign protein. In some embodiments, the present invention includes preparations of antibodies that contain contaminating proteins (also called "contaminating proteins"). In the present invention, when the antibody is recombinantly produced, it is substantially free of culture medium, e.g. For example, the culture medium may comprise less than about 20%, 15%, 10%, 5%, or 1% of the volume of the protein preparation. In certain embodiments, when an antibody is produced by chemical synthesis, it is It is substantially free of precursors or other chemicals, e.g., it is free of precursors or other chemicals involved in the synthesis of proteins. Therefore, the antibody is free from chemical precursors or other chemicals that cause the antibody to react with the antibody. The preparation contains less than about 30%, 25%, 20%, 15%, 10%, 5%, or 1% (by dry weight) of chemical precursors. Contaminants include, but are not limited to, antibodies. These may include substances that interfere with the body's therapeutic uses, and may also interfere with enzymes, hormones, and other proteins. Proteinaceous or non-proteinaceous solutes may be included. In one embodiment, antibodies (1) was determined by the Lowry method (Lowry et al., J. Bio. Chem. 193: 265-275, 1951). When the antibody is present in a solution containing more than 95% by weight of the antibody, for example, 96%, 97%, 98%, or 99% by weight of the antibody, (2) By using a spinning cup sequenator, N-terminal or internal amino acid sequences can be identified. (3) Coomassie blue staining or Preferably, the gel is purified to homogeneity by SDS-PAGE under reducing or non-reducing conditions with silver staining. The isolated antibody is purified so that at least one component of the antibody's natural environment will not be present. The term "antibody" includes antibodies in situ within recombinant cells. However, the term "antibody" generally refers to an antibody that has been isolated. The antibody is prepared by at least one purification step. The antibodies provided herein are isolated.

[0150] The four-chain antibody unit is a hetero-four-chain antibody composed of two identical light (L) chains and two identical heavy (H) chains. In the case of IgG, the four-chain unit is usually about 150,000 daltons. Each L chain is linked to an H chain by one covalent disulfide bond, but two H chains are linked to each other by one or more disulfide bonds depending on the H chain isotype Each H and L chain also has regularly spaced intrachain disulfide bridges. At the end is a variable domain (VH), followed by three constant domains for each of the α and γ chains. The L chain has four CH domains for the μ and ε isotypes. It has a variable domain (VL) at one end and then a constant domain (CL) at the other end VL aligns with VH, and CL aligns with the first constant domain of the heavy chain (CH1). The VH and VH groups are believed to form an interface between the light-chain variable domain and the heavy-chain variable domain. When VLs pair together, they form a single antigen-binding site. For the structure and properties, see, for example, Basic and Clinical Immunology ), 8th ed., Daniel P. Stites, Abba I. Terr, and Tristram G. Parslow (eds.), Appleton & Lange, Norwalk, CT, 1994, p. 71 and chapter 6.

[0151] The term "variable region" or "variable domain" typically refers to the amino-terminal region of a light or heavy chain. The heavy chain has a length of about 120 to 130 amino acids, and the light chain has a length of about 100 to 110 amino acids. , the antibodies used for the binding and specificity of each particular antibody to its particular antigen. The variable region of a heavy chain can be referred to as "VH." The variable region of a light chain can be referred to as "VH." The region can be referred to as "VL." The term "variable" refers to a particular segment of a variable region. This refers to the fact that V regions differ significantly in sequence among antibodies. The variability, however, defines the specificity of a particular antibody for its particular antigen. They are not evenly distributed across the 110 amino acid span of the variable region. However, the V regions have more variability called "hypervariable regions" each about 9 to 12 amino acids long ( For example, frames of about 15-30 amino acids separated by shorter regions (extremely variable) The stress response is more variable (e.g., relatively stable) and is called the framework region (FR). The heavy and light chain variable regions each consist of a main chain connected by three hypervariable regions. The hypervariable region comprises four FRs that form a β-sheet structure, and the hypervariable region connects the β-sheet structure. The hypervariable regions in each chain are bounded by FRs, forming loops that, depending on the chain, form part of the They are held in close proximity by the hypervariable regions from the other chains and together with the hypervariable regions from the other chains form the antigen-binding site of the antibody. Contributes to the synthesis of proteins of immunological interest (e.g., Kabat et al., Sequences of Proteins of Immunological Interest). of Proteins of Immunological Interest, 5th ed., Public Health Service, National (See Institutes of Health, Bethesda, MD, 1991). The constant region is responsible for binding the antibody to the antigen. Although it is not directly involved in the induction of cytotoxicity, it is involved in antibody-dependent cellular cytotoxicity (ADCC) and complement-dependent cytotoxicity (C The variable regions exhibit various effector functions, such as antibody engagement with DCs. The sequences are highly variable. The sequence variability is concentrated in the CDRs, while the variability within the variable regions is The smallest part of the CDRs is called the framework region (FR). The CDRs of the light and heavy chains mainly comprise the antigen In a specific embodiment, the variable region is a human variable region. It is an area.

[0152] The term "variable region residue numbering as in Kabat" or "amino acid position numbering as in Kabat" is used. The terms and their variations are described in Kabat et al., Sequences of Proteins of Immunological Interest (Sequences of Proteins of Immunological Interest), ces of Proteins of Immunological Interest, 5th ed., Public Health Service, Nation The heavy chain variable region of an antibody compilation at the University of California, San Diego, San Diego, CA (1991) refers to the numbering system used for the light chain variable region. The linear amino acid sequence of the variable domain may be a truncation of the FR or CDR of the variable domain or a truncation of the FR or CDR of the variable domain. or may contain fewer or additional amino acids corresponding to insertions into the CDRs. For example, the heavy chain variable domain contains a single amino acid insertion after residue 52 of H2 (residue 52 of H2 according to Kabat). 52a) and inserted residues after heavy chain FR residue 82 (e.g., residues 82a, 82b, 82c, 82d, 82e, 82f ... and 82c). The Kabat numbering of residues is relative to the antibody sequence and the "standard" Kabat numbering sequence. can be determined for a given antibody by alignment of regions of homology. The Kabat numbering system typically consists of a variable domain (approximately residues 1-107 of the light chain and residues 1-108 of the heavy chain). 13) (e.g., Kabat et al., immunologically interesting sequences ( Sequences of Immunological Interest), 5th edition, Public Health Service, National Ins. Titles of Health, Bethesda, Md. (1991). The "EU numbering system" or "EU index" is , usually used to refer to residues in the immunoglobulin heavy chain constant region (see, e.g., Kabat et al. (See above for the EU index reported in the paper). The "Kabat-like EU index" is Refers to the residue numbering of a human IgG1 EU antibody, including, for example, AbM, Chothia, Contact, IMGT, and AHon. Other numbering schemes are described, including:

[0153] An "intact" antibody comprises an antigen-binding site as well as a light chain constant region CL and at least a heavy chain constant region CH The antibody comprises a constant region containing a human constant region or its amino acid sequence. Mutations may be included. Preferably, the intact antibody has one or more effector functions.

[0154] An "antibody fragment" is a fragment that contains a portion of an intact antibody, preferably the antigen binding or variable region of the intact antibody. Examples of antibody fragments include, but are not limited to, Fab, Fab', F(ab')2, and Fv fragments. diabodies and di-diabodies (see, e.g., 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); WO 93 / 11161; and U.S. Pat. Nos. 5,837,242 and single chain antibody molecules (see, e.g., U.S. Pat. Nos. 4,946,778; 5,260,203; 5, dual variable domain antibodies (see, e.g., U.S. Pat. Nos. 7,612, 482,858 and 5,476,786); , 181); single variable domain antibodies (SdAbs) (see, e.g., Woolven et al., Immunogenetics 50: 98-101, 1999; Streltsov et al., Proc Natl Acad Sci USA. 101:12444-12449, 2 004); as well as multispecific antibodies formed from antibody fragments.

[0155] A "functional fragment" or "binding fragment" or "antigen-binding fragment" of a therapeutic antibody is one that does not bind to the intact antibody. exhibit at least one, if not some or all, of the biological functions resulting from the The function is at least the target antigen (e.g., a GFRAL-binding fragment or a fragment that binds to GFRAL). Includes joins.

[0156] As used herein, the term "fusion protein" refers to a protein that combines the amino acid sequence of an antibody and a heterologous A polypeptide or protein (e.g., a polypeptide or protein that is not normally part of an antibody) " refers to a polypeptide containing the amino acid sequence of a protein (e.g., a non-anti-GFRAL antigen-binding antibody)." The term "fusion" when used in connection with GFRAL or anti-GFRAL antibodies refers to a peptide or polypeptide, or fragments, variants, and / or derivatives thereof, and heterologous peptides or refers to the conjugation to the polypeptide. In some embodiments, the fusion protein comprises GFRAL or In some embodiments, the fusion protein retains the biological activity of the anti-GFRAL antibody. The VH region, VL region, VH CDRs (one, two, or three VH CDRs), and / or VL CDRs (one, two, or three VH CDRs) of the same or similar antibody. two or three VL CDRs), in which case the fusion protein contains the GFRAL epitope, Binds to GFRAL fragments and / or GFRAL polypeptides.

[0157] The term "heavy chain" when used in reference to an antibody refers to the amino terminal portion of the heavy chain consisting of about 12 amino acids. It contains a variable region of 0 to 130 or more amino acids and a constant carboxy-terminal portion. It refers to a polypeptide chain of approximately 50-70 kDa containing regions (e.g., CH1, CH2, CH3, CH4). The regions are designated alpha (α), delta (δ), epsilon ( five different types (e.g., isotypes) called ε (ε), gamma (γ), and mu (μ) The different heavy chains vary in size: α, δ, and γ are approximately 45 The light chain contains approximately 500 amino acids, while μ and ε contain approximately 550 amino acids. Combined, these different types of heavy chains form four subclasses of IgG: IgG1 IgA, IgD, IgE, IgG, and IgM antibodies, including IgG1, IgG2, IgG3, and IgG4, respectively. The heavy chain is a human heavy chain. can.

[0158] The term "light chain" when used in reference to an antibody refers to the amino-terminal portion of about 10 A variable region of 0 to about 110 or more amino acids, with the carboxy-terminal portion being constant. The light chain is a polypeptide chain of approximately 25 kDa, including the normal region. The approximate length of the light chain is 211-217 amino acids. They are called kappa (κ) or lambda (λ) based on the amino acid sequence of the constant domain. There are two different types of light chains. Light chain amino acid sequences are well known in the art. It can be a light chain.

[0159] As used herein, the term "effective amount" refers to an amount that reduces the severity and / or frequency of symptoms. to reduce symptoms and / or their underlying causes, and to prevent symptoms and / or their underlying causes from occurring and / or for example, involuntary weight loss, impaired glucose metabolism, or weight loss. ameliorate or prevent harm caused by or associated with GDF15-mediated diseases, disorders, or conditions, including is sufficient to modify the therapy (e.g., an anti-GFRAL antibody or pharmaceutical composition provided herein) See, for example, antibodies comprising the CDR, VH, and / or VL sequences shown in Tables 1-24. The term refers to the development or progression of a given GDF15-mediated disease, disorder, or condition, Reduction or amelioration of the recurrence, onset, or occurrence of vector-borne diseases, disorders, or illnesses; and and / or preventative or other therapy (e.g., a therapy other than an anti-GFRAL antibody provided herein). It also encompasses the amount necessary to improve or enhance the therapeutic effect. An effective amount may be administered in one or more doses, including intermittent doses, where one or more doses are administered over a treatment period. The treatment is administered followed by a rest period in which the antibody is not administered (e.g., one cycle of treatment and one rest period). A rest period can be followed by additional cycles, and one or more treatment periods can be followed by one or more rest periods. In some embodiments, an effective amount of an antibody provided herein is about 0 In one embodiment, the antibody concentration is between about 0.1 mg / kg (mg of antibody per kg of subject body weight) and about 100 mg / kg. Effective amounts of the antibodies provided herein are about 0.1 mg / kg, about 0.5 mg / kg, about 1 mg / kg, 3 mg / kg, 5 mg / kg, g / kg, approx. 10mg / kg, approx. 15mg / kg, approx. 20mg / kg, approx. 25mg / kg, approx. 30mg / kg, approx. 35mg / kg, approx. 40mg / k g, about 45 mg / kg, about 50 mg / kg, about 60 mg / kg, about 70 mg / kg, about 80 mg / kg, about 90 mg / kg, or about 1 00 mg / kg (or a range therein). In some embodiments, the The effective amount to be administered also achieves a particular result (e.g., (i) GFRAL; (ii) GDF15 and GFRAL; or (iii) in vitro or in vivo biomarkers induced by the binding of GDF15, GFRAL, and RET. The term "antibody" refers to the amount of an antibody provided herein that mimics or modulates a specific biological effect. For example, An effective amount includes: (i) increasing body weight; (ii) maintaining body weight; (iii) reducing weight loss; (iv) increasing the body's (v) increasing body mass (e.g., lean mass or fat mass); (vi) maintaining body mass (e.g., lean mass or fat mass); or (vi) preventing a loss of body mass (e.g., lean mass or fat mass). of an anti-GFRAL antibody described herein effective to reduce (e.g., at one or more doses) The amount of

[0160] As used herein, the term "host cell" refers to a cell that can be transfected with a nucleic acid molecule. refers to the particular subject cell and the progeny or potential progeny of such a cell. Progeny may be derived from mutations or environmental influences that may occur in subsequent generations or from the transfer of nucleic acids to the host cell genome. For integration of the molecule, the cells need not be identical to the parent cells transfected with the nucleic acid molecule. .

[0161] As used herein, the term "immunomodulatory agent" and its limitations However, variations thereof, including immunomodulatory agents, may be used to enhance the immune response of the host. In certain embodiments, the combination therapy provided herein refers to an agent that modulates the immune system. Immunomodulatory agents used in the methods do not include anti-GFRAL antibodies or antigen-binding fragments. The agents include small molecules, peptides, polypeptides, proteins, fusion proteins, antibodies, and inorganic "Small molecules" include, but are not limited to, chemical molecules, mimetics, and organic molecules. These and similar terms include peptides, peptidomimetics, amino acids, amino acid analogs, and the like. analogs, polynucleotides, polynucleotide analogs, nucleotides, nucleotide analogs organic or inorganic compounds having a molecular weight of less than about 10,000 grams per mole (i.e., heterocyclic (b) organic and / or organometallic compounds), having a molecular weight of less than about 5,000 grams per mole organic or inorganic compounds having a molecular weight of less than about 1,000 grams per mole; compounds, organic or inorganic compounds having a molecular weight of less than about 500 grams per mole, and the like These include, but are not limited to, salts, esters, and other pharmaceutically acceptable forms of such compounds. In some embodiments, the immunomodulatory agent is an immunostimulatory agent. In some embodiments, the immunomodulatory agent is an immunosuppressant. Immunomodulators are molecules known as immune checkpoint molecules (e.g., co-inhibitory or co-stimulatory). Proteins, e.g., C10orf54, CD86, CD80, PDL-1, PDL-2, CTLA-4, PD1, LAG3, BTNL2 , B7-H3, B7-H4, butyrophilin, CD48, CD244, TIM-3, CD200R, CD200, CD160, BTLA, HVEM, LAIR1, TIM1, galectin 9, TIM3, CD48, 2B4, CD155, CD112, CD113, and TIGIT ( For example, co-inhibitory), and / or CD154, TNFRSF25, GITR, 4-1BB, OX40, CD27, TMIGD2, ICOS, CD28, CD40, TL1A, GITRL, 41BBL, OX40L, CD70, HHLA2, ICOSL, cytokine, L IGHT, HVEM, CD30, CD30L, B7-H2, CD80, CD86, CD40L, TIM4, TIM1, SLAM, CD48, CD58 , modulating (e.g., inhibitory) CD155, CD112, DR3, GITR, CD2, and CD226 (e.g., costimulatory) The term "antibody" refers to an agent (e.g., an antibody) that inhibits or stimulates the growth of the target cell.

[0162] As used herein, the term "monoclonal antibody" refers to a population of substantially homogeneous antibodies. For example, the individual antibodies that make up the population may be present in minute amounts. Each monoclonal antibody is typically identical except for possible natural mutations that may occur. In a specific embodiment, the term "antigen" used herein refers to a single epitope on an antigen. A "monoclonal antibody" is an antibody produced by a single hybridoma or other cell. wherein the antibody is capable of being bound to a target antigen, e.g., by ELISA or other antigen-binding or immunoassay methods known in the art. or binds only to the GFRAL epitope as determined by competitive binding assays. The term "clonal" is not limited to any particular method of producing an antibody. For example, Monoclonal antibodies useful in this field were first described by Kohler et al. (Nature, 256:495 (1975)). They may be prepared by the hybridoma method described in, or may be prepared from bacterial, eukaryotic, or may also be produced in plant cells using recombinant DNA techniques (see, e.g., U.S. Pat. No. 4,816,567). "Monoclonal antibodies" also refer to antibodies as described, for example, in Clackson et al., Nature 352:624-6 28 (1991) and Marks et al., J. Mol. Biol., 222:581-597 (1991). The antibody may be isolated from a phage antibody library using a clonal cell line and Other methods for preparing monoclonal antibodies expressed by IgG are well known in the art (e.g., Short Protocols in Molecular Biology (2002), Vol. 5 (See Chapter 11 of the "Synopsis of the American Political Science" edition, edited by Ausubel et al., John Wiley and Sons, New York:) Methods for producing clonal antibodies are provided in the Examples herein.

[0163] The term "native" refers to a biological material, e.g., a nucleic acid molecule, a polypeptide, a host cell, When used in connection with a compound or a substance, the compound or substance is one that is found in nature and that has been manipulated, modified, or altered by humans. and / or has not been modified (e.g., isolated, purified, selected).

[0164] The antibodies provided herein may have a portion of the heavy and / or light chains derived from a particular species or derived from a particular Identical or homologous to corresponding sequences in antibodies belonging to a certain antibody class or subclass. whereas the remainder of the chain is derived from another species or belongs to another antibody class or subclass. "Chimeric" antibodies are those that are identical or homologous to the corresponding sequences in the and may include fragments of such antibodies (see, e.g., U.S. Pat. No. 4,816,567; and Morton et al., J. Immunol. 1999, 10, 144-145). (See Rison et al., Proc. Natl. Acad. Sci. USA, 81:6851-6855 (1984)).

[0165] "Humanized" forms of non-human (e.g., murine) antibodies are antibodies in which the native CDR residues are modified to conform to the desired characteristics. non-human animals such as mice, rats, rabbits, or non-human primates, with the same sex, affinity, and ability Human immunoglobulins in which residues have been replaced with residues from the corresponding CDR of a human species (e.g., donor antibody). A chimeric antibody is one that contains an immunoglobulin (e.g., recipient antibody). One or more FR region residues of the human immunoglobulin are replaced by corresponding non-human residues. Humanized antibodies may comprise residues that are not found in the recipient antibody or in the donor antibody. These modifications are made to further refine antibody performance. The light chain can include at least one or substantially all of the variable region. In the region, all or substantially all of the CDRs correspond to the CDRs of a non-human immunoglobulin and all of the FRs or substantially all of the FRs are human immunoglobulin sequences. An antibody contains at least a portion of an immunoglobulin constant region (Fc), typically the constant region of a human immunoglobulin. For further details, see Jones et al., Nature, 321. :522-525 (1986); Riechmann et al., Nature, 332:323-329 (1988); and Presta, C Urr. Op. Struct. Biol., 2:593-596 (1992); Carter et al., Proc. Natl. Acad. Sci. U SA 89:4285-4289 (1992); and U.S. Patent Nos. 6,800,738 (issued October 5, 2004), 6,719,971 No. (issued September 27, 2005), No. 6,639,055 (issued October 28, 2003), No. 6,407,213 (issued June 2002) See, for example, No. 6,054,297 (published April 18, 2000) and No. 6,054,297 (published April 25, 2000).

[0166] A "human antibody" is an antibody that contains an amino acid sequence corresponding to the amino acid sequence of an antibody produced by a human. and / or produced using any of the human antibody production techniques disclosed herein. This definition of a human antibody specifically refers to a humanized antibody that contains non-human antigen-binding residues. Human antibodies have been generated using phage display libraries (Hoogenboom and Winter, Marks et al., J. Mol. Biol., 222:581 (1991); )) and yeast display libraries (Chao et al., Nature Protocols 1:755-768 (2006 )) can be produced using a variety of techniques known in the art. , Monoclonal Antibodies and Cancer Therapy, Alan R Liss, p. 77 (1985); Boerner et al., J. Immunol., 147(1):86-95 (1991). Methods described by van Dijk and van de Winke are also available for the preparation of human monoclonal antibodies. See also, Curr. Opin. Pharmacol., 5:368-74 (2001). modified to produce such antibodies in response to a stimulus, but at its endogenous locus By administering an antigen to a transgenic animal, e.g., a mouse, in which the (See, for example, Jakobovits, A., Curr. Opin. Biotechnol. 1999 5, 6(5):561-6; Bruggemann and Taussing, Curr. Opin. Biotechnol. 1997, 8(4): 455-8; and U.S. Patent Nos. 6,075,181 and 6,150,584 relating to XENOMOUSE™ technology. See, for example, Li et al., regarding human antibodies produced by human B-cell hybridoma technology. See also Proc. Natl. Acad. Sci. USA, 103:3557-3562 (2006).

[0167] "CDR" refers to a non-framework region of an immunoglobulin (Ig or antibody) VH β-sheet framework. One of the three hypervariable regions (H1, H2, or H3) within the framework region or the β-chain of the antibody VL Three hypervariable regions (L1, L2, or L3) within the non-framework regions of the base framework Thus, a CDR refers to one of the variable regions interspersed within framework region sequences. CDR regions are well known to those skilled in the art and are, for example, the most highly variable sequences within an antibody variable (V) domain. As regions of degeneration, they are defined by Kabat (Kabat et al., J. Biol. Chem. 252: 6609-6616 (1977); Kabat, Adv. Prot. Chem. 32:1-75 (1978)). CDR region sequences are It is not part of the conserved β-sheet framework and therefore can adopt a variety of conformations. It has been structurally defined by Chothia as a residue that can J. Mol. Biol. 196:901-917 (1987)). Both terminologies are well recognized in the art. The CDR region sequences are also defined by AbM, contact, and IMGT. The positions of the CDRs within a standard antibody variable region are shown in Tables 1 to 24. (Al-Lazikani et al., J. Mol. Biol. 273:927-948 (1997); M orea et al., Methods 20:267-279 (2000). The number of residues within the hypervariable regions varies among different antibodies. Because they vary from one another, the standard variable region numbering system includes additional residues relative to the standard positions, such as Conventionally, the residue number is followed by a, b, c, etc. (Al-Lazikani et al., supra). 1997)). Such nomenclature is likewise well known to those skilled in the art.

[0168] The terms "hypervariable region," "HVR," or "HV" as used herein refer to a sequence The regions of antibody variable regions in which the sequences are hypervariable and / or form structurally defined loops are Typically, antibodies have six hypervariable regions; three in the VH (H1, H2, H3) and three in the VL (L1, L2, L3). Several hypervariable region delineations are in use and are encompassed herein. Kabat phase Complementarity determining regions (CDRs) are based on sequence variability and are the most commonly used (e.g. For example, Kabat et al., Sequences of Proteins of Immunological Interest Immunological Interest), 5th ed., Public Health Service, National Institutes of Health (See Health, Bethesda, MD. (1991)). Chothia instead refers to the location of structural loops. (See, e.g., Chothia and Lesk, J. Mol. Biol. 196:901-917 (1987)). The ends of the Chothia CDR-H1 loop, when numbered using the Kabat numbering convention, are numbered according to the length of the loop. The insertions differ between H32 and H34 depending on the sequence (because the Kabat numbering system places the insertions at H35A and H35B). If neither 35A nor 35B exists, the loop ends at 32; if only 35A exists, the loop ends at 32. (If both 35A and 35B are present, the loop ends at 34). AbM hypervariable region represents a compromise between Kabat CDRs and Chothia structural loops, and is based on Oxford Molecular's AbM antibody model. used by Ring software (e.g., Martin, Antibody Engineering ( (See Antibody Engineering, Vol. 2, Chapter 3, Springer Verlag). "Contact" hypervariable regions These hypervariable regions or CDRs are based on the analysis of available complex crystal structures. The residues from each are listed below.

[0169] Recently, a universal numbering system called ImMunoGeneTics (IMGT) Information System (registered trademark) has been developed. has been developed and widely adopted (Lafranc et al., Dev. Comp. Immunol. 27(1):55-77). 2003). IMGT is a system for identifying immunoglobulins (IGs), T cell receptors (TRs), and antigen receptors (AGRs) from humans and other vertebrates. It is a comprehensive information system specializing in the major histocompatibility complex (MHC). The references are made both to the amino acid sequence and to the position within the light or heavy chain. The "position" of the CDRs within the amino acid structure is conserved among species and exists in structures called loops. Therefore, a numbering system can be used to align variable domain sequences according to structural features. This information can be used to easily identify CDR and framework residues. CDR residues from guanine are typically transferred to an acceptor framework from a human antibody. An additional numbering system (AHon) is Honegger and For example, Kabat et al. The correspondence between the numbering systems, including the numbering system of the IMGT and the unique numbering system of the IMGT, is well known to those skilled in the art ( See, for example, Kabat (supra); Chothia and Lesk (supra); Martin (supra); Lefr anc et al. (supra) and Tables 1-24. The system combines Kabat and Chothia. [Table 1]

[0170] The hypervariable regions may be "extended hypervariable regions" such as: 24-36 or 24-34 (L1) of VL; 46-56 or 50-56 (L2), and 89-97 or 89-96 (L3) and 26-35 or 26-35A (H1) of VH, 5 H2) and 93-102, 94-102, or 95-102 (H3). As used, the terms "HVR" and "CDR" are used interchangeably.

[0171] The term "constant region" or "constant domain" refers to the region of an antibody that is not directly involved in binding to an antigen. Light and heavy chains that do not affect immune function but exhibit various effector functions, such as interaction with Fc receptors These terms refer to the carboxy-terminal portion of an immunoglobulin that contains the antigen-binding site. The immune system has a more conserved amino acid sequence than the other part of the immune system, the variable region. The constant region refers to the portion of a globulin molecule that consists of the CH1, CH2, and CH3 regions of the heavy chain and the C It may contain the L region.

[0172] The term "framework" or "FR" residues refers to the variable region residues that flank the CDRs. FR residues may be present in, for example, chimeric, humanized, human, domain antibodies, diabodies, linear antibodies, and bispecific antibodies. FR residues are hypervariable region residues or variable domain residues other than CDR residues. It is the main residue.

[0173] An "affinity matured" antibody is one in which one or more changes (e.g., modifications, additions, and / or deletions) have been made to the antibody. The affinity of the antibody for the antigen compared to the parent antibody that does not possess the amino acid sequence variation Antibodies with these changes in one or more of their HVRs that result in improved affinity. Sexually matured antibodies have nanomolar or even picomolar affinities for their target antigens. Affinity matured antibodies are produced by procedures known in the art. Hudson and Souriau, Nature Medicine 9:129-134 (2003); Hoogenboom, Nat ure Biotechnol. 23: 1105-1116 (2005); Quiroz and Sinclair, Revista Ingeneri See Biomedia 4: 39-51(2010).

[0174] A "blocking" or "antagonist" antibody is one that blocks the antigen to which it binds (e.g., Antibodies that inhibit or reduce the biological activity of GFRAL (as described herein). The locking or antagonist antibody may bind to an antigen (e.g., GFRAL as described herein). The biological activity of the compound can be substantially or completely inhibited.

[0175] An "agonist antibody" is an antibody that elicits a response, e.g., a function of a polypeptide of interest. Agonist antibodies are antibodies that mimic at least one of the activities of a ligand. For example, in this case, the ligand binds to a cell surface receptor and the binding In this case, the antibody induces cell signaling or activity via an intracellular cell signaling pathway, induce similar cell signaling or activation.

[0176] An "antagonist" of GFRAL is an agent that inhibits the biological activity of GFRAL, for example, in cells that express GFRAL. a molecule (e.g., In some embodiments, an antagonist of GFRAL (e.g., a compound according to the present invention) is used. The agonistic antibodies (as described herein) can be used to inhibit, for example, the activation and / or cell activation of GFRAL-expressing cells. They act by detectably inhibiting or otherwise reducing signal transduction pathways, thereby and the GFRAL-mediated biological activity of the cells compared to the GFRAL-mediated biological activity in the absence of an antagonist. In some embodiments, the genotypes provided herein can detectably reduce the genotype. The antibody inhibits signal transduction of a complex containing GFRAL, GDF15, and / or RET. The present invention relates to an antagonistic anti-GFRAL antibody, which is an anti-GFRAL antibody comprising the compound The reduction need not be complete, as long as it is detectable using an assay. For example, Analyzing the biological activity of GFRAL using the cell-based assay described in the Examples. can be done.

[0177] "Binding affinity" typically refers to the ability of a molecule (e.g., a binding protein, e.g., an antibody) to bind to a single molecule. It refers to the strength of the total non-covalent interactions between a site and its binding partner (e.g., an antigen). Unless otherwise indicated, as used herein, "binding affinity" refers to the affinity of the members of a binding pair. It refers to the intrinsic binding affinity that reflects the 1:1 interaction between a target molecule (e.g., antibody and antigen). The affinity of a binding molecule X for its binding partner Y is usually expressed as the dissociation constant (K D ) Affinity can be measured by general methods known in the art, including the methods described herein. Low affinity antibodies usually bind antigens slowly and release their antigens rapidly. High affinity antibodies usually bind antigens faster and tend to dissociate more quickly, whereas high affinity antibodies usually bind antigens faster and tend to dissociate more quickly. Various methods for measuring binding affinity are known in the art. are known in the art, any of which may be used for the purposes of this disclosure. Exemplary embodiments include the following: D " or "K D The "value" is measured by assays known in the art, for example, by binding assays. It is possible. D is carried out, for example, using a Fab-type antibody of interest and its antigen. , can be measured by radiolabeled antigen binding assay (RIA) (Chen et al., 1999, J. Mo l Biol 293:865-881). K D or K D The values ​​are, for example, BIAcore™-2000 or BIAcore™-3000 Surface plasmon resonance assays were performed using Biacore (BIAcore, Piscataway, NJ). or by using, for example, the OctetQK384 system (ForteBio, Menlo Park, CA). This can be measured by biolayer interferometry. The "rate of association" or "association rate" or "k on" may also be used in, for example, BIAcore™-2000 or BIAcore TM-3000 (BIAcore, Piscataway, NJ) or OctetQK384 system (ForteBio, Menlo Park, Determined using the same surface plasmon resonance or biolayer interferometry as above using CA) It is possible.

