Anti-CD26 proteins and their uses

Abstract

Provided herein are proteins comprising an anti-CD26 antigen-binding domain. Also provided herein are methods for treating an age-related or inflammatory disease in a subject, comprising administering to the subject one of these proteins or a cell expressing one of these proteins.

Description

[Technical Field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims priority to U.S. Provisional Patent Application No. 63 / 017,467, filed April 29, 2020, which is incorporated herein by reference in its entirety.

[0002] Technical Field The present disclosure relates to the field of biotechnology, and more particularly to anti-CD26 antibodies and their uses. [Background technology]

[0003] background CD26 (also known as DPP4, dipeptidyl-peptidase-4, DDP4) is a transmembrane glycoprotein that is membrane-anchored by its signal peptide and forms homodimers or tetramers on the plasma membrane. CD26 is an aminopeptidase that cleaves N-terminal dipeptides primarily from peptides or small proteins (e.g., less than 80–100 amino acid residues) using proline or alanine as the penultimate amino acid.

[0004] CD26 is expressed in numerous tissues, including the brush border membrane of the intestine and kidney, vascular endothelium, liver and pancreas, glandular epithelial cells, and by cells of the immune system (Gutschmidt et al., Histochemistry 73(2):285-304, 1982 (Non-Patent Document 1); Gorrell et al., Cell. Immunol. 134(1):205-215, 1991 (Non-Patent Document 2); Tanaka et al., J. Immunol. 149(2):481-486, 1992 (Non-Patent Document 3); Abbott et al., Immunogentics 40(5):331-338, 1994 (Non-Patent Document 4); Buhling et al., Immunol. Lett. 45(1-2):47-51, 1995 (Non-Patent Document 5); Dikov et al., Cell. Mol. Biol. 50 Online Pub:OL565-568, 2004 (Non-Patent Document 6); Broxmeyer et al., Stem Cells Dev. 25(8):575-585, 2016 (Non-Patent Document 7); Hollande et al., Nat. Immunol. 20(3):257-264, 2019 (Non-Patent Document 8). CD26 has also been found to function as a binding site for the chemokine CXCR4 receptor, the T cell differentiation antigen CD45, and the sodium-hydrogen exchanger-3 (Mentlein et al., Regul. Pept. 85(1):9-24, 1999 (Non-Patent Document 9); Lambeir et al., Crit. Rev. Clin. Lab. Sci. 40(3):209-294, 2003 (Non-Patent Document 10)). Therefore, CD26 can be considered a multifunctional protein with various functions beyond its role as a proteinase. Its role as a receptor or ligand for a variety of different molecules, alone or in combination with its enzymatic activity, enables it to influence physiological processes such as the interactions between cells and the extracellular matrix involved in cell migration, activation, and proliferation.

[0005] CD26 also plays a key role in glucose metabolism. Incretin peptides, such as gastric inhibitory polypeptide (GIP) and glucagon-like peptide (GLP-1), stimulate insulin secretion from pancreatic β cells and, through the vasoconstrictive effects of glucagon, are involved in the regulation of postprandial blood glucose. These peptides are rapidly inactivated by CD26, resulting in a short half-life. In addition to incretin peptides, CD26 also cleaves many other proteins. Physiological targets include GLP1, GLP2, brain natriuretic peptide, peptide YY, stromal cell-derived factor, erythropoietin, granulocyte colony-stimulating factor, and substance P. Pharmacological targets include gastric releasing peptide, growth hormone-releasing factor, macrophage-derived chemokine, eotaxin, IFN-γ-inducible protein-10, granulocyte-macrophage colony-stimulating factor, erythropoietin, IL-3, neuropeptide Y, B-type natriuretic peptide, and peptide YY (Mulvihill et al., Endocr. Rev. 35(6):992-1019, 2014). Furthermore, CD26 is known to regulate the function of chemokines such as CXCR3 through post-translational cleavage of X-Pro or X-Ala motifs, resulting in amino-terminal dipeptide cleavage of the chemokine and altering its biological function (Broxmeyer et al., Stem Cells Dev. 25(8):575-585, 2016). CD26 also mediates amino-terminal cleavage of the chemokine CXCL10, restricting the migration of CXCR3 natural killer (NK) cells and T cells and limiting the decline of antitumor immunity in preclinical models of melanoma and colorectal cancer (Barreira et al., Nat. Immunol. 16(8):850-858, 2015). Combination immunotherapy using checkpoint blockade in the presence of the CD26 inhibitor sitagliptin has also been shown to reduce tumor growth by enhancing the antitumor activity of T cells and eosinophils in preclinical mouse models of hepatocellular carcinoma and breast cancer (Hollande et al., Nat. Immunol. 20(3):257-264, 2019).

[0006] In summary, CD26 exerts its physiological role through its enzymatic activity by regulating many peptides or through interactions with various binding partners. Consequently, changes in CD26 expression and / or activity have been implicated in several pathological processes, including inflammation, viral infection, immune-mediated diseases, tumor growth, cellular senescence, and metabolic diseases (Mentlein et al., Regul. Pept. 85(1):9-24, 1999; Lambeir et al., Crit. Rev. Clin. Lab. Sci. 40(3):209-294, 2003; Yu et al., FEBS J. 277(5):1126-1144, 2010; Kim et al., Genes Dev. 31(15):1529-1534, 2017 (Non-Patent Document 16), Deacon, Front. Endocrinol. 10:80, 2019 (Non-Patent Document 17). Therefore, CD26 is a cell surface target protein for drug development to treat various diseases, including viral infections and age-related pathologies. [Prior art documents] [Non-patent literature]

[0007] [Non-Patent Document 1] Gutschmidt et al., Histochemistry 73(2):285-304,1982 [Non-patent document 2] Gorrell et al.,Cell.Immunol.134(1):205-215,1991 [Non-patent document 3] Tanaka et al., J.Immunol.149(2):481-486,1992 [Non-patent document 4] Abbott et al., Immunogentics 40(5):331-338,1994 [Non-Patent Document 5] Buhling et al.,Immunol.Lett.45(1-2):47-51,1995

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Non-Patent Document 16

Non-Patent Document 17

Summary of the Invention

[0008] overview Provided herein is a protein comprising an anti-CD26 antigen-binding domain, the anti-CD26 antigen-binding domain comprising: (a) a heavy chain variable domain comprising CDR1 comprising SEQ ID NO: 1, CDR2 comprising SEQ ID NO: 2, and CDR3 comprising SEQ ID NO: 3, and a light chain variable domain comprising CDR1 comprising SEQ ID NO: 4, CDR2 comprising SEQ ID NO: 5, and CDR3 comprising SEQ ID NO: 6; (b) a heavy chain variable domain comprising CDR1 comprising SEQ ID NO: 7, CDR2 comprising SEQ ID NO: 8, and CDR3 comprising SEQ ID NO: 9, and a CDR1 comprising SEQ ID NO: 10, CDR2 comprising SEQ ID NO: 11, and a light chain variable domain comprising CDR1 comprising SEQ ID NO: 12, CDR2 comprising SEQ ID NO: 13, and a CDR3 comprising SEQ ID NO: 14; (c) a light chain variable domain comprising CDR1 comprising SEQ ID NO: 13, CDR2 comprising SEQ ID NO: 14, and CDR3 comprising SEQ ID NO: 15, and a light chain variable domain comprising CDR1 comprising SEQ ID NO: 16, CDR2 comprising SEQ ID NO: 17, and CDR3 comprising SEQ ID NO: 18; (d) a heavy chain variable domain comprising CDR1 comprising SEQ ID NO: 19, CDR2 comprising SEQ ID NO: 20, and CDR3 comprising SEQ ID NO: 21, and a CDR1 comprising SEQ ID NO: 22, CDR2 comprising SEQ ID NO: 23, and CDR3 comprising SEQ ID NO: 24. (e) a light chain variable domain comprising a CDR1 comprising SEQ ID NO: 25, a CDR2 comprising SEQ ID NO: 26, and a CDR3 comprising SEQ ID NO: 27, and a light chain variable domain comprising a CDR1 comprising SEQ ID NO: 28, a CDR2 comprising SEQ ID NO: 29, and a CDR3 comprising SEQ ID NO: 30; (f) a heavy chain variable domain comprising a CDR1 comprising SEQ ID NO: 31, a CDR2 comprising SEQ ID NO: 32, and a CDR3 comprising SEQ ID NO: 33, and a CDR1 comprising SEQ ID NO: 34, a CDR2 comprising SEQ ID NO: 35, and a CDR3 comprising SEQ ID NO: 36. (g) a heavy chain variable domain comprising CDR1 comprising SEQ ID NO: 37, CDR2 comprising SEQ ID NO: 38, and CDR3 comprising SEQ ID NO: 39, and a light chain variable domain comprising CDR1 comprising SEQ ID NO: 40, CDR2 comprising SEQ ID NO: 41, and CDR3 comprising SEQ ID NO: 42; (h) a heavy chain variable domain comprising CDR1 comprising SEQ ID NO: 43, CDR2 comprising SEQ ID NO: 44, and CDR3 comprising SEQ ID NO: 45, and a light chain variable domain comprising CDR1 comprising SEQ ID NO: 46, CDR2 comprising SEQ ID NO: 47, and CDR3 comprising SEQ ID NO: 48;(i) a heavy chain variable domain comprising CDR1 comprising SEQ ID NO: 49, CDR2 comprising SEQ ID NO: 50, and CDR3 comprising SEQ ID NO: 51, and a light chain variable domain comprising CDR1 comprising SEQ ID NO: 52, CDR2 comprising SEQ ID NO: 53, and CDR3 comprising SEQ ID NO: 54; or (j) a heavy chain variable domain comprising CDR1 comprising SEQ ID NO: 55, CDR2 comprising SEQ ID NO: 56, and CDR3 comprising SEQ ID NO: 57, and a light chain variable domain comprising CDR1 comprising SEQ ID NO: 58, CDR2 comprising SEQ ID NO: 59, and CDR3 comprising SEQ ID NO: 60.

[0009] In some embodiments, the antigen-binding domain comprises a heavy chain variable domain comprising CDR1 comprising SEQ ID NO: 1, CDR2 comprising SEQ ID NO: 2, and CDR3 comprising SEQ ID NO: 3, and a light chain variable domain comprising CDR1 comprising SEQ ID NO: 4, CDR2 comprising SEQ ID NO: 5, and CDR3 comprising SEQ ID NO: 6. In some embodiments, the heavy chain variable domain comprises a sequence at least 80% identical to SEQ ID NO: 61. In some embodiments, the heavy chain variable domain comprises a sequence at least 90% identical to SEQ ID NO: 61. In some embodiments, the heavy chain variable domain comprises a sequence at least 95% identical to SEQ ID NO: 61. In some embodiments, the light chain variable domain comprises a sequence at least 80% identical to SEQ ID NO: 62. In some embodiments, the light chain variable domain comprises a sequence at least 90% identical to SEQ ID NO: 62. In some embodiments, the light chain variable domain comprises a sequence at least 95% identical to SEQ ID NO: 62.

[0010] In some embodiments, the antigen binding domain comprises a heavy chain variable domain comprising CDR1 comprising SEQ ID NO:7, CDR2 comprising SEQ ID NO:8, and CDR3 comprising SEQ ID NO:9, and a light chain variable domain comprising CDR1 comprising SEQ ID NO:10, CDR2 comprising SEQ ID NO:11, and CDR3 comprising SEQ ID NO:12. In some embodiments, the heavy chain variable domain comprises a sequence at least 80% identical to SEQ ID NO:63. In some embodiments, the heavy chain variable domain comprises a sequence at least 90% identical to SEQ ID NO:63. In some embodiments, the heavy chain variable domain comprises a sequence at least 95% identical to SEQ ID NO:63. In some embodiments, the light chain variable domain comprises a sequence at least 80% identical to SEQ ID NO:64. In some embodiments, the light chain variable domain comprises a sequence at least 90% identical to SEQ ID NO:64. In some embodiments, the light chain variable domain comprises a sequence at least 95% identical to SEQ ID NO:64.