[0178] The phrases "substantially similar" or "substantially the same" are used by those skilled in the art to distinguish between two values. The difference between the values ​​(e.g., K D biological characteristics measured by the Two numbers that are high enough to be considered of little or no scientific and / or statistical significance between values ​​(e.g., one value associated with an antibody of the disclosure and another value associated with a reference antibody) For example, the difference between the two values ​​is expressed as a function of the value for the reference antibody. , less than about 50%, less than about 40%, less than about 30%, less than about 20%, less than about 10%, or less than about 5%. do.

[0179] As used herein, the phrases "substantially reduced" or "substantially different" mean that A vendor may interpret the difference between the two values ​​in relation to the biological characteristic measured by the values. and two numbers high enough to be considered statistically significant (e.g., associated with the antibodies of the present disclosure). The difference between the two values ​​is expressed as the degree of difference between the two values ​​(one value associated with the corresponding antibody and the other value associated with the reference antibody). The difference between the two values ​​is preferably greater than about 10%, about 2%, as a function of the value for the reference antibody. It may be greater than 0%, greater than about 30%, greater than about 40%, or greater than about 50%.

[0180] Antibody "effector functions" are determined by the Fc region (e.g., a native sequence Fc region or amino acid sequence) of an antibody. This refers to the biological activity resulting from the amino acid sequence variants (Fc region) and varies depending on the antibody isotype. Examples of antibody effector functions include: C1q binding and complement-dependent cytotoxicity; Fc receptor binding; Antibody-dependent cell-mediated cytotoxicity (ADCC); phagocytosis; cell surface receptors (e.g. B-cell receptors) downregulation of IL-1; and B cell activation.

[0181] The term "Fc region" as used herein refers to, for example, a native sequence Fc region, a recombinant Fc Used to define the C-terminal region of an immunoglobulin heavy chain, including the Fc region and variant Fc regions Although the boundaries of the Fc region of an immunoglobulin heavy chain can vary, the human IgG heavy chain Fc region , often from amino acid residues at positions Cys226 or Pro230 to its carboxyl terminus The C-terminal lysine of the Fc region (residue 447 according to the EU numbering system) is defined as extending to during the production or purification of the antibody, or by recombinantly manipulating the nucleic acid encoding the antibody heavy chain. Thus, compositions of intact antibodies can be prepared by removing all K447 residues. a population of antibodies in which the K447 residue has been removed, a population of antibodies in which the K447 residue has not been removed, and an antibody population in which the K447 residue has been removed. The antibody population may comprise a mixture of antibodies lacking the K447 residue and antibodies lacking the K447 residue.

[0182] A "functional Fc region" possesses an "effector function" of a native sequence Fc region. Typical "effector functions" include C1q binding; complement-dependent cytotoxicity (CDC); Fc receptor binding; antibody-dependent cell-mediated cytotoxicity (ADCC); phagocytosis; cell surface receptors (e.g. B cell receptors); Such effector functions include down-regulation of the Fc region (the BCR). It is intended to be combined with a binding region or domain (e.g., an antibody variable region or domain). requires the use of various assays disclosed herein and / or known in the art. can be evaluated including

[0183] A "native sequence Fc region" is an Fc region found in nature and that has not been engineered, modified, and / or modified by man. has not been modified (e.g., not isolated, purified, selected, or otherwise modified, such as by other sequences such as variable region sequences) an amino acid sequence identical to the amino acid sequence of the Fc region (not including or combined with said sequence); The native sequence human Fc region contains the amino acid sequence of a native sequence human IgG1 Fc region (non- A and A allotypes); native sequence human IgG2 Fc region; native sequence human IgG3 Fc region; and native sequence human IgG4 Fc regions, as well as naturally occurring variants thereof.

[0184] A "variant Fc region" is a variant Fc region that contains at least one amino acid modification (e.g., substitution, addition, or deletion). , preferably by one or more amino acid substitutions, to the amino acid sequence of a native sequence Fc region. Preferably, the variant Fc region comprises a native sequence Fc region or a parent Fc region that contains a different amino acid sequence. At least one amino acid substitution, e.g., about 1 to about 10, compared to the Fc region of the polypeptide. amino acid substitutions, preferably about 1 to about 5 amino acid substitutions, in a native sequence Fc region or The variant Fc region herein preferably has a nucleotide sequence similar to that of the parent polypeptide. at least about 80% homology with the native sequence Fc region and / or the Fc region of the parent polypeptide; More preferably, there is at least about 90% homology therewith, e.g., at least about 90% homology therewith. It shares approximately 95% homology with the human IgG1 Fc sequence. For example, it has two amino acids at two positions in the human IgG1 Fc sequence that are substituted with alanine. Variants with amino acid changes are shown in bold in the amino acid sequences provided below. R: [ka]

[0185] For example, two sequences in a truncated human IgG1 Fc sequence (IgG1ΔK Fc) lacking the C-terminal lysine residue A variant having two amino acid changes to alanine at positions Shown in bold: [ka]

[0186] For example, a truncated mutant of the human IgG1 Fc sequence (IgG1ΔK Fc) lacking the C-terminal lysine residue is shown in the amino acid sequence provided below: [ka]

[0187] For example, a variant having an amino acid change to proline at a position in the human IgG4 Fc sequence is Shown in bold in the amino acid sequence provided below: [ka]

[0188] For example, a proline at a position in the human IgG4 Fc sequence (IgG4ΔK Fc) where there is no C-terminal lysine Variants with amino acid changes to are shown in bold in the amino acid sequences provided below. Currently: [ka]

[0189] For example, a variant having an amino acid change to glutamine at position in the human IgG1 Fc sequence is Shown in bold in the amino acid sequence provided below: [ka]

[0190] Any of the VH domains in Tables 1 to 24 and Figures 4A, 5A, 5C, 5E, 6A, 7A, 8A, 9A, and 10A It can be combined with the variant Fc regions described herein. Examples containing variant Fc regions Exemplary heavy chain constructs include the following constructs, designated as shown below: variable region sequences are in bold and CDR sequences are underlined. Currently:

[0191] 3P10 Fab Hc: [ka]

[0192] 25M22 Fab Hc: [ka]

[0193] 8D8 Fab Hc: [ka]

[0194] 5F12 Fab Hc: [ka]

[0195] "Light chain constant region" includes kappa and lambda constant regions. Tables 1-24 and Figures 4B, 5 Any of the VL domains of 5B, 5D, 5F, 6B, 7B, 8B, 9B, and 10B can be used in combination with any of the VL domains of 5B, 5D, 5F, 6B, 7B, 8B, 9B, and 10B described herein. The polypeptides may be combined with the αpα or λ constant regions.

[0196] Exemplary kappa constant regions are provided below: [ka]

[0197] Exemplary lambda constant regions are provided below: [ka]

[0198] Exemplary light chain constructs containing constant regions include those designated as shown below: The following constructs can be mentioned; variable region sequences are in bold and CDR sequences are in bold. The columns are underlined:

[0199] 3P10 Fab Lc: [ka]

[0200] 25M22 Fab Lc: [ka]

[0201] 8D8 Fab Lc: [ka]

[0202] 5F12 Fab Lc: [ka]

[0203] The term "variant" when used in connection with GFRAL or anti-GFRAL antibodies 1 or more (e.g., about 1 to about 25, about 1 to about 20) amino acids compared to the native or unmodified GFRAL sequence about 1 to about 15, about 1 to about 10, or about 1 to about 5 amino acid sequence substitutions, deletions, and / or deletions or an addition. For example, a GFRAL variant can be , 1 or more (e.g., about 1 to about 25, about 1 to about 20) amino acids relative to the amino acid sequence of native GFRAL , about 1 to about 15, about 1 to about 10, or about 1 to about 5, etc. Also by way of example, variants of anti-GFRAL antibodies may be native or previously unmodified anti-GFRAL antibodies. One or more (e.g., about 1 to about 25, about 1 to about 20, about 1 to about 15) amino acids of the amino acid sequence of the FRAL antibody Mutants can be generated by alterations of at least one, about 1 to about 10, or about 1 to about 5 amino acids. It may be a naturally occurring one, such as an allelic variant or splice variant, or The polypeptide variant may be artificially constructed. In some embodiments, the GFRAL variant may be prepared from a corresponding nucleic acid molecule. or anti-GFRAL antibody variants, which retain at least the functional activity of GFRAL or anti-GFRAL antibody, respectively. In some embodiments, the anti-GFRAL antibody variant binds to GFRAL and / or In some embodiments, the anti-GFRAL antibody variant is In some embodiments, the agonist binds to GFRAL and / or is agonistic to GFRAL activity. wherein the variant is a VH or VL region or subregion of GFRAL or an anti-GFRAL antibody, e.g. , encoded by single nucleotide polymorphism (SNP) variants of a nucleic acid molecule encoding one or more CDRs will be done.

[0204] The term "vector" refers to any vector that is used to transfer nucleic acids, including, for example, nucleic acid sequences into a host cell. Refers to a substance used to carry or contain a sequence. Vectors applicable for use Examples of such vectors include expression vectors, plasmids, phage vectors, viral vectors, These include episomes and artificial chromosomes, which are capable of stable integration into the chromosomes of host cells. Furthermore, the vector may contain a selection sequence or marker operable for The above selectable marker genes and appropriate expression control sequences can be included. Selectable marker genes that can be used include those that provide resistance to antibiotics or toxins, for example. They provide nutrients, complement autotrophic deficiencies, or supply important nutrients that are not present in the culture medium. The columns include constitutive and inducible promoters, transcriptional enhancers, transcriptional It may contain a terminator, etc. Two or more nucleic acid molecules (e.g., heavy and light chains of an antibody) If both VH and VL of an antibody are to be co-expressed, both nucleic acid molecules can be expressed, e.g. For example, they can be inserted into a single expression vector or into separate expression vectors. For expression, the encoding nucleic acids may be operably linked to one common expression control sequence or may be linked to different The expression control sequences may be linked to one or more expression control sequences, e.g., one inducible promoter and one constitutive promoter. The nucleic acid molecule can be introduced into a host cell using methods well known in the art. Such a method can be performed by, for example, nucleic acid analysis, for example, mRNA sequencing. for either ELISA blot or polymerase chain reaction (PCR) amplification, or expression of the gene product. Immunoblotting of the entire sample or expression of the introduced nucleic acid sequence or its corresponding gene product Other suitable analytical methods for testing the desired product (e.g., the nucleic acid molecule of the present invention) include: It will be understood by those skilled in the art that the vectors can be expressed in sufficient amounts to produce the anti-GFRAL antibodies described herein. It is understood that expression levels can be optimized using methods well known in the art to ensure sufficient expression. It is further understood that the present invention can be obtained.

[0205] "Antibody-dependent cell-mediated cytotoxicity" or "ADCC" refers to the use of specific cytotoxic cells (e.g. Fc receptors (FcRs) present on immune cells (e.g., natural killer (NK) cells, neutrophils, and macrophages) These cytotoxic effector cells target antigen-bearing target cells via secretory Ig bound to the and subsequently allowing the target cells to be killed by a cytotoxin. Antibodies "arm" cytotoxic cells and are completely necessary for such killing. NK cells, the primary cells mediating ADCC, express only FcγRIII, whereas Monocytes express FcγRI, FcγRII, and FcγRIII. FcR expression on hematopoietic cells is known. (See, e.g., Ravetch and Kinet, Annu. Rev. Immunol. 9:457-92 (1991), p. 464. To assess the ADCC activity of a molecule of interest, an in vitro ADCC assay is used. (See, for example, U.S. Patent Nos. 5,500,362 or 5,821,337). Useful effector cells for such assays include peripheral blood mononuclear cells (PBMCs) and natural Alternatively, or in addition, ADCC activity of a molecule of interest can be achieved by using a cell line that expresses a specific antigen. , may be assayed 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 are selected for use. That's fine.

[0206] "Fc receptor" or "FcR" refers to a receptor that binds to the Fc region of an antibody. A preferred FcR is a native sequence human FcR. Furthermore, a preferred FcR is one that binds to IgG antibodies. receptors (e.g., gamma receptors) that are FcγRI, FcγRII, and FcγRIII subclasses receptors, including allelic variants and alternatively spliced ​​forms of these receptors. FcγRII receptors include FcγRIIA (an "activating receptor") and FcγRIIB (an "inhibitory receptor"). Rarely, they have similar amino acid sequences that differ primarily in their cytoplasmic domains (e.g. (See review by Daeron, Annu. Rev. Immunol. 15:203-234 (1997)). FcRs are known (e.g., See, e.g., 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. Med. 126:330-41 (1995). Other FcRs, including those to be identified in the future, are referred to herein as "FcRs." These are encompassed by the term "FcR." This term refers to the new receptors involved in the transfer of maternal IgG to the fetus. and the fetal receptor FcRn (see, e.g., Guyer et al., J. Immunol. 117:587 (1976) and K (See Immunol. 24:249 (1994)). Antibodies with improved or reduced binding to FcRs Isotropic mutants have been described (e.g., WO 2000 / 42072; U.S. Pat. Nos. 7,183,387, 7,332, Shields et al., J. Biol. Chem. 9(2):6591-6604 (2001). reference).

[0207] "Complement dependent cytotoxicity" or "CDC" refers to the lysing of a target cell in the presence of complement. Activation of the classical complement pathway occurs when the first component of the complement system (C1q) binds to its cognate antigen. Complement activation is initiated by binding to antibodies (of the appropriate subclass) that are To do this, a CDC assay (e.g., Gazzano-Santoro et al., J. Immunol. Methods 202:1 63 (1996)) may be performed. variants (polypeptides having variant Fc regions) and increased or decreased C1q binding ability Polypeptide variants have been described (e.g., U.S. Pat. No. 6,194,551, WO 1999 / 51642 (See Idusogie et al., J. Immunol. 164: 4178-4184 (2000)). Or no antibody at all may be selected for use.

[0208] When calculating percent identity, the sequences being compared are sorted by the number of sequences that give the largest match between the sequences. The percent identity can be determined by aligning the sequences. The computer program is the GCG program package, which includes GAP (Devereux et al., 1984). ) Nucl. Acid Res. 12:387; Genetics Computer Group, University of Wisconsin, Madi The computer algorithm GAP should be used to determine percent sequence identity. The two polypeptides or polynucleotides are aligned. The sequences are then compared to their respective amino acids. The optimal match of an acid or nucleotide (determined by the algorithm) The gap opening penalty ( It is calculated as three times the average diagonal, where "average diagonal" is the average diagonal of the comparison matrix being used. the average of the diagonals of the matrix; the "diagonal" is the average of each complete matrix by a particular comparison matrix. A score or number assigned to a perfect amino acid match and a gap extension penalty (This is usually 1 / 10 of the gap opening penalty), as well as PAM 250 or BLOSUM A comparison matrix such as 62 is used with the algorithm. For the standard comparison matrix (PAM 250 comparison matrix, see Dayhoff et al. 1978) Atlas of Protein Sequence and Structure 5:345-352; BLOSUM 62 comparative matrix For more information, see Henikoff et al. (1992) Proc. Natl. Acad. Sci. USA 89:10915-1091. 9) is also used by the algorithm.

[0209] Percent identity for polypeptide or nucleotide sequences using the GAP program Exemplary parameters for determining gender are: (i) Algorithm: Needle Man et al., 1970, J. Mol. Biol. 48:443-453; (ii) Comparison matrix: Henikoff et al. BLOSUM 62 from JSPS, 1992 (supra); (iii) gap penalty: 12 (except for end gaps) (iv) gap length penalty: 4; and (v) similarity threshold: 0.

[0210] A particular alignment scheme for aligning two amino acid sequences results in two Only a short region of the sequence may be matched, and this small aligned region may Even if there is no significant relationship between two full-length sequences, they may have very high sequence identity. Therefore, the alignment method chosen (e.g., the GAP program) If desired, several amino acids of the target polypeptide, for example, at least 50 consecutive amino acids, may be added. The alignment can be adjusted to result in an alignment over consecutive amino acids. do.

[0211] "Percent (%) amino acid sequence identity" with respect to a reference polypeptide sequence is used to align a sequence. and, if necessary, introducing gaps to achieve the maximum percent sequence identity. Then, the reference polypeptide sequence is compared without taking into account any conservative substitutions as part of the sequence identity. Defined as the percentage of amino acid residues in the candidate sequence that are identical to the amino acid residues in the string Alignment to determine percent amino acid sequence identity can be performed, for example, using publicly available Publicly available computer software, such as BLAST, BLAST-2, ALIGN, or Megal This can be accomplished in a variety of ways within the skill of the art using ign (DNASTAR) software. Those skilled in the art will find that maximal alignment is achieved over the entire length of the sequences being compared. Suitable methods for aligning sequences, including any algorithms required to achieve this alignment, are also known. The appropriate parameters can be determined.

[0212] A "modification" of an amino acid residue / position refers to a change in the primary amino acid sequence compared to the starting amino acid sequence. refers to a change in sequence, where the change is caused by an alteration of the sequence that includes the amino acid residue / position. For example, typical modifications include substitution of a residue with another amino acid (e.g., conservative or indicates a non-conservative substitution), one or more (e.g., usually less than 5, 4, or 3) adjacent to the residue / position and / or deletion of said residues / positions.

[0213] An "epitope" is a site on the surface of an antigen molecule to which a single antibody molecule binds; e.g., and capable of binding to one or more antigen-binding regions of an animal, e.g., a mammal (e.g., Possessing antigenic or immunogenic activity in humans, i.e., capable of eliciting an immune response , antigens, e.g., GFRAL polypeptides, GFRAL polypeptide fragments, or GFRAL epitopes Epitopes with immunogenic activity are those that elicit an antibody response in animals. An epitope having antigenic activity is a portion of a polypeptide that induces an immune response. The antibody binding activity is determined by any method known in the art, including by immunoassay. An antigenic epitope is a portion of a polypeptide that binds to the target protein. An antigenic epitope is not necessarily immunogenic. Epitopes are often chemically active surface molecules such as amino acids or sugar side chains. It consists of a group of molecules and has specific three-dimensional structural characteristics and specific charge characteristics. The term specifically includes linear epitopes and conformational epitopes. The contributing region of the polypeptide may be consecutive amino acids of the polypeptide, or An epitope may consist of two or more non-contiguous regions of a polypeptide. An epitope may or may not be a three-dimensional surface feature of an antigen. In embodiments, a GFRAL epitope is a three-dimensional surface feature of a GFRAL polypeptide. In an embodiment, a GFRAL epitope is a linear feature of a GFRAL polypeptide. has several or many different epitopes and reacts with many different antibodies It is possible.

[0214] When two antibodies recognize identical, overlapping, or adjacent epitopes in three-dimensional space, The antibody may target "essentially the same epitope" or "the same epitope" as the reference antibody. Two antibodies bind to identical, overlapping, or adjacent epitopes in three-dimensional space. The most widely used and rapid method for determining binding is a competitive assay. This can be done using several different methods, for example, using either labeled antigens or labeled antibodies. In some assays, antigens are plated onto a 96-well plate. The antibody is immobilized on a substrate or expressed on the cell surface, and unlabeled antibody blocks the binding of labeled antibody. The capacity is measured using radioactive, fluorescent, or enzymatic labels.

[0215] "Epitope mapping" refers to mapping the binding site, or epitope, of an antibody to its target antigen. The process of identifying antibody epitopes can be linear or conformational. A linear epitope is formed by a continuous sequence of amino acids in a protein. Conformational epitopes are discontinuous in the protein sequence, but are present when the protein is in its tertiary form. It is formed from amino acids that are brought together when folding into the parent structure. The epitope is a protein that binds to another protein or ligand, for example, after activation or binding. They are formed when the three-dimensional structure of a protein changes to a different three-dimensional structure.

[0216] "Epitope binning" is a process that classifies antibodies based on the epitopes they recognize. More specifically, epitope binning is the process of identifying the epitope recognition characteristics of various antibodies. A computational process for identifying and clustering antibodies based on their epitope recognition properties. A competitive assay combined with a denaturing process is used to identify antibodies with different binding specificities. The present invention includes a method and system for

[0217] "GFRAL-mediated disease," "GFRAL-mediated disorder," and "GFRAL-mediated illness" are used interchangeably. These proteins are mediated either completely or partially by GFRAL or by the interaction of GFRAL with GDF15. any mutations that are caused by or are the result of GFRAL or the interaction of GFRAL with GDF15 and / or alternatively inhibiting the in vivo effects of GDF15. refers to any disease, disorder, or illness in which it is desirable to treat a GFRAL-mediated disease, disorder, or illness. These include GDF15-mediated diseases, disorders, or conditions.

[0218] “GDF15-mediated disease,” “GDF15-mediated disorder,” and “GDF15-mediated disease” are used interchangeably. (i) GDF15 protein (e.g., activity of GDF15 protein, e.g., GDF15 signaling) or increased levels of GDF15 protein) or GFRAL protein and GDF15 protein It is caused entirely or in part by interaction with the protein and / or the RET protein. or (ii) a GDF15 protein (e.g., an activity of the GDF15 protein, e.g., a GDF15 signaling or increased levels of GDF15 protein) or GFRAL protein and GDF15 protein any disease, disorder, or condition that is the result of an interaction with a protein and / or RET protein Alternatively, any disease, disorder, or condition in which it is desirable to inhibit the in vivo effects of GDF15. refers to a disease. GDF15-mediated diseases, disorders, or conditions include involuntary weight loss, cachexia, , sarcopenia, muscle wasting, bone wasting, cardiovascular disease, chronic inflammatory diseases (e.g., chronic kidney disease, Chronic obstructive pulmonary disease), and GDF15 protein-induced or GDF15 protein-dependent Sensitivity (e.g., to relevant chemotherapeutic agents) (e.g., antitumor antibodies, e.g., trastuzumab) For example, cancers, including cancers with reduced resistance.

[0219] As used herein, the term "therapeutically effective amount" refers to a therapeutically effective amount of a compound administered to treat a given disease, disorder, or condition. reducing and / or ameliorating the severity and / or duration of the disease and / or symptoms associated therewith; an agent (e.g., an antibody described herein or a compound described herein) sufficient to improve A therapeutically effective amount of a drug, including a therapeutic agent, refers to the amount of (i) a drug or any other drug that is effective in treating a given disease, (ii) a reduction or amelioration of the development or progression of a given disease, disorder, or condition; or (iii) a reduction or amelioration of disease recurrence, onset, or occurrence; and / or (iv) a reduction or amelioration of disease recurrence, onset, or occurrence; and / or (v) a reduction or amelioration of disease recurrence, onset, or occurrence; For example, improving the prophylactic or therapeutic efficacy of a therapy other than administration of an antibody provided herein. The amount of the substance / molecule / drug (e.g., anti-GFRAL antibody) of the present disclosure may be an amount necessary for enhancing or enhancing the effect of the substance / molecule / drug (e.g., anti-GFRAL antibody) of the present disclosure. A "therapeutically effective amount" of a substance / molecule / drug (or a combination thereof) is determined based on the individual's condition, age, sex, and weight, and the amount of the substance / molecule / drug. Therapeutic efficacy may vary depending on factors such as the ability of the agent to elicit a desired response in an individual. Amounts include those amounts in which any toxic or detrimental effects of the substance / molecule / agent are outweighed by the therapeutically beneficial effects. In certain embodiments, the term "therapeutically effective amount" refers to a therapeutically effective amount for treating a disease in a subject or mammal. The amount of an antibody or other agent (e.g., or drug) effective to "treat" a disease, disorder, or condition. Refers to...

[0220] A "prophylactically effective amount" refers to the dosage necessary to achieve the desired prophylactic result and to It refers to an amount effective for a period of time necessary to achieve that result. Generally, a prophylactic dose is used to treat a disease. The present invention relates to a method for treating a disease, disorder, or disease, and is therefore useful for preventing or treating the disease. A therapeutically effective amount may, but is not necessarily, less than a therapeutically effective amount. stomach.

[0221] "Chronic" administration is the administration of acute-like substances in order to maintain the initial therapeutic effect (activity) over an extended period of time. Administering a drug in a continuous manner (e.g., over a period of days, weeks, months, or years), rather than in a single dose "Intermittent" administration refers to the administration of a drug in short bursts rather than continuously without interruption. Rather, it is a periodic procedure in nature.

[0222] Administration "in combination with" one or more further agents includes simultaneous (e.g., concurrent) administration and any "in the context of administration of other therapies (e.g., other drugs)" The term "in combination with" 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 a subject. The first therapy (e.g., drug) is administered to a patient who has or is currently suffering from a GDF15-mediated disease, disorder, or condition. to a subject having, susceptible to, or at risk for the disease; Prior to administration of the second therapy (e.g., drug) (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 Between, 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 8 weeks, 9 weeks, 10 weeks, 11 weeks or 12 weeks before), concurrently with or after the administration (e.g., 1 minute, 15 minutes, 3 0 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 The administration can be made within 1 week, 9 weeks, 10 weeks, 11 weeks, or 12 weeks after the initial administration.

[0223] Any additional therapy (e.g., drug) may be administered in any order with other additional therapies (e.g., drugs). In certain embodiments, the antibody can be administered in combination with one or more therapies, e.g., Administration in combination with a drug (e.g., a therapy, including a drug other than a currently administered antibody) to prevent, treat, manage, or treat a GDF15-mediated disease, disorder, or condition, or a symptom thereof. and / or improve the efficacy and safety of therapies that can be administered in combination with the antibody (e.g., Non-limiting examples of the therapeutic agent (drug) include, for example, analgesics, anesthetics, antibiotics, or immunomodulators. or the US Pharmacopoeia and / or Physician's Desk Reference. Drugs useful in combination therapy include any other drug listed in the FDA's Medical Journal Desk Reference. Examples of agents include: nonsteroidal anti-inflammatory drugs (NSAIDs), such as aspirin, ibuprofen, Buprofen and other propionic acid derivatives (alminoprofen, benoxaprofen bucloxic acid, carprofen, fenbufen, fenoprofen, fluprofen Flurbiprofen, Indoprofen, Ketoprofen, Miroprofen, Napro Xene, Oxaprozin, Pirprofen, Pranoprofen, Suprofen, Thiapro phenic acid, and tioxaprofen), acetic acid derivatives (indomethacin, acemetacin, alginic acid, Clofenac, clidanac, diclofenac, fenclofenac, fenclozic acid, Fentiazac, fuirofenac, ibufenac, isoxepac, oxypi nacoxib, sulindac, tiopinac, tolmetin, zidometacin, and zomepirac), phenacin flufenamic acid derivatives (flufenamic acid, meclofenamic acid, mefenamic acid, niflumic acid, and t fenamic acid), biphenylcarboxylic acid derivatives (diflunisal and flufenisal), oxy cams (isoxicam, piroxicam, sudoxicam, and tenoxican), Salicylates (acetylsalicylic acid, sulfasalazine), and pyrazolones (Apazone, Bezpiperylon, feprazone, mofebutazone, oxyphenbutazone Other combinations include, but are not limited to, cyclohexyl benzoate ... Other drugs for combination use include cyclooxygenase-2 (COX-2) inhibitors. Donisolone, prednisone, methylprednisolone, betamethasone, dexamethasone, or Examples of such combinations include steroids such as hydrocortisone. It is possible that the steroids required when treating a subject in combination with the antibody may be By gradually tapering the steroid dose, one or more side effects of steroids can be reduced or eliminated. Further examples of drugs for combination include cytokine suppressive anti-inflammatory drugs. 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. Antibodies that inhibit the activity of the antibody or their antagonists are also included. Humanized or human TNF antibodies, REMICADE, anti-TNF antibody fragments (e.g., CDP870), and soluble p55 or p75 TNF receptor, their derivatives, p75TNFRIgG (ENBREL®) or p55TNFR1gG (LE NERCEPT®), soluble IL-13 receptor (sIL-13), and TNFα-converting enzyme (TACE) inhibition as well as IL-1 inhibitors (e.g., interleukin-1 inhibitors). Other combinations include interleukin-1 converting enzyme inhibitors. These include anti-p7, anti-p-selectin glycoprotein ligand (PSGL), and anti-p-selectin 11. Other examples of useful drugs include interferon-β1a (AVONEX); interferon-β1b (BETA) TASERON®; Copaxone; Hyperbaric oxygen; Intravenous immunoglobulin; Clabrib ine); and antibodies to other human cytokines or growth factors or to these other human cytokine or growth factor antagonists (e.g., against CD40 ligand and CD80) antibodies).