[0011] In some embodiments, the antigen-binding domain comprises a heavy chain variable domain comprising CDR1 comprising SEQ ID NO: 13, CDR2 comprising SEQ ID NO: 14, and CDR3 comprising SEQ ID NO: 15, and a light chain variable domain comprising CDR1 comprising SEQ ID NO: 16, CDR2 comprising SEQ ID NO: 17, and CDR3 comprising SEQ ID NO: 18. In some embodiments, the heavy chain variable domain comprises a sequence at least 80% identical to SEQ ID NO: 65. In some embodiments, the heavy chain variable domain comprises a sequence at least 90% identical to SEQ ID NO: 65. In some embodiments, the heavy chain variable domain comprises a sequence at least 95% identical to SEQ ID NO: 65. In some embodiments, the light chain variable domain comprises a sequence at least 80% identical to SEQ ID NO: 66. In some embodiments, the light chain variable domain comprises a sequence at least 90% identical to SEQ ID NO: 66. In some embodiments, the light chain variable domain comprises a sequence at least 95% identical to SEQ ID NO: 66.

[0012] In some embodiments, the antigen-binding domain comprises a heavy chain variable domain comprising CDR1 comprising SEQ ID NO: 19, CDR2 comprising SEQ ID NO: 20, and CDR3 comprising SEQ ID NO: 21, and a light chain variable domain comprising CDR1 comprising SEQ ID NO: 22, CDR2 comprising SEQ ID NO: 23, and CDR3 comprising SEQ ID NO: 24. In some embodiments, the heavy chain variable domain comprises a sequence at least 80% identical to SEQ ID NO: 67. In some embodiments, the heavy chain variable domain comprises a sequence at least 90% identical to SEQ ID NO: 67. In some embodiments, the heavy chain variable domain comprises a sequence at least 95% identical to SEQ ID NO: 67. In some embodiments, the light chain variable domain comprises a sequence at least 80% identical to SEQ ID NO: 68. In some embodiments, the light chain variable domain comprises a sequence at least 90% identical to SEQ ID NO: 68. In some embodiments, the light chain variable domain comprises a sequence at least 95% identical to SEQ ID NO: 68.

[0013] In some embodiments, the antigen binding domain comprises a heavy chain variable domain comprising CDR1 comprising SEQ ID NO:25, CDR2 comprising SEQ ID NO:26, and CDR3 comprising SEQ ID NO:27, and a light chain variable domain comprising CDR1 comprising SEQ ID NO:28, CDR2 comprising SEQ ID NO:29, and CDR3 comprising SEQ ID NO:30. In some embodiments, the heavy chain variable domain comprises a sequence at least 80% identical to SEQ ID NO:69. In some embodiments, the heavy chain variable domain comprises a sequence at least 90% identical to SEQ ID NO:69. In some embodiments, the heavy chain variable domain comprises a sequence at least 95% identical to SEQ ID NO:69. In some embodiments, the light chain variable domain comprises a sequence at least 80% identical to SEQ ID NO:70. In some embodiments, the light chain variable domain comprises a sequence at least 90% identical to SEQ ID NO:70. In some embodiments, the light chain variable domain comprises a sequence at least 95% identical to SEQ ID NO:70.

[0014] In some embodiments, the antigen binding domain comprises a heavy chain variable domain comprising CDR1 comprising SEQ ID NO: 31, CDR2 comprising SEQ ID NO: 32, and CDR3 comprising SEQ ID NO: 33, and a light chain variable domain comprising CDR1 comprising SEQ ID NO: 34, CDR2 comprising SEQ ID NO: 35, and CDR3 comprising SEQ ID NO: 36. In some embodiments, the heavy chain variable domain comprises a sequence at least 80% identical to SEQ ID NO: 71. In some embodiments, the heavy chain variable domain comprises a sequence at least 90% identical to SEQ ID NO: 71. In some embodiments, the heavy chain variable domain comprises a sequence at least 95% identical to SEQ ID NO: 71. In some embodiments, the light chain variable domain comprises a sequence at least 80% identical to SEQ ID NO: 72. In some embodiments, the light chain variable domain comprises a sequence at least 90% identical to SEQ ID NO: 72. In some embodiments, the light chain variable domain comprises a sequence at least 95% identical to SEQ ID NO: 72.

[0015] In some embodiments, the antigen-binding domain comprises a heavy chain variable domain comprising CDR1 comprising SEQ ID NO: 37, CDR2 comprising SEQ ID NO: 38, and CDR3 comprising SEQ ID NO: 39, and a light chain variable domain comprising CDR1 comprising SEQ ID NO: 40, CDR2 comprising SEQ ID NO: 41, and CDR3 comprising SEQ ID NO: 42. In some embodiments, the heavy chain variable domain comprises a sequence at least 80% identical to SEQ ID NO: 73. In some embodiments, the heavy chain variable domain comprises a sequence at least 90% identical to SEQ ID NO: 73. In some embodiments, the heavy chain variable domain comprises a sequence at least 95% identical to SEQ ID NO: 73. In some embodiments, the light chain variable domain comprises a sequence at least 80% identical to SEQ ID NO: 74. In some embodiments, the light chain variable domain comprises a sequence at least 90% identical to SEQ ID NO: 74. In some embodiments, the light chain variable domain comprises a sequence at least 95% identical to SEQ ID NO: 74.

[0016] In some embodiments, the antigen binding domain comprises a heavy chain variable domain comprising CDR1 comprising SEQ ID NO: 43, CDR2 comprising SEQ ID NO: 44, and CDR3 comprising SEQ ID NO: 45, and a light chain variable domain comprising CDR1 comprising SEQ ID NO: 46, CDR2 comprising SEQ ID NO: 47, and CDR3 comprising SEQ ID NO: 48. In some embodiments, the heavy chain variable domain comprises a sequence at least 80% identical to SEQ ID NO: 75. In some embodiments, the heavy chain variable domain comprises a sequence at least 90% identical to SEQ ID NO: 75. In some embodiments, the heavy chain variable domain comprises a sequence at least 95% identical to SEQ ID NO: 75. In some embodiments, the light chain variable domain comprises a sequence at least 80% identical to SEQ ID NO: 76. In some embodiments, the light chain variable domain comprises a sequence at least 90% identical to SEQ ID NO: 76. In some embodiments, the light chain variable domain comprises a sequence at least 95% identical to SEQ ID NO: 76.

[0017] In some embodiments, the antigen binding domain comprises a heavy chain variable domain comprising CDR1 comprising SEQ ID NO:49, CDR2 comprising SEQ ID NO:50, and CDR3 comprising SEQ ID NO:51, and a light chain variable domain comprising CDR1 comprising SEQ ID NO:52, CDR2 comprising SEQ ID NO:53, and CDR3 comprising SEQ ID NO:54. In some embodiments, the heavy chain variable domain comprises a sequence at least 80% identical to SEQ ID NO:77. In some embodiments, the heavy chain variable domain comprises a sequence at least 90% identical to SEQ ID NO:77. In some embodiments, the heavy chain variable domain comprises a sequence at least 95% identical to SEQ ID NO:77. In some embodiments, the light chain variable domain comprises a sequence at least 80% identical to SEQ ID NO:78. In some embodiments, the light chain variable domain comprises a sequence at least 90% identical to SEQ ID NO:78. In some embodiments, the light chain variable domain comprises a sequence at least 95% identical to SEQ ID NO:78.

[0018] In some embodiments, the antigen binding domain comprises a heavy chain variable domain comprising CDR1 comprising SEQ ID NO:55, CDR2 comprising SEQ ID NO:56, and CDR3 comprising SEQ ID NO:57, and a light chain variable domain comprising CDR1 comprising SEQ ID NO:58, CDR2 comprising SEQ ID NO:59, and CDR3 comprising SEQ ID NO:60. In some embodiments, the heavy chain variable domain comprises a sequence at least 80% identical to SEQ ID NO:79. In some embodiments, the heavy chain variable domain comprises a sequence at least 90% identical to SEQ ID NO:79. In some embodiments, the heavy chain variable domain comprises a sequence at least 95% identical to SEQ ID NO:79. In some embodiments, the light chain variable domain comprises a sequence at least 80% identical to SEQ ID NO:80. In some embodiments, the light chain variable domain comprises a sequence at least 90% identical to SEQ ID NO:80. In some embodiments, the light chain variable domain comprises a sequence at least 95% identical to SEQ ID NO:80.

[0019] In some embodiments, the protein is a multi-chain protein. In some embodiments, the protein is a single-chain protein. In some embodiments, the protein is an antibody or an antigen-binding antibody fragment. In some embodiments, the antibody is an IgG1 antibody, an IgG2 antibody, an IgG3 antibody, or an IgG4 antibody. In some embodiments, the antibody is humanized. In some embodiments, the antibody is human. In some embodiments, the protein is an scFv. In some embodiments, the protein is a chimeric antigen receptor (CAR).

[0020] Also provided herein is a pharmaceutical composition comprising any one of the proteins described herein and a pharmaceutically acceptable carrier. Also provided herein is a kit comprising any one of the pharmaceutical compositions described herein. Also provided herein is a nucleic acid encoding any one of the proteins described herein. Also provided herein is a vector comprising any one of the nucleic acids described herein. Also provided herein is a pharmaceutical composition comprising any one of the nucleic acids described herein or any one of the vectors described herein. Also provided herein is a kit comprising any one of the pharmaceutical compositions described herein.

[0021] Provided herein is a cell comprising any one of the nucleic acids described herein or any one of the vectors described herein. In some embodiments, the cell is an immune cell. In some embodiments, the immune cell is a T cell, a B cell, or a natural killer (NK) cell. In some embodiments, the immune cell is a regulatory T (Treg) cell. In some embodiments, the immune cell is an autologous cell. In some embodiments, the immune cell is an allogeneic cell.

[0022] Also provided herein is a pharmaceutical composition comprising any one of the cells described herein and a pharmaceutically acceptable carrier. Also provided herein is a kit comprising any one of the pharmaceutical compositions described herein.

[0023] Provided herein are methods for treating an age-related or inflammatory disease in a subject, comprising administering to the subject a therapeutically effective amount of any one of the proteins described herein or any one of the pharmaceutical compositions described herein. Also provided herein are methods for treating an age-related or inflammatory disease in a subject, comprising administering to the subject a therapeutically effective amount of any one of the nucleic acids described herein, any one of the vectors described herein, or any one of the pharmaceutical compositions described herein. Also provided herein are methods for treating an age-related or inflammatory disease in a subject, comprising administering to the subject a therapeutically effective amount of any one of the cells described herein or any one of the pharmaceutical compositions described herein.

[0024] In some embodiments, the age-related disease is inflammation-aging associated. In some embodiments, the subject is further administered (i) a therapeutically effective amount of an NK cell activator and / or NK cells and / or a monoclonal antibody, and / or (ii) a therapeutically effective amount of a Treg cell activator and / or Treg cells and / or a monoclonal antibody and / or an advanced glycation end products (AGE) inhibitor.

[0025] In some embodiments, the method includes administering to the subject a therapeutically effective amount of NK cells. In some embodiments, the NK cells are autologous, haploidentical, or allogeneic NK cells isolated from peripheral blood, umbilical cord blood, or isolated and differentiated from iPSCs. In some embodiments, the method further includes isolating NK cells from the subject and culturing the isolated NK cells in a liquid culture medium under conditions sufficient to induce or increase the proliferation of the NK cells, wherein after the isolation and culturing steps, the NK cells are administered to the subject. In some embodiments, the liquid culture medium comprises a multi-chain chimeric polypeptide. In some embodiments, the NK cells comprise a chimeric antigen receptor. In some embodiments, the protein is any one of the chimeric antigen receptors described herein.

[0026] In some embodiments, the method comprises administering to the subject a therapeutically effective amount of an NK cell activator and / or a monoclonal antibody. In some embodiments, the NK cell activator is one or more multi-chain chimeric polypeptides. In some embodiments, the monoclonal antibody is an anti-tissue factor antibody or any one of the antibodies described herein. In some embodiments, the NK cell activator comprises one or more multi-chain chimeric polypeptides and the monoclonal antibody comprises one or more of the anti-tissue factor antibodies and / or any one of the antibodies described herein.

[0027] In some embodiments, the method comprises administering to the subject a therapeutically effective amount of Treg cells. In some embodiments, the Treg cells are autologous, haploidentical, or allogeneic Treg cells isolated from peripheral blood or umbilical cord blood. In some embodiments, the method further comprises culturing the isolated Treg cells in a liquid culture medium under conditions sufficient to induce or increase the proliferation of Treg cells, wherein after the isolating and culturing steps, the Treg cells are administered to the subject. In some embodiments, the liquid culture medium comprises one or more single-chain chimeric polypeptides. In some embodiments, the Treg cells comprise a chimeric antigen receptor. In some embodiments, the chimeric antigen receptor comprises an extracellular domain that specifically binds to tissue factor, to CD26 (e.g., any of the anti-CD26 antigen-binding domains described herein), or to CD36. In some embodiments, the method comprises administering to the subject a therapeutically effective amount of a Treg cell activator and / or a monoclonal antibody and / or an AGE inhibitor. In some embodiments, the Treg cell activator is one or more single-chain chimeric polypeptides. In some embodiments, the monoclonal antibody is one or both of an anti-tissue factor antibody, an anti-CD26 antibody (e.g., any of the anti-CD26 antibodies described herein), and / or an anti-CD36 antibody. In some embodiments, the AGE inhibitor is a soluble RAGE trap. In some embodiments, the Treg cell activator comprises one or more single-chain chimeric polypeptides, the monoclonal antibody comprises one or more of an anti-tissue factor antibody, an anti-CD26 antibody (e.g., any of the anti-CD26 antibodies described herein), and / or an anti-CD36 antibody, and the AGE inhibitor comprises one or more soluble RAGE traps.

[0028] In some embodiments, the multi-chain chimeric polypeptide comprises: (a) a first chimeric polypeptide, the first chimeric polypeptide comprising (i) a first target binding domain, (ii) a soluble tissue factor domain, and (iii) a first domain of a pair of affinity domains; and (b) a second chimeric polypeptide, the second chimeric polypeptide comprising (i) a second domain of the pair of affinity domains, and (ii) a second target binding domain, wherein the first chimeric polypeptide and the second chimeric polypeptide are associated via binding between the first domain and the second domain of the pair of affinity domains.

[0029] In some embodiments, the single-chain chimeric polypeptide comprises (i) a first target binding domain, (ii) a soluble tissue factor domain, and (iii) a second target binding domain.

[0030] In some embodiments, the age-related disorder is selected from the group consisting of Alzheimer's disease, aneurysms, cystic fibrosis, fibrosis in pancreatitis, glaucoma, hypertension, idiopathic pulmonary fibrosis, inflammatory bowel disease, intervertebral disc degeneration, macular degeneration, osteoarthritis, type 2 diabetes mellitus, lipoatrophy, lipodystrophy, atherosclerosis, cataracts, COPD, idiopathic pulmonary fibrosis, renal transplant failure, liver fibrosis, bone loss, myocardial infarction, sarcopenia, wound healing, alopecia, cardiomyocyte hypertrophy, osteoarthritis, Parkinson's disease, age-related loss of lung tissue elasticity, macular degeneration, cachexia, glomerulosclerosis, liver cirrhosis, NAFLD, osteoporosis, amyotrophic lateral sclerosis, Huntington's disease, spinocerebellar ataxia, multiple sclerosis, neurodegeneration, stroke, cancer, dementia, vascular disease, susceptibility to infection, chronic inflammation, and renal dysfunction.

[0031] In some embodiments, the inflammatory disease is selected from the group consisting of rheumatoid arthritis, inflammatory bowel disease, lupus erythematosus, lupus nephritis, diabetic nephropathy, CNS injury, Alzheimer's disease, Parkinson's disease, amyotrophic lateral sclerosis, Crohn's disease, multiple sclerosis, Guillain-Barré syndrome, psoriasis, Graves' disease, ulcerative colitis, non-alcoholic steatohepatitis, and mood disorders.

[0032] In some embodiments, the age-related disease is a cancer selected from the group consisting of a solid tumor, a hematological tumor, a sarcoma, an osteosarcoma, a glioblastoma, a neuroblastoma, a melanoma, a rhabdomyosarcoma, an Ewing's sarcoma, an osteosarcoma, a B-cell neoplasm, multiple myeloma, a B-cell lymphoma, a B-cell non-Hodgkin's lymphoma, a Hodgkin's lymphoma, a chronic lymphocytic leukemia (CLL), an acute myeloid leukemia (AML), a chronic myeloid leukemia (CML), an acute lymphocytic leukemia (ALL), a myelodysplastic syndrome (MDS), a cutaneous T-cell lymphoma, a retinoblastoma, a gastric cancer, a urothelial cancer, a lung cancer, a renal cell carcinoma, a gastroesophageal cancer, a pancreatic cancer, a prostate cancer, a breast cancer, a colorectal cancer, an ovarian cancer, a non-small cell lung cancer, a squamous cell head and neck cancer, an endometrial cancer, a cervical cancer, a liver cancer, and a hepatocellular carcinoma.

[0033] Also provided herein are methods of treating cancer in a subject, comprising administering to the subject a therapeutically effective amount of any one of the proteins described herein.

[0034] Also provided herein are methods of treating an infectious disease in a subject, comprising administering to the subject a therapeutically effective amount of any one of the proteins described herein.

[0035] Also provided herein are methods of treating an infectious disease in a subject, comprising administering to the subject a therapeutically effective amount of any one of the proteins described herein or any one of the pharmaceutical compositions described herein.

[0036] An "antigen-binding domain" is one or more protein domains (e.g., formed from amino acids from a single polypeptide, or formed from amino acids from two or more polypeptides (e.g., the same or different polypeptides)) that can specifically bind to an antigen. In some examples, an antigen-binding domain can bind to an antigen or epitope with similar specificity and affinity as a naturally occurring antibody. In some embodiments, the antigen-binding domain may be an antibody or a fragment thereof. In some embodiments, the antigen-binding domain may comprise an alternative scaffold. Non-limiting examples of antigen-binding domains are described herein. Additional examples of antigen-binding domains are known in the art.

[0037] The term "antibody" is used herein in its broadest sense and includes certain types of immunoglobulin molecules that contain one or more antigen-binding domains that specifically bind to an antigen or epitope. Antibodies specifically include, for example, intact antibodies (e.g., intact immunoglobulins, such as human IgG (e.g., human IgG1, human IgG2, human IgG3, human IgG4)), antibody fragments, and multispecific antibodies. An example of an antigen-binding domain is an antigen-binding domain formed by a VH-VL dimer. Additional examples of antibodies are described herein. Additional examples of antibodies are known in the art.

[0038] "Affinity" refers to the strength of the sum of non-covalent interactions between an antigen-binding site and its binding partner (e.g., antigen or epitope). Unless otherwise indicated, as used herein, "affinity" refers to the intrinsic binding affinity, which reflects a 1:1 interaction between a member of the antigen-binding domain and the antigen or epitope. The affinity of a molecule X for its partner Y is determined by the dissociation equilibrium constant (K D) The kinetic components that contribute to the dissociation equilibrium constant are described in more detail below. Affinity can be measured by common methods known in the art, including those described herein. Affinity can be determined, for example, using surface plasmon resonance (SPR) technology (e.g., BIACORE®) or biolayer interferometry (e.g., FORTEBIO®). Additional methods for determining the affinity of an antigen-binding domain for its corresponding antigen or epitope are known in the art.

[0039] As used herein, a "single-chain protein" refers to a single protein chain.

[0040] As used herein, a "multi-chain protein" refers to a polypeptide that includes two or more (e.g., 3, 4, 5, 6, 7, 8, 9, or 10) protein chains (e.g., at least a first chimeric polypeptide and a second polypeptide), where the two or more protein chains associate via non-covalent bonds to form a quaternary structure.

[0041] The term "pair of affinity domains" refers to 1 × 10 -7 Less than M (e.g., 1 × 10 -8 Less than M, 1 x 10 -9 Less than M, 1 x 10 -10 Less than M or 1 x 10 -11 K (less than M) D A pair of affinity domains is two different protein domains that specifically bind to each other at a specific site. In some examples, the pair of affinity domains can be a pair of naturally occurring proteins. In some embodiments, the pair of affinity domains can be a pair of synthetic proteins. Non-limiting examples of affinity domain pairs are described herein.

[0042] The term "epitope" refers to a portion of an antigen that specifically binds to an antigen-binding domain. An epitope may consist of, for example, surface-accessible amino acid residues and / or sugar side chains and may have specific three-dimensional structural characteristics, as well as specific charge characteristics. Conformational and nonconformational epitopes are distinguished in that the binding to the former, but not the latter, may be lost in the presence of denaturing solvents. An epitope may include amino acid residues directly involved in binding and other amino acid residues not directly involved in binding. Methods for identifying the epitope to which an antigen-binding domain binds are known in the art.

[0043] The term "treatment" means ameliorating at least one symptom of a disorder. In some examples, the disorder being treated is cancer, and ameliorating at least one symptom of cancer includes reducing abnormal proliferation, gene expression, signaling, translation, and / or secretion of a factor. Generally, treatment methods involve administering to a subject in need of, or determined to be in need of, such treatment a therapeutically effective amount of a composition that alleviates at least one symptom of the disorder.

[0044] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Methods and materials are described herein for use in the present invention; other suitable methods and materials known in the art can also be used. The materials, methods, and examples are illustrative only and are not intended to be limiting. All publications, patent applications, patents, sequences, database entries, and other references mentioned herein are incorporated by reference in their entirety. In case of conflict, the present specification, including definitions, will control.