[0224] As used herein, a "carrier" refers to a substance that is exposed to the agent at the dosage and concentration employed. a pharmaceutically acceptable carrier, excipient, or stabilizer that is non-toxic to the cells or mammals in which it is administered; Often the physiologically acceptable carrier is an aqueous pH buffered solution. Examples of commercially acceptable carriers include buffers, e.g., phosphate, citrate, and other organic acids; antioxidants, including ascorbic acid; low molecular weight (e.g., less than about 10 amino acid residues) polypeptides proteins, such as serum albumin, gelatin, or immunoglobulins; hydrophilic polymers, e.g., polyvinylpyrrolidone; amino acids, e.g., glycine, glutamine, asparagine, arginine, or lysine; monosaccharides, disaccharides, and glucose, mannose or other carbohydrates, including dextrin; chelating agents, e.g., EDTA; sugar alcohols , for example, mannitol or sorbitol; salt-forming counterions, for example, sodium; and and / or non-ionic surfactants, e.g., TWEEN™, polyethylene glycol (PEG) and PLURONICS™. The term "carrier" refers to the agent with which a therapeutic agent is administered. The diluent administered, adjuvant (e.g., Freund's adjuvant (complete or incomplete) )), excipient, or vehicle. Such carriers, including pharmaceutical carriers, The bacterial liquid can be, for example, water and oil, including oils of petroleum, animal, vegetable, or synthetic origin. Oils such as peanut oil, soybean oil, mineral oil, sesame oil, etc. Water may be present in the composition (e.g., For example, saline solution and ethanol are exemplary carriers when the pharmaceutical composition is administered intravenously. Aqueous dextrose and glycerol solutions are useful liquid carriers, particularly for injectable solutions. Suitable excipients (e.g., pharmaceutical excipients) include starch, glucose, sugar, lactose, sucrose, gelatin, malt, rice, flour, chalk, silica gel, Sodium stearate, glycerol monostearate, talc, sodium chloride, dehydrated Examples include dry milk, glycerol, propylene, glycol, water, and ethanol. The composition, if desired, can also contain minor amounts of wetting or emulsifying agents, or pH buffering agents. The compositions may be in the form of solutions, suspensions, emulsions, tablets, pills, capsules, powders, sustained release formulations, or the like. Oral compositions can take the form of a pharmaceutical formulation, including standard carriers, e.g. Pharmaceutical grade mannitol, lactose, starch, magnesium stearate, saccharin Suitable pharmaceutical carriers include sodium, cellulose, magnesium carbonate, etc. See, for example, Remington's Pharmaceutical Sciences (1990) Mack Publishing Compositions containing the pharmaceutical compounds are described in the U.S. Pat. No. 6,229,133, filed on Dec. 1, 2002, at the University of Illinois, Easton, PA. An effective amount of anti-GFRAL antibody, e.g., in isolated or purified form, is administered to a subject (e.g., a patient). The formulation may contain a suitable amount of carrier so as to provide the form for proper administration. It should be appropriate for the mode of administration.

[0225] As used herein, the term "pharmaceutically acceptable" refers to an animal, more particularly Approved by a federal or state regulatory agency for human use or It is intended to be listed in the European Pharmacopoeia, the European Pharmacopoeia, or any other generally recognized pharmacopoeia. Taste.

[0226] The term "pharmaceutical formulation" refers to a pharmaceutical composition in which the biological activity of the active ingredient (e.g., an anti-GFRAL antibody) is effective. and is in a form that allows for the administration of the formulation without being unacceptable to the subject to whom it is administered. Refers to a preparation that does not contain any additional toxic ingredients. Such a preparation may be sterile. stomach.

[0227] A "sterile" preparation is aseptic or free of all living microorganisms and their spores.

[0228] As used herein, a "polyclonal antibody" refers to a protein having many epitopes. are generated in an immunogenic response to a protein, and therefore, are likely to be associated with the same epitope and Polyclonal refers to a population of antibodies that contain a variety of different antibodies directed against different epitopes. Methods for producing antibodies are known in the art (see, e.g., Chapter 11: Short Protocols in Molecular Biology). Short Protocols in Molecular Biology (2002), 5th ed., edited by Ausubel et al., John Wiley and Sons, New York).

[0229] An "isolated nucleic acid" is a nucleic acid that is naturally associated with other genomic DNA sequences, as well as with the native sequence. Substantially separated from proteins or complexes, e.g., ribosomes and polymerases An "isolated" nucleic acid molecule is a nucleic acid, e.g., RNA, DNA, or mixed polymers. A nucleic acid molecule that is separated from other nucleic acid molecules that are present in the natural source of the molecule. An "isolated" nucleic acid molecule, e.g., a cDNA molecule, is one that has been isolated from other cellular material or produced by recombinant techniques. If produced, it is substantially free of culture medium, or if chemically synthesized, , and can be substantially free of chemical precursors or other chemicals. In one embodiment, one or more nucleic acid molecules encoding the antibodies described herein are isolated. or purified. This term includes a nucleic acid sequence that has been removed from its natural environment and is not intended to be recombinant. DNA isolates or cloned DNA isolates and chemically synthesized analogs or heterologous systems A substantially pure molecule includes an isolated form of the molecule. The state may be included.

[0230] "Polynucleotide" or "nucleic acid," as used interchangeably herein, refers to a nucleic acid of any length. A nucleotide refers to a polymer of nucleotides, including DNA and RNA. xylribonucleotides, ribonucleotides, modified nucleotides or bases, and / or These analogs, or polymers produced by DNA or RNA polymerase or by synthetic reactions, The polynucleotide can be any substrate that can be incorporated into a modified mer. Nucleotides may include, for example, methylated nucleotides and their analogs. The term "oligonucleotide" as used herein generally refers to a short, usually single-stranded, usually synthetic polymer. A nucleotide refers to a nucleotide sequence that is usually, but not necessarily, less than about 200 nucleotides in length. The terms "oligonucleotide" and "polynucleotide" are used interchangeably. The above description of polynucleotides is not mutually exclusive. The cells producing the anti-GFRAL antibodies of the present disclosure include parent hybrids. and bacterial and eukaryotic host cells into which nucleic acid encoding the antibody has been introduced. Suitable host cells are disclosed below.

[0231] As used herein, "treat," "treatment," and "treatment" are used interchangeably. The term "treating" refers to the administration of one or more therapies (one or more prophylactic or therapeutic agents, e.g., GDFs resulting from the administration of antibodies provided herein, including but not limited to, 15 Refers to a reduction or amelioration of the progression, severity, and / or duration of a vector-borne disease, disorder, or illness. In some embodiments, the agent is an anti-GFRAL antibody. Treatments include (i) increasing weight; (ii) maintaining weight; and (iii) reducing weight loss. (iv) increasing body mass (e.g., lean mass or fat mass); (v) increasing body mass; (vi) maintaining body mass (e.g., lean mass or fat mass); However, the present invention also includes reducing the loss of fat mass or fat mass, or any combination thereof. , but not limited to these.

[0232] As used herein, the term "prophylactic agent" refers to a drug or composition that is effective in treating a GDF15-mediated disease in a subject. the onset, recurrence, occurrence, or spread of any disease, disorder, or illness, and / or symptoms associated therewith; In some embodiments, the term "agent" refers to any agent capable of fully or partially inhibiting In some embodiments, the term "prophylactic agent" refers to the antibodies provided herein. In the present specification, the term "prophylactic agent" refers to an agent other than an antibody provided herein. In some embodiments, the prophylactic agent is a drug that treats a GDF15-mediated disease, disorder, or condition, and / or Preventing symptoms associated with or treating GDF15-mediated diseases, disorders, or conditions, and / or are useful for interfering with the onset, development, progression, and / or severity of symptoms associated therewith. is known to be, has been used for, or is currently being used for In some embodiments, the prophylactic agent is a fully human anti-GFRAL antibody. For example, a fully human anti-GFRAL monoclonal antibody.

[0233] The term "prophylactic agent" refers to a drug or method for treating a GDF15-mediated disease, disorder, or condition in a subject, or The onset, recurrence, development, or spread of these symptoms can be completely or partially inhibited. In certain embodiments, the term "prophylactic agent" refers to any agent described herein. In certain other embodiments, the term "prophylactic agent" refers to an anti-GFRAL antibody. In certain embodiments, the prophylactic agent is a GDF15 antibody. or a GDF15-mediated disease, disorder, or condition, or a symptom thereof. Interfering with the onset, development, progression, and / or severity of a disorder, disease, or symptoms thereof is known to be useful for, or has been used for, or is In a specific embodiment, the prophylactic agent is a humanized The anti-GFRAL antibody is, for example, a humanized anti-GFRAL monoclonal antibody.

[0234] The term "package insert" refers to a material commonly included in commercial packaging for therapeutic products. In rare cases, the indications, usage, dosage, administration, contraindications, and / or is used to refer to instructions that contain warning information.

[0235] "Prevent," "preventing," and "prevention" The term refers to a therapy or combination of therapies (e.g., prophylactic or therapeutic agents, e.g., a combination of prophylactic and therapeutic agents) provided herein. and GDF15-mediated diseases mediated by the administration of antibodies (e.g., antibody combinations provided herein). the complete or complete progression of any disease, disorder, or illness, or the onset, recurrence, occurrence, or spread of any of their symptoms Refers to partial inhibition.

[0236] The term "recombinant antibody" refers to an antibody that is prepared, expressed, produced, or isolated by recombinant means. Recombinant antibodies are antibodies produced by transfecting a recombinant expression vector into a host cell. antibodies expressed using recombinant combinatorial antibody libraries, antibodies isolated from recombinant combinatorial antibody libraries, Transgenic and / or transchromosomal human immunoglobulin genes Antibodies isolated from normal animals (e.g., mice or bovine) (see, e.g., Taylor, L. (See, e.g., D. et al. (1992) Nucl. Acids Res. 20:6287-6295), or immunoglobulin genetic It may be prepared, expressed, or produced by any other means involving splicing of the DNA sequence into other DNA sequences. Such recombinant antibodies can be human germline derived or isolated antibodies. The antibody can have variable and constant regions, including those derived from immunoglobulin sequences (K abat, EA et al. (1991), Sequences of Immunologically Interesting Proteins ins of Immunological Interest), 5th edition, US Department of Health and Human Servi. ces, NIH Publication No. 91-3242). In some embodiments, however, Such recombinant antibodies can be produced by in vitro mutagenesis (or transgenes for human Ig sequences). When using nicked animals, they may be subjected to in vivo somatic mutagenesis (somatic mutagenesis), which Therefore, the amino acid sequences of the VH and VL regions of the recombinant antibody are derived from human germline VH and VL sequences. derived from and related to, but not naturally occurring in the human antibody germline repertoire in vivo. It is an array that may not be

[0237] The term "side effect" refers to an undesirable and harmful effect of a therapy (e.g., a prophylactic or therapeutic agent). Unwanted effects are not necessarily adverse. An adverse effect from a prophylactic or therapeutic agent might be harmful or uncomfortable or dangerous. Examples of side effects These include diarrhea, cough, gastroenteritis, wheezing, nausea, vomiting, loss of appetite, abdominal cramps, fever, pain, Weight loss, dehydration, hair loss, difficulty breathing, insomnia, dizziness, mucositis, neuromuscular effects, fatigue, dry skin, loss of appetite, rash or swelling at the site of administration, flu-like symptoms, e.g. Symptoms experienced by patients include fever, chills, and fatigue, gastrointestinal upset, and allergic reactions. Additional undesirable effects that can occur are numerous and known in the art. This is described in the Physician's Desk Reference (60th ed., 2006).

[0238] The terms "subject" and "patient" are used interchangeably herein and are used herein to refer to In the context of the disclosed methods, it refers to an animal that is the recipient of a therapeutic or prophylactic case. As used herein, in some embodiments, a subject is a mammal, e.g., a non-spirit. Long-legged animals (e.g., cattle, pigs, horses, cats, dogs, rats, etc.) or primates (e.g., monkeys and and humans). In a specific embodiment, the subject is a human. The subject is a mammal (eg, a human) having a GDF15-mediated disease, disorder, or condition. In another embodiment, the subject is at risk of developing a GDF15-mediated disease, disorder, or condition. A mammal (eg, a human).

[0239] "Substantially all" means at least about 60%, at least about 65%, at least about 70%, at least at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or indicates approximately 100%.

[0240] The term "therapeutic agent" refers to a therapeutic agent that treats a GDF15-mediated disease, disorder, or condition, or a symptom thereof. to treat, prevent, or alleviate one or more symptoms of a disease, disorder, or illness, It refers to any agent that can be used in the treatment, prevention, or amelioration of In certain other embodiments, the therapeutic agent refers to an anti-GFRAL antibody described herein. In this context, a therapeutic agent refers to an agent other than an antibody provided herein. The therapeutic agent is a compound used to treat a GDF15-mediated disease, disorder, or condition, or one or more symptoms of these conditions. known to be useful in, or has been used to, prevent, or mitigate or a drug currently used for that purpose.

[0241] A combination of therapies (e.g., prophylactic or For example, the synergistic effect of a combination of prophylactic and / or therapeutic agents may be due to GDF15-mediated administration of one or more of the drugs to a subject with a disease, disorder, or condition This allows for the use of lower dosages and / or less frequent administration of the drug. the ability to utilize lower amounts of prophylactic or therapeutic therapy and / or administer said therapy less frequently; The present invention relates to the use of such a therapy in the prevention, management, treatment, or amelioration of a GDF15-mediated disease, disorder, or condition. and reducing the toxicity associated with administering the therapy to a subject without reducing the efficacy of the therapy. In addition, the synergy may be beneficial in the prevention or management or treatment of a GDF15 mediated disease, disorder, or condition. Finally, the efficacy of a therapy (e.g., The synergistic effects of a combination of agents (e.g., prophylactic or therapeutic agents) may be expected to reduce the adverse effects associated with the use of any single agent. In some embodiments, adverse or unwanted side effects can be avoided or reduced. Thus, combination therapy may include the use of an antibody provided herein and insulin (e.g., insulin supplementation). In some embodiments, the combination therapy comprises an anti-GFRAL antibody and insulin. wherein the combination therapy is administered at a lower daily dose than is typically administered in insulin monotherapy. In some embodiments, the combination therapy includes, for example, a daily dose of insulin. , less than 90%, less than 80%, and 70% of the usual daily insulin dosage for insulin monotherapy Less than, Less than 60%, Less than 50%, Less than 40%, Less than 30%, Less than 20%, Less than 15%, Less than 10%, 5% Including less than, less than 3%, or less than 1%.

[0242] The term "therapy" refers to a treatment for a GDF15-mediated disease, disorder, or condition (e.g., type 1 diabetes or type 2 diabetes). Any protocol that can be used in the prevention, management, treatment, and / or amelioration of diabetes In some embodiments, "therapies" refers to and the term "therapy" refers to biotherapy, supportive therapy, and / or other therapeutic approaches known to those skilled in the art, e.g. , the prevention, management, treatment, and / or administration of GDF15-mediated diseases, disorders, or conditions known to the medical profession. or other therapies that may be useful in improving the condition.

[0243] The term "detectable probe" refers to a composition that provides a detectable signal. The term includes, but is not limited to, any molecule that provides a detectable signal through its activity. This includes any fluorophore, chromophore, radiolabel, enzyme, antibody, or antibody fragment, and the like.

[0244] The term "diagnostic agent" refers to an agent administered to a subject that aids in the diagnosis of a disease, disorder, or condition. Such materials can be used to localize and pinpoint disease-causing processes. In certain embodiments, the diagnostic agent can be administered to a subject. or a GDF15-mediated disease, disorder, or condition when administered to or contacted with a sample from a subject. and agents conjugated to the anti-GFRAL antibodies described herein that aid in diagnosis. nothing.

[0245] The term "detectable agent" refers to a molecule of interest in a sample or subject, e.g., a molecule as defined herein. Used to confirm the existence or presence of the described anti-GFRAL antibodies A detectable agent refers to a substance that can be visualized or otherwise detected. The substance can be a substance that is determined and / or measured by a method (eg, by quantification).

[0246] The term "encode" or its grammatical equivalents is used when referring to a nucleic acid molecule. When engineered, the resulting product may be in its native state or, when engineered by methods well known to those skilled in the art, can be transcribed to produce mRNA that is translated into polypeptides and / or fragments thereof. The antisense strand refers to a nucleic acid molecule that can The code sequence can then be deduced.

[0247] The term "excipient" refers to any substance that acts as a diluent, vehicle, preservative, binder, or stabilizer. Refers to commonly used inactive substances, such as proteins (e.g., serum albumin), amino acids, etc. (e.g., aspartic acid, glutamic acid, lysine, arginine, glycine, histidine etc.), fatty acids and phospholipids (e.g., alkyl sulfonates, caprylates, etc.), interfaces surfactants (e.g., SDS, polysorbates, non-ionic surfactants, etc.), sugars (e.g., sugars, sugars, maltose, trehalose, etc.), and polyols (e.g., mannitol, These include, but are not limited to, sorbitol, etc., which are incorporated herein by reference in their entirety. Remington's Pharmaceutical Sciences (1990) Mack Publishing See also lishing Co., Easton, PA.

[0248] The term "fragment" as used herein in the context of a peptide or polypeptide The term refers to a peptide or polypeptide that contains less than the full-length amino acid sequence. Fragments can be, for example, amino-terminal truncations, carboxy-terminal truncations, and / or amino acid sequences. Fragments can arise from internal deletion of residues from a sequence, e.g., by alternative RNA splicing. In some embodiments, the fragments may be generated by protease activity in vivo or by protease activity in vivo. The fragment contains at least one amino acid sequence of a GFRAL polypeptide or an antibody that binds to a GFRAL polypeptide. At least 5 consecutive amino acid residues, at least 10 consecutive amino acid residues, at least 1 5 consecutive amino acid residues, at least 20 consecutive amino acid residues, at least 25 at least 40 consecutive amino acid residues, at least 50 consecutive amino acid residues at least 60 consecutive amino acid residues, at least 70 consecutive amino acid residues amino acid residues, at least 80 consecutive amino acid residues, at least 90 consecutive amino acid residues amino acid residues, at least 100 consecutive amino acid residues, at least 125 consecutive amino acid residues a group consisting of at least 150 contiguous amino acid residues, or 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 consecutive amino acid residues In some embodiments, the GFRAL polypeptide comprises a polypeptide comprising the amino acid sequence of the group. A fragment of an antibody that binds to a peptide may comprise at least one, at least two, or at least two of the peptides of the antibody. It has at least three or more functions.

[0249] The words "manage," "managing," and "management" The term refers to the beneficial effect a subject derives from a therapy (e.g., a prophylactic or therapeutic agent), which therapy is effective in treating a disease. In some embodiments, the therapeutic agent may be used to prevent the progression or worsening of the disease. To achieve this, the subject is administered one or more therapies (e.g., prophylactic or therapeutic agents, e.g., those provided herein). and administering a GDF15-mediated disease, disorder, or condition, or a symptom thereof, to "manage" the disease, disorder, or condition. Understand."

[0250] "Administering" or "administration" refers to administering a substance as it exists outside the body (e.g., and / or Injected by any other physical delivery method described herein or known in the art. refers to the act of shooting or otherwise physically delivering an illness, disorder, or disease; When the condition is being treated, administration of the substance usually involves treating the disease, disorder, or illness, or after the onset of symptoms thereof. When administered, administration of the substance typically results in a reduction in the onset of the disease, disorder, or condition, or a symptom thereof. It is performed before the disease.

[0251] The term "analog" as used herein in the context of a polypeptide refers to a GFRA Similar or identical to the L polypeptide, a fragment of the GFRAL polypeptide, or an anti-GFRAL antibody The GFRAL polypeptide, a fragment of the GFRAL polypeptide, or a polypeptide that possesses the function of the GFRAL polypeptide, but does not necessarily does not contain the same or identical amino acid sequence as the anti-GFRAL antibody, and a polypeptide having a structure similar to or identical to that of an anti-GFRAL antibody, a fragment of a GFRAL polypeptide, or an anti-GFRAL antibody Polypeptides with similar amino acid sequences are also referred to as refers to a polypeptide that satisfies at least one of the following: (a) a GFR as described herein; AL polypeptide (e.g., SEQ ID NO: 500), a fragment of a GFRAL polypeptide, or an anti-GFRAL antibody The amino acid sequence and 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 at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or (b) a polypeptide having an amino acid sequence that is at least 99% identical to acid residues, at least 10 amino acid residues, at least 15 amino acid residues, at least 20 amino acids residues, at least 25 amino acid residues, at least 40 amino acid residues, at least 50 amino acid residues a group, 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 a GFRAL polypeptide of at least 150 amino acid residues as described herein, A fragment of a peptide or a nucleotide sequence encoding an anti-GFRAL antibody (or its VH or VL region) The encoded sequence is a nucleotide sequence that hybridizes under stringent conditions to the The polypeptide to be cloned (see, e.g., Sambrook et al. (2001), Molecular Cloning: Laboratory Manual) Al (Molecular Cloning: A Laboratory Manual), Cold Spring Harbor Laboratory Pres. s, Cold Spring Harbor, NY; Maniatis et al. (1982), Molecular Cloning: A Laboratory Manual. Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Press, Cold Sprin (c) a GFRAL polypeptide as described herein, see e.g., GFRAL polypeptide. a nucleotide sequence encoding a fragment of a nucleotide sequence encoding an anti-GFRAL antibody (or its VH or VL region); Column and 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 A polypeptide encoded by a nucleotide sequence that is at least 99% identical to the polypeptide described herein. GFRAL polypeptides, GFRAL polypeptide fragments, or anti-GFRAL antibodies having a structure similar to that of the GFRAL polypeptides, The polypeptide may be a GFRAL polypeptide, a fragment of GFRAL, or a GFRAL polypeptide described herein. It refers to a polypeptide that has the same secondary, tertiary, or quaternary structure as the RAL antibody. The structure can be characterized by a variety of techniques, including, but not limited to, X-ray crystallography, nuclear magnetic resonance, and crystallographic electron microscopy. It can be determined by methods known to those skilled in the art.

[0252] The term "composition" refers to a composition containing specified ingredients (e.g., and optionally, a specified amount of a specified ingredient. It is intended to encompass any product resulting directly or indirectly from the combination.

[0253] The term "derivative" as used herein in relation to polypeptides means an amino acid sequence. GFRAL polypeptides, which have been modified by the introduction of substitutions, deletions, or additions of amino acid residues, A fragment of a GFRAL polypeptide or an antibody that binds to a GFRAL polypeptide, comprising the amino acid sequence of the antibody. The term "derivative" as used herein refers to, for example, a polypeptide GFRAL polynucleotides that are chemically modified by covalent attachment of any type of molecule to the peptide. It also refers to a peptide, a fragment of a GFRAL polypeptide, or an antibody that binds to a GFRAL polypeptide. For example, but not limited to, a GFRAL polypeptide, a fragment of a GFRAL polypeptide, or a GF The RAL antibody may be modified, for example, by glycosylation, acetylation, pegylation, phosphorylation, amidation, or other modifications known in the art. Derivatization with protecting / blocking groups, proteolytic cleavage, cellular ligands or other proteins The derivative may be chemically modified, such as by conjugation to a molecule. Either the type or position of the molecule is a naturally occurring peptide or polypeptide or a starting peptide. Derivatives are peptides or polypeptides that are modified differently from the peptides or polypeptides. The GFRAL polypeptide further includes deletion of one or more chemical groups naturally occurring on the peptide. Fragments of the L polypeptide or derivatives of the GFRAL antibody include, but are not limited to, specific chemical cleavage. techniques known to those skilled in the art, including acetylation, formylation, metabolic synthesis of tunicamycin, etc. Furthermore, the GFRAL polypeptide may be chemically modified by chemical modification using A derivative of a GFRAL polypeptide, a fragment of a GFRAL polypeptide, or a GFRAL antibody may contain one or more atypical amino acids. The polypeptide derivative may comprise a GFRAL polypeptide, GFRAL polypeptide, or the like, as described herein. A fragment of a FRAL polypeptide or a GFRAL antibody possesses a similar or identical function.

[0254] The term "involuntary weight loss" refers to conditions such as cachexia, cirrhosis, hyperthyroidism, and chronic kidney disease. , intention observed in many diseases such as Parkinson's disease, cancer, eating disorders, and sarcopenia. This refers to weight loss that does not occur.

[0255] The term "cachexia" refers to weight loss, muscle atrophy, and other conditions in people who are not actively trying to lose weight. Cachexia refers to a wasting syndrome characterized by atrophy, fatigue, weakness, and marked loss of appetite. Some chronic diseases (e.g., cancer, chronic kidney disease, chronic inflammatory diseases, muscle wasting, e.g., muscular dystrophy) These conditions can contribute significantly to the morbidity of patients suffering from anorexia nervosa (e.g., anorexia nervosa, and dysphagia). In terminal cancer, cachexia is common (occurring in the most terminally ill cancer patients) and accounts for approximately 100% of all cancer-related deaths. It accounts for about a quarter of all deaths.

[0256] (Composition and method for producing same) Binding proteins, e.g., antibodies that bind to GFRAL (e.g., human GFRAL), are provided. The disclosed antibodies are useful, for example, for diagnosing or treating GDF15-mediated diseases, disorders, or conditions. In certain embodiments, the antibodies of the present disclosure are used to treat, but are not limited to, cachexia or chronic diseases ( For example, cirrhosis of the liver, hyperthyroidism, Parkinson's disease, cancer, chronic kidney disease, and chronic obstructive pulmonary disease (AIDS, tuberculosis, chronic inflammatory diseases, sepsis, muscle wasting, and anorexia nervosa) Involuntary weight loss, including involuntary weight loss in subjects with a disease, disorder, or The present invention relates to a method for treating a disease, or broadly any disease, disorder, or condition in which it is desirable to inhibit the in vivo action of GDF15. Or it is useful for diagnosing or treating a disease.

[0257] Provided herein are GFRAL polypeptides, GFRAL polypeptide fragments, GFRAL polypeptide fragments, and GFRAL polypeptide fragments. The antibody is an antibody (e.g., a monoclonal antibody) that binds to the GFRAL epitope or the GFRAL epitope. In some embodiments, the anti-GFRAL antibody binds to the extracellular domain (ECD) of GFRAL. Also provided are antibodies that bind to GFRAL polypeptides, such as those described herein. The anti-GFRAL antibodies provided herein are useful as diagnostic agents, detectable antibodies, and the like. The antibody may also be conjugated or recombinantly fused to a suitable drug or therapeutic agent. Provided are compositions comprising anti-GFRAL antibodies.

[0258] Also provided herein are GFRAL polypeptides, GFRAL polypeptide fragments, GFRAL Peptide or immunoglobulin heavy chain, light chain, VH of anti-GFRAL antibody that binds to GFRAL epitope region, VL region, VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2, and / or VL CDR3. Also provided are isolated nucleic acid molecules encoding the GFRAL polypeptide, GFRAL. The present invention relates to a method for producing a polypeptide fragment, a GFRAL peptide, or an anti-GFRAL antibody that binds to a GFRAL epitope. Also provided are vectors and host cells containing the nucleic acid molecules encoding the GFRAL polypeptide. Antibodies that bind to GFRAL peptides, GFRAL polypeptide fragments, GFRAL peptides, or GFRAL epitopes This is a method for producing the above.

[0259] Methods of using anti-GFRAL antibodies are provided herein. The methods include administering to a subject (e.g., a patient) and treating, preventing, or alleviating one or more symptoms of a GDF15-mediated disease, disorder, or condition in a subject. and GDF15-mediated diseases, disorders, or conditions, including: Non-limiting examples of DF15-mediated diseases, disorders, or conditions include involuntary weight loss, wasting disorders, Disease, cachexia or chronic diseases (e.g., cirrhosis of the liver, hyperthyroidism, Parkinson's disease, cancer, chronic Renal disease, chronic obstructive pulmonary disease, AIDS, tuberculosis, chronic inflammatory diseases, sepsis, muscle wasting, and neurological disorders. Involuntary weight loss in subjects suffering from anorexia nervosa. Other illnesses, disorders, or diseases that can cause weight loss include eating disorders, muscle strophies, or multiple sclerosis.

[0260] (Anti-GFRAL antibody) In some embodiments, the present disclosure provides compounds that find use herein as therapeutic agents. Exemplary antibodies include polyclonal, monoclonal, and Humanized, human, bispecific, and heteroconjugate antibodies and improved affinity These variants may have different sex or other properties.

[0261] In some embodiments, provided herein is a GFRAL polypeptide, GFRAL Antibodies that bind to GFRAL, including polypeptide fragments, GFRAL peptides, or GFRAL epitopes In some embodiments, the anti-GFRAL antibody is a GFRAL polypeptide. A humanized antibody that binds to GFRAL, comprising a peptide fragment, a GFRAL peptide, or a GFRAL epitope. The antibody may be a human (e.g., one containing a human constant region).

[0262] In some embodiments, the anti-GFRAL antibody is a monoclonal antibody described herein. Antibodies (e.g., 1C1, 3P10, 12A3, 5F12, 5A20, 8D8, 17J16, 25M22, 2B8, 22N5, 2I23, 6N 16, 1B3, 19K19, 2B3, 8C10, 2A9, 24G2, 6G9, 2B11, 1A3, P1B6, P1H8, or P8G4) Any one of a VH region, a VL region, a VH CDR1, a VH CDR2, a VH CDR3, a VL CDR1, a VL CDR2, and / or The VL CDR3 comprises, for example, the amino acid sequences shown in Tables 1 to 24. In embodiments, the isolated antibodies or functional fragments thereof provided herein are selected from the group consisting of antibodies, antibodies, and antibodies described in Tables 1-24. As shown: (a) antibody designated 1C1; (b) antibody designated 3P10; (c) antibody designated 12A3 (d) the antibody designated 5F12; (e) the antibody designated 5A20; (f) the antibody designated 8D8; (g) the antibody designated 5F12 ...A20; (g) the antibody designated 8D8; (h) the antibody designated 25M22; (i) the antibody designated 2B8; (j) the antibody designated 22N5 (k) antibody designated as 2I23; (l) antibody designated as 6N16; (m) antibody designated as 1B3 (n) antibody designated as 19K19; (o) antibody designated as 2B3; (p) antibody designated as 8C10; ( (q) antibody designated as 2A9; (r) antibody designated as 24G2; (s) antibody designated as 6G9; (t) antibody designated as 2B11 (u) antibody designated 1A3; (v) antibody designated P1B6; (w) antibody designated P1H8 (x) an antibody designated P8G4; or one, two, and / or a combination of antibodies designated 1C1 to P8G4 or three heavy chain CDRs and / or one, two, and / or three light chain CDRs.

[0263] The antibody designated 1C1 comprises a VH sequence that is SEQ ID NO:1 and a VL sequence that is SEQ ID NO:2.

[0264] The antibody designated 3P10 comprises a VH sequence that is SEQ ID NO:3 and a VL sequence that is SEQ ID NO:4.