[0045] [The present invention 1001] A protein comprising an anti-CD26 antigen-binding domain, the anti-CD26 antigen-binding domain (a) a heavy chain variable domain comprising a CDR1 comprising SEQ ID NO: 1, a CDR2 comprising SEQ ID NO: 2, and a CDR3 comprising SEQ ID NO: 3; and a light chain variable domain, comprising CDR1 comprising SEQ ID NO: 4, CDR2 comprising SEQ ID NO: 5, and CDR3 comprising SEQ ID NO: 6; (b) a heavy chain variable domain comprising a CDR1 comprising SEQ ID NO: 7, a CDR2 comprising SEQ ID NO: 8, and a CDR3 comprising SEQ ID NO: 9; and a light chain variable domain comprising CDR1 comprising SEQ ID NO: 10, CDR2 comprising SEQ ID NO: 11, and CDR3 comprising SEQ ID NO: 12; (c) a heavy chain variable domain comprising a CDR1 comprising SEQ ID NO: 13, a CDR2 comprising SEQ ID NO: 14, and a CDR3 comprising SEQ ID NO: 15; and a light chain variable domain, comprising CDR1 comprising SEQ ID NO: 16, CDR2 comprising SEQ ID NO: 17, and CDR3 comprising SEQ ID NO: 18; (d) a heavy chain variable domain comprising a CDR1 comprising SEQ ID NO: 19, a CDR2 comprising SEQ ID NO: 20, and a CDR3 comprising SEQ ID NO: 21; and a light chain variable domain, comprising a CDR1 comprising SEQ ID NO: 22, a CDR2 comprising SEQ ID NO: 23, and a CDR3 comprising SEQ ID NO: 24; (e) a heavy chain variable domain comprising a CDR1 comprising SEQ ID NO: 25, a CDR2 comprising SEQ ID NO: 26, and a CDR3 comprising SEQ ID NO: 27; and a light chain variable domain, comprising CDR1 comprising SEQ ID NO: 28, CDR2 comprising SEQ ID NO: 29, and CDR3 comprising SEQ ID NO: 30; (f) a heavy chain variable domain comprising a CDR1 comprising SEQ ID NO: 31, a CDR2 comprising SEQ ID NO: 32, and a CDR3 comprising SEQ ID NO: 33; and a light chain variable domain, comprising a CDR1 comprising SEQ ID NO: 34, a CDR2 comprising SEQ ID NO: 35, and a CDR3 comprising SEQ ID NO: 36; (g) a heavy chain variable domain comprising a CDR1 comprising SEQ ID NO: 37, a CDR2 comprising SEQ ID NO: 38, and a CDR3 comprising SEQ ID NO: 39; and a light chain variable domain, comprising CDR1 comprising SEQ ID NO: 40, CDR2 comprising SEQ ID NO: 41, and CDR3 comprising SEQ ID NO: 42; (h) a heavy chain variable domain comprising a CDR1 comprising SEQ ID NO: 43, a CDR2 comprising SEQ ID NO: 44, and a CDR3 comprising SEQ ID NO: 45; and a light chain variable domain, comprising CDR1 comprising SEQ ID NO: 46, CDR2 comprising SEQ ID NO: 47, and CDR3 comprising SEQ ID NO: 48; (i) a heavy chain variable domain comprising a CDR1 comprising SEQ ID NO: 49, a CDR2 comprising SEQ ID NO: 50, and a CDR3 comprising SEQ ID NO: 51; and a light chain variable domain comprising a CDR1 comprising SEQ ID NO: 52, a CDR2 comprising SEQ ID NO: 53, and a CDR3 comprising SEQ ID NO: 54; or (j) a heavy chain variable domain comprising a CDR1 comprising SEQ ID NO: 55, a CDR2 comprising SEQ ID NO: 56, and a CDR3 comprising SEQ ID NO: 57; and a light chain variable domain comprising CDR1 comprising SEQ ID NO: 58, CDR2 comprising SEQ ID NO: 59, and CDR3 comprising SEQ ID NO: 60 The protein comprising: [The present invention 1002] the antigen-binding domain a heavy chain variable domain comprising CDR1 comprising SEQ ID NO: 1, CDR2 comprising SEQ ID NO: 2, and CDR3 comprising SEQ ID NO: 3, and a light chain variable domain comprising CDR1 comprising SEQ ID NO: 4, CDR2 comprising SEQ ID NO: 5, and CDR3 comprising SEQ ID NO: 6. 1001. The protein of the present invention, comprising: [The present invention 1003] 1002. The protein of the present invention, wherein said heavy chain variable domain comprises a sequence that is at least 80% identical to SEQ ID NO:61. [The present invention 1004] 1003. The protein of the present invention, wherein said heavy chain variable domain comprises a sequence that is at least 90% identical to SEQ ID NO:61. [The present invention 1005] 1004. The protein of the present invention, wherein said heavy chain variable domain comprises a sequence that is at least 95% identical to SEQ ID NO:61. [The present invention 1006] The protein of any of claims 1002 to 1005, wherein the light chain variable domain comprises a sequence that is at least 80% identical to SEQ ID NO: 62. [The present invention 1007] 1006. The protein of the present invention, wherein said light chain variable domain comprises a sequence that is at least 90% identical to SEQ ID NO:62. [The present invention 1008] 1007. The protein of the present invention, wherein said light chain variable domain comprises a sequence that is at least 95% identical to SEQ ID NO:62. [The present invention 1009] the antigen-binding domain a heavy chain variable domain comprising CDR1 comprising SEQ ID NO:7, CDR2 comprising SEQ ID NO:8, and CDR3 comprising SEQ ID NO:9, and a light chain variable domain comprising CDR1 comprising SEQ ID NO:10, CDR2 comprising SEQ ID NO:11, and CDR3 comprising SEQ ID NO:12. 1001. The protein of the present invention, comprising: [The present invention 1010] 1009. The protein of the present invention, wherein said heavy chain variable domain comprises a sequence that is at least 80% identical to SEQ ID NO:63. [The present invention 1011] 10. The protein of claim 10, wherein said heavy chain variable domain comprises a sequence that is at least 90% identical to SEQ ID NO:63. [The present invention 1012] 1011. The protein of the present invention, wherein said heavy chain variable domain comprises a sequence that is at least 95% identical to SEQ ID NO:63. [The present invention 1013] The protein of any of claims 1009 to 1012, wherein the light chain variable domain comprises a sequence that is at least 80% identical to SEQ ID NO: 64. [The present invention 1014] 1013. The protein of the present invention, wherein said light chain variable domain comprises a sequence that is at least 90% identical to SEQ ID NO:64. [The present invention 1015] 1014. The protein of the present invention, wherein said light chain variable domain comprises a sequence that is at least 95% identical to SEQ ID NO:64. [The present invention 1016] the antigen-binding domain a heavy chain variable domain comprising CDR1 comprising SEQ ID NO: 13, CDR2 comprising SEQ ID NO: 14, and CDR3 comprising SEQ ID NO: 15, and a light chain variable domain comprising CDR1 comprising SEQ ID NO: 16, CDR2 comprising SEQ ID NO: 17, and CDR3 comprising SEQ ID NO: 18. 1001. The protein of the present invention, comprising: [The present invention 1017] 1016. The protein of the present invention, wherein said heavy chain variable domain comprises a sequence that is at least 80% identical to SEQ ID NO:65. [The present invention 1018] 1017. The protein of the present invention, wherein said heavy chain variable domain comprises a sequence that is at least 90% identical to SEQ ID NO:65. [The present invention 1019] 1018. The protein of the present invention, wherein said heavy chain variable domain comprises a sequence that is at least 95% identical to SEQ ID NO:65. [The present invention 1020] The protein of any of claims 1016 to 1019, wherein the light chain variable domain comprises a sequence that is at least 80% identical to SEQ ID NO: 66. [The present invention 1021] 1020. The protein of the present invention, wherein said light chain variable domain comprises a sequence that is at least 90% identical to SEQ ID NO:66. [The present invention 1022] 1021. The protein of the present invention, wherein said light chain variable domain comprises a sequence that is at least 95% identical to SEQ ID NO:66. [The present invention 1023] the antigen-binding domain a heavy chain variable domain comprising CDR1 comprising SEQ ID NO: 19, CDR2 comprising SEQ ID NO: 20, and CDR3 comprising SEQ ID NO: 21, and a light chain variable domain comprising CDR1 comprising SEQ ID NO: 22, CDR2 comprising SEQ ID NO: 23, and CDR3 comprising SEQ ID NO: 24. 1001. The protein of the present invention, comprising: [The present invention 1024] The protein of the present invention 1023, wherein said heavy chain variable domain comprises a sequence that is at least 80% identical to SEQ ID NO:67. [The present invention 1025] 1024. The protein of the present invention, wherein said heavy chain variable domain comprises a sequence that is at least 90% identical to SEQ ID NO:67. [The present invention 1026] 1025. The protein of the present invention, wherein said heavy chain variable domain comprises a sequence that is at least 95% identical to SEQ ID NO:67. [The present invention 1027] The protein of any of claims 1023 to 1026, wherein the light chain variable domain comprises a sequence that is at least 80% identical to SEQ ID NO: 68. [The present invention 1028] 1027. The protein of the present invention, wherein said light chain variable domain comprises a sequence that is at least 90% identical to SEQ ID NO:68. [The present invention 1029] 1028. The protein of the present invention, wherein said light chain variable domain comprises a sequence at least 95% identical to SEQ ID NO:68. [The present invention 1030] the antigen-binding domain a heavy chain variable domain comprising CDR1 comprising SEQ ID NO: 25, CDR2 comprising SEQ ID NO: 26, and CDR3 comprising SEQ ID NO: 27, and a light chain variable domain comprising CDR1 comprising SEQ ID NO: 28, CDR2 comprising SEQ ID NO: 29, and CDR3 comprising SEQ ID NO: 30. 1001. The protein of the present invention, comprising: [The present invention 1031] 1030. The protein of the present invention, wherein said heavy chain variable domain comprises a sequence that is at least 80% identical to SEQ ID NO:69. [The present invention 1032] 1031. The protein of the present invention, wherein said heavy chain variable domain comprises a sequence that is at least 90% identical to SEQ ID NO:69. [The present invention 1033] 1032. The protein of the present invention, wherein said heavy chain variable domain comprises a sequence at least 95% identical to SEQ ID NO:69. [The present invention 1034] The protein of any of claims 1030 to 1033, wherein the light chain variable domain comprises a sequence that is at least 80% identical to SEQ ID NO: 70. [This invention 1035] 1034. The protein of the present invention, wherein said light chain variable domain comprises a sequence at least 90% identical to SEQ ID NO:70. [The present invention 1036] 1035. The protein of the present invention, wherein said light chain variable domain comprises a sequence at least 95% identical to SEQ ID NO:70. [This invention 1037] the antigen-binding domain a heavy chain variable domain comprising CDR1 comprising SEQ ID NO: 31, CDR2 comprising SEQ ID NO: 32, and CDR3 comprising SEQ ID NO: 33, and a light chain variable domain comprising CDR1 comprising SEQ ID NO: 34, CDR2 comprising SEQ ID NO: 35, and CDR3 comprising SEQ ID NO: 36. 1001. The protein of the present invention, comprising: [The present invention 1038] 1037. The protein of the present invention, wherein said heavy chain variable domain comprises a sequence that is at least 80% identical to SEQ ID NO: 71. [This invention 1039] 1038. The protein of the present invention, wherein said heavy chain variable domain comprises a sequence at least 90% identical to SEQ ID NO: 71. [The present invention 1040] 1039. The protein of the present invention, wherein said heavy chain variable domain comprises a sequence at least 95% identical to SEQ ID NO:71. [This invention 1041] The protein of any of claims 1037 to 1040, wherein the light chain variable domain comprises a sequence that is at least 80% identical to SEQ ID NO: 72. [The present invention 1042] 1041. The protein of the present invention, wherein said light chain variable domain comprises a sequence at least 90% identical to SEQ ID NO: 72. [This invention 1043] 1042. The protein of the present invention, wherein said light chain variable domain comprises a sequence at least 95% identical to SEQ ID NO: 72. [This invention 1044] the antigen-binding domain a heavy chain variable domain comprising CDR1 comprising SEQ ID NO: 37, CDR2 comprising SEQ ID NO: 38, and CDR3 comprising SEQ ID NO: 39, and a light chain variable domain comprising CDR1 comprising SEQ ID NO: 40, CDR2 comprising SEQ ID NO: 41, and CDR3 comprising SEQ ID NO: 42. 1001. The protein of the present invention, comprising: [This invention 1045] 1044. The protein of the present invention, wherein said heavy chain variable domain comprises a sequence that is at least 80% identical to SEQ ID NO: 73. [The present invention 1046] 1045. The protein of the present invention, wherein said heavy chain variable domain comprises a sequence that is at least 90% identical to SEQ ID NO:73. [This invention 1047] 1046. The protein of the present invention, wherein said heavy chain variable domain comprises a sequence at least 95% identical to SEQ ID NO: 73. [This invention 1048] The protein of any of claims 1044 to 1047, wherein the light chain variable domain comprises a sequence that is at least 80% identical to SEQ ID NO: 74. [This invention 1049] 1048. The protein of the present invention, wherein said light chain variable domain comprises a sequence at least 90% identical to SEQ ID NO: 74. [The present invention 1050] 1049. The protein of the present invention, wherein said light chain variable domain comprises a sequence at least 95% identical to SEQ ID NO:74. [This invention 1051] the antigen-binding domain a heavy chain variable domain comprising CDR1 comprising SEQ ID NO: 43, CDR2 comprising SEQ ID NO: 44, and CDR3 comprising SEQ ID NO: 45, and a light chain variable domain comprising CDR1 comprising SEQ ID NO: 46, CDR2 comprising SEQ ID NO: 47, and CDR3 comprising SEQ ID NO: 48. 1001. The protein of the present invention, comprising: [This invention 1052] 1051. The protein of the present invention, wherein said heavy chain variable domain comprises a sequence that is at least 80% identical to SEQ ID NO: 75. [This invention 1053] 1052. The protein of the present invention, wherein said heavy chain variable domain comprises a sequence at least 90% identical to SEQ ID NO: 75. [This invention 1054] 1053. The protein of the present invention, wherein said heavy chain variable domain comprises a sequence at least 95% identical to SEQ ID NO: 75. [This invention 1055] The protein of any of claims 1051 to 1054, wherein the light chain variable domain comprises a sequence that is at least 80% identical to SEQ ID NO: 76. [The present invention 1056] 1055. The protein of the present invention, wherein said light chain variable domain comprises a sequence at least 90% identical to SEQ ID NO:76. [This invention 1057] 1056. The protein of the present invention, wherein said light chain variable domain comprises a sequence at least 95% identical to SEQ ID NO:76. [This invention 1058] the antigen-binding domain a heavy chain variable domain comprising CDR1 comprising SEQ ID NO: 49, CDR2 comprising SEQ ID NO: 50, and CDR3 comprising SEQ ID NO: 51, and a light chain variable domain comprising CDR1 comprising SEQ ID NO: 52, CDR2 comprising SEQ ID NO: 53, and CDR3 comprising SEQ ID NO: 54. 1001. The protein of the present invention, comprising: [This invention 1059] 1058. The protein of the present invention, wherein said heavy chain variable domain comprises a sequence at least 80% identical to SEQ ID NO:77. [The present invention 1060] 1059. The protein of the present invention, wherein said heavy chain variable domain comprises a sequence at least 90% identical to SEQ ID NO:77. [The present invention 1061] 1060. The protein of the present invention, wherein said heavy chain variable domain comprises a sequence at least 95% identical to SEQ ID NO:77. [The present invention 1062] The protein of any of claims 1058 to 1061, wherein the light chain variable domain comprises a sequence that is at least 80% identical to SEQ ID NO: 78. [The present invention 1063] 1062. The protein of the present invention, wherein said light chain variable domain comprises a sequence at least 90% identical to SEQ ID NO: 78. [This invention 1064] 1063. The protein of the present invention, wherein said light chain variable domain comprises a sequence at least 95% identical to SEQ ID NO: 78. [This invention 1065] the antigen-binding domain a heavy chain variable domain comprising CDR1 comprising SEQ ID NO: 55, CDR2 comprising SEQ ID NO: 56, and CDR3 comprising SEQ ID NO: 57, and a light chain variable domain comprising CDR1 comprising SEQ ID NO: 58, CDR2 comprising SEQ ID NO: 59, and CDR3 comprising SEQ ID NO: 60. 1001. The protein of the present invention, comprising: [The present invention 1066] 1065. The protein of the present invention, wherein said heavy chain variable domain comprises a sequence that is at least 80% identical to SEQ ID NO:79. [This invention 1067] 1066. The protein of the present invention, wherein said heavy chain variable domain comprises a sequence that is at least 90% identical to SEQ ID NO:79. [The present invention 1068] 1067. The protein of the present invention, wherein said heavy chain variable domain comprises a sequence that is at least 95% identical to SEQ ID NO:79. [This invention 1069] The protein of any of claims 1065 to 1068, wherein the light chain variable domain comprises a sequence that is at least 80% identical to SEQ ID NO: 80. [The present invention 1070] 1069. The protein of the present invention, wherein said light chain variable domain comprises a sequence at least 90% identical to SEQ ID NO:80. [This invention 1071] 1070. The protein of the present invention, wherein said light chain variable domain comprises a sequence at least 95% identical to SEQ ID NO:80. [This invention 1072] Any one of the proteins of the present inventions 1001 to 1071, which is a multi-chain protein. [This invention 1073] Any one of the proteins of the present invention 1001 to 1071, which is a single-chain protein. [This invention 1074] The protein of any one of claims 1001 to 1071, which is an antibody or an antigen-binding antibody fragment. [This invention 1075] The protein of the present invention 1074, wherein the antibody is an IgG1 antibody, an IgG2 antibody, an IgG3 antibody, or an IgG4 antibody. [This invention 1076] The protein of the present invention 1074 or 1075, wherein said antibody is humanized. [This invention 1077] The protein of the present invention 1074 or 1075, wherein the antibody is human. [This invention 1078] Any one of the proteins of the present invention 1001 to 1071, which is an scFv. [This invention 1079] The protein of any one of claims 1001 to 1071, which is a chimeric antigen receptor (CAR). [The present invention 1080] A pharmaceutical composition comprising any one of the proteins of the present inventions 1001 to 1079 and a pharmaceutically acceptable carrier. [This invention 1081] A kit comprising the pharmaceutical composition of the present invention. [This invention 1082] A nucleic acid encoding any one of the proteins of the present invention 1001 to 1079. [This invention 1083] A vector comprising the nucleic acid of the present invention. [This invention 1084] A pharmaceutical composition comprising the nucleic acid of the present invention 1082 or the vector of the present invention 1083. [This invention 1085] A kit comprising the pharmaceutical composition of the present invention. [This invention 1086] A cell comprising the nucleic acid of the present invention 1082 or the vector of the present invention 1083. [This invention 1087] The cell of the present invention 1086, which is an immune cell. [This invention 1088] The cell of the present invention 1087, wherein the immune cell is a T cell, a B cell, or a natural killer (NK) cell. [This invention 1089] The cell of the present invention 1087, wherein the immune cell is a regulatory T (Treg) cell. [The present invention 1090] The cell of any one of 1087 to 1089 of the present invention, wherein the immune cell is an autologous cell. [This invention 1091] The cell of any one of 1087 to 1089 of the present invention, wherein the immune cell is an allogeneic cell. [This invention 1092] A pharmaceutical composition comprising the cells of any one of the present inventions 1086 to 1091 and a pharmaceutically acceptable carrier. [This invention 1093] A kit comprising the pharmaceutical composition of the present invention. [This invention 1094] A method for treating an age-related disease or an inflammatory disease in a subject, the method comprising administering to the subject a therapeutically effective amount of any one of the proteins of the present inventions 1001 to 1079 or the pharmaceutical composition of the present invention 1080. [This invention 1095] A method for treating an age-related disease or an inflammatory disease in a subject, the method comprising administering to the subject a therapeutically effective amount of a nucleic acid of the present invention 1082, a vector of the present invention 1083, or a pharmaceutical composition of the present invention 1084. [This invention 1096] A method for treating an age-related disease or an inflammatory disease in a subject, the method comprising administering to the subject a therapeutically effective amount of any one of the cells of the present inventions 1086 to 1091 or the pharmaceutical composition of the present invention 1092. [This invention 1097] The method of any one of claims 1094 to 1096, wherein the age-related disease is inflammation-aging-related. [This invention 1098] The object is (i) a therapeutically effective amount of an NK cell activator and / or NK cells and / or a monoclonal antibody, and / or (ii) a therapeutically effective amount of a Treg cell activator and / or Treg cells and / or a monoclonal antibody and / or an advanced glycation end product (AGE) inhibitor The method of any one of claims 1094 to 1097, further comprising administering [This invention 1099] The method of claim 1098, comprising administering to said subject a therapeutically effective amount of NK cells. [The present invention 1100] The NK cells Autologous, haploidentical, or allogeneic NK cells isolated from peripheral blood, isolated from umbilical cord blood, or isolated and differentiated from iPSCs The method of the present invention 1099. [The present invention 1101] The method comprises: isolating the NK cells from the subject; and culturing the isolated NK cells in a liquid culture medium under conditions sufficient to induce or increase proliferation of the NK cells; Further comprising: After the isolating and culturing steps, the NK cells are administered to the subject. The method of the present invention 1100. [The present invention 1102] 1101. The method of claim 1101, wherein said liquid culture medium comprises a multi-chain chimeric polypeptide. [The present invention 1103] 13. The method of any one of claims 1099 to 1102, wherein said NK cells comprise a chimeric antigen receptor. [The present invention 1104] 1103. The method of claim 1103, wherein said protein is a chimeric antigen receptor of claim 1079. [This invention 1105] The method of claim 1098, comprising administering to said subject a therapeutically effective amount of an NK cell activator and / or a monoclonal antibody. [The present invention 1106] 1105. The method of claim 1105, wherein said NK cell activator is one or more multi-chain chimeric polypeptides. [This invention 1107] The method of the present invention 1105, wherein the monoclonal antibody is an anti-tissue factor antibody or any one of the antibodies of the present inventions 1074 to 1077. [This invention 1108] The method of the present invention 1105, wherein the NK cell activator comprises one or more multi-chain chimeric polypeptides, and the monoclonal antibody comprises an anti-tissue factor antibody and / or one or more of the antibodies of the present invention 1074 to 1077. [This invention 1109] The method of any of claims 1098 to 1108, comprising administering to said subject a therapeutically effective amount of Treg cells. [The present invention 1110] The Treg cells Autologous, haploidentical, or allogeneic Treg cells isolated from peripheral or umbilical cord blood The method of the present invention 1109. [The present invention 1111] The method comprises: isolating the Treg cells from the subject; culturing the isolated Treg cells in a liquid culture medium under conditions sufficient to induce or increase proliferation of the Treg cells; Further comprising: After the isolating and culturing steps, the Treg cells are administered to the subject. The method of the present invention 1110. [The present invention 1112] 1111. The method of claim 1111, wherein said liquid culture medium comprises one or more single-chain chimeric polypeptides. [The present invention 1113] The method of any one of claims 1109 to 1112, wherein said Treg cells comprise a chimeric antigen receptor. [This invention 1114] 1113. The method of claim 1113, wherein said chimeric antigen receptor comprises an extracellular domain that specifically binds to tissue factor, to CD26, or to CD36. [This invention 1115] The method of any one of claims 1098 to 1114, comprising administering to the subject a therapeutically effective amount of a Treg cell activator and / or a monoclonal antibody and / or an AGE inhibitor. [The present invention 1116] 1115. The method of claim 1115, wherein said Treg cell activator is one or more single-chain chimeric polypeptides. [This invention 1117] 1115. The method of claim 1115, wherein said monoclonal antibody is one or both of an anti-tissue factor antibody, an anti-CD26 antibody, and / or an anti-CD36 antibody. [This invention 1118] 1115. The method of claim 1115, wherein said AGE inhibitor is a soluble RAGE trap. [This invention 1119] The method of the present invention 1115, wherein the Treg cell activator comprises one or more single-chain chimeric polypeptides, the monoclonal antibody comprises one or more of an anti-tissue factor antibody, an anti-CD26 antibody, and / or an anti-CD36 antibody, and the AGE inhibitor comprises one or more soluble RAGE traps. [The present invention 1120] the multi-chain chimeric polypeptide (a) a first chimeric polypeptide, (i) a first target binding domain; (ii) a soluble tissue factor domain, and (iii) the first domain of the pair of affinity domains the first chimeric polypeptide comprising: (b) a second chimeric polypeptide, (i) the second domain of the pair of affinity domains, and (ii) a second target-binding domain and the second chimeric polypeptide comprising Including, the first chimeric polypeptide and the second chimeric polypeptide are associated via binding between the first domain and the second domain of the pair of affinity domains; The method of the present invention 1102, 1106, or 1108. [This invention 1121] the single-chain chimeric polypeptide (i) a first target binding domain; (ii) a soluble tissue factor domain, and (iii) a second target-binding domain The method of the present invention 1112, 1116, or 1119, comprising: [This invention 1122] 1098-1121, wherein the age-related disorder is selected from the group consisting of Alzheimer's disease, aneurysm, cystic fibrosis, fibrosis in pancreatitis, glaucoma, hypertension, idiopathic pulmonary fibrosis, inflammatory bowel disease, intervertebral disc degeneration, macular degeneration, osteoarthritis, type 2 diabetes mellitus, lipoatrophy, lipodystrophy, atherosclerosis, cataracts, COPD, idiopathic pulmonary fibrosis, kidney transplant failure, liver fibrosis, bone loss, myocardial infarction, sarcopenia, wound healing, alopecia, cardiomyocyte hypertrophy, osteoarthritis, Parkinson's disease, age-related loss of lung tissue elasticity, macular degeneration, cachexia, glomerulosclerosis, liver cirrhosis, NAFLD, osteoporosis, amyotrophic lateral sclerosis, Huntington's disease, spinocerebellar ataxia, multiple sclerosis, neurodegeneration, stroke, cancer, dementia, vascular disease, susceptibility to infection, chronic inflammation, and renal dysfunction. [This invention 1123] 1098-1121. Any of the methods of claims 1098-1121, wherein the inflammatory disease is selected from the group consisting of rheumatoid arthritis, inflammatory bowel disease, lupus erythematosus, lupus nephritis, diabetic nephropathy, CNS injury, Alzheimer's disease, Parkinson's disease, amyotrophic lateral sclerosis, Crohn's disease, multiple sclerosis, Guillain-Barré syndrome, psoriasis, Graves' disease, ulcerative colitis, non-alcoholic steatohepatitis, and mood disorders. [This invention 1124] 1122. The method of claim 1122, wherein the age-related disease is a cancer selected from the group consisting of solid tumors, hematological tumors, sarcoma, osteosarcoma, glioblastoma, neuroblastoma, melanoma, rhabdomyosarcoma, Ewing's sarcoma, osteosarcoma, B-cell neoplasms, multiple myeloma, B-cell lymphoma, B-cell non-Hodgkin's lymphoma, Hodgkin's lymphoma, chronic lymphocytic leukemia (CLL), acute myeloid leukemia (AML), chronic myeloid leukemia (CML), acute lymphocytic leukemia (ALL), myelodysplastic syndrome (MDS), cutaneous T-cell lymphoma, retinoblastoma, gastric cancer, urothelial cancer, lung cancer, renal cell carcinoma, gastroesophageal cancer, pancreatic cancer, prostate cancer, breast cancer, colorectal cancer, ovarian cancer, non-small cell lung cancer, squamous cell head and neck cancer, endometrial cancer, cervical cancer, liver cancer, and hepatocellular carcinoma. [This invention 1125] A method for treating cancer in a subject, comprising administering to the subject a therapeutically effective amount of any one of the proteins of the present inventions 1074 to 1077. [Invention 1126] A method for treating an infectious disease in a subject, the method comprising administering to the subject a therapeutically effective amount of any one of the proteins of the present inventions 1074 to 1077. [This invention 1127] A method for treating an infectious disease in a subject, comprising administering to the subject a therapeutically effective amount of any of the proteins of present inventions 1001 to 1079 or the pharmaceutical composition of present invention 1080. Other features and advantages of the invention will be apparent from the following detailed description and figures, and from the claims. [Brief explanation of the drawings]