[0265] The antibody designated 12A3 comprises a VH sequence that is SEQ ID NO:5 and a VL sequence that is SEQ ID NO:6.

[0266] The antibody designated 5F12 comprises a VH sequence that is SEQ ID NO:7 and a VL sequence that is SEQ ID NO:8.

[0267] The antibody designated 5A20 comprises a VH sequence that is SEQ ID NO: 9 and a VL sequence that is SEQ ID NO: 10. .

[0268] The antibody designated 8D8 comprises a VH sequence of SEQ ID NO: 11 and a VL sequence of SEQ ID NO: 12. .

[0269] The antibody designated 17J16 contains the VH sequence of SEQ ID NO: 13 and the VL sequence of SEQ ID NO: 14. nothing.

[0270] The antibody designated 25M22 contains the VH sequence of SEQ ID NO: 15 and the VL sequence of SEQ ID NO: 16. nothing.

[0271] The antibody designated 2B8 comprises a VH sequence that is SEQ ID NO: 17 and a VL sequence that is SEQ ID NO: 18. .

[0272] The antibody designated 22N5 comprises a VH sequence of SEQ ID NO: 19 and a VL sequence of SEQ ID NO: 20. .

[0273] The antibody designated 2I23 comprises a VH sequence of SEQ ID NO: 21 and a VL sequence of SEQ ID NO: 22. .

[0274] The antibody designated 6N16 comprises a VH sequence of SEQ ID NO: 23 and a VL sequence of SEQ ID NO: 24. .

[0275] The antibody designated 1B3 comprises a VH sequence that is SEQ ID NO: 25 and a VL sequence that is SEQ ID NO: 26. .

[0276] The antibody designated 19K19 contains the VH sequence of SEQ ID NO:27 and the VL sequence of SEQ ID NO:28. nothing.

[0277] The antibody designated 2B3 comprises a VH sequence that is SEQ ID NO: 29 and a VL sequence that is SEQ ID NO: 30. .

[0278] The antibody designated 8C10 comprises a VH sequence that is SEQ ID NO: 31 and a VL sequence that is SEQ ID NO: 32. .

[0279] The antibody designated 2A9 comprises a VH sequence of SEQ ID NO: 33 and a VL sequence of SEQ ID NO: 34. .

[0280] The antibody designated 24G2 comprises a VH sequence that is SEQ ID NO: 35 and a VL sequence that is SEQ ID NO: 36. .

[0281] The antibody designated 6G9 comprises a VH sequence of SEQ ID NO: 37 and a VL sequence of SEQ ID NO: 38. .

[0282] The antibody designated 2B11 comprises a VH sequence that is SEQ ID NO: 39 and a VL sequence that is SEQ ID NO: 40. .

[0283] The antibody designated 1A3 comprises the VH sequence of SEQ ID NO: 480 and the VL sequence of SEQ ID NO: 481. nothing.

[0284] The antibody designated P1B6 comprises the VH sequence of SEQ ID NO: 482 and the VL sequence of SEQ ID NO: 483. nothing.

[0285] The antibody designated P1H8 comprises the VH sequence of SEQ ID NO: 484 and the VL sequence of SEQ ID NO: 485. nothing.

[0286] The antibody designated P8G4 comprises the VH sequence of SEQ ID NO: 486 and the VL sequence of SEQ ID NO: 487. nothing. Table 1: CDR sequences of antibody 1C [Table 2] Table 2: CDR sequences of antibody 3P10 [Table 3] Table 3: CDR sequences of antibody 12A3 [Table 4] Table 4: CDR sequences of antibody 5F12 [Table 5] Table 5: CDR sequences of antibody 5A20 [Table 6] Table 6: CDR sequences of antibody 8D8 [Table 7] Table 7: CDR sequences of antibody 17J16 [Table 8] Table 8: CDR sequences of antibody 25M22 [Table 9] Table 9: CDR sequences of antibody 2B8 [Table 10] Table 10: CDR sequences of antibody 22N5 [Table 11] Table 11: CDR sequences of antibody 2I23 [Table 12] Table 12: CDR sequences of antibody 6N16 [Table 13] Table 13: CDR sequences of antibody 1B3 [Table 14] Table 14: CDR sequences of antibody 19K19 [Table 15] Table 15: CDR sequences of antibody 2B3 [Table 16] Table 16: CDR sequences of antibody 8C10 [Table 17] Table 17: CDR sequences of antibody 2A9 [Table 18] Table 18: CDR sequences of antibody 24G2 [Table 19] Table 19: CDR sequences of antibody 6G9 [Table 20] Table 20: CDR sequences of antibody 2B11 [Table 21] Table 21: CDR sequences of antibody 1A3 [Table 22] Table 22: CDR sequences of antibody P1B6 [Table 23] Table 23: CDR sequences of antibody P1H8 [Table 24] Table 24: CDR sequences of antibody P8G4 [Table 25]

[0287] 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 comprise: (i) a VH domain or VH region; and / or (ii) a VL domain or combination of VL regions.

[0288] In some embodiments, the antibodies provided herein are selected from the six antibodies identified in Tables 1-24. CDRs, e.g., VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2, and / or VL CDR3 In some embodiments, the antibodies provided herein comprise or consist of the CDRs: In some embodiments, the antibody comprises one of the CDRs listed in Tables 1-24. VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2, and / or VL CDR3 as identified in and / or comprising one, two, three, four, or five CDRs selected from the group consisting of: In some embodiments, the antibody is described herein as: (a) 1C1 (b) the antibody designated 3P10; (c) the antibody designated 12A3; (d) the antibody designated 5F12; (e) antibody designated 5A20; (f) antibody designated 8D8; (g) antibody designated 17J16; (h) t25M2 (i) the antibody designated 2B8; (j) the antibody designated 22N5; (k) the antibody designated 2I23 (l) the antibody designated as 6N16; (m) the antibody designated as 1B3; (n) the antibody designated as 19K19 (o) antibody designated 2B3; (p) antibody designated 8C10; (q) antibody designated 2A9; (r) 24G2 (s) antibody designated as 6G9; (t) antibody designated as 2B11; (u) antibody designated as 1A3 (v) the antibody designated P1B6; (w) the antibody designated P1H8; or (x) the antibody designated P8G4 VH CDR1, VH CDR2, and VH CDR3 of a mouse monoclonal antibody selected from the group consisting of antibodies , VL CDR1, VL CDR2, and / or VL CDR3. Thus, in some embodiments, the CDRs comprise or consist of one or five CDRs. The antibody comprises a VH CDR1, a VH CDR2, a VH CDR3, a VL CDR1, a VL CDR2, a VL CDR3, a VL CDR4, a VL CDR5, a VL CDR6, a VL CDR7, a VL CDR8, a VL CDR9, a VL CDR10, a VL CDR11, a VL CDR12, a VL CDR13, a VL CDR14, and / or one, two, three, four, or five CDRs of any one of the VL CDR3 comprises or consists of said CDRs.

[0289] In some embodiments, the antibodies provided herein are selected from the group consisting of one or more of the antibodies listed in Tables 1-24. comprising or consisting of the above (e.g., one, two, or three) VH CDRs. In the present specification, the antibodies can be selected from one or more (e.g., one, two, three, or four) of the antibodies listed in Tables 1 to 24. In yet another embodiment, the antibody provided herein comprises one or three VL CDRs. comprises one or more (e.g., one, two, or three) VH CDRs and a CDR sequence as set forth in Tables 1-24. Thus, in certain embodiments, the antibody comprises one or more VL CDRs as set forth in , 57, 49, 221–296, 515–530, 488–493, 1795, 1796, 488–493, 1795, 1796 In another embodiment, the antibody comprises a VH CDR1 having any one of the amino acid sequences: VH CDR2 having any one of the amino acid sequences of SEQ ID NOs: 132 to 220 and 497 to 514 In another embodiment, the antibody is selected from the group consisting of SEQ ID NOs: 57, 49, 57, 49, 221-296, 515-530, 48 8 to 493, 1795, 1796, 488 to 493, 1795, 1796 In some embodiments, the antibody comprises a VH CDR3 having an amino acid sequence shown in Tables 1 to 24. VH CDR1 and / or VH CDR3 independently selected from VH CDR1, VH CDR2, and VH CDR3 shown in any one of In some embodiments, the antibody comprises a VH CDR2 and / or a VH CDR3. 2, 531 to 546. The antibody comprises a VL CDR2 having the amino acid sequence of any one of SEQ ID NOs: 373 to 422. In another embodiment, the antibody comprises an amino acid sequence selected from the group consisting of any one of SEQ ID NOs: 423 to 479, 558 to 569. In some embodiments, the antibody comprises a VL CDR3 having an amino acid sequence shown in Tables 1-24. VL CDR1, VL CDR2, and VL CDR3 independently selected from any one of the amino acid sequences It comprises CDR1 and / or VL CDR2 and / or VL CDR3.

[0290] Also provided herein are compounds that contain one or more (e.g., one, two, or more) of the compounds listed in Tables 1-24. An antibody that comprises three (or more) VH CDRs and one or more (e.g., one, two, or three) VL CDRs. Provided herein are: VH CDR1 (SEQ ID NOs: 57, 49, 221-296, 515-530, 488-499, 591-600, 592-593, 593-604, 594-605, 595-606, 596-607, 597-608, 598-610, 59 93, 1795, 1796) and VL CDR1 (SEQ ID NOs: 297-372, 531-546); VH CDR1 (SEQ ID NOs: 57, 49, 221 to 296, 515 to 530, 488 to 493, 1795, 1796) and VL CDR2 (SEQ ID NOs: 373 to 422); VH CDR1 ( SEQ ID NOs: 57, 49, 221-296, 515-530, 488-493, 1795, 1796) and VL CDR3 (SEQ ID NO: 423 VH CDR2 (SEQ ID NOs: 132 to 220, 497 to 514) and VL CDR1 (SEQ ID NOs: 297 to 37 2, 531-546); VH CDR2 (SEQ ID NOs: 132-220, 497-514) and VL CDR2 (SEQ ID NOs: 373-422); VH CDR2 (SEQ ID NOs: 132 to 220, 497 to 514) and VL CDR3 (SEQ ID NOs: 423 to 479, 558 to 569); VH C DR3 (SEQ ID NOs: 57, 49, 221-296, 515-530, 488-493, 1795, 1796) and VL CDR1 (SEQ ID NOs: VH CDR3 (SEQ ID NOs: 57, 49, 221-296, 515-530, 488-493, 179 5, 1796) and VL CDR2 (SEQ ID NOs: 373 to 422); VH CDR3 (SEQ ID NOs: 57, 49, 221 to 296, 515 to 53 0, 488-493, 1795, 1796) and VL CDR3 (SEQ ID NOs: 423-479, 558-569); VH CDR1 (SEQ ID NOs: Nos. 57, 49, 221-296, 515-530, 488-493, 1795, 1796), VH CDR2 (SEQ ID NOs. 132-220, 4 97 to 514), and VL CDR1 (SEQ ID NOs: 297 to 372, 531 to 546); VH CDR1 (SEQ ID NOs: 57, 49, 221 to 296, 515-530, 488-493, 1795, 1796), VH CDR2 (SEQ ID NOs: 132-220, 497-514), and V L CDR2 (SEQ ID NOs: 373 to 422); VH CDR1 (SEQ ID NOs: 57, 49, 221 to 296, 515 to 530, 488 to 493, 1795, 1796), VH CDR2 (SEQ ID NOs: 132 to 220, 497 to 514), and VL CDR3 (SEQ ID NOs: 423 to 479, VH CDR2 (SEQ ID NOs: 132 to 220, 497 to 514), VH CDR3 (SEQ ID NOs: 57, 49, 221 to 296) , 515-530, 488-493, 1795, 1796), and VL CDR1 (SEQ ID NOs: 297-372, 531-546), VH C DR2 (SEQ ID NOs: 132 to 220, 497 to 514), VH CDR3 (SEQ ID NOs: 57, 49, 221 to 296, 515 to 530, 488 VL CDR2 (SEQ ID NOs: 373 to 422); VH CDR2 (SEQ ID NOs: 132 to 220, 497 to 514), VH CDR3 (SEQ ID NOs: 57, 49, 221 to 296, 515 to 530, 488 to 493, 1795, 1796), and V L CDR3 (SEQ ID NOs: 423 to 479, 558 to 569); VH CDR1 (SEQ ID NOs: 57, 49, 221 to 296, 515 to 530, 488 to 493, 1795, 1796), VL CDR1 (SEQ ID NOs: 297 to 372, 531 to 546), and VL CDR2 (SEQ ID NOs: 373-422); VH CDR1 (SEQ ID NOs: 57, 49, 221-296, 515-530, 488-493, 1795, 1796), VL CDR1 (SEQ ID NOs: 297 to 372, 531 to 546), and VL CDR3 (SEQ ID NOs: 423 to 479, 558 to 569); VH C DR1 (SEQ ID NOs: 57, 49, 221-296, 515-530, 488-493, 1795, 1796), VL CDR2 (SEQ ID NO: 3 73 to 422), and VL CDR3 (SEQ ID NOs: 423 to 479, 558 to 569); VH CDR2 (SEQ ID NOs: 132 to 220, 497 VH CDR1 (SEQ ID NOs: 297 to 372, 531 to 546), and VL CDR2 (SEQ ID NOs: 373 to 422); DR2 (SEQ ID NOs: 132 to 220, 497 to 514), VL CDR1 (SEQ ID NOs: 297 to 372, 531 to 546), and VL CDR 3 (SEQ ID NOs: 423 to 479, 558 to 569); VH CDR2 (SEQ ID NOs: 132 to 220, 497 to 514), VL CDR2 (SEQ ID NOs: VL CDR3 (SEQ ID NOs: 423-479, 558-569); VH CDR3 (SEQ ID NOs: 57, 49, 221 to 296, 515 to 530, 488 to 493, 1795, 1796), VL CDR1 (SEQ ID NOs: 297 to 372, 531 to 546), and VL CDR2 (SEQ ID NOs: 373 to 422); VH CDR3 (SEQ ID NOs: 57, 49, 221 to 296, 515 to 530, 488 to 590) 493, 1795, 1796), VL CDR1 (SEQ ID NOs: 297 to 372, 531 to 546), and VL CDR3 (SEQ ID NOs: 423 to 429). 479, 558-569); VH CDR3 (SEQ ID NOs: 57, 49, 221-296, 515-530, 488-493, 1795, 1796 ), VL CDR2 (SEQ ID NOs: 373 to 422), and VL CDR3 (SEQ ID NOs: 423 to 479, 558 to 569); VH CDR1 ( SEQ ID NOs: 57, 49, 221-296, 515-530, 488-493, 1795, 1796), VH CDR2 (SEQ ID NOs: 132- 220, 497-514), VH CDR3 (SEQ ID NOs: 57, 49, 221-296, 515-530, 488-493, 1795, 1796 ), and VL CDR1 (SEQ ID NOs: 297 to 372, 531 to 546); VH CDR1 (SEQ ID NOs: 57, 49, 221 to 296, 51 5 to 530, 488 to 493, 1795, 1796), VH CDR2 (SEQ ID NOs: 132 to 220, 497 to 514), VH CDR3 (SEQ ID NOs: Nos. 57, 49, 221-296, 515-530, 488-493, 1795, 1796), and VL CDR2 (SEQ ID NOs. 373- 422); VH CDR1 (SEQ ID NOs: 57, 49, 221-296, 515-530, 488-493, 1795, 1796), VH CDR2 (SEQ ID NOs: 132 to 220, 497 to 514), VH CDR3 (SEQ ID NOs: 57, 49, 221 to 296, 515 to 530, 488 to 4 93, 1795, 1796), and VL CDR3 (SEQ ID NOs: 423 to 479, 558 to 569); VH CDR1 (SEQ ID NOs: 57, 49 , 221 to 296, 515 to 530, 488 to 493, 1795, 1796), VH CDR2 (SEQ ID NOs: 132 to 220, 497 to 514) VL CDR1 (SEQ ID NOs: 297 to 372, 531 to 546), and VL CDR2 (SEQ ID NOs: 373 to 422); VH CDR1 (SEQ ID NOs: 373 to 422); Sequence numbers 57, 49, 221 to 296, 515 to 530, 488 to 493, 1795, 1796), VH CDR2 (SEQ ID NOs: 132 to 22 0, 497 to 514), VL CDR1 (SEQ ID NOs: 297 to 372, 531 to 546), and VL CDR3 (SEQ ID NOs: 423 to 479 , 558-569); VH CDR1 (SEQ ID NOs: 57, 49, 221-296, 515-530, 488-493, 1795, 1796), VH CDR2 (SEQ ID NOs: 132 to 220, 497 to 514), VL CDR2 (SEQ ID NOs: 373 to 422), and VL CDR3 (SEQ ID NOs: VH CDR1 (SEQ ID NOs: 57, 49, 221-296, 515-530, 488-493, 1 795, 1796), VH CDR3 (SEQ ID NOs: 57, 49, 221-296, 515-530, 488-493, 1795, 1796), V L CDR1 (SEQ ID NOs: 297 to 372, 531 to 546), and VL CDR2 (SEQ ID NOs: 373 to 422); VH CDR1 (SEQ ID NOs: Nos. 57, 49, 221-296, 515-530, 488-493, 1795, 1796), VH CDR3 (SEQ ID NOs. 57, 49, 2 21 to 296, 515 to 530, 488 to 493, 1795, 1796), VL CDR1 (SEQ ID NOs: 297 to 372, 531 to 546), and VL CDR3 (SEQ ID NOs: 423 to 479, 558 to 569); VH CDR1 (SEQ ID NOs: 57, 49, 221 to 296, 515 to 520) 530, 488-493, 1795, 1796), VH CDR3 (SEQ ID NOs: 57, 49, 221-296, 515-530, 488-493 , 1795, 1796), VL CDR2 (SEQ ID NOs: 373 to 422), and VL CDR3 (SEQ ID NOs: 423 to 479, 558 to 569 VH CDR2 (SEQ ID NOs: 132 to 220, 497 to 514), VH CDR3 (SEQ ID NOs: 57, 49, 221 to 296, 515 to 5 30, 488-493, 1795, 1796), VL CDR1 (SEQ ID NOs: 297-372, 531-546), and VL CDR2 (SEQ ID NOs: VH CDR2 (SEQ ID NOs: 132 to 220, 497 to 514), VH CDR3 (SEQ ID NOs: 57, 49, 221) ~296, 515~530, 488~493, 1795, 1796), VL CDR1 (SEQ ID NO: 297~372, 531~546), and and VL CDR3 (SEQ ID NOs: 423 to 479, 558 to 569); VH CDR2 (SEQ ID NOs: 132 to 220, 497 to 514), VH CDR3 (SEQ ID NOs: 132 to 220, 497 to 514), DR3 (SEQ ID NOs: 57, 49, 221-296, 515-530, 488-493, 1795, 1796), VL CDR2 (SEQ ID NO: 3 73 to 422), and VL CDR3 (SEQ ID NOs: 423 to 479, 558 to 569); VH CDR1 (SEQ ID NOs: 57, 49, 221 to 222) 296, 515-530, 488-493, 1795, 1796), VH CDR2 (SEQ ID NOs: 132-220, 497-514), VH CD R3 (SEQ ID NOs: 57, 49, 221-296, 515-530, 488-493, 1795, 1796), VL CDR1 (SEQ ID NO: 29 VL CDR2 (SEQ ID NOs: 373-422); VH CDR1 (SEQ ID NOs: 57, 49, 221-222) 96, 515-530, 488-493, 1795, 1796), VH CDR2 (SEQ ID NOs: 132-220, 497-514), VH CDR 3 (SEQ ID NOs: 57, 49, 221-296, 515-530, 488-493, 1795, 1796), VL CDR1 (SEQ ID NO: 297 VL CDR3 (SEQ ID NOs: 423-479, 558-569); VH CDR1 (SEQ ID NOs: 57, 4 9, 221-296, 515-530, 488-493, 1795, 1796), VH CDR2 (SEQ ID NOs: 132-220, 497-514 ), VH CDR3 (SEQ ID NOs: 57, 49, 221-296, 515-530, 488-493, 1795, 1796), VL CDR2 (SEQ ID NOs: 57, 49, 221-296, 515-530, 488-493, 1795, 1796), VL CDR3 (SEQ ID NOs: 423-479, 558-569); VH CDR1 (SEQ ID NOs: 57, 49 , 221 to 296, 515 to 530, 488 to 493, 1795, 1796), VH CDR2 (SEQ ID NOs: 132 to 220, 497 to 514) VL CDR1 (SEQ ID NOs: 297 to 372, 531 to 546), VL CDR2 (SEQ ID NOs: 373 to 422), and VL CDR3 (SEQ ID NOs: 374 to 376). VH CDR1 (SEQ ID NOs: 57, 49, 221-296, 515-530, 488-493) , 1795, 1796), VH CDR3 (SEQ ID NOs: 57, 49, 221-296, 515-530, 488-493, 1795, 1796) VL CDR1 (SEQ ID NOs: 297 to 372, 531 to 546), VL CDR2 (SEQ ID NOs: 373 to 422), and VL CDR3 (SEQ ID NOs: 374 to 376). VH CDR2 (SEQ ID NOs: 132-220, 497-514), VH CDR3 (SEQ ID NOs: 132-220, 497-514), VH CDR4 (SEQ ID NOs: 132-220, 497-514), VH CDR5 (SEQ ID NOs: 132-220, 497-514), VH CDR6 (S 57, 49, 221-296, 515-530, 488-493, 1795, 1796), VL CDR1 (SEQ ID NOs: 297-372, 531 to 546), VL CDR2 (SEQ ID NOs: 373 to 422), and VL CDR3 (SEQ ID NOs: 423 to 479, 558 to 569); or Any of the VH CDRs (SEQ ID NOs: 41 to 296) and VL CDRs (SEQ ID NOs: 297 to 477) listed in Tables 1 to 24 It is an antibody containing a combination.

[0291] In certain embodiments, the antibodies or fragments thereof described herein are derived from humanized frameworks. In one embodiment, the antibodies or fragments thereof described herein comprise a fragment sequence of a target gene. The fragment comprises a VH region comprising the VH FR1, VH FR2, VH FR3, and VH FR4 amino acid sequences shown in Table 25; and / or (b) comprising the VL FR1, VL FR2, VL FR3, and VL FR4 amino acid sequences set forth in Table 25. Contains the VL region. Table 25: Exemplary framework sequences of humanized anti-GFRAL antibodies [Table 26] TIFF0007610541000066.tif244170TIFF0007610541000067.tif244170TIFF0007610541000068.tif245170TIFF0007610541000069.tif246170TIFF0007610541000070.tif245170TIFF0007610541000071.tif245170TIFF0007610541000072.tif245170TIFF0007610541000073.tif245170TIFF0007610541000074.tif246170TIFF0007610541000075.tif245170TIFF0007610541000076.tif245170TIFF0007610541000077.tif246170TIFF0007610541000078.tif245170TIFF0007610541000079.tif246170TIFF0007610541000080.tif245170TIFF0007610541000081.tif245170TIFF0007610541000082.tif245170TIFF0007610541000083.tif245170TIFF0007610541000084.tif245170TIFF0007610541000085.tif245170TIFF0007610541000086.tif245170TIFF0007610541000087.tif245170TIFF0007610541000088.tif246170TIFF0007610541000089.tif245170TIFF0007610541000090.tif245170TIFF0007610541000091.tif245170TIFF0007610541000092.tif246170TIFF0007610541000093.tif246170TIFF0007610541000094.tif246170TIFF0007610541000095.tif247170TIFF0007610541000096.tif246170TIFF0007610541000097.tif245170TIFF0007610541000098.tif245170TIFF0007610541000099.tif242170TIFF0007610541000100.tif246170TIFF0007610541000101.tif24 6170TIFF0007610541000102.tif245170TIFF0007610541000103.tif248170TIFF0007610541000104.tif245170.

[0292] In some embodiments, the antibody or fragment thereof described herein is selected from the group consisting of: (1) SEQ ID NOs: 570-5 (2) a VH FR1 having an amino acid sequence selected from SEQ ID NOs: 579 to 602; (3) VH FR2 having an amino acid sequence selected from SEQ ID NOs: 603 to 1726; and / or (4) a VH FR4 having an amino acid sequence selected from SEQ ID NOs: 1727 to 1728. Thus, in some embodiments, the humanized antibody comprises a VH region of SEQ ID NOs: 570-578. In some embodiments, the VH region comprises a VH FR1 having an amino acid sequence selected from the group consisting of: The humanized antibody comprises a VH FR2 having an amino acid sequence selected from SEQ ID NOs: 579 to 602. In some embodiments, the humanized antibody comprises a VH region selected from SEQ ID NOs: 603-1726. In some embodiments, the humanized VH region comprises a VH FR3 having an amino acid sequence The antibody has a VH region including a VH FR4 having an amino acid sequence selected from SEQ ID NOs: 1727 to 1728. include.

[0293] In some embodiments, the antibody or fragment thereof described herein is selected from the group consisting of: (1) SEQ ID NO: 1729 (2) a VL FR1 having an amino acid sequence selected from SEQ ID NOs: 1751 to 1777; (3) VL FR2 having an amino acid sequence selected from SEQ ID NOs: 1778 to 1792; and / or (4) a VL FR3 having an amino acid sequence selected from SEQ ID NOs: 1793 to 1794. Thus, in some embodiments, the humanized antibody comprises a VL region comprising SEQ ID NO: 17 In some embodiments, the VL region includes a VL FR1 having an amino acid sequence selected from the group consisting of 29 to 1750. In such a case, the humanized antibody has a VL having an amino acid sequence selected from SEQ ID NOs: 1751 to 1777. In some embodiments, the humanized antibody comprises a VL region comprising FR2. In some embodiments, the VL region comprises a VL FR3 having an amino acid sequence selected from the group consisting of: The humanized antibody comprises a VL FR4 having an amino acid sequence selected from SEQ ID NOs: 1793 to 1794. The VL region includes

[0294] In certain embodiments, the antibodies or fragments thereof described herein comprise a VH region and a VL region. wherein the VH region comprises: (1) an amino acid sequence selected from SEQ ID NOs: 570 to 578; VH FR1; (2) VH FR2 having an amino acid sequence selected from SEQ ID NOs: 579 to 602; (3) SEQ ID NO: 60 (3) a VH FR3 having an amino acid sequence selected from SEQ ID NOs: 1727 to 1728; and / or (4) a VH FR3 having an amino acid sequence selected from SEQ ID NOs: 1729 to 1729. and the VL region further comprises a VH FR4 having an amino acid sequence of: (1) SEQ ID NOs: 1729-1750 (2) a VL FR1 having an amino acid sequence selected from SEQ ID NOs: 1751 to 1777; (3) VL FR2 having an amino acid sequence selected from SEQ ID NOs: 1778 to 1792; and / or (4) further comprising a VL FR4 having an amino acid sequence selected from SEQ ID NOs: 1793-1794. .

[0295] Also provided herein are compounds containing one or more (e.g., one, two, three, or A VH FR is an antibody comprising one or more (e.g., one, two, three, or four) VL FRs. In particular, provided herein are: VH FR1 (SEQ ID NOs: 570-578) and VL FR1 (SEQ ID NO: 1729) VH FR1 (SEQ ID NOs: 570 to 578) and VL FR2 (SEQ ID NOs: 1751 to 1777); VH FR1 (SEQ ID NOs: 570-578) and VL FR3 (SEQ ID NOs: 1778-1792); VH FR1 (SEQ ID NOs: 570-578) and VL FR4 (SEQ ID NOs: VH FR2 (SEQ ID NOs: 579-602) and VL FR1 (SEQ ID NOs: 1729-1750); VH FR2 ( VH FR2 (SEQ ID NOs: 579 to 602) and VL FR2 (SEQ ID NOs: 1751 to 1777); VH FR2 (SEQ ID NOs: 579 to 602) and VL FR3 (SEQ ID NOs: 1778 to 1792); VH FR2 (SEQ ID NOs: 579 to 602) and VL FR4 (SEQ ID NOs: 1793 to 1794); VH FR3 (SEQ ID NOs: 603 to 1726) and VL FR1 (SEQ ID NOs: 1729 to 1750); VH FR3 (SEQ ID NOs: 603 to 17 26) and VL FR2 (SEQ ID NOs: 1751 to 1777); VH FR3 (SEQ ID NOs: 603 to 1726) and VL FR3 (SEQ ID NO: 1 VH FR3 (SEQ ID NOs: 603 to 1726) and VL FR4 (SEQ ID NOs: 1793 to 1794); VH FR4 (SEQ ID NOs: VH FR4 (SEQ ID NOs: 1727-1728) and VL FR1 (SEQ ID NOs: 1729-1750); VH FR4 (SEQ ID NOs: 1727-1728) and VL FR2 (SEQ ID NOs: 1751 to 1777); VH FR4 (SEQ ID NOs: 1727 to 1728) and VL FR3 (SEQ ID NOs: 1778 to 1792) VH FR4 (SEQ ID NOs: 1727-1728) and VL FR4 (SEQ ID NOs: 1793-1794); VH FR1 (SEQ ID NO: 570 VH FR2 (SEQ ID NOs: 579 to 602), and VL FR1 (SEQ ID NOs: 1729 to 1750); VH FR1 (SEQ ID NOs: VH FR1 (SEQ ID NOs: 570 to 578), VH FR2 (SEQ ID NOs: 579 to 602), and VL FR2 (SEQ ID NOs: 1751 to 1777); VH FR1 (SEQ ID NOs: 1751 to 1777); VH FR2 (SEQ ID NOs: 579-602), and VL FR3 (SEQ ID NOs: 1778-1792); VH FR 1 (SEQ ID NOs: 570 to 578), VH FR2 (SEQ ID NOs: 579 to 602), and VL FR4 (SEQ ID NOs: 1793 to 1794); V VH FR2 (SEQ ID NOs: 579 to 602), VH FR3 (SEQ ID NOs: 603 to 1726), and VL FR1 (SEQ ID NOs: 1729 to 175 0); VH FR2 (SEQ ID NOs: 579 to 602), VH FR3 (SEQ ID NOs: 603 to 1726), and VL FR2 (SEQ ID NO: 1751 VH FR2 (SEQ ID NOs: 579 to 602), VH FR3 (SEQ ID NOs: 603 to 1726), and VL FR3 (SEQ ID NOs: Nos. 1778 to 1792); VH FR2 (SEQ ID NOs. 579 to 602), VH FR3 (SEQ ID NOs. 603 to 1726), and VL FR4 ( SEQ ID NOs: 1793 to 1794); VH FR1 (SEQ ID NOs: 570 to 578), VL FR1 (SEQ ID NOs: 1729 to 1750), and V L FR2 (SEQ ID NOs: 1751 to 1777); VH FR1 (SEQ ID NOs: 570 to 578), VL FR1 (SEQ ID NOs: 1729 to 1750) , and VL FR3 (SEQ ID NOs: 1778 to 1792); VH FR1 (SEQ ID NOs: 570 to 578), VL FR1 (SEQ ID NO: 1729 VH FR1 (SEQ ID NOs: 570 to 578), VL FR2 (SEQ ID NOs: 570 to 578), and VL FR4 (SEQ ID NOs: 1793 to 1794). VH FR1 (SEQ ID NOs: 570-578), VL FR2 (SEQ ID NOs: 1751-1777), and VL FR3 (SEQ ID NOs: 1778-1792); VH FR1 (SEQ ID NOs: 570-578), VL FR2 ( VL FR4 (SEQ ID NOs: 1751 to 1777), and VH FR2 (SEQ ID NOs: 579 to 602), V VL FR1 (SEQ ID NOs: 1729 to 1750), and VL FR2 (SEQ ID NOs: 1751 to 1777); VH FR2 (SEQ ID NOs: 579 to 6 02), VL FR1 (SEQ ID NOs: 1729 to 1750), and VL FR3 (SEQ ID NOs: 1778 to 1792); VH FR2 (SEQ ID NOs: VL FR1 (SEQ ID NOs: 1729-1750), and VL FR4 (SEQ ID NOs: 1793-1794); VH FR2 (SEQ ID NOs: 1794-1795); VL FR2 (SEQ ID NOs: 1751 to 1777), and VL FR3 (SEQ ID NOs: 1778 to 1792); VH FR2 (SEQ ID NOs: 579 to 602), VL FR2 (SEQ ID NOs: 1751 to 1777), and VL FR4 (SEQ ID NOs: 1793 to 1794) VH FR3 (SEQ ID NOs: 603 to 1726), VL FR1 (SEQ ID NOs: 1729 to 1750), and VL FR2 (SEQ ID NO: 175 VH FR3 (SEQ ID NOs: 603 to 1726), VL FR1 (SEQ ID NOs: 1729 to 1750), and VL FR3 (SEQ ID NOs: VH FR3 (SEQ ID NOs: 603 to 1726), VL FR1 (SEQ ID NOs: 1729 to 1750), and VL F R4 (SEQ ID NOs: 1793 to 1794); VH FR3 (SEQ ID NOs: 603 to 1726), VL FR2 (SEQ ID NOs: 1751 to 1777), and VL FR3 (SEQ ID NOs: 1778 to 1792); VH FR3 (SEQ ID NOs: 603 to 1726), VL FR2 (SEQ ID NOs: 1751 to 1759), 1777), and VL FR4 (SEQ ID NOs: 1793 to 1794); VH FR4 (SEQ ID NOs: 1727 to 1728), VL FR1 (SEQ ID NOs: VH FR4 (SEQ ID NOs. 1727-1728), VL FR5 (SEQ ID NOs. 1729-1750), and VL FR2 (SEQ ID NOs. 1751-1777); VH FR4 (SEQ ID NOs. 1727-1728), VL FR 1 (SEQ ID NOs: 1729 to 1750), and VL FR3 (SEQ ID NOs: 1778 to 1792); VH FR4 (SEQ ID NOs: 1727 to 1728 ), VL FR1 (SEQ ID NOs: 1729 to 1750), and VL FR4 (SEQ ID NOs: 1793 to 1794); VH FR4 (SEQ ID NO: 17 VL FR2 (SEQ ID NOs: 1751 to 1777), and VL FR3 (SEQ ID NOs: 1778 to 1792); VH FR4 (SEQ ID NOs: 1751 to 1777); VL FR2 (SEQ ID NOs: 1751-1777), and VL FR4 (SEQ ID NOs: 1793-1794); V VH FR1 (SEQ ID NOs: 570 to 578), VH FR2 (SEQ ID NOs: 579 to 602), VH FR3 (SEQ ID NOs: 603 to 1726), and and VL FR1 (SEQ ID NOs: 1729 to 1750); VH FR1 (SEQ ID NOs: 570 to 578), VH FR2 (SEQ ID NOs: 579 to 602) , VH FR3 (SEQ ID NOs: 603 to 1726), and VL FR2 (SEQ ID NOs: 1751 to 1777); VH FR1 (SEQ ID NO: 570 to 578), VH FR2 (SEQ ID NOs: 579 to 602), VH FR3 (SEQ ID NOs: 603 to 1726), and VL FR3 (SEQ ID NOs: 1778 to 1792); VH FR1 (SEQ ID NOs: 570 to 578), VH FR2 (SEQ ID NOs: 579 to 602), VH FR3 (SEQ ID NOs: VH FR1 (SEQ ID NOs: 570-578), VH FR2 (SEQ ID NOs: 603-1726), and VL FR4 (SEQ ID NOs: 1793-1794); VH FR4 (SEQ ID NOs: 1727-1728), and VL FR1 (SEQ ID NOs: 1729-1750); VH FR1 (SEQ ID NOs: 570 to 578), VH FR2 (SEQ ID NOs: 579 to 602), VH FR4 (SEQ ID NOs: 1727 to 1728), and and VL FR2 (SEQ ID NOs: 1751 to 1777); VH FR1 (SEQ ID NOs: 570 to 578), VH FR2 (SEQ ID NOs: 579 to 602) , VH FR4 (SEQ ID NOs: 1727 to 1728), and VL FR3 (SEQ ID NOs: 1778 to 1792); VH FR1 (SEQ ID NO: 570 to 578), VH FR2 (SEQ ID NOs: 579 to 602), VH FR4 (SEQ ID NOs: 1727 to 1728), and VL FR4 (SEQ ID NOs: Nos. 1793 to 1794); VH FR1 (SEQ ID NOs. 570 to 578), VH FR3 (SEQ ID NOs. 603 to 1726), VH FR4 (SEQ ID NOs. VL FR1 (SEQ ID NOs: 1727-1728), and VL FR1 (SEQ ID NOs: 1729-1750); VH FR1 (SEQ ID NOs: 570-578), VH FR 3 (SEQ ID NOs: 603 to 1726), VH FR4 (SEQ ID NOs: 1727 to 1728), and VL FR2 (SEQ ID NOs: 1751 to 1777) VH FR1 (SEQ ID NOs: 570 to 578), VH FR3 (SEQ ID NOs: 603 to 1726), VH FR4 (SEQ ID NOs: 1727 to 1728) ), and VL FR3 (SEQ ID NOs: 1778 to 1792); VH FR1 (SEQ ID NOs: 570 to 578), VH FR3 (SEQ ID NO: 603 VH FR2 (sequence numbers 1726-1727), VH FR4 (sequence numbers 1727-1728), and VL FR4 (sequence numbers 1793-1794); VH FR2 (sequence numbers 1794-1795); VH FR3 (SEQ ID NOs: 603 to 1726), VH FR4 (SEQ ID NOs: 1727 to 1728), and VL FR1 (SEQ ID NOs: 1729 to 1750); VH FR2 (SEQ ID NOs: 579 to 602), VH FR3 (SEQ ID NOs: 603 to 1726), VH FR 4 (SEQ ID NOs: 1727 to 1728), and VL FR2 (SEQ ID NOs: 1751 to 1777); VH FR2 (SEQ ID NOs: 579 to 602) , VH FR3 (SEQ ID NOs: 603 to 1726), VH FR4 (SEQ ID NOs: 1727 to 1728), and VL FR3 (SEQ ID NO: 1778 VH FR2 (SEQ ID NOs: 579 to 602), VH FR3 (SEQ ID NOs: 603 to 1726), VH FR4 (SEQ ID NO: 172 VH FR1 (SEQ ID NOs: 570 to 578), VH FR2 (SEQ ID NOs: 570 to 578), and VL FR4 (SEQ ID NOs: 1793 to 1794). VL FR1 (SEQ ID NOs: 1729-1750), and VL FR2 (SEQ ID NOs: 1751-1777); VH FR 1 (SEQ ID NOs: 570 to 578), VH FR2 (SEQ ID NOs: 579 to 602), VL FR1 (SEQ ID NOs: 1729 to 1750), and V L FR3 (SEQ ID NOs: 1778 to 1792); VH FR1 (SEQ ID NOs: 570 to 578), VH FR2 (SEQ ID NOs: 579 to 602), V VL FR1 (SEQ ID NOs: 1729 to 1750), and VL FR4 (SEQ ID NOs: 1793 to 1794); VH FR1 (SEQ ID NOs: 570 to 575 78), VH FR3 (SEQ ID NOs: 603 to 1726), VL FR1 (SEQ ID NOs: 1729 to 1750), and VL FR2 (SEQ ID NO: 1 VH FR1 (SEQ ID NOs: 570 to 578), VH FR3 (SEQ ID NOs: 603 to 1726), VL FR1 (SEQ ID NOs: 751 to 1777); VH FR1 (SEQ ID NOs: 570-578), VH FR3 (SEQ ID NOs: 1729-1750), and VL FR3 (SEQ ID NOs: 1778-1792); VL FR1 (SEQ ID NOs: 1729-1750), and VL FR4 (SEQ ID NOs: 1793-1794); VH FR1 (SEQ ID NOs: 570 to 578), VH FR4 (SEQ ID NOs: 1727 to 1728), VL FR1 (SEQ ID NOs: 1729 to 1750), and VL FR2 (SEQ ID NOs: 1751 to 1777); VH FR1 (SEQ ID NOs: 570 to 578), VH FR4 (SEQ ID NOs: 1727 to 1729) 728), VL FR1 (SEQ ID NOs: 1729 to 1750), and VL FR3 (SEQ ID NOs: 1778 to 1792); VH FR1 (SEQ ID NOs: Nos. 570 to 578), VH FR4 (SEQ ID NOs. 1727 to 1728), VL FR1 (SEQ ID NOs. 1729 to 1750), and VL FR4 ( SEQ ID NOs: 1793 to 1794); VH FR2 (SEQ ID NOs: 579 to 602), VH FR3 (SEQ ID NOs: 603 to 1726), VL FR1 (SEQ ID NOs: 1729 to 1750), and VL FR2 (SEQ ID NOs: 1751 to 1777); VH FR2 (SEQ ID NOs: 579 to 602), VH FR3 (SEQ ID NOs: 603 to 1726), VL FR1 (SEQ ID NOs: 1729 to 1750), and VL FR3 (SEQ ID NOs: 1778 to 1780). 1792); VH FR2 (SEQ ID NOs: 579 to 602), VH FR3 (SEQ ID NOs: 603 to 1726), VL FR1 (SEQ ID NO: 1729 VH FR2 (SEQ ID NOs: 579 to 602), VH FR4 (SEQ ID NOs: 1793 to 1794), and VL FR4 (SEQ ID NOs: 1794 to 1750). VL FR1 (SEQ ID NOs: 1727-1728), VL FR1 (SEQ ID NOs: 1729-1750), and VL FR2 (SEQ ID NOs: 1751-1777); VH FR 2 (SEQ ID NOs: 579 to 602), VH FR4 (SEQ ID NOs: 1727 to 1728), VL FR1 (SEQ ID NOs: 1729 to 1750), and and VL FR3 (SEQ ID NOs: 1778 to 1792); VH FR2 (SEQ ID NOs: 579 to 602), VH FR4 (SEQ ID NOs: 1727 to 1728) 8), VL FR1 (SEQ ID NOs: 1729 to 1750), and VL FR4 (SEQ ID NOs: 1793 to 1794); VH FR3 (SEQ ID NO: 6 VH FR4 (SEQ ID NOs: 1727-1728), VL FR1 (SEQ ID NOs: 1729-1750), and VL FR2 (SEQ ID NOs: 1730-1732). Sequence numbers 1751 to 1777); VH FR3 (SEQ ID NOs: 603 to 1726), VH FR4 (SEQ ID NOs: 1727 to 1728), VL FR1 (SEQ ID NOs: 1729 to 1750), and VL FR3 (SEQ ID NOs: 1778 to 1792); VH FR3 (SEQ ID NOs: 603 to 1726) , VH FR4 (SEQ ID NOs: 1727 to 1728), VL FR1 (SEQ ID NOs: 1729 to 1750), and VL FR4 (SEQ ID NO: 179 VH FR1 (SEQ ID NOs: 570 to 578), VH FR2 (SEQ ID NOs: 579 to 602), VL FR2 (SEQ ID NOs: 175 VH FR1 (SEQ ID NOs: 570 to 578), VH FR2 (SEQ ID NOs: 571 to 573), and VL FR3 (SEQ ID NOs: 1778 to 1792); VH FR1 (SEQ ID NOs: 570 to 578), VH FR2 (SEQ ID NOs: 571 to 573), and VL FR3 (SEQ ID NOs: VL FR2 (SEQ ID NOs: 1751 to 1777), and VL FR4 (SEQ ID NOs: 1793 to 1794); VH FR 1 (SEQ ID NOs: 570 to 578), VH FR3 (SEQ ID NOs: 603 to 1726), VL FR2 (SEQ ID NOs: 1751 to 1777), and VL FR3 (SEQ ID NOs: 1778 to 1792); VH FR1 (SEQ ID NOs: 570 to 578), VH FR3 (SEQ ID NOs: 603 to 1726) , VL FR2 (SEQ ID NOs: 1751 to 1777), and VL FR4 (SEQ ID NOs: 1793 to 1794); VH FR1 (SEQ ID NO: 570 to 578), VH FR4 (SEQ ID NOs: 1727 to 1728), VL FR2 (SEQ ID NOs: 1751 to 1777), and VL FR3 (SEQ ID NOs: VH FR1 (SEQ ID NOs: 570 to 578), VH FR4 (SEQ ID NOs: 1727 to 1728), VL FR2 (SEQ ID NOs: 1730 to 1731), VL FR4 (SEQ ID NOs: 1793-1794); VH FR2 (SEQ ID NOs: 579-602); VH FR3 (SEQ ID NOs: 603 to 1726), VL FR2 (SEQ ID NOs: 1751 to 1777), and VL FR3 (SEQ ID NOs: 1778 to 179 2); VH FR2 (SEQ ID NOs: 579 to 602), VH FR3 (SEQ ID NOs: 603 to 1726), VL FR2 (SEQ ID NOs: 1751 to 1757) 77), and VL FR4 (SEQ ID NOs: 1793 to 1794); VH FR2 (SEQ ID NOs: 579 to 602), VH FR4 (SEQ ID NO: 17 VL FR2 (SEQ ID NOs: 1751 to 1777), and VL FR3 (SEQ ID NOs: 1778 to 1792); VH FR2 (SEQ ID NOs: 1751 to 1777), and VL FR3 (SEQ ID NOs: 1778 to 1792); VH FR4 (SEQ ID NOs: 1727-1728), VL FR2 (SEQ ID NOs: 1751-1777), and VL FR4 (SEQ ID NOs: 1793 to 1794); VH FR3 (SEQ ID NOs: 603 to 1726), VH FR4 (SEQ ID NOs: 1727 to 1728), VL FR2 (SEQ ID NOs: 1751 to 1777), and VL FR3 (SEQ ID NOs: 1778 to 1792); VH FR3 (SEQ ID NOs: 603 to 609); 1726), VH FR4 (SEQ ID NOs: 1727 to 1728), VL FR2 (SEQ ID NOs: 1751 to 1777), and VL FR4 (SEQ ID NOs: Nos. 1793 to 1794); VH FR1 (SEQ ID NOs. 570 to 578), VH FR2 (SEQ ID NOs. 579 to 602), VL FR3 (SEQ ID NOs. VL FR4 (SEQ ID NOs: 1793-1794); VH FR1 (SEQ ID NOs: 570-578), VH FR3 ( VL FR3 (SEQ ID NOs: 603-1726), VL FR3 (SEQ ID NOs: 1778-1792), and VL FR4 (SEQ ID NOs: 1793-1794); VH FR1 (SEQ ID NOs: 570 to 578), VH FR4 (SEQ ID NOs: 1727 to 1728), VL FR3 (SEQ ID NOs: 1778 to 1792) and VL FR4 (SEQ ID NOs: 1793 to 1794); VH FR2 (SEQ ID NOs: 579 to 602), VH FR3 (SEQ ID NOs: 603 to 604), 1726), VL FR3 (SEQ ID NOs: 1778 to 1792), and VL FR4 (SEQ ID NOs: 1793 to 1794); VH FR2 (SEQ ID NOs: Nos. 579 to 602), VH FR4 (SEQ ID NOs. 1727 to 1728), VL FR3 (SEQ ID NOs. 1778 to 1792), and VL FR4 ( SEQ ID NOs: 1793 to 1794); VH FR3 (SEQ ID NOs: 603 to 1726), VH FR4 (SEQ ID NOs: 1727 to 1728), VL F R3 (SEQ ID NOs: 1778 to 1792), and VL FR4 (SEQ ID NOs: 1793 to 1794); VH FR1 (SEQ ID NOs: 570 to 578) , VL FR1 (SEQ ID NOs: 1729 to 1750), VL FR2 (SEQ ID NOs: 1751 to 1777), and VL FR3 (SEQ ID NO: 177 VH FR1 (SEQ ID NOs: 570 to 578), VL FR1 (SEQ ID NOs: 1729 to 1750), VL FR2 (SEQ ID NOs: 1 VH FR1 (SEQ ID NOs: 570-578), VL FR1 (SEQ ID NOs: 751-1777), and VL FR4 (SEQ ID NOs: 1793-1794); VH FR1 (SEQ ID NOs: 570-578), VL FR1 (SEQ ID NOs: 570-578), VL FR3 (SEQ ID NOs: 1778-1792), and VL FR4 (SEQ ID NOs: 1793-1794); V H FR1 (SEQ ID NOs: 570 to 578), VL FR2 (SEQ ID NOs: 1751 to 1777), VL FR3 (SEQ ID NOs: 1778 to 1792) , and VL FR4 (SEQ ID NOs: 1793 to 1794); VH FR2 (SEQ ID NOs: 579 to 602), VL FR1 (SEQ ID NO: 1729 VL FR2 (SEQ ID NOs: 1751 to 1777), and VL FR3 (SEQ ID NOs: 1778 to 1792); VH FR2 (SEQ ID NOs: 1751 to 1777), and VL FR3 (SEQ ID NOs: 1778 to 1792); VL FR1 (SEQ ID NOs: 1729 to 1750), VL FR2 (SEQ ID NOs: 1751 to 1777), and VL FR 4 (SEQ ID NOs: 1793 to 1794); VH FR2 (SEQ ID NOs: 579 to 602), VL FR1 (SEQ ID NOs: 1729 to 1750), VL VL FR3 (SEQ ID NOs: 1778 to 1792), and VH FR4 (SEQ ID NOs: 1793 to 1794); VH FR2 (SEQ ID NOs: 579 to 602) ), VL FR2 (SEQ ID NOs: 1751 to 1777) , VL FR3 (SEQ ID NOs: 1778 to 1792), and VL FR4 (SEQ ID NOs: 1793 to 1794); VH FR3 (SEQ ID NO: 603 to 1726), VL FR1 (SEQ ID NOs: 1729 to 1750), VL FR2 (SEQ ID NOs: 1751 to 1777), and VL FR3 (SEQ ID NOs: VH FR3 (SEQ ID NOs: 603 to 1726), VL FR1 (SEQ ID NOs: 1729 to 1750), VL FR2 ( VL FR4 (SEQ ID NOs: 1751 to 1777), and VH FR3 (SEQ ID NOs: 603 to 1726); VL FR1 (SEQ ID NOs: 1729 to 1750), VL FR3 (SEQ ID NOs: 1778 to 1792), and VL FR4 (SEQ ID NO: 1793 VH FR3 (SEQ ID NOs: 603 to 1726), VL FR2 (SEQ ID NOs: 1751 to 1777), VL FR3 (SEQ ID NO: 1 778 to 1792), and VL FR4 (SEQ ID NOs: 1793 to 1794); VH FR4 (SEQ ID NOs: 1727 to 1728), VL FR1 ( VL FR2 (SEQ ID NOs: 1729-1750), VL FR2 (SEQ ID NOs: 1751-1777), and VL FR3 (SEQ ID NOs: 1778-1792); VH FR4 (SEQ ID NOs: 1727 to 1728), VL FR1 (SEQ ID NOs: 1729 to 1750), VL FR2 (SEQ ID NOs: 1751 to 1752) 77), and VL FR4 (SEQ ID NOs: 1793 to 1794); VH FR4 (SEQ ID NOs: 1727 to 1728), VL FR1 (SEQ ID NOs: 1729 to 1750), VL FR3 (SEQ ID NOs: 1778 to 1792), and VL FR4 (SEQ ID NOs: 1793 to 1794); VH FR4 ( VL FR2 (SEQ ID NOs: 1751 to 1777), VL FR3 (SEQ ID NOs: 1778 to 1792), and and VL FR4 (SEQ ID NOs: 1793 to 1794); VH FR1 (SEQ ID NOs: 570 to 578), VH FR2 (SEQ ID NOs: 579 to 602) , VH FR3 (SEQ ID NOs: 603 to 1726), VH FR4 (SEQ ID NOs: 1727 to 1728), and VL FR1 (SEQ ID NO: 1729) VH FR1 (SEQ ID NOs: 570 to 578), VH FR2 (SEQ ID NOs: 579 to 602), VH FR3 (SEQ ID NO: 603) VH FR1 (sequence numbers 1751 to 1757); VH FR4 (sequence numbers 1727 to 1728); and VL FR2 (sequence numbers 1751 to 1777); VH FR1 (sequence numbers 1751 to 1757); VH FR2 (SEQ ID NOs: 570 to 578), VH FR3 (SEQ ID NOs: 603 to 1726), VH FR4 (SEQ ID NOs: 579 to 602), VH FR5 (SEQ ID NOs: 603 to 1726), VH FR6 (SEQ ID NOs: 579 to 602), VH FR7 (SEQ ID NOs: 57 VL FR3 (SEQ ID NOs: 1778-1792); VH FR1 (SEQ ID NOs: 570-578), VH FR 2 (SEQ ID NOs: 579 to 602), VH FR3 (SEQ ID NOs: 603 to 1726), VH FR4 (SEQ ID NOs: 1727 to 1728), and VL FR4 (SEQ ID NOs: 1793 to 1794); VH FR1 (SEQ ID NOs: 570 to 578), VH FR2 (SEQ ID NOs: 579 to 602), VH FR3 (SEQ ID NOs: 603 to 1726), VL FR1 (SEQ ID NOs: 1729 to 1750), and VL FR2 (SEQ ID NOs: 1751 to 1754). 1777); VH FR1 (SEQ ID NOs: 570 to 578), VH FR2 (SEQ ID NOs: 579 to 602), VH FR3 (SEQ ID NOs: 603 to 604), 726), VL FR1 (SEQ ID NOs: 1729 to 1750), and VL FR3 (SEQ ID NOs: 1778 to 1792); VH FR1 (SEQ ID NOs: Nos. 570 to 578), VH FR2 (SEQ ID NOs. 579 to 602), VH FR3 (SEQ ID NOs. 603 to 1726), VL FR1 (SEQ ID NOs. VL FR4 (SEQ ID NOs: 1729-1750), and VH FR1 (SEQ ID NOs: 570-578), VH FR2 ( VH FR3 (SEQ ID NOs: 579 to 602), VL FR2 (SEQ ID NOs: 1751 to 1777), and VL VH FR3 (SEQ ID NOs: 1778 to 1792); VH FR1 (SEQ ID NOs: 570 to 578), VH FR2 (SEQ ID NOs: 579 to 602), VH VL FR3 (SEQ ID NOs: 603 to 1726), VL FR2 (SEQ ID NOs: 1751 to 1777), and VL FR4 (SEQ ID NOs: 1793 to 1798). 94); VH FR1 (SEQ ID NOs: 570 to 578), VH FR2 (SEQ ID NOs: 579 to 602), VH FR3 (SEQ ID NOs: 603 to 172) 6), VL FR3 (SEQ ID NOs: 1778 to 1792), and VL FR4 (SEQ ID NOs: 1793 to 1794); VH FR1 (SEQ ID NO: 5 70 to 578), VH FR2 (SEQ ID NOs: 579 to 602), VH FR4 (SEQ ID NOs: 1727 to 1728), VL FR1 (SEQ ID NO: 1 VH FR1 (SEQ ID NOs: 570-578), VH FR2 (SEQ ID NOs: 729-1750), and VL FR2 (SEQ ID NOs: 1751-1777); VH FR4 (SEQ ID NOs: 1727 to 1728), VL FR1 (SEQ ID NOs: 1729 to 1750), and VL VH FR3 (SEQ ID NOs: 1778 to 1792); VH FR1 (SEQ ID NOs: 570 to 578), VH FR2 (SEQ ID NOs: 579 to 602), VH FR4 (SEQ ID NOs: 1727 to 1728), VL FR1 (SEQ ID NOs: 1729 to 1750), and VL FR4 (SEQ ID NOs: 1793 to 1794). 94); VH FR1 (SEQ ID NOs: 570 to 578), VH FR2 (SEQ ID NOs: 579 to 602), VH FR4 (SEQ ID NOs: 1727 to 1728) 28), VL FR2 (SEQ ID NOs: 1751 to 1777), and VL FR3 (SEQ ID NOs: 1778 to 1792); VH FR1 (SEQ ID NO: 570 to 578), VH FR2 (SEQ ID NOs: 579 to 602), VH FR4 (SEQ ID NOs: 1727 to 1728), VL FR2 (SEQ ID NOs: VH FR1 (SEQ ID NOs: 570-578), VH FR2 (SEQ ID NOs: 1751-1777), and VL FR4 (SEQ ID NOs: 1793-1794); VH FR1 (SEQ ID NOs: 570-578), VH FR2 (SEQ ID NOs: 1751-1777), and VL FR4 (SEQ ID NOs: 1 VH FR4 (SEQ ID NOs: 1727-1728), VL FR3 (SEQ ID NOs: 1778-1792), and VL FR4 (SEQ ID NOs: 1793 to 1794); VH FR2 (SEQ ID NOs: 579 to 602), VH FR3 (SEQ ID NOs: 603 to 1726), VH FR4 (SEQ ID NOs: 1727 to 1728), VL FR1 (SEQ ID NOs: 1729 to 1750), and VL FR2 (SEQ ID NOs: 1751 to 1752). VH FR2 (SEQ ID NOs: 579 to 602), VH FR3 (SEQ ID NOs: 603 to 1726), VH FR4 (SEQ ID NOs: 1727 to 1728), 1728), VL FR1 (SEQ ID NOs: 1729 to 1750), and VL FR3 (SEQ ID NOs: 1778 to 1792); VH FR2 (SEQ ID NOs: Nos. 579 to 602), VH FR3 (SEQ ID NOs. 603 to 1726), VH FR4 (SEQ ID NOs. 1727 to 1728), VL FR1 (SEQ ID NOs. VL FR4 (SEQ ID NOs: 1729-1750), and VH FR5 (SEQ ID NOs: 1793-1794); VH FR2 (SEQ ID NOs: 579-602), VH FR 3 (SEQ ID NOs: 603 to 1726), VH FR4 (SEQ ID NOs: 1727 to 1728), VL FR2 (SEQ ID NOs: 1751 to 1777), and and VL FR3 (SEQ ID NOs: 1778 to 1792); VH FR2 (SEQ ID NOs: 579 to 602), VH FR3 (SEQ ID NOs: 603 to 1726 ), VH FR4 (SEQ ID NOs: 1727 to 1728), VL FR2 (SEQ ID NOs: 1751 to 1777), and VL FR4 (SEQ ID NO: 17 VH FR2 (SEQ ID NOs: 579 to 602), VH FR3 (SEQ ID NOs: 603 to 1726), VH FR4 (SEQ ID NO: 1 727 to 1728), VL FR3 (SEQ ID NOs: 1778 to 1792), and VL FR4 (SEQ ID NOs: 1793 to 1794); VH FR1 ( SEQ ID NOs: 570 to 578), VH FR2 (SEQ ID NOs: 579 to 602), VL FR1 (SEQ ID NOs: 1729 to 1750), VL FR2 ( VL FR3 (SEQ ID NOs: 1751 to 1777), and VH FR1 (SEQ ID NOs: 570 to 578), V H FR2 (SEQ ID NOs: 579 to 602), VL FR1 (SEQ ID NOs: 1729 to 1750), VL FR2 (SEQ ID NOs: 1751 to 1777) and VL FR4 (SEQ ID NOs: 1793 to 1794); VH FR1 (SEQ ID NOs: 570 to 578), VH FR2 (SEQ ID NOs: 579 to 580), 602), VL FR1 (SEQ ID NOs: 1729 to 1750), VL FR3 (SEQ ID NOs: 1778 to 1792), and VL FR4 (SEQ ID NOs: Nos. 1793 to 1794); VH FR1 (SEQ ID NOs. 570 to 578), VH FR2 (SEQ ID NOs. 579 to 602), VL FR2 (SEQ ID NOs. VL FR3 (SEQ ID NOs: 1778-1792), and VL FR4 (SEQ ID NOs: 1793-1794); VH FR 1 (SEQ ID NOs: 570 to 578), VH FR3 (SEQ ID NOs: 603 to 1726), VL FR1 (SEQ ID NOs: 1729 to 1750), VL F R2 (SEQ ID NOs: 1751 to 1777), and VL FR3 (SEQ ID NOs: 1778 to 1792); VH FR1 (SEQ ID NOs: 570 to 578) , VH FR3 (SEQ ID NOs: 603 to 1726), VL FR1 (SEQ ID NOs: 1729 to 1750), VL FR2 (SEQ ID NOs: 1751 to 1752) 77), and VL FR4 (SEQ ID NOs: 1793 to 1794); VH FR1 (SEQ ID NOs: 570 to 578), VH FR3 (SEQ ID NO: 60 VL FR1 (SEQ ID NOs: 1729 to 1750), VL FR3 (SEQ ID NOs: 1778 to 1792), and VL FR4 (SEQ ID NOs: 1781 to 1784). Sequence numbers 1793 to 1794; VH FR1 (SEQ ID NOs: 570 to 578), VH FR3 (SEQ ID NOs: 603 to 1726), VL FR2 ( VL FR3 (SEQ ID NOs: 1751 to 1777), VL FR3 (SEQ ID NOs: 1778 to 1792), and VL FR4 (SEQ ID NOs: 1793 to 1794); VH FR1 (SEQ ID NOs: 570 to 578), VH FR4 (SEQ ID NOs: 1727 to 1728), VL FR1 (SEQ ID NOs: 1729 to 1750) ), VL FR2 (SEQ ID NOs: 1751 to 1777), and VL FR3 (SEQ ID NOs: 1778 to 1792); VH FR1 (SEQ ID NO: 57 0 to 578), VH FR4 (SEQ ID NOs: 1727 to 1728), VL FR1 (SEQ ID NOs: 1729 to 1750), VL FR2 (SEQ ID NOs: VH FR1 (SEQ ID NOs: 570-578), VH FR4 (SEQ ID NOs: 1751-1777), and VL FR4 (SEQ ID NOs: 1793-1794); VH FR1 (SEQ ID NOs: 570-578), VH FR4 (SEQ ID NOs: 1751-1777), and VL FR4 (SEQ ID NOs: 1 VL FR1 (SEQ ID NOs: 1729-1750), VL FR3 (SEQ ID NOs: 1778-1792), and V L FR4 (SEQ ID NOs: 1793 to 1794); VH FR1 (SEQ ID NOs: 570 to 578), VH FR4 (SEQ ID NOs: 1727 to 1728) , VL FR2 (SEQ ID NOs: 1751 to 1777), VL FR3 (SEQ ID NOs: 1778 to 1792), and VL FR4 (SEQ ID NO: 179 VH FR2 (SEQ ID NOs: 579 to 602), VH FR3 (SEQ ID NOs: 603 to 1726), VL FR1 (SEQ ID NO: 17 29 to 1750), VL FR2 (SEQ ID NOs: 1751 to 1777), and VL FR3 (SEQ ID NOs: 1778 to 1792); VH FR2 (SEQ ID NOs: 1779 to 1792); VH FR3 (SEQ ID NOs: 603 to 1726), VL FR1 (SEQ ID NOs: 1729 to 1750), VL FR2 ( VL FR4 (SEQ ID NOs: 1751 to 1777), and VH FR2 (SEQ ID NOs: 579 to 602), V H FR3 (SEQ ID NOs: 603 to 1726), VL FR1 (SEQ ID NOs: 1729 to 1750), VL FR3 (SEQ ID NOs: 1778 to 1792) and VL FR4 (SEQ ID NOs: 1793 to 1794); VH FR2 (SEQ ID NOs: 579 to 602), VH FR3 (SEQ ID NOs: 603 to 604), 1726), VL FR2 (SEQ ID NOs: 1751 to 1777), VL FR3 (SEQ ID NOs: 1778 to 1792), and VL FR4 (SEQ ID NOs: Nos. 1793-1794); VH FR2 (SEQ ID NOs. 579-602), VH FR4 (SEQ ID NOs. 1727-1728), VL FR1 (SEQ ID NOs. VL FR2 (SEQ ID NOs: 1729-1750), VL FR3 (SEQ ID NOs: 1778-1792), and VL FR4 (SEQ ID NOs: 1751-1777). FR2 (SEQ ID NOs: 579 to 602), VH FR4 (SEQ ID NOs: 1727 to 1728), VL FR1 (SEQ ID NOs: 1729 to 1750), V VL FR2 (SEQ ID NOs: 1751 to 1777), and VL FR4 (SEQ ID NOs: 1793 to 1794); VH FR2 (SEQ ID NOs: 579 to 6 02), VH FR4 (SEQ ID NOs: 1727 to 1728), VL FR1 (SEQ ID NOs: 1729 to 1750), VL FR3 (SEQ ID NO: 1778 VH FR2 (SEQ ID NOs: 579 to 602), VH FR4 (SEQ ID NOs: 579 to 602), and VL FR4 (SEQ ID NOs: 1793 to 1794). VL FR2 (SEQ ID NOs: 1751 to 1777), VL FR3 (SEQ ID NOs: 1778 to 1792), and VL FR 4 (SEQ ID NOs: 1793 to 1794); VH FR3 (SEQ ID NOs: 603 to 1726), VH FR4 (SEQ ID NOs: 1727 to 1728), VL VL FR1 (SEQ ID NOs: 1729 to 1750), VL FR2 (SEQ ID NOs: 1751 to 1777), and VL FR3 (SEQ ID NOs: 1778 to 1779). 792); VH FR3 (SEQ ID NOs: 603 to 1726), VH FR4 (SEQ ID NOs: 1727 to 1728), VL FR1 (SEQ ID NO: 1729) VL FR2 (SEQ ID NOs: 1751 to 1777), and VL FR4 (SEQ ID NOs: 1793 to 1794); VH FR3 (SEQ ID NOs: VH FR4 (SEQ ID NOs: 1727-1728), VL FR1 (SEQ ID NOs: 1729-1750), VL FR3 ( VL FR4 (SEQ ID NOs: 1778 to 1792), and VH FR3 (SEQ ID NOs: 603 to 1726); VH FR4 (SEQ ID NOs: 1727 to 1728), VL FR2 (SEQ ID NOs: 1751 to 1777), VL FR3 (SEQ ID NOs: 1778 to 179 2), and VL FR4 (SEQ ID NOs: 1793 to 1794); VH FR1 (SEQ ID NOs: 570 to 578), VL FR1 (SEQ ID NO: 172 VL FR2 (SEQ ID NOs: 1751 to 1777), VL FR3 (SEQ ID NOs: 1778 to 1792), and VL FR4 (SEQ ID NOs: 1793 to 1794). Sequence numbers 1793 to 1794; VH FR2 (SEQ ID NOs: 579 to 602), VL FR1 (SEQ ID NOs: 1729 to 1750), VL FR2 ( VL FR3 (SEQ ID NOs: 1751 to 1777), VL FR3 (SEQ ID NOs: 1778 to 1792), and VL FR4 (SEQ ID NOs: 1793 to 1794); VH FR3 (SEQ ID NOs: 603 to 1726), VL FR1 (SEQ ID NOs: 1729 to 1750), VL FR2 (SEQ ID NOs: 1751 to 177 7), VL FR3 (SEQ ID NOs: 1778 to 1792), and VL FR4 (SEQ ID NOs: 1793 to 1794); VH FR4 (SEQ ID NO: 1 727 to 1728), VL FR1 (SEQ ID NOs: 1729 to 1750), VL FR2 (SEQ ID NOs: 1751 to 1777), VL FR3 (SEQ ID NOs: VL FR4 (SEQ ID NOs: 1778-1792), and VL FR4 (SEQ ID NOs: 1793-1794); VH FR1 (SEQ ID NOs: 570-578), VH FR 2 (SEQ ID NOs: 579 to 602), VH FR3 (SEQ ID NOs: 603 to 1726), VH FR4 (SEQ ID NOs: 1727 to 1728), VL F R1 (SEQ ID NOs: 1729 to 1750), and VL FR2 (SEQ ID NOs: 1751 to 1777); VH FR1 (SEQ ID NOs: 570 to 578) , VH FR2 (SEQ ID NOs: 579 to 602), VH FR3 (SEQ ID NOs: 603 to 1726), VH FR4 (SEQ ID NOs: 1727 to 1728) ), VL FR1 (SEQ ID NOs: 1729 to 1750), and VL FR3 (SEQ ID NOs: 1778 to 1792); VH FR1 (SEQ ID NO: 57 0 to 578), VH FR2 (SEQ ID NOs: 579 to 602), VH FR3 (SEQ ID NOs: 603 to 1726), VH FR4 (SEQ ID NO: 172 VL FR1 (SEQ ID NOs: 1729-1750), and VL FR4 (SEQ ID NOs: 1793-1794); VH FR1 (SEQ ID NOs: 1729-1750), and VL FR4 (SEQ ID NOs: 1793-1794); VH FR2 (SEQ ID NOs: 579 to 602), VH FR3 (SEQ ID NOs: 603 to 1726), VH FR4 (SEQ ID NOs: 570 to 578), VH FR5 (SEQ ID NOs: 579 to 602), VH FR6 (SEQ ID NOs: 603 to 1726), VH FR7 (SEQ ID NOs: 57 VL FR2 (SEQ ID NOs: 1751-1777), and VL FR3 (SEQ ID NOs: 1778-1792); V VH FR1 (SEQ ID NOs: 570 to 578), VH FR2 (SEQ ID NOs: 579 to 602), VH FR3 (SEQ ID NOs: 603 to 1726), VH FR4 (SEQ ID NOs: 1727 to 1728), VL FR2 (SEQ ID NOs: 1751 to 1777), and VL FR4 (SEQ ID NOs: 1793 to 1798). 794); VH FR1 (SEQ ID NOs: 570 to 578), VH FR2 (SEQ ID NOs: 579 to 602), VH FR3 (SEQ ID NOs: 603 to 607) 26), VH FR4 (SEQ ID NOs: 1727 to 1728), VL FR3 (SEQ ID NOs: 1778 to 1792), and VL FR4 (SEQ ID NOs: 1793 to 1794); VH FR1 (SEQ ID NOs: 570 to 578), VH FR2 (SEQ ID NOs: 579 to 602), VH FR3 (SEQ ID NOs: 603 to 1726), VL FR1 (SEQ ID NOs: 1729 to 1750), VL FR2 (SEQ ID NOs: 1751 to 1777), and VL FR3 ( SEQ ID NOs: 1778 to 1792); VH FR1 (SEQ ID NOs: 570 to 578), VH FR2 (SEQ ID NOs: 579 to 602), VH FR3 ( VL FR1 (SEQ ID NOs: 1729 to 1750), VL FR2 (SEQ ID NOs: 1751 to 1777), and VL FR4 (SEQ ID NOs: 1793 to 1794); VH FR1 (SEQ ID NOs: 570 to 578), VH FR2 (SEQ ID NOs: 579 to 602), VH FR3 (SEQ ID NOs: 603 to 1726), VL FR1 (SEQ ID NOs: 1729 to 1750), VL FR3 (SEQ ID NOs: 1778 to 1792 ), and VL FR4 (SEQ ID NOs: 1793 to 1794); VH FR1 (SEQ ID NOs: 570 to 578), VH FR2 (SEQ ID NO: 579 to 602), VH FR3 (SEQ ID NOs: 603 to 1726), VL FR2 (SEQ ID NOs: 1751 to 1777), VL FR3 (SEQ ID NO: 17 VH FR1 (SEQ ID NOs: 570 to 578), VH FR2 (SEQ ID NOs: 570 to 579), and VL FR4 (SEQ ID NOs: 1793 to 1794). VH FR4 (SEQ ID NOs: 1727-1728), VL FR1 (SEQ ID NOs: 1729-1750), VL FR2 (SEQ ID NOs: 1730-1732), VL FR3 (SEQ ID NOs: 1733-1734), VL FR4 (SEQ ID NOs: 1734-1735), VL FR5 (SEQ ID NOs: VL FR3 (SEQ ID NOs: 1778-1792); VH FR1 (SEQ ID NOs: 570-578), VH FR2 (SEQ ID NOs: 579 to 602), VH FR4 (SEQ ID NOs: 1727 to 1728), VL FR1 (SEQ ID NOs: 1729 to 1750), V VL FR2 (SEQ ID NOs: 1751 to 1777), and VL FR4 (SEQ ID NOs: 1793 to 1794); VH FR1 (SEQ ID NOs: 570 to 575) 78), VH FR2 (SEQ ID NOs: 579 to 602), VH FR4 (SEQ ID NOs: 1727 to 1728), VL FR1 (SEQ ID NOs: 1729 to 1730), 1750), VL FR3 (SEQ ID NOs: 1778 to 1792), and VL FR4 (SEQ ID NOs: 1793 to 1794); VH FR1 (SEQ ID NOs: Nos. 570-578), VH FR2 (SEQ ID NO. Nos. 579 to 602), VH FR4 (SEQ ID NOs. 1727 to 1728), VL FR2 (SEQ ID NOs. 1751 to 1777), VL FR3 (SEQ ID NOs. VL FR4 (SEQ ID NOs: 1778-1792), and VL FR4 (SEQ ID NOs: 1793-1794); VH FR1 (SEQ ID NOs: 570-578), VH FR 3 (SEQ ID NOs: 603 to 1726), VH FR4 (SEQ ID NOs: 1727 to 1728), VL FR1 (SEQ ID NOs: 1729 to 1750), VL VL FR2 (SEQ ID NOs: 1751 to 1777), and VL FR3 (SEQ ID NOs: 1778 to 1792); VH FR1 (SEQ ID NOs: 570 to 57 8), VH FR3 (SEQ ID NOs: 603 to 1726), VH FR4 (SEQ ID NOs: 1727 to 1728), VL FR1 (SEQ ID NOs: 1729 to 1730), 1750), VL FR2 (SEQ ID NOs: 1751 to 1777), and VL FR4 (SEQ ID NOs: 1793 to 1794); VH FR1 (SEQ ID NOs: Nos. 570 to 578), VH FR3 (SEQ ID NOs. 603 to 1726), VH FR4 (SEQ ID NOs. 1727 to 1728), VL FR1 (SEQ ID NOs. VL FR3 (SEQ ID NOs: 1778-1792), and VL FR4 (SEQ ID NOs: 1793-1794); VH FR1 (SEQ ID NOs: 570 to 578), VH FR3 (SEQ ID NOs: 603 to 1726), VH FR4 (SEQ ID NOs: 1727 to 1728), VL VL FR2 (SEQ ID NOs: 1751 to 1777), VL FR3 (SEQ ID NOs: 1778 to 1792), and VL FR4 (SEQ ID NOs: 1793 to 1794). VH FR2 (SEQ ID NOs: 579 to 602), VH FR3 (SEQ ID NOs: 603 to 1726), VH FR4 (SEQ ID NOs: 1727 to 1729), 1728), VL FR1 (SEQ ID NOs: 1729 to 1750), VL FR2 (SEQ ID NOs: 1751 to 1777), and VL FR3 (SEQ ID NOs: Nos. 1778 to 1792); VH FR2 (SEQ ID NOs. 579 to 602), VH FR3 (SEQ ID NOs. 603 to 1726), VH FR4 (SEQ ID NOs. VL FR1 (SEQ ID NOs: 1727-1728), VL FR1 (SEQ ID NOs: 1729-1750), VL FR2 (SEQ ID NOs: 1751-1777), and VL FR4 (SEQ ID NOs: 1793 to 1794); VH FR2 (SEQ ID NOs: 579 to 602), VH FR3 (SEQ ID NOs: 603 to 1726), VH FR4 (SEQ ID NOs: 1727 to 1728), VL FR1 (SEQ ID NOs: 1729 to 1750), VL FR3 (SEQ ID NOs: 1778 to 1792) and VL FR4 (SEQ ID NOs: 1793 to 1794); VH FR2 (SEQ ID NOs: 579 to 602), VH FR3 (SEQ ID NOs: 603 to 604), 1726), VH FR4 (SEQ ID NOs: 1727 to 1728), VL FR2 (SEQ ID NOs: 1751 to 1777), VL FR3 (SEQ ID NO: 17 VH FR1 (SEQ ID NOs: 570 to 578), VH FR2 (SEQ ID NOs: 570 to 579), and VL FR4 (SEQ ID NOs: 1793 to 1794). VL FR1 (SEQ ID NOs: 1729 to 1750), VL FR2 (SEQ ID NOs: 1751 to 1777), VL FR3 (SEQ ID NOs: 1752 to 1753), VL FR4 (SEQ ID NOs: 1754 to 1755), VL FR5 (SEQ ID NOs: 1756 to 1757), VL FR6 (S VL FR4 (SEQ ID NOs: 1793-1794); VH FR1 (SEQ ID NOs: 570-578), VH FR3 (SEQ ID NOs: 603 to 1726), VL FR1 (SEQ ID NOs: 1729 to 1750), VL FR2 (SEQ ID NOs: 1751 to 1777), VL FR3 (SEQ ID NOs: 1778 to 1792), and VL FR4 (SEQ ID NOs: 1793 to 1794); VH FR1 (SEQ ID NOs: 570 to 5796); 578), VH FR4 (SEQ ID NOs: 1727 to 1728), VL FR1 (SEQ ID NOs: 1729 to 1750), VL FR2 (SEQ ID NO: 175 VL FR3 (SEQ ID NOs: 1778-1792), and VL FR4 (SEQ ID NOs: 1793-1794); VH FR2 (SEQ ID NOs: 1795-1796); VH FR3 (SEQ ID NOs: 603 to 1726), VL FR1 (SEQ ID NOs: 1729 to 1750), VL FR2 ( VL FR3 (SEQ ID NOs: 1751 to 1777), VL FR3 (SEQ ID NOs: 1778 to 1792), and VL FR4 (SEQ ID NOs: 1793 to 1794); VH FR2 (SEQ ID NOs: 579 to 602), VH FR4 (SEQ ID NOs: 1727 to 1728), VL FR1 (SEQ ID NOs: 1729 to 1750) ), VL FR2 (SEQ ID NOs: 1751 to 1777), VL FR3 (SEQ ID NOs: 1778 to 1792), and VL FR4 (SEQ ID NO: 17 VH FR3 (SEQ ID NOs: 603 to 1726), VH FR4 (SEQ ID NOs: 1727 to 1728), VL FR1 (SEQ ID NOs: VL FR2 (SEQ ID NOs: 1729-1750), VL FR3 (SEQ ID NOs: 1778-1792), and VL FR 4 (SEQ ID NOs: 1793 to 1794); VH FR1 (SEQ ID NOs: 570 to 578), VH FR2 (SEQ ID NOs: 579 to 602), VH FR 3 (SEQ ID NOs: 603 to 1726), VH FR4 (SEQ ID NOs: 1727 to 1728), VL FR1 (SEQ ID NOs: 1729 to 1750), VL VL FR2 (SEQ ID NOs: 1751 to 1777), and VL FR3 (SEQ ID NOs: 1778 to 1792); VH FR1 (SEQ ID NOs: 570 to 57 8), VH FR2 (SEQ ID NOs: 579 to 602), VH FR3 (SEQ ID NOs: 603 to 1726), VH FR4 (SEQ ID NOs: 1727 to 1728) 28), VL FR1 (SEQ ID NOs: 1729 to 1750), VL FR2 (SEQ ID NOs: 1751 to 1777), and VL FR4 (SEQ ID NO: 1793 to 1794); VH FR1 (SEQ ID NOs: 570 to 578), VH FR2 (SEQ ID NOs: 579 to 602), VH FR3 (SEQ ID NOs: 603 to 1726), VH FR4 (SEQ ID NOs: 1727 to 1728), VL FR2 (SEQ ID NOs: 1751 to 1777), VL FR3 (SEQ ID NOs: VL FR4 (SEQ ID NOs: 1778-1792), and VL FR4 (SEQ ID NOs: 1793-1794); VH FR1 (SEQ ID NOs: 570-578), VH FR 2 (SEQ ID NOs: 579 to 602), VH FR3 (SEQ ID NOs: 603 to 1726), VL FR1 (SEQ ID NOs: 1729 to 1750), VL F R2 (SEQ ID NOs: 1751 to 1777), VL FR3 (SEQ ID NOs: 1778 to 1792), and VL FR4 (SEQ ID NOs: 1793 to 1799) 4); VH FR1 (SEQ ID NOs: 570 to 578), VH FR2 (SEQ ID NOs: 579 to 602), VH FR4 (SEQ ID NOs: 1727 to 1728) 8), VL FR1 (SEQ ID NOs: 1729 to 1750), VL FR2 (SEQ ID NOs: 1751 to 1777), VL FR3 (SEQ ID NO: 1778 VH FR1 (SEQ ID NOs: 570 to 578), VH FR2 (SEQ ID NOs: 571 to 573), and VL FR4 (SEQ ID NOs: 1793 to 1794). Nos. 579 to 602), VH FR3 (SEQ ID NOs. 603 to 1726), VH FR4 (SEQ ID NOs. 1727 to 1728), VL FR1 (SEQ ID NOs. VL FR2 (SEQ ID NOs: 1729 to 1750), VL FR3 (SEQ ID NOs: 1778 to 1792), and VL FR4 (SEQ ID NOs: 1793 to 1794); or VH FR (SEQ ID NOs: 478 to 1636) and VL FR (SEQ ID NOs: 1795 to 1796) listed in Table 25 and any combination thereof (numbers 1637 to 1702).