[0046] [Figure 1] ELISA binding analysis of selected scFv clones is shown, where plates of scFv clones were tested to test their binding to CD26, Fc, and proA / L, and DNA was prepared for the scFv constructs and sent for DNA sequencing to determine the LC / HC variable domain sequences. [Figure 2] Sequence analysis of five unique scFv-binding clones is shown, where unique clones were identified and sequencing results show that their LC and HC variable domains are intact and that the sequences of CDR-L1, CDR-L2, CDR-L3, CDR-H1, CDR-H2, and CDR-H3 are unique to each other. [Figure 3] Binding of scFv supernatants to CD26 is shown by serial dilutions of scFv supernatants. [Figure 4]The concentration of scFv was measured, and the ELISA binding data was concentration-corrected to show the concentration-corrected binding of scFv to CD26. [Figure 5] Screening of CD26-binding scFvs is shown, with clones CD26-03G and CD26-04E indicated by circles. [Figure 6] Screening of CD26-binding scFvs in the YCM screen is shown, with scFv clones CD26-01F, CD26-01G, and CD26-07H indicated by circles. [Figure 7] The sequences of the selected scFvs are shown. [Figure 8] Figure 1 shows the human CD26 binding activity of anti-CD26 monoclonal antibodies. Anti-CD26 IgG1 kappa monoclonal antibodies were constructed based on the selected scFv sequences. CD26 binding of purified anti-CD26 monoclonal antibodies was determined by ELISA using 96-well Maxisorp plates coated with (A) human CD26-Fc fusion protein or (B) goat anti-human IgG. The plates were blocked with blocking buffer. Purified anti-CD26 monoclonal antibodies were diluted in blocking buffer and added to wells of plates coated with CD26-Fc or goat anti-human IgG. Anti-CD26 monoclonal antibodies were probed with goat anti-human kappa-HRP / ABTS and read at 405 nM using an ELISA plate reader. The results show that CD26Ab-01D and CD26Ab-04A can bind to CD26, with CD26Ab-01D having superior binding activity to CD26Ab-04A. CD26Ab-12D and CD26Ab-03B have weak CD26 binding activity, but CD26Ab-10B does not have significant CD26 binding activity. [Figure 9]Figure 1 shows the human CD26 binding activity of anti-CD26 monoclonal antibodies. CHO cells transfected with human CD26 were stained with (A) 50 nM of five different anti-CD26 monoclonal antibodies or (B) 1 μg / test of five different biotinylated anti-CD26 antibodies (production of 26Ab-10B was very low and not biotinylated). Probes were used with goat anti-human IgG-PE for non-biotinylated antibodies or streptavidin-PE for biotinylated antibodies. Data were analyzed using a BD FACSCelesta equipped with BD FACSDiva software. An anti-tissue factor antibody (anti-TF Ab) was used as a negative control, and PE-conjugated anti-CD26 (BioLegend) was used as a positive control. The results show that CD26Ab-01D and CD26-04A bound well to CD26-expressing cells, while the other three of the five antibodies showed weaker binding. [Figure 10] Figure 1 shows the ADCC activity of different anti-CD26 monoclonal antibodies. Human CD26-transfected CHO cells (CHO26) were labeled with CellTrace Violet and used as target cells, and fresh human NK cells (left: donor-1, right: donor-2) were used as effector cells. Effector cells were plated with violet-labeled target cells containing 26Ab-01D or 26Ab-04A at 5 nM concentrations at the indicated effector:target (E:T) ratio. Anti-tissue factor antibody (anti-TF Ab) was used as a control. Target cell inhibition (%) was calculated using the following formula: (1 - [number of viable CHO26 cells in the experimental sample / number of viable CHO26 cells in the sample without splenocytes]) × 100. This represents anti-CD26 antibody-mediated ADCC, assessed by flow cytometry on day 2. The results demonstrate CD26Ab-01D and CD26Ab-04A-dependent and NK cell-mediated cytotoxicity against CD26-positive CHO cells. [Figure 11]This figure shows the interaction between human CD26 and adenosine deaminase (ADA). Human CD26-Fc fusion protein was used to coat a 96-well Maxisorp plate. The plate was blocked with blocking buffer. Human ADA (R&D systems) was diluted in blocking buffer and added to the wells of the CD26-Fc-coated plate. Two biotinylated anti-CD26 monoclonal antibodies (CD26Ab-01D and CD26Ab-04A) were added to the plate. The anti-CD26 monoclonal antibodies were probed with SAHRP / ABTS and read at 405 nM on an ELISA plate reader. The results show that ADA was able to block the binding of CD26Ab-01D and CD26Ab-04A to CD26. [Figure 12] Schematic diagram of the nucleic acid encoding the anti-CD26 CAR in a lentiviral vector. [Figure 13] 1 is a series of images showing total Treg cells and anti-CD26CAR Treg cells stimulated with specific antigen (CD26 / beads), non-specific antigen (TF / beads), or TCR (CD3 / CD28 / beads) for 3 days. [Figure 14] Graph showing fold expansion of anti-CD26CAR Treg cells after stimulation with specific antigen (CD26 / beads), non-specific antigen (TF / beads), or TCR (CD3 / CD28 / beads) for 3 days. [Figure 15] Fluorescence-assisted cell sorting (FACS) data series showing staining of anti-CD26CAR Treg cells with CD26-Fc or tissue factor (TF). [Figure 16] Graph showing the expression of cell markers in anti-CD26CAR Treg cells and non-transduced Treg cells. [Figure 17] Graph of suppressive activity of anti-CD26CAR Treg cells and non-transduced Treg cells. [Figure 18] 10 is a graph showing the suppression of IFNg production by Tresp cells by anti-CD26CAR Treg cells or non-transduced Treg cells. [Figure 19]Graph showing IL-10 production by anti-CD26CAR Treg cells and non-transduced Treg cells. DETAILED DESCRIPTION OF THE INVENTION