[0296] In yet another embodiment, among the antibodies or functional fragments exemplified for binding to GFRAL, Antibodies that compete with one of the antibodies exemplified herein are provided. It may also bind to the same epitope as one of the exemplified antibodies, or to an overlapping epitope. Antibodies and fragments that compete with or bind to the same epitope as the antibody have similar functional characteristics. Exemplary antigen-binding proteins and fragments include those in Tables 1 to 24. These include those having the VH and VL regions and CDRs provided herein, including those having the VH and VL regions and CDRs. Thus, as specific examples, the antibodies provided include: (a) antibodies described in Tables 1 to 24; (b) one, two, three, four, five, or all six of the CDRs listed in Tables 1-24; or (c) a VH and VL selected from the VH and VL regions described for the antibody; or The antibody competes with an antibody containing two light chains and two heavy chains containing the designated VH and VL. This includes things that

[0297] In certain embodiments, the antibodies of the compositions and methods of using the antibodies provided herein includes the anti-GFRAL monoclonal antibodies described herein, for example in the Examples section. Any anti-GFRAL antibody can be used in any of the methods provided herein. In some embodiments of the various methods provided herein, the antibody comprises SEQ ID NO: The VH region comprises the amino acid sequence of SEQ ID NO: 1, or a humanized variant thereof. In some embodiments of the various methods, the antibody comprises a V In some embodiments of the various methods provided herein, In embodiments, the antibody comprises a VH region comprising the amino acid sequence of SEQ ID NO: 5, or a humanized variant thereof. In some embodiments of the various methods provided herein, the antibody , a VH region comprising the amino acid sequence of SEQ ID NO: 7, or a humanized variant thereof. In some embodiments of the various methods provided, the antibody has the amino acid sequence of SEQ ID NO:9. The VH region comprising the sequence of any of the various methods provided herein may be a humanized variant thereof. In some embodiments, the antibody comprises a VH region comprising the amino acid sequence of SEQ ID NO: 11, or In some embodiments of the various methods provided herein, The antibody comprises a VH region comprising the amino acid sequence of SEQ ID NO: 13, or a humanized variant thereof. In some embodiments of the various methods provided herein, the antibody comprises the antibody of SEQ ID NO: 15. The VH region includes a VH region comprising the amino acid sequence of the VH region, or a humanized variant thereof. In some embodiments of the method, the antibody comprises a VH region comprising the amino acid sequence of SEQ ID NO: 17; or humanized variants thereof. wherein the antibody comprises a VH region comprising the amino acid sequence of SEQ ID NO: 17, or a humanized variant thereof. In some embodiments of the various methods provided herein, the antibody comprises SEQ ID NO: No. 19, or a humanized variant thereof. In some embodiments of the various methods, the antibody comprises the amino acid sequence of SEQ ID NO: 21. The VH region, or a humanized variant thereof, may be used in some embodiments of the various methods provided herein. In embodiments, the antibody comprises a VH region comprising the amino acid sequence of SEQ ID NO: 23, or a humanized variant thereof. In some embodiments of the various methods provided herein, the antibody , a VH region comprising the amino acid sequence of SEQ ID NO: 25, or a humanized variant thereof. In some embodiments of the various methods provided, the antibody has the amino acid sequence of SEQ ID NO:27. The VH region comprising the sequence of any of the various methods provided herein may be a humanized variant thereof. In some embodiments, the antibody comprises a VH region comprising the amino acid sequence of SEQ ID NO: 29, or In some embodiments of the various methods provided herein, The antibody comprises a VH region comprising the amino acid sequence of SEQ ID NO: 31, or a humanized variant thereof. In some embodiments of the various methods provided herein, the antibody comprises the antibody of SEQ ID NO: 33. The VH region includes a VH region comprising the amino acid sequence of the VH region, or a humanized variant thereof. In some embodiments of the method, the antibody comprises a VH region comprising the amino acid sequence of SEQ ID NO: 35; or humanized variants thereof. wherein the antibody comprises a VH region comprising the amino acid sequence of SEQ ID NO: 37, or a humanized variant thereof. In some embodiments of the various methods provided herein, the antibody comprises SEQ ID NO: No. 39, or a humanized variant thereof. In some embodiments of the various methods, the antibody comprises the amino acid sequence of SEQ ID NO: 480. Some of the various methods provided herein include a VH region comprising a VH region containing ... In one embodiment, the antibody comprises a VH region comprising the amino acid sequence of SEQ ID NO: 482, or In some embodiments of the various methods provided herein, the The antibody comprises a VH region comprising the amino acid sequence of SEQ ID NO: 484, or a humanized variant thereof. In some embodiments of the various methods provided herein, the antibody is selected from the group consisting of SEQ ID NO: 486 The VH region comprises the amino acid sequence of, or a humanized variant thereof.

[0298] In some embodiments of the various methods provided herein, the antibody is selected from the group consisting of SEQ ID NO: 2, or a humanized variant thereof. In some embodiments of the various methods, the antibody comprises a VL comprising the amino acid sequence of SEQ ID NO:4. The present invention relates to a method for the preparation of a humanized or humanized variant ... In embodiments, the antibody comprises a VL region comprising the amino acid sequence of SEQ ID NO: 6, or a humanized variant thereof. In some embodiments of the various methods provided herein, the antibody comprises: The VL region comprises the amino acid sequence of SEQ ID NO: 8, or a humanized variant thereof. In some embodiments of the various methods, the antibody has the amino acid sequence of SEQ ID NO: 10. or a humanized variant thereof. In some embodiments, the antibody comprises a VL region comprising the amino acid sequence of SEQ ID NO: 12, or a VL region thereof. In some embodiments of the various methods provided herein, the The antibody comprises a VL region comprising the amino acid sequence of SEQ ID NO: 14, or a humanized variant thereof. In some embodiments of the various methods provided herein, the antibody is an amino acid sequence of SEQ ID NO: 16. The VL region includes a VL region comprising the amino acid sequence of α-amino acid ... In some embodiments, the antibody comprises a VL region comprising the amino acid sequence of SEQ ID NO: 18; and humanized variants thereof. In some embodiments of the various methods provided herein, wherein the antibody comprises a VL region comprising the amino acid sequence of SEQ ID NO: 20, or a humanized variant thereof. In some embodiments of the various methods provided herein, the antibody is selected from the group consisting of SEQ ID NO: The VL region comprises a sequence of 22 amino acids, or a humanized variant thereof. In some embodiments of the various methods, the antibody comprises a VL comprising the amino acid sequence of SEQ ID NO:24. The present invention relates to a method for the preparation of a humanized or humanized variant ... In embodiments, the antibody comprises a VL region comprising the amino acid sequence of SEQ ID NO: 26, or a humanized variant thereof. In some embodiments of the various methods provided herein, the antibody comprises: The VL region comprises the amino acid sequence of SEQ ID NO: 28, or a humanized variant thereof. In some embodiments of the various methods, the antibody has the amino acid sequence of SEQ ID NO: 30. or a humanized variant thereof. In some embodiments, the antibody comprises a VL region comprising the amino acid sequence of SEQ ID NO: 32, or a VL region thereof. In some embodiments of the various methods provided herein, the The antibody comprises a VL region comprising the amino acid sequence of SEQ ID NO: 34, or a humanized variant thereof. In some embodiments of the various methods provided herein, the antibody is an amino acid sequence of SEQ ID NO: 36. The VL region includes a VL region comprising the amino acid sequence of α-amino acid ... In some embodiments, the antibody comprises a VL region comprising the amino acid sequence of SEQ ID NO: 38; or and humanized variants thereof. In some embodiments of the various methods provided herein, wherein the antibody comprises a VL region comprising the amino acid sequence of SEQ ID NO: 40, or a humanized variant thereof. In some embodiments of the various methods provided herein, the antibody is selected from the group consisting of SEQ ID NO: The VL region comprises a VL region comprising the amino acid sequence of 481, or a humanized variant thereof. In some embodiments of the various methods, the antibody comprises the amino acid sequence of SEQ ID NO: 483. Some of the various methods provided herein include a VL region comprising a VL region containing a VL region of a humanized variant thereof. In one embodiment, the antibody comprises a VL region comprising the amino acid sequence of SEQ ID NO: 485, or a human In some embodiments of the various methods provided herein, the antibody The antibody comprises a VL region comprising the amino acid sequence of SEQ ID NO: 487, or a humanized variant thereof.