[0047] Detailed Description Provided herein are proteins comprising an anti-CD26 antigen-binding domain, nucleic acids encoding same, cells comprising any of these nucleic acids or proteins, compositions comprising any of these proteins, nucleic acids, and cells, and methods of treating a subject with an age-related or inflammatory disease using any of the compositions described herein.

[0048] Non-limiting aspects of these proteins, nucleic acids, cells, compositions, and methods are described below.

[0049] CD26 CD26 (also known as DPP4, dipeptidyl-peptidase-4, or DDP4) is a transmembrane glycoprotein that is membrane-anchored by its signal peptide and forms homodimers or tetramers on the plasma membrane. CD26 is an aminopeptidase that cleaves N-terminal dipeptides primarily from peptides or small proteins (fewer than 80–100 amino acid residues) using proline or alanine as the penultimate amino acid. It can also cleave protein substrates containing glycine, serine, valine, or leucine, but at a slower rate. The enzyme cannot cleave substrates with proline at the 3 position.

[0050] CD26 is expressed in numerous tissues, including the brush border membrane of the intestine and kidney, vascular endothelium, liver and pancreas, glandular epithelial cells, and by cells of the immune system (Gutschmidt et al., Histochemistry 73(2):285-304, 1981; Gorrell et al., Cell Immunol. 134(1):205-215, 1991; Tanaka et al., J. Immunol. 149(2):481-486, 1992; Abbott et al., Immunogenetics 40(5):331-338, 1994; Buhling et al., Immunol. Lett. 45(1-2):47-51, 1995; Dikov et al., Cell. Mol. Biol. 50 Online Pub:OL565-568, 2004; Broxmeyer et al., Stem Cells Dev. 25(8):575-585, 2016; Hollande et al., Nat. Immunol. 20(3):257-264, 2019). The primary structure of CD26 consists of a 6-amino acid cytoplasmic domain, a 22-amino acid transmembrane domain, and a 738-amino acid extracellular portion. The extracellular portion contains the catalytically active site triad Ser 630 , Asp 708 , and His 740It consists of a C-terminal catalytic region with a cysteine-rich domain, a cysteine-rich region, and a large glycosylation-rich region connected to the transmembrane segment by a flexible stalk (Klemann et al., Clin. Exp. Immunol. 185(1):1-21, 2016). The crystal structure of human CD26 reveals two domains: an eight-bladed propeller and an α / β-hydrolase domain (Engel et al., Proc. Natl. Acad. Sci. USA 100(9):5063-5068, 2003). The propeller is open and consists of subdomains composed of blades II-V and VI-VIII, which are the glycosylation-rich and cysteine-rich regions, respectively. Adenosine deaminase (ADA) and caveolin-1 bind to the glycosylation-rich domain of human CD26, whereas collagen, fibronectin, plasminogen, and streptokinase bind to the cysteine-rich region (Klemann et al., Clin. Exp. Immunol. 185(1):1-21, 2016). Two openings exist for substrate interaction: the side opening and the propeller tunnel (Rasmussen et al., Nat. Struct. Biol. 10(1):19-25, 2003; Weihofen et al., J. Biol. Chem. 279(41):43330-43335, 2004). The CD26 substrate neuropeptide Y was found to enter the side opening of CD26 (Aertgeerts et al., Protein Sci. 13(2):412-421, 2004). CD26 has also been found to function as a binding site for the chemokine receptor CXCR4, the T cell differentiation antigen CD45, and the sodium-hydrogen exchanger-3 (Mentlein et al., Regul. Pept. 85(1):9-24, 1999; Lambeir et al., Crit. Rev. Clin. Lab. Sci. 40(3):209-294, 2003). Therefore, CD26 can be considered a multifunctional protein with various functions beyond its role as a proteinase.Its role as a receptor or ligand for a variety of different molecules, alone or in combination with its enzymatic activity, enables it to influence physiological processes such as the interactions between cells and the extracellular matrix involved in cell migration, activation, and proliferation.

[0051] CD26 plays a key role in glucose metabolism. Incretin peptides, such as gastric inhibitory polypeptide (GIP) and glucagon-like peptide (GLP-1), promote insulin secretion from pancreatic β-cells and, through the static effects of glucagon, are involved in the regulation of postprandial blood glucose. These peptides are rapidly inactivated by CD26, resulting in a short half-life. - / - Mice are protected from the development of diet-induced obesity and have improved postprandial glycemic control due to the extended half-life of incretin peptides. - / -Mice also show improved insulin sensitivity, reduced islet hypertrophy, and protection against streptozotocin-induced beta-cell loss and hyperglycemia (Marguet et al., Proc. Natl. Acad. Sci. USA 97(12):6874-6879, 2000; Conarello et al., Proc. Natl. Acad. Sci. USA 100(11):6825-6830, 2003). Several CD26 inhibitors have been approved by the US FDA as antidiabetic drugs, including sitagliptin, saxagliptin, linagliptin, vildagliptin, and alogliptin. Most clinical trials using CD26 inhibitors have shown a reduction in HbA1C of approximately 0.6–0.8% in patients with baseline levels of approximately 8% (Inzucchi et al., Circulation 117(4):574-584, 2008). In general, most studies have also demonstrated improvements in the homeostasis model assessment beta cell function index (HOMA-β) and fasting proinsulin:insulin ratio, suggesting improved beta cell function (Raz et al., Diabetologia 49(11):2564-2571, 2006). Side effects and the incidence of hypoglycemia with these medications are very low (Raz et al., Diabetologia 49(11):2564-2571, 2006; Lambeir et al., Crit. Rev. Clin. Lab. Sci. 40(3):209-294, 2003).

[0052] In addition to incretin peptides, CD26 also cleaves many other proteins. Physiological targets include GLP1, GLP2, brain natriuretic peptide, peptide YY, interstitial cell-derived factor, erythropoietin, granulocyte-colony-stimulating factor, and substance P. Pharmacological targets include gastric-releasing peptide, growth hormone-releasing factor, macrophage-derived chemokine, eotaxin, IFN-γ-inducible protein-10, granulocyte-macrophage colony-stimulating factor, erythropoietin, IL-3, neuropeptide Y, B-type natriuretic peptide, and peptide YY (Mulvihill et al., Endocr. Rev. 35(6):992-1019, 2014).

[0053] CD26 is known to regulate the function of chemokines, such as CXCR3, through post-translational cleavage of X-Pro or X-Ala motifs, resulting in amino-terminal dipeptide cleavage of the chemokine and altering its biological function (Broxmeyer et al., Stem Cells Dev. 25(8):575-585, 2016). CD26 also mediates amino-terminal cleavage of the chemokine CXCL10, limiting the migration of CXCR3 natural killer (NK) cells and T cells and limiting the decline of antitumor immunity in preclinical models of melanoma and colorectal cancer (Barreira da Silva et al., Nat. Immunol. 16(8):850-858, 2015). Combination immunotherapy using checkpoint blockade in the presence of the CD26 inhibitor sitagliptin has also been shown to reduce tumor growth by enhancing the antitumor activity of T cells and eosinophils in preclinical mouse models of hepatocellular carcinoma and breast cancer (Hollande et al., Nat. Immunol. 20(3):257-264, 2019).

[0054] As mentioned above, CD26 also interacts with various ligands. Through its interaction with these ligands, CD26 plays a role in various processes, including promoting T cell activation and functional regulation of antigen-presenting cells (APCs). CD26 can directly activate and proliferate T cells through CARMA1-mediated nuclear factor κB (NF-κB) expression in T cells (Ohnuma et al., J. Immunol. 167(12):6745-6755, 2001; Ohnuma et al., Proc. Natl. Acad. Sci. USA 101(39):14186-14191, 2004). CD26 on T cells directly interacts with APCs via caveolin-1. Upon binding, Tollip and interleukin-1 receptor-associated kinase 1 (1RAK-1) are displaced from caveolin-1, resulting in subsequent phosphorylation of 1RAK-1 (Ohnuma et al., Mol. Cell. Biol. 25(17):7743-7757, 2005; Ohnuma et al., Front. Biosci. 13:2299-2310, 2008). This leads to upregulation of the costimulatory molecule CD86, strengthening the binding of T cells and APCs at the immunological synapse (Ohnuma et al., Proc. Natl. Acad. Sci. USA 101(39):14186-14191, 2004). Blocking CD26-mediated T cell costimulation with a soluble caveolin-1-Ig fusion protein induces anergy in CD4+ T cells (Ohhuma et al., Biochem. Biophys. Res. Comm. 386(2):327-332, 2009).

[0055] The interaction of CD26 and ADA also promotes T cell activation by providing a microenvironment suitable for T cell proliferation. Extracellular ATP or ADP is first converted to AMP by CD39 and CD73, generating adenosine (Deaglio et al., J. Exp. Med. 204(6):1257-1265, 2007). Adenosine is then processed by ADA and converted to inosine (Resta et al., Immunol. Rev. 161:95-109, 1998). Adenosine is involved in the central nervous system, immune system, and immune responses to A1, A2, and A3. 2A , A 2B CD26 exerts multiple physiological effects both in the periphery and in peripheral tissues mediated by a G-protein-coupled adenosine receptor identified as A3 (Borea et al., Physiol. Rev. 98(3):1591-1625, 2018). By anchoring ADA to its surface, CD26 regulates pericellular adenosine levels and modulates T cell activation. Lack of ADA activity leads to the accumulation of adenosine, which inhibits T cell proliferation in a dose-dependent manner. Jurkat cells expressing a CD26 mutant lacking ADA-binding activity are sensitive to adenosine-mediated inhibition of T cell proliferation (Dong et al., J. Immunol. 159(12):6070-6076, 1997). Cells expressing ADA and CD26 on their surface are much more resistant to the inhibitory effects of adenosine (Dong et al., J. Immunol. 156(4):1349-1355, 1996; Dong et al., J. Immunol. 159(12):6070-6076, 1997; Zhong et al., Diabetes 62(1):149-157, 2013). Evidence indicates that ADA colocalizes with adenosine receptors on dendritic cells and interacts with CD26 expressed on lymphocytes (Moreno et al., Front. Pharmacol. 9:106, 2018). This ability of ADA functions as a costimulatory signal to enhance T cell activation and induce the production of T helper cell (Th1) proinflammatory cytokines.

[0056] CD26 binds to multiple components of the extracellular matrix, including collagen, fibronectin, and the HIV-1 Tat protein (Loster et al., Biochem. Biophys. Res. Comm. 217(1):341-348, 1995; Zhong et al., Diabetes 62(1):149-157, 2013). Interactions with these matrix components may play a role in sequestration of CD26, enabling additional functions such as matrix remodeling, metastasis, and chemotaxis. Research on anti-CD26 antibodies currently in development represents a promising approach for cancer treatment. Preclinical studies have demonstrated that a humanized monoclonal antibody targeting human CD26 is effective against multiple myeloma in vitro and in vivo via the mechanism of antibody-dependent cell-mediated cytotoxicity (ADCC) (Nishida et al., Blood Cancer J. 8(11):99, 2018). A humanized monoclonal antibody against CD26 showed promising antitumor effects and was well tolerated in a recently reported phase I clinical trial in patients with advanced malignant pleural mesothelioma (Takeda et al., Lung Cancer 137:64-70, 2019).

[0057] CD26 is also involved in cellular senescence, a hallmark of aging. The accumulation of senescent cells in tissues is strongly associated with age-related pathologies (Childs et al., Nat. Rev. Drug Discov. 16(10):718-735, 2017; Kirkland et al., EBioMedicine 21:21-28, 2017). Mass spectrometry analysis revealed that CD26 is upregulated on the surface of human senescent diploid fibroblasts (Kim et al., Genes Dev. 31(15):1529-1534, 2017). Increased CD26 expression on non-dividing but senescent fibroblasts sensitized them to NK-mediated ADCC by anti-CD26 antibodies (Kim et al., Genes Dev. 31(15):1529-1534, 2017).

[0058] Senescence is a form of irreversible growth arrest accompanied by phenotypic changes, resistance to apoptosis, and activation of signaling pathways that sense damage. Cellular senescence was first described in cultured human fibroblasts, which lost the ability to proliferate and reached permanent arrest after approximately 50 population doublings (referred to as the Hayflick limit). Senescence is considered a stress response that can be induced by a wide range of endogenous and exogenous insults, including oxidative and genotoxic stress, DNA damage, telomere attrition, oncogenic activation, mitochondrial dysfunction, or chemotherapeutic agents.

[0059] Senescent cells remain metabolically active and can influence tissue hemostasis, disease, and aging through their secretory phenotype. Senescence is considered a physiological process and is important in promoting wound healing, tissue homeostasis, regeneration, and fibrosis regulation. For example, transient induction of senescent cells is observed during healing and contributes to wound resolution. Perhaps one of the most important roles of senescence is its role in tumor suppression. However, the accumulation of senescent cells also promotes aging and age-related diseases and conditions. The senescent phenotype also triggers chronic inflammatory responses, which may enhance chronic inflammatory conditions and promote tumor growth. The link between senescence and aging was initially based on the observation that senescent cells accumulate in aging tissues. The use of transgenic models has made it possible to systematically detect senescent cells in many age-related pathologies. Strategies for selective elimination of senescent cells have demonstrated that senescent cells can indeed play a causal role in aging and related pathologies.

[0060] Senescent cells exhibit important distinctive characteristics, including changes in morphology, chromatin organization, gene expression, and metabolism. Several biochemical and functional features associated with cellular senescence exist, including (i) increased expression of cyclin-dependent kinase inhibitors p16 and p21, (ii) the presence of senescence-associated β-galactosidase, a marker of lysosomal activity, (iii) the appearance of senescence-associated heterochromatin foci and downregulation of lamin B1 levels, (iv) resistance to apoptosis caused by increased expression of anti-apoptotic BCL family proteins, and (v) upregulation of CD26 (DPP4), CD36 (scavenger receptor), forkhead box 4 (FOXO4), and secretory carrier membrane protein 4 (SCAMP4). Senescent cells also exhibit inflammatory properties, known as the senescence-associated secretory phenotype (SASP). Through the SASP, senescent cells operate in a cell-autonomous manner to reinforce senescence (autocrine effect) and communicate with and modify the microenvironment (paracrine effect), producing a wide range of proinflammatory cytokines (IL-6, IL-8), growth factors (TGF-β), chemokines (CCL-2), and matrix metalloproteinases (MMP-3, MMP-9). SASP factors can contribute to tumor suppression by triggering senescence surveillance, the immune-mediated clearance of senescent cells. However, chronic inflammation is also a known factor in tumorigenesis, and accumulating evidence indicates that chronic SASP may also drive cancer and age-related diseases.

[0061] The secretory profile of senescent cells is context-dependent. For example, mitochondrial dysfunction-associated senescence (MiDAS), induced by various mitochondrial dysfunctions in human fibroblasts, resulted in the emergence of a SASP lacking IL-1-dependent inflammatory factors. A decrease in the NAD+ / NADH ratio activated AMPK signaling, which induced MiDAS through p53 activation. As a result, p53 inhibited NF-κB signaling, a key inducer of pro-inflammatory SASP. In contrast, cellular senescence induced by persistent DNA damage in human cells induced an inflammatory SASP, which depended on the activation of ataxia-telangiectasia mutated (ATM) kinase but not p53 activation. Specifically, the expression and secretion levels of IL-6 and IL-8 were increased. It has also been demonstrated that ectopic expression of p16INK4a and p21CIP1 induced a senescent phenotype in human fibroblasts without the inflammatory SASP, indicating that growth arrest itself did not stimulate the SASP.

[0062] One of the most defining characteristics of senescence is stable growth arrest. This is achieved through two key pathways, p16 / Rb and p53 / p21, both of which are central to tumor suppression. DNA damage leads to (1) increased deposition of γH2Ax (a histone-coding gene) and 53BP1 (involved in DNA damage response) in chromatin, which ultimately activates a kinase cascade that leads to p53 activation, and (2) activation of p16INK4a and ARF (both encoded by CDKN2A) and p15INK4b (encoded by CDKN2B). p53 induces the transcription of cyclin-dependent kinase inhibitor (p21), which, together with p16INK4a and p15INK4b, blocks genes for cell cycle progression (CDK4 and CDK6). This ultimately leads to hypophosphorylation of the retinoblastoma protein (Rb) and cell cycle arrest at G1 phase.

[0063] Selective killing of senescent cells has been shown to significantly improve the health span of mice relative to normal aging and to improve the outcome of age-related disease or cancer therapy (Ovadya et al., J. Clin. Invest. 128(4):1247-1254, 2018). In nature, senescent cells are normally eliminated by innate immune cells. Induction of senescence not only prevents the potential proliferation and transformation of damaged / altered cells, but also supports tissue repair through the production of SASP factors, which function primarily as chemoattractants for NK cells (e.g., IL-15 and CCL2) and macrophages (e.g., CFS-1 and CCL2). These innate immune cells mediate immune surveillance mechanisms to eliminate stressed cells. Senescent cells typically upregulate the NK cell-activating receptors NKG2D and DNAM-1, which belong to the family of stress-inducible ligands and are key components of the frontline immune defense against infectious diseases and malignancies. Upon receptor activation, NK cells can specifically induce the death of senescent cells through their cytolytic mechanism. The role of NK cells in immune surveillance of senescent cells has been demonstrated in liver fibrosis (Sagiv et al., Oncogene 32(15):1971-1977, 2013), hepatocellular carcinoma (Iannello et al., J. Exp. Med. 210(10):2057-2069, 2013), multiple myeloma (Soriani et al., Blood 113(15):3503-3511, 2009), and glioma cells stressed by mevalonate pathway dysfunction (Ciaglia et al., Int. J. Cancer 142(1):176-190, 2018). Endometrial cells undergo acute cellular senescence and fail to differentiate into decidual cells. Differentiated decidual cells secrete IL-15, which recruits uterine NK cells to target and eliminate undifferentiated senescent cells, helping to remodel and rejuvenate the endometrium (Brighton et al., Elife 6:e31274, 2017).Using a similar mechanism, during liver fibrosis, p53-expressing senescent hepatic satellite cells skewed the polarization of resident Kupfer's macrophages and newly infiltrated macrophages, which exhibit senolytic activity, toward a pro-inflammatory M1 phenotype. F4 / 80+ macrophages have been shown to play an important role in the clearance of senescent cells in the mouse uterus to maintain uterine function after parturition.

[0064] Senescent cells primarily recruit NK cells by upregulating ligands for NKG2D (expressed on NK cells), chemokines, and other SASP factors. In vivo models of liver fibrosis have demonstrated effective clearance of senescent cells by activated NK cells (Krizhanovsky et al., Cell 134(4):657-667, 2008). Studies have described various models for studying aging, including liver fibrosis (Krizhanovsky et al., Cell 134(4):657-667, 2008), osteoarthritis (Xu et al., J. Gerontol. A Biol. Sci. Med. Sci. 72(6):780-785, 2017), and Parkinson's disease (Chinta et al., Cell Rep. 22(4):930-940, 2018). Animal models for studying senescent cells are described in Krizhanovsky et al., Cell 134(4):657-667, 2008; Baker et al., Nature 479(7372):232-236, 2011; Farr et al., Nat. Med. 23(9):1072-1079, 2017; Bourgeois et al., FEBS Lett. 592(12):2083-2097, 2018; and Xu et al., Nat. Med. 24(8):1246-1256, 2018.