[0299] In some embodiments of the various methods provided herein, the antibody is selected from the group consisting of SEQ ID NO: a VH region comprising the amino acid sequence of SEQ ID NO: 1, or a humanized variant thereof, and the amino acid sequence of SEQ ID NO: 2 Some of the various methods provided herein include a VL region comprising the VL region, or a humanized variant thereof. In one embodiment, the antibody comprises a VH region comprising the amino acid sequence of SEQ ID NO: 3, or a human and a VL region comprising the amino acid sequence of SEQ ID NO: 4, or a humanized variant thereof. In some embodiments of the various methods provided herein, the antibody is selected from the group consisting of SEQ ID NO:5 or a humanized variant thereof, and a VH region comprising the amino acid sequence of SEQ ID NO: 6. Some of the various methods provided herein include a VL region comprising the VL region, or a humanized variant thereof. In one embodiment, the antibody comprises a VH region comprising the amino acid sequence of SEQ ID NO: 7, or a human and a VL region comprising the amino acid sequence of SEQ ID NO: 8, or a humanized variant thereof. In some embodiments of the various methods provided herein, the antibody is selected from the group consisting of SEQ ID NO:9 or a humanized variant thereof, and a VH region comprising the amino acid sequence of SEQ ID NO: 10. Some of the various methods provided herein include a VL region comprising the VL region, or a humanized variant thereof. In one embodiment, the antibody comprises a VH region comprising the amino acid sequence of SEQ ID NO: 11, or a human and a VL region comprising the amino acid sequence of SEQ ID NO: 12, or a humanized variant thereof. In some embodiments of the various methods provided herein, the antibody is selected from the group consisting of SEQ ID NO: 13 or a humanized variant thereof, and a VH region comprising the amino acid sequence of SEQ ID NO: 14. Some of the various methods provided herein include a VL region comprising the VL region, or a humanized variant thereof. In one embodiment, the antibody comprises a VH region comprising the amino acid sequence of SEQ ID NO: 15, or a human and a VL region comprising the amino acid sequence of SEQ ID NO: 16, or a humanized variant thereof. In some embodiments of the various methods provided herein, the antibody is SEQ ID NO: 17 or a humanized variant thereof, and a VH region comprising the amino acid sequence of SEQ ID NO: 18. Some of the various methods provided herein include a VL region comprising the VL region, or a humanized variant thereof. In one embodiment, the antibody comprises a VH region comprising the amino acid sequence of SEQ ID NO: 19, or a human and a VL region comprising the amino acid sequence of SEQ ID NO: 20, or a humanized variant thereof. In some embodiments of the various methods provided herein, the antibody is selected from the group consisting of SEQ ID NO: 21 or a humanized variant thereof, and a VH region comprising the amino acid sequence of SEQ ID NO: 22. Some of the various methods provided herein include a VL region comprising the VL region, or a humanized variant thereof. In one embodiment, the antibody comprises a VH region comprising the amino acid sequence of SEQ ID NO: 23, or a human and a VL region comprising the amino acid sequence of SEQ ID NO: 24, or a humanized variant thereof. In some embodiments of the various methods provided herein, the antibody is selected from the group consisting of SEQ ID NO: 25 or a humanized variant thereof, and a VH region comprising the amino acid sequence of SEQ ID NO: 26. Some of the various methods provided herein include a VL region comprising the VL region, or a humanized variant thereof. In one embodiment, the antibody comprises a VH region comprising the amino acid sequence of SEQ ID NO: 27, or a human and a VL region comprising the amino acid sequence of SEQ ID NO: 28, or a humanized variant thereof. In some embodiments of the various methods provided herein, the antibody is selected from the group consisting of SEQ ID NO: 29 or a humanized variant thereof, and a VH region comprising the amino acid sequence of SEQ ID NO: 30. Some of the various methods provided herein include a VL region comprising the VL region, or a humanized variant thereof. In one embodiment, the antibody comprises a VH region comprising the amino acid sequence of SEQ ID NO: 31, or a human and a VL region comprising the amino acid sequence of SEQ ID NO: 32, or a humanized variant thereof. In some embodiments of the various methods provided herein, the antibody is selected from the group consisting of SEQ ID NO: 33 or a humanized variant thereof, and a VH region comprising the amino acid sequence of SEQ ID NO: 34. Some of the various methods provided herein include a VL region comprising the VL region, or a humanized variant thereof. In one embodiment, the antibody comprises a VH region comprising the amino acid sequence of SEQ ID NO: 35, or a human and a VL region comprising the amino acid sequence of SEQ ID NO: 36, or a humanized variant thereof. In some embodiments of the various methods provided herein, the antibody is selected from the group consisting of SEQ ID NO: 37 or a humanized variant thereof, and a VH region comprising the amino acid sequence of SEQ ID NO: 38. Some of the various methods provided herein include a VL region comprising the VL region, or a humanized variant thereof. In one embodiment, the antibody comprises a VH region comprising the amino acid sequence of SEQ ID NO: 39, or a human and a VL region comprising the amino acid sequence of SEQ ID NO: 40, or a humanized variant thereof. In some embodiments of the various methods provided herein, the antibody is selected from the group consisting of SEQ ID NO: 48 0, or a humanized variant thereof, and the amino acid sequence of SEQ ID NO: 481. or a humanized variant thereof. In some embodiments, the antibody comprises a VH region comprising the amino acid sequence of SEQ ID NO: 482, or a humanized variant thereof, and a VL region comprising the amino acid sequence of SEQ ID NO: 483, or a humanized variant thereof; In some embodiments of the various methods provided herein, the antibody comprises the sequence a VH region comprising the amino acid sequence of SEQ ID NO: 484, or a humanized variant thereof, and an amino acid sequence of SEQ ID NO: 485; The VL region comprises a VL region comprising the VL sequence, or a humanized variant thereof. In some embodiments, the antibody comprises a VH region comprising the amino acid sequence of SEQ ID NO: 486; a VL region comprising the amino acid sequence of SEQ ID NO: 487, or a humanized variant thereof; Includes variants.

[0300] In some embodiments of the various methods provided herein, the antibody is selected from the group consisting of SEQ ID NO: VH CDR1, VH CDR2, and VH CDR3 of a VH domain having the amino acid sequence of SEQ ID NO: 1, and The VL region comprises VL CDR1, VL CDR2, and VL CDR3, each of which comprises two amino acid sequences. In some embodiments of the various methods, the antibody has the amino acid sequence of SEQ ID NO:3. and the amino acid sequence of SEQ ID NO: 4. The VL region includes VL CDR1, VL CDR2, and VL CDR3. In some embodiments of the method, the antibody comprises a VH domain having the amino acid sequence of SEQ ID NO:5. and a VL region comprising the amino acid sequence of SEQ ID NO: 6. VL CDR1, VL CDR2, and VL CDR3. In one embodiment, the antibody comprises a VH domain having the amino acid sequence of SEQ ID NO: 7, a VH CDR1 of the VH domain having the amino acid sequence of SEQ ID NO: 7, VH CDR2 and VH CDR3, and VL CDR1 and VL CDR of the VL region comprising the amino acid sequence of SEQ ID NO: 8 In some embodiments of the various methods provided herein, The antibody comprises a VH domain having the amino acid sequence of SEQ ID NO:9, a VH CDR1, a VH CDR2, and a VH CDR3. CDR3, and VL CDR1, VL CDR2, and VL CDR3 of the VL region comprising the amino acid sequence of SEQ ID NO: 10 In some embodiments of the various methods provided herein, the antibody comprises a sequence VH CDR1, VH CDR2, and VH CDR3 of a VH domain having the amino acid sequence of sequence number 11; and The VL region comprises VL CDR1, VL CDR2, and VL CDR3, each of which comprises the amino acid sequence of SEQ ID NO: 12. In some embodiments of the various methods provided herein, the antibody comprises the antibody of SEQ ID NO: 13. VH CDR1, VH CDR2, and VH CDR3 of the VH domain having the amino acid sequence of SEQ ID NO: 14 The VL region comprises the amino acid sequences VL CDR1, VL CDR2, and VL CDR3. In some embodiments of the various methods, the antibody has the amino acid sequence of SEQ ID NO: 15. VH CDR1, VH CDR2, and VH CDR3 of the VH domain having the amino acid sequence of SEQ ID NO: 16 The VL region includes VL CDR1, VL CDR2, and VL CDR3. In some embodiments of the method, the antibody comprises a VH domain having the amino acid sequence of SEQ ID NO: 17. a VL region comprising the VH CDR1, VH CDR2, and VH CDR3 of SEQ ID NO: 17 and the amino acid sequence of SEQ ID NO: 18; Some of the various methods provided herein include the VL CDR1, VL CDR2, and VL CDR3 of In one embodiment, the antibody comprises a VH CDR of a VH domain having the amino acid sequence of SEQ ID NO: 19. 1, VH CDR2, and VH CDR3, and VL CDR1, VL CDR2, and VL CDR3 of the VL region comprising the amino acid sequence of SEQ ID NO: 20. In some embodiments of the various methods provided herein, The antibody comprises a VH domain having the amino acid sequence of SEQ ID NO: 21, a VH CDR1, a VH CDR2, and a VH CDR3. and VH CDR3, and VL CDR1, VL CDR2, and VL CDR3 of the VL region comprising the amino acid sequence of SEQ ID NO: 22. In some embodiments of the various methods provided herein, the antibody comprises a CDR3. VH CDR1, VH CDR2, and VH CDR3 of the VH domain having the amino acid sequence of SEQ ID NO: 23, as well as and a VL region comprising VL CDR1, VL CDR2, and VL CDR3 having the amino acid sequence of SEQ ID NO:24. In some embodiments of the various methods provided herein, the antibody is selected from the group consisting of SEQ ID NO: 25 VH CDR1, VH CDR2, and VH CDR3 of a VH domain having the amino acid sequence of SEQ ID NO: 2 The VL region comprises a VL CDR1, a VL CDR2, and a VL CDR3, each of which comprises a sequence of six amino acids. In some embodiments of the various methods, the antibody has the amino acid sequence of SEQ ID NO:27. VH CDR1, VH CDR2, and VH CDR3 of a VH domain having the amino acid sequence of SEQ ID NO: 28 The VL region includes VL CDR1, VL CDR2, and VL CDR3. In some embodiments of the method, the antibody comprises a VH domain having the amino acid sequence of SEQ ID NO:29. The main VH CDR1, VH CDR2, and VH CDR3, and the VL region comprising the amino acid sequence of SEQ ID NO: 30 In some of the various methods provided herein, the VL CDR1, VL CDR2, and VL CDR3 regions are included. In some embodiments, the antibody comprises a VH domain having the amino acid sequence of SEQ ID NO: 31. DR1, VH CDR2, and VH CDR3, and VL CDR1 of the VL region comprising the amino acid sequence of SEQ ID NO: 32; In some embodiments of the various methods provided herein, wherein the antibody comprises a VH domain having the amino acid sequence of SEQ ID NO: 33, a VH CDR1, a VH CDR2, and VH CDR3, and VL CDR1, VL CDR2, and VL CDR3 of the VL region comprising the amino acid sequence of SEQ ID NO: 34. In some embodiments of the various methods provided herein, the antibody comprises a L CDR3. VH CDR1, VH CDR2, and VH CDR3 of a VH domain having the amino acid sequence of SEQ ID NO: 35; and a VL region comprising VL CDR1, VL CDR2, and VL CDR3 having the amino acid sequence of SEQ ID NO: 36. In some embodiments of the various methods provided herein, the antibody is selected from the group consisting of SEQ ID NO: VH CDR1, VH CDR2, and VH CDR3 of the VH domain having a sequence of 37 amino acids, and SEQ ID NO: The VL region comprises VL CDR1, VL CDR2, and VL CDR3 of the VL region comprising the amino acid sequence of SEQ ID NO:38. In some embodiments of the various methods provided, the antibody comprises the amino acid sequence of SEQ ID NO: 39. VH CDR1, VH CDR2, and VH CDR3 of the VH domain having the sequence The VL region includes VL CDR1, VL CDR2, and VL CDR3, each of which comprises a sequence similar to that provided herein. In some embodiments of the various methods, the antibody has the amino acid sequence of SEQ ID NO: 480. VH domain VH CDR1, VH CDR2, and VH CDR3, and the amino acid sequence of SEQ ID NO: 481. The VL region includes VL CDR1, VL CDR2, and VL CDR3. In some embodiments, the antibody comprises a VH domain having the amino acid sequence of SEQ ID NO: 482. and a VL region comprising the amino acid sequence of SEQ ID NO: 483. VL CDR1, VL CDR2, and VL CDR3. In one embodiment, the antibody comprises a VH CDR1 of a VH domain having the amino acid sequence of SEQ ID NO: 484. , VH CDR2, and VH CDR3, and a VL region comprising the amino acid sequence of SEQ ID NO: 485. In some embodiments of the various methods provided herein, The antibody comprises a VH domain having the amino acid sequence of SEQ ID NO: 486, VH CDR1, VH CDR2, and VH CDR3, and VL CDR1, VL CDR2, and VL region comprising the amino acid sequence of SEQ ID NO: 487. Other VH domains, VL domains, VH CDR1, VH CDR2, and VL CDR3 described herein. The VH CDR3, VL CDR1, VL CDR2, and VL CDR3 sequences can also be used in the various methods provided herein. It is intended for use in

[0301] (1. Polyclonal antibodies) The antibodies of the present disclosure may include polyclonal antibodies. Polyclonal antibodies can be generated, for example, by immunizing and, if desired, by injecting azido-. The vaccine can be raised in a mammal by one or more injections of an immunizing agent. Alternatively, the adjuvant is injected into the mammal by multiple subcutaneous or intraperitoneal injections. The immunizing agent may comprise a GFRAL polypeptide or a fusion protein thereof. Conjugate to a protein known to be immunogenic in the mammalian host or where it is useful to immunize a mammal with the protein and one or more adjuvants. Examples of such immunogenic proteins include keyhole limpet hemocyanin (KLM) and hemocyanin (HEM). These include thrombocytopenic acid, serum albumin, bovine thyroglobulin, and soybean trypsin inhibitor. Examples of adjuvants that can be used include, but are not limited to, Ribi, CpG, and polysaccharides. Ri1C, Freund's complete adjuvant, and MPL-TDM adjuvant (monophosphoryl lipid A , synthetic trehalose dicorynomycolate). The immunization protocol is designed to minimize excessive experimental The mammal is then bled and the serum is extracted with GFAP. RAL antibody titers can be assayed. If desired, antibody titers are elevated. Additionally or alternatively, the mammal can be boosted until a plateau is reached. , fusion of lymphocytes and preparation of monoclonal antibodies from hybridomas as described below. For example, the antibody may be obtained from an immunized animal.

[0302] (2. Monoclonal antibodies) The antibodies of the present disclosure may alternatively include monoclonal antibodies. Using the hybridoma method first described by Kohler et al. (Nature, 256:495 (1975)) They may be produced by recombinant DNA methods (see, e.g., U.S. Pat. No. 4,816,567). It may also be manufactured.

[0303] In the hybridoma method, mice or other suitable host animals, e.g., hamsters, are cultured in the manner described above. Immunize the animal so that it produces or produces antibodies that specifically bind to the protein used for immunization. Alternatively, lymphocytes may be immunized in vitro. After immunization, lymphocytes are isolated and then fused with a suitable fusing agent, e.g., polyethylene glycol. The resulting hybridoma cells are then fused with a myeloma cell line using the fusion protein to form hybridoma cells (Goding, 1999). , Monoclonal Antibodies: Principles and Practice ), pp. 59-103 (Academic Press, 1986).

[0304] The hybridoma cells thus prepared are seeded into a suitable culture medium and grown therein. The medium is preferably maintained in the presence of unfused parental myeloma cells (also called fusion partners). For example, the parent myeloma cells contain one or more substances that inhibit the growth or survival of the cells that are to be treated with the anti-myeloma agent. lacking the enzyme xanthine guanine phosphoribosyltransferase (HGPRT or HPRT) In this case, the selective culture medium for hybridomas usually contains hypoxanthine, aminopterin, and thymidine (HAT medium), which inhibit the growth of HGPRT-deficient cells.

[0305] Preferred fusion partner myeloma cells are those that fuse efficiently and are expressed by the selected antibody-producing cells. Supports stable, high-level antibody production from fused cells and selects against unfused parental cells. Cells that are sensitive to a selective medium. Preferred myeloma cell lines are murine myeloma lines, e.g. , SP-2 and derivatives, e.g., American Type Culture Collection X63-Ag8-653 cells available from the Type Culture Collection), Manassas, Virginia, USA; and the Salk Institute Cell Distribution Center, San Derived from MOPC-21 and MPC-11 mouse tumors available from San Diego, California, USA Human myeloma and mouse-human heteromyeloma for the production of human monoclonal antibodies Cell lines have also been described (Kozbor, J., Immunol., 133:3001 (1984); and Brodeur et al. Literature on Monoclonal Antibody Production Techniques and Applications Ques and Applications, pp. 51-63 (Marcel Dekker, New York, 1987).

[0306] The culture medium in which the hybridoma cells are growing is used to produce monoclonal antibodies against the antigen. Assayed for viability. Monoclonal antibodies produced by hybridoma cells The binding specificity of the antibody can be determined by immunoprecipitation or by in vitro binding assays, e.g., radioimmunoassays. Determined by radioimmunoassay (RIA) or enzyme-linked immunosorbent assay (ELISA). The binding affinity of a cloned antibody is determined, for example, by the method described in Munson et al., Anal. Biochem., 107:220 (1980). This can be determined by Scatchard analysis as described.

[0307] Once hybridoma cells produce antibodies of desired specificity, affinity, and / or activity Once identified, clones were subcloned by limiting dilution and analyzed by standard methods. (Goding, Monoclonal Antibodies: Principles and Practice) nal Antibodies: Principles and Practice), pp. 59-103 (Academic Press, 1986)). Suitable culture media for this purpose include, for example, D-MEM or RPMI-1640 medium. Furthermore, the hybridoma cells can be cultured, for example, by intraperitoneal injection of the cells into mice. and can be grown in vivo as ascites tumors in animals.

[0308] The monoclonal antibodies secreted by the subclones can be purified, for example, by affinity chromatography. (e.g., those using protein A or protein G-Sepharose) or Ion exchange chromatography, hydroxylapatite chromatography, gel electrolysis The antibody is suitably isolated from the culture medium, ascites fluid, or serum by conventional antibody purification procedures such as electrophoresis, dialysis, etc. Be separated.

[0309] DNA encoding monoclonal antibodies can be isolated using conventional procedures (e.g., by PCR of murine antibodies). An oligonucleotide probe capable of specifically binding to genes encoding the low and light chains The hybridomas are easily isolated and sequenced (using a probe). Mammalian cells serve as a preferred source of such DNA. Once isolated, D The NA can be placed into an expression vector, which then has to be transfected. E. coli cells, monkey COS cells, Chinese flu virus, and other bacteria do not produce antibody proteins. transfecting the vector into host cells, such as Chinese hamster ovary (CHO) cells or myeloma cells; Obtaining monoclonal antibody synthesis in recombinant host cells. Intracellular expression of antibody-encoding DNA. For a review of recombinant expression in humans, see Skerra et al., Curr. Opinion in Immunol., 5 :256-262 (1993) and Pluckthun, Immunol. Revs. 130:151-188 (1992). .

[0310] In some embodiments, antibodies that bind to the GFRAL epitope are selected from those that: (1) are regulated by stringency; DNA bound to the filter under mild conditions (e.g., 6× sodium chloride / sodium citrate) Hybridization in 0.2x SSC / 0.1% SDS at approximately 45°C followed by hybridization in 0.2x SSC / 0.1% SDS. one or more washes at approximately 50-65°C), under very stringent conditions (e.g., filter Hybridization to nucleic acid bound to 0.1% ATP in 6×SSC at about 45° C., followed by 0.1% ATP. 1×SSC / 0.2% SDS at about 68° C.), or other stringency washes known to those skilled in the art. Under favorable hybridization conditions, the VH and / or VL domains described herein A nucleotide sequence that hybridizes to the complement of a nucleotide sequence encoding any one of the The amino acid sequence of the VH domain and / or the VL domain encoded by the nucleotide sequence Contains amino acid sequences (see, e.g., Ausubel, FM et al., eds., 1989, Current Protocols in Molecular Biology (C Current Protocols in Molecular Biology, Volume I, Green Publishing Associates, Inc. John Wiley & Sons, New York, see pages 6.3.1 to 6.3.6 and 2.10.3).

[0311] In some embodiments, antibodies that bind to the GFRAL epitope are identified by stringent Hybridization to filter-bound DNA under suitable conditions (e.g., in 6×SSC at about 45° C.) (incubation, followed by one or more washes in 0.2x SSC / 0.1% SDS at approximately 50-65°C), Under stringent conditions (e.g., in 6×SSC at about 45° C. to the filter-bound nucleic acid) hybridization at 0.1×SSC / 0.2% SDS at about 68° C., followed by one or more cycles of hybridization at about 68° C. in 0.1×SSC / 0.2% SDS. Washing), or other stringent hybridization conditions known to those of skill in the art. and a nucleic acid encoding any one of the VH CDRs and / or VL CDRs shown in Tables 1 to 24. VH CDRs encoded by nucleotide sequences that hybridize to the complement of the nucleotide sequence or the amino acid sequence of the VL CDR (see, for example, Ausubel, FM et al., eds., 1989, pp. 131-135, 1989). Current Protocols in Molecular Biology, Vol. I, Green P Publishing Associates and John Wiley & Sons, New York, 6.3.1-6.3.6 and 2.10.3 page).

[0312] In further embodiments, the monoclonal antibody or antibody fragment is, for example, an antibody fragment. Antibody Phage Display: Methods and Protocols , edited by PM O'Brien and R. Aitken, Humana Press, Totawa NJ, 2002. It can be isolated from an antibody phage library produced using the method. The synthetic antibody clones are prepared by combining various antibody variable regions (Fv) fused to phage coat proteins. The fragments are selected by screening a phage library containing phages displaying the fragments. Such a phage library is then screened against the desired antigen. A clone expressing an Fv fragment capable of binding to a desired antigen is adsorbed to the antigen, thereby The binding clones are then separated from the non-binding clones in the library by dissolving them from the antigen. The antigen adsorption / elution cycles can be further enriched.

[0313] Variable domains can be prepared using the methods described, for example, in Winter et al., Ann. Rev. Immunol., 12: 433-455 (1994). As described in

[2004] , VH and VL are covalently linked via a short, flexible peptide. as single-chain Fv (scFv) fragments in which the VH and VL are each fused to a constant domain and non-covalently linked. The Fab fragments are functionally displayed on the phage either as interacting Fab fragments or as Fab fragments interacting with each other. It is possible.

[0314] The VH and VL gene repertoires were cloned separately by polymerase chain reaction (PCR). can be randomly recombined in a phage library, which can then be As described in [Illegible], antigen-binding clones can be searched for. Libraries from immunized sources can be used to generate immunized strains without the need to construct hybridomas. Provide high affinity antibodies against the antigen. Alternatively, clone the naive repertoire. As described by Griffiths et al. (EMBO J, 12: 725-734 (1993)), A single source of human antibodies against a wide range of non-self antigens and also self antigens without immunization. Finally, naive repertoire can be provided, for example, as described by Hoogenboom and Wint. As described by [J. Mol. Biol., 227: 381-388 (1992)], stem cell-derived Unrearranged V-gene segments were cloned and PCR-enhanced containing random sequences. Primers are used to encode highly variable CDR3 regions and achieve in vitro rearrangement. It can also be prepared synthetically by

[0315] Screening of the library can be accomplished by a variety of techniques known in the art. For example, GFRAL (e.g., a GFRAL polypeptide, fragment, or epitope) can be administered to a mammalian cell line by inhalation. It may be used to coat the wells of the adsorption plate and fixed to the adsorption plate. or expressed on host cells used in cell sorting or streptavidin coating. Conjugated to biotin for capture by modified beads or display live Any other method for panning the array can be used. Slow dissociation kinetics Selection of antibodies with good binding affinity (e.g., good binding affinity) can be performed using methods such as those described in Bass et al., Proteins, 8: 309-313. 14 (1990) and WO 92 / 09690. The use of ray and the reference in Marks et al., Biotechnol., 10: 779-783 (1992) This can be facilitated by a low coating density of the antigen.

[0316] Anti-GFRAL antibodies are suitable antigen screens for selection of phage clones of interest. A screening procedure is designed to subsequently identify VH and / or VH fragments from the phage clones of interest. VL sequences (e.g., Fv sequences), or various CDR sequences derived from VH and VL sequences, and Kabat et al. , Sequences of Proteins of Immunological Interest rest), 5th edition, NIH Publication 91-3242, Bethesda MD (1991), Vol. 1 to Vol. 3 A full-length anti-GFRAL antibody clone is constructed using a suitable constant region (e.g., Fc) sequence. This can be obtained by:

[0317] (3.Antibody fragment) The present disclosure provides antibodies and antibody fragments that bind to GFRAL. There are advantages to using antibody fragments rather than whole bodies due to the smaller size of the fragments. , allowing for rapid clearance and improved access to cells, tissues, or organs. For a review of specific antibody fragments, see Hudson et al. (2003) Nat. Med. 9:129-134. Please refer to.

[0318] Various techniques have been developed for the production of antibody fragments. Traditionally, these fragments are produced from intact antibodies. obtained by proteolytic digestion of the body (see, e.g., Morimoto et al., Journal of Biology Biochemical and Biophysical Methods 24:107-117 (1992); and Brennan et al., Science, 229:81 (1985). However, these fragments are not currently expressed by recombinant host cells. Fab, Fv, and ScFv antibody fragments can all be produced directly in E. coli or yeast cells. The fragments are expressed in and secreted from Escherichia coli, thereby allowing for the facile production of large amounts 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 the Fab'-SH fragments. to form F(ab')2 fragments (Carter et al., Bio / Technology 10:163-166). According to another approach, F(ab')2 fragments can be isolated directly from recombinant host cell culture. It is possible to produce a Fa with an extended in vivo half-life containing salvage receptor binding epitope residues. F(ab')2 and F(ab')2 fragments are described, for example, in U.S. Patent No. 5,869,046. Other techniques for producing Fv fragments will be apparent to those skilled in the art. In certain embodiments, the antibody is a single chain Fv fragment. and scFv (see, e.g., WO 93 / 16185; U.S. Pat. Nos. 5,571,894; and 5,587,458). Fv and scFv have intact binding sites that lack constant regions; therefore, they are scFv fusion proteins may be suitable for reducing non-specific binding when used in vivo. , fusion of an effector protein at either the amino or carboxy terminus of the scFv It may be constructed to produce a nucleotide sequence (e.g., antibody engineering, B See Orrebaeck, ed. (supra). Antibody fragments may also be used, for example, as described in the above-cited references. Such linear antibodies may be, for example, "linear antibodies" as described above. It may be monospecific or multispecific, eg bispecific.

[0319] Smaller antibody-derived binding structures contain individual variable domains, also called single variable domain antibodies (SdAbs). Certain types of organisms, such as camelids and cartilaginous fish, have As part of their immune system, high-affinity single V domains mounted on Fc-equivalent domain structures It possesses a nucleotide-like domain (Woolven et al., Immunogenetics 50: 98-101, 1999; Streltsov (Proc Natl Acad Sci USA. 101:12444-12449, 2004). V-like domain (Camelidae) These primates (called VhH in humans and V-NAR in sharks) typically exhibit long surface loops, which allow These allow penetration of the cavity of the target antigen. They also mask hydrophobic surface patches. This stabilizes the isolated VH domain.

[0320] These VhH and V-NAR domains have been used to artificially generate sdAbs. The V domain variants were selected from a phage library and synthesized into stable, high-binding VL and V It has been designed using other techniques resulting in H-derived domains.

[0321] The antibodies that bind to GFRAL provided herein include synthetic antibodies, monoclonal antibodies, and the like. , recombinantly produced antibodies, multispecific antibodies (including bispecific antibodies), human antibodies, humanized antibodies, cloned antibodies, chimeric antibodies, intrabodies, anti-idiotypic (anti-Id) antibodies, and the like and the like. Non-limiting examples of functional fragments (e.g., fragments that bind to GFRAL) include single-chain Fv (scFv) ) (including, for example, monospecific, bispecific, etc.), Fab fragments, F(ab') fragments, F(ab)2 fragments, F( ab')2 fragments, disulfide-linked Fv (sdFv), Fd fragments, Fv fragments, diabodies, triabodies , tetrabodies, and minibodies.

[0322] The antibodies provided herein include immunoglobulin molecules and immunologically related molecules of immunoglobulin molecules. The therapeutically active portion includes, for example, a molecule containing an antigen binding site that binds to the GFRAL epitope. The immunoglobulin molecules provided herein include, but are not limited to, immunoglobulins. Any type (e.g., IgG, IgE, IgM, IgD, IgA, and IgY) or class (e.g., IgG The antibodies may be of any of the following classes: IgG1, IgG2, IgG3, IgG4, IgA1, and IgA2, or any of the subclasses.

[0323] Antibody variants and derivatives include antibody functional groups that retain the ability to bind to the GFRAL epitope. Exemplary functional fragments include Fab fragments (e.g., fragments containing the antigen-binding domain). an antibody fragment having a light chain and a portion of a heavy chain cross-linked by a disulfide bond; F ab' (e.g., a single antigen-binding domain comprising Fab and an additional portion of the heavy chain via the hinge region) antibody fragments containing F(ab')2 (e.g., interchain disulfide bonds in the hinge region of the heavy chain); two Fab' molecules connected by a Fab' bond; the Fab' molecules may be directed against the same or different epitopes. bispecific Fabs (e.g., each of which is directed against a different epitope); Fab molecules with two antigen-binding domains; single-chain Fab chains containing the variable region, also known as sFv (e.g. For example, a single light and heavy chain variable antigen of an antibody linked by a chain of 10 to 25 amino acids. intrinsic binding determining region; disulfide-linked Fv, or dsFv (e.g., linked by disulfide bonds) single light and heavy chain variable antigen-binding determining regions of the antibody; camelized VH (e.g., VH binding region); A single antibody fragment in which some amino acids on the contact surface are those found in the heavy chains of natural camel antibodies. variable antigen-binding determining regions of the heavy chain of a sFv; bispecific sFv (e.g., each of which binds a different epitope); an sFv or dsFv molecule having two antigen-binding domains that can be directed against a diabody; (For example, the VH domain of a first sFv associates with the VL domain of a second sFv, and the VL domain of the first sFv Dimerized sFv formed when the VH domain of a second sFv associates with the VH domain of a second sFv; The antigen-binding regions may be directed against the same or different epitopes; and diabodies (e.g., formed in a manner similar to diabodies, but with three antigen-binding domains in a single complex) The three antigen-binding domains can bind to the same or different epitopes. Derivatives of antibodies include those that modify one or more of the antibody binding sites. CDR sequences are also included. When two or more CDR sequences are present, the CDR sequences are In certain embodiments, the antibody comprises a single chain Fv ("scFv"). and antibody fragments comprising the VH and VL domains of the antibody, wherein these domains are bound to a single polypeptide. The scFv polypeptide is present in a peptide chain, and the scFv has the desired structure for antigen binding. The VH domain and the VL domain may further comprise a polypeptide linker that enables the formation of For a review of scFvs, see Pluckthun, The Pharmacology of Monoclonal Antibodies. in the field of Monoclonal Antibodies, Vol. 113, edited by Rosenburg and Moore, Springer-Verlag See rlag, New York, pp. 269-315 (1994).

[0324] 4. Humanized Antibodies The present disclosure provides humanized antibodies that bind to GFRAL, including human GFRAL. The antibody may comprise one or more CDRs shown in Tables 1 to 24. Various methods for humanizing non-human antibodies are available. For example, humanized antibodies can be prepared by cleaving an antibody from a source that is non-human. These non-human amino acid residues can have one or more amino acid residues inserted. These are often referred to as "import" residues, which are typically taken from an "import" variable domain. Humanization can be carried out, for example, by the method of Winter and coworkers (Jones et al., 1986) Nature 321:522-525; Riechmann et al. (1988) Nature 332:323-327; Verhoeyen et al. (1988) The hypervariable region sequences were substituted for the corresponding sequences of a human antibody according to the method of the present invention (Science 239:1534-1536). This can be carried out by using

[0325] In some cases, a humanized antibody retains all six complementarity-determining regions of the parent non-human (e.g., rodent) antibody. The amino acid sequences of the CDR regions (CDRs) are grafted onto a human antibody framework. For example, Padlan et al. (FASEB J. 9:133-139, 1995) reported that only about one-third of the residues in the CDRs are substituted. These were identified as "specificity-determining residues," or SDRs. In the SDR grafting technique, only the SDR residues are grafted onto a human antibody framework (e.g., See, for example, Kashmiri et al., Methods 36: 25-34, 2005).

[0326] Human variable domains of both the light and heavy chains to be used in making humanized antibodies The choice of antibody can be important to reduce antigenicity. For example, the so-called "best fit" method According to the method of the present invention, the sequence of the variable domain of a non-human (e.g., rodent) antibody is compared with a known human variable domain. Screen against the entire library of sequences. The human sequence that is closest to the rodent sequence can be selected as the human framework for a humanized antibody (Sims et al., 1993). J. Immunol. 151:2296; Chothia et al. (1987) J. Mol. Biol. 196:901). A unique sequence derived from the consensus sequence of all human antibodies of a particular subgroup of light or heavy chains. Use a fixed framework. The same framework can be used for several different humanized antibodies. (Carter et al. (1992) Proc. Natl. Acad. Sci. USA, 89:4285; Pre (1993) J. Immunol., 151:2623). In some cases, the framework The abundant human subclass V L 6 Subgroup I(V L 6I) and V H Subgroup III(V H III) consensus sequence. Alternatively, a human germline gene may be used as a framework. Use it as a source of disk space.

[0327] In an alternative paradigm based on CDR comparison, called hyper-humanization, FR homology is important. This method is based on the comparison of non-human sequences with the functional human germline gene repertoire. Then, a gene encoding a canonical structure that is the same as or closely related to the mouse sequence is selected. Next, among the genes that share the standard structure with non-human antibodies, the genes with the most homology within the CDRs are selected. Finally, non-human CDRs are grafted onto these FRs (e.g., See Tan et al., J. Immunol. 169: 1119-1125, 2002).

[0328] Generally, antibodies are synthesized by the method described above while retaining their affinity for the antigen and other favorable biological properties. It is even more desirable to humanize it in a manner that achieves this goal. , the parental sequences and various conceptual humanized products using three-dimensional models of the parental and humanized sequences. Humanized antibodies are prepared through a process of analysis. Three-dimensional immunoglobulin models are generally The potential of the selected candidate immunoglobulin sequences is readily available and well known to those skilled in the art. Computer programs are available to illustrate and display possible 3D structures. These include, for example, WAM (Whitelegg and Rees, Protein Eng. 13: 819-824, 200 0), Modeller (Sali and Blundell, J. Mol. Biol. 234: 779-815, 1993), and Sw Includes the iss PDB Viewer (Guex and Peitsch, Electrophoresis 18: 2714-2713, 1997). Inspection of these representations will provide insight into the role of residues in the function of the candidate immunoglobulin sequence. Analysis of the role played by the candidate immunoglobulin, e.g., in influencing its ability to bind to its antigen. In this way, desired properties such as increased affinity for the target antigen can be identified. FR residues are selected from the recipient and import sequences so that desired antibody characteristics are achieved. In general, hypervariable region residues are directly involved in influencing antigen binding. is directly and most substantially involved.

[0329] Another method for antibody humanization is to humanize the antibody, called human string content (HSC). This method is based on a measure of humanness. The target sequence is then compared to the offspring repertoire and the difference is scored as an HSC. Humanize by maximizing its HSC rather than using a global identity measure. Multiple diverse humanized variants are generated (Lazar et al., Mol. Immunol. 44: 1986-1998, 2 007).

[0330] In addition to the methods described above, experimental methods can be used to generate and select humanized antibodies. These ...

Claims

1. An antibody or a fragment thereof that specifically binds to a human GDNF family receptor alpha-like (GFRAL) protein, the antibody or the fragment thereof comprising a heavy chain variable (VH) region and a light chain variable (VL) region; (a) the VH region comprises a VH complementarity determining region (CDR) 1, a VH CDR2, and a VH CDR3 from the amino acid sequence set forth in SEQ ID NO:11; and the VL region comprises a VL CDR1, a VL CDR2, and a VL CDR3 from the amino acid sequence set forth in SEQ ID NO:12; (b) the VH region comprises a VH CDR1, a VH CDR2, and a VH CDR3 from the amino acid sequence set forth in SEQ ID NO: 15; and the VL region comprises a VL CDR1, a VL CDR2, and a VL CDR3 from the amino acid sequence set forth in SEQ ID NO: 16; (c) the VH region comprises a VH CDR1, a VH CDR2, and a VH CDR3 from the amino acid sequence set forth in SEQ ID NO:1; and the VL region comprises a VL CDR1, a VL CDR2, and a VL CDR3 from the amino acid sequence set forth in SEQ ID NO:2; (d) the VH region comprises a VH CDR1, a VH CDR2, and a VH CDR3 from the amino acid sequence set forth in SEQ ID NO:5; and the VL region comprises a VL CDR1, a VL CDR2, and a VL CDR3 from the amino acid sequence set forth in SEQ ID NO:6; (e) the VH region comprises a VH CDR1, a VH CDR2, and a VH CDR3 from the amino acid sequence set forth in SEQ ID NO:9; and the VL region comprises a VL CDR1, a VL CDR2, and a VL CDR3 from the amino acid sequence set forth in SEQ ID NO:10; (f) the VH region comprises a VH CDR1, a VH CDR2, and a VH CDR3 from the amino acid sequence set forth in SEQ ID NO: 13; and the VL region comprises a VL CDR1, a VL CDR2, and a VL CDR3 from the amino acid sequence set forth in SEQ ID NO: 14; (g) the VH region comprises a VH CDR1, a VH CDR2, and a VH CDR3 from the amino acid sequence set forth in SEQ ID NO: 17; and the VL region comprises a VL CDR1, a VL CDR2, and a VL CDR3 from the amino acid sequence set forth in SEQ ID NO: 18; (h) the VH region comprises a VH CDR1, a VH CDR2, and a VH CDR3 from the amino acid sequence set forth in SEQ ID NO: 19; and the VL region comprises a VL CDR1, a VL CDR2, and a VL CDR3 from the amino acid sequence set forth in SEQ ID NO: 20; (i) the VH region comprises a VH CDR1, a VH CDR2, and a VH CDR3 from the amino acid sequence set forth in SEQ ID NO:27; and the VL region comprises a VL CDR1, a VL CDR2, and a VL CDR3 from the amino acid sequence set forth in SEQ ID NO:28; (j) the VH region comprises a VH CDR1, a VH CDR2, and a VH CDR3 from the amino acid sequence set forth in SEQ ID NO:29; and the VL region comprises a VL CDR1, a VL CDR2, and a VL CDR3 from the amino acid sequence set forth in SEQ ID NO:30; (k) the VH region comprises a VH CDR1, a VH CDR2, and a VH CDR3 from the amino acid sequence set forth in SEQ ID NO: 31; and the VL region comprises a VL CDR1, a VL CDR2, and a VL CDR3 from the amino acid sequence set forth in SEQ ID NO: 32; (l) the VH region comprises a VH CDR1, a VH CDR2 and a VH CDR3 from the amino acid sequence set forth in SEQ ID NO: 33; and the VL region comprises a VL CDR1, a VL CDR2 and a VL CDR3 from the amino acid sequence set forth in SEQ ID NO: 34; or (m) the VH region comprises a VH CDR1, a VH CDR2, and a VH CDR3 from the amino acid sequence set forth in SEQ ID NO: 35; and the VL region comprises a VL CDR1, a VL CDR2, and a VL CDR3 from the amino acid sequence set forth in SEQ ID NO: 36; The antibody or a fragment thereof.

2. (a)(i) the VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2 and VL CDR3 comprise the amino acid sequences set forth in SEQ ID NOs: 64, 157, 241, 317, 388, and 438, respectively; (ii) the VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2 and VL CDR3 comprise the amino acid sequences set forth in SEQ ID NOs: 65, 220, 242, 318, 389 and 438, respectively; (iii) the VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2 and VL CDR3 comprise the amino acid sequences set forth in SEQ ID NOs: 66, 157, 241, 317, 388, and 438, respectively; (iv) the VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2 and VL CDR3 comprise the amino acid sequences set forth in SEQ ID NOs: 67, 158, 243, 319, 389 and 439, respectively; (v) the VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2 and VL CDR3 comprise the amino acid sequences set forth in SEQ ID NOs: 68, 159, 244, 320, 390 and 440, respectively; or (vi) the VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2 and VL CDR3 comprise the amino acid sequences set forth in SEQ ID NOs: 64, 160, 241, 317, 388, and 438, respectively; (b)(i) the VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2 and VL CDR3 comprise the amino acid sequences set forth in SEQ ID NOs: 72, 166, 249, 325, 393, and 444, respectively; (ii) the VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2 and VL CDR3 comprise the amino acid sequences set forth in SEQ ID NOs: 73, 167, 250, 326, 386, and 444, respectively; (iii) the VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2 and VL CDR3 comprise the amino acid sequences set forth in SEQ ID NOs: 74, 166, 249, 325, 393 and 444, respectively; (iv) the VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2 and VL CDR3 comprise the amino acid sequences set forth in SEQ ID NOs: 75, 168, 251, 327, 386, and 445, respectively; (v) the VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2 and VL CDR3 comprise the amino acid sequences set forth in SEQ ID NOs: 76, 169, 252, 328, 394 and 446, respectively; or (vi) the VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2 and VL CDR3 comprise the amino acid sequences set forth in SEQ ID NOs: 72, 170, 249, 325, 393, and 444, respectively; (c)(i) the VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2 and VL CDR3 comprise the amino acid sequences set forth in SEQ ID NOs: 41, 132, 221, 297, 373, and 423, respectively; (ii) the VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2 and VL CDR3 comprise the amino acid sequences set forth in SEQ ID NOs: 42, 133, 222, 298, 374, and 423, respectively; (iii) the VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2 and VL CDR3 comprise the amino acid sequences set forth in SEQ ID NOs: 43, 132, 221, 297, 373, and 423, respectively; (iv) the VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2 and VL CDR3 comprise the amino acid sequences set forth in SEQ ID NOs: 44, 134, 223, 299, 374 and 424, respectively; (v) the VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2 and VL CDR3 comprise the amino acid sequences set forth in SEQ ID NOs: 45, 135, 224, 300, 375 and 425, respectively; or (vi) the VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2 and VL CDR3 comprise the amino acid sequences set forth in SEQ ID NOs: 41, 136, 221, 297, 373, and 423, respectively; (d)(i) the VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2 and VL CDR3 comprise the amino acid sequences set forth in SEQ ID NOs: 51, 142, 229, 305, 379 and 429, respectively; (ii) the VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2 and VL CDR3 comprise the amino acid sequences set forth in SEQ ID NOs: 52, 143, 230, 306, 380, and 429, respectively; (iii) the VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2 and VL CDR3 comprise the amino acid sequences set forth in SEQ ID NOs: 53, 142, 229, 305, 379 and 429, respectively; (iv) the VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2 and VL CDR3 comprise the amino acid sequences set forth in SEQ ID NOs: 54, 144, 231, 307, 380, and 430, respectively; (v) the VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2 and VL CDR3 comprise the amino acid sequences set forth in SEQ ID NOs: 55, 145, 232, 308, 381 and 431, respectively; or (vi) the VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2 and VL CDR3 comprise the amino acid sequences set forth in SEQ ID NOs: 51, 146, 229, 305, 379 and 429, respectively; (e)(i) the VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2 and VL CDR3 comprise the amino acid sequences set forth in SEQ ID NOs: 60, 152, 237, 313, 385, and 435, respectively; (ii) the VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2 and VL CDR3 comprise the amino acid sequences set forth in SEQ ID NOs: 61, 153, 238, 314, 386, and 435, respectively; (iii) the VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2 and VL CDR3 comprise the amino acid sequences set forth in SEQ ID NOs: 62, 152, 237, 313, 385 and 435, respectively; (iv) the VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2 and VL CDR3 comprise the amino acid sequences set forth in SEQ ID NOs: 49, 154, 239, 315, 386, and 436, respectively; (v) the VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2 and VL CDR3 comprise the amino acid sequences set forth in SEQ ID NOs: 63, 155, 240, 316, 387 and 437, respectively; or (vi) the VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2 and VL CDR3 comprise the amino acid sequences set forth in SEQ ID NOs: 60, 156, 237, 313, 385 and 435, respectively; (f)(i) the VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2 and VL CDR3 comprise the amino acid sequences set forth in SEQ ID NOs: 69, 161, 245, 321, 391, and 441, respectively; (ii) the VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2 and VL CDR3 comprise the amino acid sequences set forth in SEQ ID NOs: 61, 162, 246, 322, 386, and 441, respectively; (iii) the VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2 and VL CDR3 comprise the amino acid sequences set forth in SEQ ID NOs: 70, 161, 245, 321, 391, and 441, respectively; (iv) the VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2 and VL CDR3 comprise the amino acid sequences set forth in SEQ ID NOs: 49, 163, 247, 323, 386, and 442, respectively; (v) the VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2 and VL CDR3 comprise the amino acid sequences set forth in SEQ ID NOs: 71, 164, 248, 324, 392 and 443, respectively; or (vi) the VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2 and VL CDR3 comprise the amino acid sequences set forth in SEQ ID NOs: 69, 165, 245, 321, 391 and 441, respectively; (g)(i) the VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2 and VL CDR3 comprise the amino acid sequences set forth in SEQ ID NOs: 77, 171, 253, 329, 395, and 447, respectively; (ii) the VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2 and VL CDR3 comprise the amino acid sequences set forth in SEQ ID NOs: 78, 143, 254, 330, 396, and 447, respectively; (iii) the VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2 and VL CDR3 comprise the amino acid sequences set forth in SEQ ID NOs: 79, 171, 253, 329, 395, and 447, respectively; (iv) the VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2 and VL CDR3 comprise the amino acid sequences set forth in SEQ ID NOs: 80, 144, 255, 331, 396, and 448, respectively; (v) the VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2 and VL CDR3 comprise the amino acid sequences set forth in SEQ ID NOs: 81, 145, 256, 332, 397 and 449, respectively; or (vi) the VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2 and VL CDR3 comprise the amino acid sequences set forth in SEQ ID NOs: 77, 146, 253, 329, 395, and 447, respectively; (h)(i) the VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2 and VL CDR3 comprise the amino acid sequences set forth in SEQ ID NOs: 82, 172, 257, 333, 398, and 450, respectively; (ii) the VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2 and VL CDR3 comprise the amino acid sequences set forth in SEQ ID NOs: 83, 173, 258, 334, 399, and 450, respectively; (iii) the VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2 and VL CDR3 comprise the amino acid sequences set forth in SEQ ID NOs: 84, 172, 257, 333, 398, and 450, respectively; (iv) the VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2 and VL CDR3 comprise the amino acid sequences set forth in SEQ ID NOs: 49, 174, 259, 335, 399 and 451, respectively; (v) the VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2 and VL CDR3 comprise the amino acid sequences set forth in SEQ ID NOs: 85, 175, 260, 336, 400 and 452, respectively; or (vi) the VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2 and VL CDR3 comprise the amino acid sequences set forth in SEQ ID NOs: 82, 176, 257, 333, 398, and 450, respectively; (i) the VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2 and VL CDR3 comprise the amino acid sequences set forth in SEQ ID NOs: 100, 166, 249, 325, 393 and 444, respectively; (ii) the VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2 and VL CDR3 comprise the amino acid sequences set forth in SEQ ID NOs: 101, 167, 250, 326, 386, and 444, respectively; (iii) the VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2 and VL CDR3 comprise the amino acid sequences set forth in SEQ ID NOs: 102, 166, 249, 325, 393 and 444, respectively; (iv) the VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2 and VL CDR3 comprise the amino acid sequences set forth in SEQ ID NOs: 103, 168, 251, 327, 386, and 445, respectively; (v) the VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2 and VL CDR3 comprise the amino acid sequences set forth in SEQ ID NOs: 104, 169, 252, 328, 407, and 446, respectively; or (vi) the VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2 and VL CDR3 comprise the amino acid sequences set forth in SEQ ID NOs: 100, 170, 249, 325, 393 and 444, respectively; (j)(i) the VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2 and VL CDR3 comprise the amino acid sequences set forth in SEQ ID NOs: 105, 191, 273, 349, 408, and 462, respectively; (ii) the VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2 and VL CDR3 comprise the amino acid sequences set forth in SEQ ID NOs: 106, 192, 274, 350, 389, and 462, respectively; (iii) the VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2 and VL CDR3 comprise the amino acid sequences set forth in SEQ ID NOs: 107, 191, 273, 349, 408, and 462, respectively; (iv) the VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2 and VL CDR3 comprise the amino acid sequences set forth in SEQ ID NOs: 108, 193, 275, 351, 389 and 463, respectively; (v) the VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2 and VL CDR3 comprise the amino acid sequences set forth in SEQ ID NOs: 109, 194, 276, 352, 409 and 464, respectively; or (vi) the VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2 and VL CDR3 comprise the amino acid sequences set forth in SEQ ID NOs: 105, 195, 273, 349, 408, and 462, respectively; (k)(i) the VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2 and VL CDR3 comprise the amino acid sequences set forth in SEQ ID NOs: 110, 196, 277, 353, 410, and 465, respectively; (ii) the VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2 and VL CDR3 comprise the amino acid sequences set forth in SEQ ID NOs: 111, 197, 278, 354, 411, and 465, respectively; (iii) the VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2 and VL CDR3 comprise the amino acid sequences set forth in SEQ ID NOs: 112, 196, 277, 353, 410 and 465, respectively; (iv) the VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2 and VL CDR3 comprise the amino acid sequences set forth in SEQ ID NOs: 113, 198, 279, 355, 411, and 466, respectively; (v) the VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2 and VL CDR3 comprise the amino acid sequences set forth in SEQ ID NOs: 114, 199, 280, 356, 412 and 467, respectively; or (vi) the VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2 and VL CDR3 comprise the amino acid sequences set forth in SEQ ID NOs: 110, 200, 277, 353, 410 and 465, respectively; (l)(i) the VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2 and VL CDR3 comprise the amino acid sequences set forth in SEQ ID NOs: 115, 201, 281, 357, 413, and 468, respectively; (ii) the VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2 and VL CDR3 comprise the amino acid sequences set forth in SEQ ID NOs: 116, 202, 282, 358, 414 and 468, respectively; (iii) the VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2 and VL CDR3 comprise the amino acid sequences set forth in SEQ ID NOs: 117, 201, 281, 357, 413, and 468, respectively; (iv) the VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2 and VL CDR3 comprise the amino acid sequences set forth in SEQ ID NOs: 118, 134, 283, 359, 414 and 469, respectively; (v) the VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2 and VL CDR3 comprise the amino acid sequences set forth in SEQ ID NOs: 119, 203, 284, 360, 415 and 470, respectively; or (vi) the VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2 and VL CDR3 comprise the amino acid sequences set forth in SEQ ID NOs: 115, 204, 281, 357, 413 and 468, respectively; or (m)(i) the VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2 and VL CDR3 comprise the amino acid sequences set forth in SEQ ID NOs: 120, 205, 285, 361, 416, and 471, respectively; (ii) the VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2 and VL CDR3 comprise the amino acid sequences set forth in SEQ ID NOs: 121, 206, 286, 362, 411, and 471, respectively; (iii) the VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2 and VL CDR3 comprise the amino acid sequences set forth in SEQ ID NOs: 122, 205, 285, 361, 416, and 471, respectively; (iv) the VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2 and VL CDR3 comprise the amino acid sequences set forth in SEQ ID NOs: 80, 207, 287, 363, 411, and 472, respectively; (v) the VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2 and VL CDR3 comprise the amino acid sequences set forth in SEQ ID NOs: 123, 208, 288, 364, 417 and 473, respectively; or (vi) the VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2 and VL CDR3 comprise the amino acid sequences set forth in SEQ ID NOs: 120, 209, 285, 361, 416, and 471, respectively; The antibody or fragment thereof described in claim 1.

3. (i) the VH region is at least 80% identical to the amino acid sequence set forth in SEQ ID NO:11, and the VL region is at least 80% identical to the amino acid sequence set forth in SEQ ID NO:12; (ii) the VH region is at least 80% identical to the amino acid sequence set forth in SEQ ID NO: 15, and the VL region is at least 80% identical to the amino acid sequence set forth in SEQ ID NO: 16; (iii) the VH region is at least 80% identical to the amino acid sequence set forth in SEQ ID NO:1, and the VL region is at least 80% identical to the amino acid sequence set forth in SEQ ID NO:2; (iv) the VH region is at least 80% identical to the amino acid sequence set forth in SEQ ID NO:5, and the VL region is at least 80% identical to the amino acid sequence set forth in SEQ ID NO:6; (v) the VH region is at least 80% identical to the amino acid sequence set forth in SEQ ID NO:9, and the VL region is at least 80% identical to the amino acid sequence set forth in SEQ ID NO:10; (vi) the VH region is at least 80% identical to the amino acid sequence set forth in SEQ ID NO: 13, and the VL region is at least 80% identical to the amino acid sequence set forth in SEQ ID NO: 14; (vii) the VH region is at least 80% identical to the amino acid sequence set forth in SEQ ID NO: 17, and the VL region is at least 80% identical to the amino acid sequence set forth in SEQ ID NO: 18; (viii) the VH region is at least 80% identical to the amino acid sequence set forth in SEQ ID NO: 19, and the VL region is at least 80% identical to the amino acid sequence set forth in SEQ ID NO: 20; (ix) the VH region is at least 80% identical to the amino acid sequence set forth in SEQ ID NO:27, and the VL region is at least 80% identical to the amino acid sequence set forth in SEQ ID NO:28; (x) the VH region is at least 80% identical to the amino acid sequence set forth in SEQ ID NO:29, and the VL region is at least 80% identical to the amino acid sequence set forth in SEQ ID NO:30; (xi) the VH region is at least 80% identical to the amino acid sequence set forth in SEQ ID NO: 31, and the VL region is at least 80% identical to the amino acid sequence set forth in SEQ ID NO: 32; (xii) the VH region is at least 80% identical to the amino acid sequence set forth in SEQ ID NO: 33, and the VL region is at least 80% identical to the amino acid sequence set forth in SEQ ID NO: 34; or (xiii) the VH region is at least 80% identical to the amino acid sequence set forth in SEQ ID NO: 35, and the VL region is at least 80% identical to the amino acid sequence set forth in SEQ ID NO: 36; 3. The antibody or fragment thereof according to claim 1 or 2.

4. (i) the VH region comprises the amino acid sequence set forth in SEQ ID NO:11, and the VL region comprises the amino acid sequence set forth in SEQ ID NO:12; (ii) the VH region comprises an amino acid sequence set forth in any one of SEQ ID NOs: 15 and 1936-1941, and the VL region comprises an amino acid sequence set forth in any one of SEQ ID NOs: 16 and 1943-1947; (iii) the VH region comprises an amino acid sequence set forth in any one of SEQ ID NOs:1 and 1919-1927, and the VL region comprises an amino acid sequence set forth in any one of SEQ ID NOs:2 and 1929-1934; (iv) the VH region comprises the amino acid sequence set forth in SEQ ID NO:5, and the VL region comprises the amino acid sequence set forth in SEQ ID NO:6; (v) the VH region comprises the amino acid sequence set forth in SEQ ID NO:9, and the VL region comprises the amino acid sequence set forth in SEQ ID NO:10; (vi) the VH region comprises an amino acid sequence set forth in any one of SEQ ID NOs: 13 and 1949-1952, and the VL region comprises an amino acid sequence set forth in any one of SEQ ID NOs: 14 and 1954-1956; (vii) the VH region comprises the amino acid sequence set forth in SEQ ID NO: 17, and the VL region comprises the amino acid sequence set forth in SEQ ID NO: 18; (viii) the VH region comprises the amino acid sequence set forth in SEQ ID NO: 19, and the VL region comprises the amino acid sequence set forth in SEQ ID NO: 20; (ix) the VH region comprises the amino acid sequence set forth in SEQ ID NO:27, and the VL region comprises the amino acid sequence set forth in SEQ ID NO:28; (x) the VH region comprises the amino acid sequence set forth in SEQ ID NO:29, and the VL region comprises the amino acid sequence set forth in SEQ ID NO:30; (xi) the VH region comprises the amino acid sequence set forth in SEQ ID NO: 31, and the VL region comprises the amino acid sequence set forth in SEQ ID NO: 32; (xii) the VH region comprises the amino acid sequence set forth in SEQ ID NO: 33, and the VL region comprises the amino acid sequence set forth in SEQ ID NO: 34; or (xiii) the VH region comprises the amino acid sequence set forth in SEQ ID NO: 35, and the VL region comprises the amino acid sequence set forth in SEQ ID NO: 36; 3. The antibody or fragment thereof according to claim 1 or 2.

5. The antibody or fragment thereof according to any one of claims 1 to 4, which is (a) a human IgG1 antibody, a human IgG2 antibody, or a human IgG4 antibody; and (b) comprises a human kappa light chain constant region or a human lambda light chain constant region.

6. The fragment, wherein the fragment is a Fab, Fab', F(ab') fragment formed from an antibody fragment. 2 , Fv, scFv, (scFv) 2 5. The antibody or fragment thereof according to any one of claims 1 to 4, which is a single chain antibody molecule, a dual variable region antibody, or a multispecific antibody.

7. An isolated polynucleotide or polynucleotides encoding the antibody or fragment thereof of any one of claims 1 to 6.

8. 8. A vector or vectors comprising the polynucleotide or polynucleotides of claim 7.

9. A host cell comprising the polynucleotide or polynucleotides of claim 7, or the vector or vectors of claim 8.

10. A host cell producing the antibody or fragment thereof according to any one of claims 1 to 6.

11. A pharmaceutical composition comprising the antibody or fragment thereof according to any one of claims 1 to 6.

12. A pharmaceutical composition for treating involuntary weight loss in a human subject, comprising the antibody or fragment thereof according to any one of claims 1 to 6.

13. A pharmaceutical composition for preventing involuntary weight loss in a human subject, comprising the antibody or fragment thereof according to any one of claims 1 to 6.

14. 14. The pharmaceutical composition of claim 12 or 13, wherein the human subject has cachexia, anorexia nervosa, or a chronic disease.

15. 15. The pharmaceutical composition of claim 14, wherein the chronic disease is selected from the group consisting of cirrhosis of the liver, hyperthyroidism, Parkinson's disease, cancer, chronic kidney disease, chronic obstructive pulmonary disease, AIDS, tuberculosis, chronic inflammatory diseases, sepsis, muscle wasting, and anorexia nervosa.

16. A pharmaceutical composition for treating cachexia, anorexia nervosa, or a GDF15-mediated disease or disorder in a human subject, comprising the antibody or fragment thereof according to any one of claims 1 to 6.

17. 17. The pharmaceutical composition of claim 16, wherein the GDF15-mediated disease or disorder is sarcopenia, muscle wasting, bone wasting, cardiovascular disease, or chronic inflammatory disease.

18. A pharmaceutical composition for treating cancer in a human subject with elevated GDF15 levels, comprising an antibody or fragment thereof according to any one of claims 1 to 6.

19. 19. The pharmaceutical composition of claim 18, wherein the cancer is melanoma, gastric cancer, pancreatic cancer or prostate cancer.

20. 19. The pharmaceutical composition of claim 18, wherein the cancer is pancreatic cancer.

21. 21. The pharmaceutical composition of any one of claims 12 to 20, comprising a combination therapy with one or more additional therapeutic agents.

22. A pharmaceutical composition for use in diagnosing a disorder associated with expression of GFRAL in a human subject, comprising the antibody or fragment thereof according to any one of claims 1 to 6, wherein the antibody or fragment thereof is conjugated to a detectable agent, and the diagnosis comprises (a)(i) administering the pharmaceutical composition to the human subject, or (ii) contacting a sample obtained from the human subject with the pharmaceutical composition, and (b) confirming the presence or localization of the antibody or fragment thereof in the human subject or sample.

23. A method for producing an antibody or fragment thereof according to any one of claims 1 to 6, comprising culturing a host cell according to claim 9 or 10, and purifying the antibody or fragment thereof.

24. 24. A method for producing a sterile pharmaceutical composition comprising formulating an antibody or fragment thereof produced by the method of claim 23 as a sterile pharmaceutical composition.

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