[0065] Studies have also shown that CD26 plays a role in infectious diseases. Middle East Respiratory Syndrome (MERS) is a viral respiratory disease caused by infection with the coronavirus MERS-CoV. The mortality rate from MERS is approximately 30% (CDC Coronavirus / MERS website). CD26 is a functional receptor for MERS-CoV entry in humans (Raj et al., J. Virol. 88(3):1834-1838, 2014). Binding of the MERS-CoA spike protein S to CD26 mediates viral attachment and internalization. Residues involved in CD26 virus binding are identical to those in the ADA-binding domain, indicating potential competition for CD26 binding (Lu et al., Nature 500(7461):227-231, 2013). The S1 domain of the COVID-19 spike glycoprotein has also been suggested to interact with human CD26 (Vankadari et al., Emerg. Microbes Infect. 9(1):601-604, 2020).

[0066] In summary, CD26 exerts its physiological role through its enzymatic activity by regulating numerous peptides or through interactions with various binding partners. Consequently, alterations in CD26 expression and / or activity have been implicated in several pathological processes, including inflammation, viral infection, immune-mediated diseases, tumor growth, cellular senescence, and metabolic disorders (Mentlein et al., Regul. Pept. 85(1):9-24, 1999; Lambeir et al., Crit. Rev. Clin. Lab. Sci. 40(3):209-294, 2003; Yu et al., FEBS J. 277(5):1126-1144, 2010; Kim et al., Genes Dev. 31(15):1529-1534, 2017; Deacon et al., Front. Endocrinol. 10:80, 2019). Therefore, CD26 is a cell surface target protein for drug development to treat a variety of diseases, including viral infections and age-related pathologies.

[0067] This application describes the identification of novel human-derived monoclonal antibodies and antigen-binding domains that specifically bind to human CD26. Data provided herein demonstrate that these antibodies and their derivatives can be used as full-length antibodies, scFvs, and scFvs in chimeric antigen receptors that specifically recognize CD26. These antibodies and their derivatives have been applied to the treatment of human diseases.

[0068] protein Provided herein is a protein comprising an anti-CD26 antigen-binding domain, the anti-CD26 antigen-binding domain comprising: (a) a heavy chain variable domain comprising CDR1 comprising SEQ ID NO: 1, CDR2 comprising SEQ ID NO: 2, and CDR3 comprising SEQ ID NO: 3, and a light chain variable domain comprising CDR1 comprising SEQ ID NO: 4, CDR2 comprising SEQ ID NO: 5, and CDR3 comprising SEQ ID NO: 6; (b) a heavy chain variable domain comprising CDR1 comprising SEQ ID NO: 7, CDR2 comprising SEQ ID NO: 8, and CDR3 comprising SEQ ID NO: 9, and a CDR1 comprising SEQ ID NO: 10, CDR2 comprising SEQ ID NO: 11, and a light chain variable domain comprising CDR1 comprising SEQ ID NO: 12, CDR2 comprising SEQ ID NO: 13, and a CDR3 comprising SEQ ID NO: 14; (c) a light chain variable domain comprising CDR1 comprising SEQ ID NO: 13, CDR2 comprising SEQ ID NO: 14, and CDR3 comprising SEQ ID NO: 15, and a light chain variable domain comprising CDR1 comprising SEQ ID NO: 16, CDR2 comprising SEQ ID NO: 17, and CDR3 comprising SEQ ID NO: 18; (d) a heavy chain variable domain comprising CDR1 comprising SEQ ID NO: 19, CDR2 comprising SEQ ID NO: 20, and CDR3 comprising SEQ ID NO: 21, and a CDR1 comprising SEQ ID NO: 22, CDR2 comprising SEQ ID NO: 23, and CDR3 comprising SEQ ID NO: 24. (e) a light chain variable domain comprising a heavy chain variable domain comprising a CDR1 comprising SEQ ID NO: 25, a CDR2 comprising SEQ ID NO: 26, and a CDR3 comprising SEQ ID NO: 27, and a light chain variable domain comprising a CDR1 comprising SEQ ID NO: 28, a CDR2 comprising SEQ ID NO: 29, and a CDR3 comprising SEQ ID NO: 30; (f) a heavy chain variable domain comprising a CDR1 comprising SEQ ID NO: 31, a CDR2 comprising SEQ ID NO: 32, and a CDR3 comprising SEQ ID NO: 33, and a CDR1 comprising SEQ ID NO: 34, a CDR2 comprising SEQ ID NO: 35, and a CDR3 comprising SEQ ID NO: 36. (g) a heavy chain variable domain comprising CDR1 comprising SEQ ID NO: 37, CDR2 comprising SEQ ID NO: 38, and CDR3 comprising SEQ ID NO: 39, and a light chain variable domain comprising CDR1 comprising SEQ ID NO: 40, CDR2 comprising SEQ ID NO: 41, and CDR3 comprising SEQ ID NO: 42; (h) a heavy chain variable domain comprising CDR1 comprising SEQ ID NO: 43, CDR2 comprising SEQ ID NO: 44, and CDR3 comprising SEQ ID NO: 45, and a light chain variable domain comprising CDR1 comprising SEQ ID NO: 46, CDR2 comprising SEQ ID NO: 47, and CDR3 comprising SEQ ID NO: 48;(i) a heavy chain variable domain comprising CDR1 comprising SEQ ID NO: 49, CDR2 comprising SEQ ID NO: 50, and CDR3 comprising SEQ ID NO: 51, and a light chain variable domain comprising CDR1 comprising SEQ ID NO: 52, CDR2 comprising SEQ ID NO: 53, and CDR3 comprising SEQ ID NO: 54; or (j) a heavy chain variable domain comprising CDR1 comprising SEQ ID NO: 55, CDR2 comprising SEQ ID NO: 56, and CDR3 comprising SEQ ID NO: 57, and a light chain variable domain comprising CDR1 comprising SEQ ID NO: 58, CDR2 comprising SEQ ID NO: 59, and CDR3 comprising SEQ ID NO: 60.

[0069] In some examples of any of the proteins described herein, the antigen-binding domain comprises a heavy chain variable domain comprising a CDR1 comprising SEQ ID NO: 1, a CDR2 comprising SEQ ID NO: 2, and a CDR3 comprising SEQ ID NO: 3, and a light chain variable domain comprising a CDR1 comprising SEQ ID NO: 4, a CDR2 comprising SEQ ID NO: 5, and a CDR3 comprising SEQ ID NO: 6. In some examples of any of the proteins described herein, the heavy chain variable domain comprises a sequence that is at least 80% (e.g., at least 85%, at least 90%, at least 92%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) identical to SEQ ID NO: 61. In some examples of any of the proteins described herein, the light chain variable domain comprises a sequence that is at least 80% (e.g., at least 85%, at least 90%, at least 92%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) identical to SEQ ID NO: 62.

[0070] In some examples of any of the proteins described herein, the antigen-binding domain comprises a heavy chain variable domain comprising CDR1 comprising SEQ ID NO:7, CDR2 comprising SEQ ID NO:8, and CDR3 comprising SEQ ID NO:9, and a light chain variable domain comprising CDR1 comprising SEQ ID NO:10, CDR2 comprising SEQ ID NO:11, and CDR3 comprising SEQ ID NO:12. In some examples of any of the proteins described herein, the heavy chain variable domain comprises a sequence that is at least 80% (e.g., at least 85%, at least 90%, at least 92%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) identical to SEQ ID NO:63. In some examples of any of the proteins described herein, the light chain variable domain comprises a sequence that is at least 80% (e.g., at least 85%, at least 90%, at least 92%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) identical to SEQ ID NO:64.

[0071] In some examples of any of the proteins described herein, the antigen-binding domain comprises a heavy chain variable domain comprising CDR1 comprising SEQ ID NO: 13, CDR2 comprising SEQ ID NO: 14, and CDR3 comprising SEQ ID NO: 15, and a light chain variable domain comprising CDR1 comprising SEQ ID NO: 16, CDR2 comprising SEQ ID NO: 17, and CDR3 comprising SEQ ID NO: 18. In some examples of any of the proteins described herein, the heavy chain variable domain comprises a sequence that is at least 80% (e.g., at least 85%, at least 90%, at least 92%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) identical to SEQ ID NO: 65. In some examples of any of the proteins described herein, the light chain variable domain comprises a sequence that is at least 80% (e.g., at least 85%, at least 90%, at least 92%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) identical to SEQ ID NO: 66.

[0072] In some examples of any of the proteins described herein, the antigen-binding domain comprises a heavy chain variable domain comprising a CDR1 comprising SEQ ID NO: 19, a CDR2 comprising SEQ ID NO: 20, and a CDR3 comprising SEQ ID NO: 21, and a light chain variable domain comprising a CDR1 comprising SEQ ID NO: 22, a CDR2 comprising SEQ ID NO: 23, and a CDR3 comprising SEQ ID NO: 24. In some examples of any of the proteins described herein, the heavy chain variable domain comprises a sequence that is at least 80% (e.g., at least 85%, at least 90%, at least 92%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) identical to SEQ ID NO: 67. In some examples of any of the proteins described herein, the light chain variable domain comprises a sequence that is at least 80% (e.g., at least 85%, at least 90%, at least 92%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) identical to SEQ ID NO: 68.

[0073] In some examples of any of the proteins described herein, the antigen-binding domain comprises a heavy chain variable domain comprising a CDR1 comprising SEQ ID NO:25, a CDR2 comprising SEQ ID NO:26, and a CDR3 comprising SEQ ID NO:27, and a light chain variable domain comprising a CDR1 comprising SEQ ID NO:28, a CDR2 comprising SEQ ID NO:29, and a CDR3 comprising SEQ ID NO:30. In some examples of any of the proteins described herein, the heavy chain variable domain comprises a sequence that is at least 80% (e.g., at least 85%, at least 90%, at least 92%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) identical to SEQ ID NO:69. In some examples of any of the proteins described herein, the light chain variable domain comprises a sequence that is at least 80% (e.g., at least 85%, at least 90%, at least 92%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) identical to SEQ ID NO:70.

[0074] In some examples of any protein described herein, the antigen-binding domain comprises a heavy chain variable domain comprising a CDR1 comprising SEQ ID NO: 31, a CDR2 comprising SEQ ID NO: 32, and a CDR3 comprising SEQ ID NO: 33, and a light chain variable domain comprising a CDR1 comprising SEQ ID NO: 34, a CDR2 comprising SEQ ID NO: 35, and a CDR3 comprising SEQ ID NO: 36. In some examples of any protein described herein, the heavy chain variable domain comprises a sequence that is at least 80% (e.g., at least 85%, at least 90%, at least 92%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) identical to SEQ ID NO: 71. In some examples of any protein described herein, the light chain variable domain comprises a sequence that is at least 80% (e.g., at least 85%, at least 90%, at least 92%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) identical to SEQ ID NO: 72.

[0075] In some examples of any protein described herein, the antigen-binding domain comprises a heavy chain variable domain comprising CDR1 comprising SEQ ID NO: 37, CDR2 comprising SEQ ID NO: 38, and CDR3 comprising SEQ ID NO: 39, and a light chain variable domain comprising CDR1 comprising SEQ ID NO: 40, CDR2 comprising SEQ ID NO: 41, and CDR3 comprising SEQ ID NO: 42. In some examples of any protein described herein, the heavy chain variable domain comprises a sequence that is at least 80% (e.g., at least 85%, at least 90%, at least 92%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) identical to SEQ ID NO: 73. In some examples of any protein described herein, the light chain variable domain comprises a sequence that is at least 80% (e.g., at least 85%, at least 90%, at least 92%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) identical to SEQ ID NO: 74.

[0076] In some examples of any of the proteins described herein, the antigen-binding domain comprises a heavy chain variable domain comprising a CDR1 comprising SEQ ID NO: 43, a CDR2 comprising SEQ ID NO: 44, and a CDR3 comprising SEQ ID NO: 45, and a light chain variable domain comprising a CDR1 comprising SEQ ID NO: 46, a CDR2 comprising SEQ ID NO: 47, and a CDR3 comprising SEQ ID NO: 48. In some examples of any of the proteins described herein, the heavy chain variable domain comprises a sequence that is at least 80% (e.g., at least 85%, at least 90%, at least 92%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) identical to SEQ ID NO: 75. In some examples of any of the proteins described herein, the light chain variable domain comprises a sequence that is at least 80% (e.g., at least 85%, at least 90%, at least 92%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) identical to SEQ ID NO: 76.

[0077] In some examples of any protein described herein, the antigen-binding domain comprises a heavy chain variable domain comprising CDR1 comprising SEQ ID NO:49, CDR2 comprising SEQ ID NO:50, and CDR3 comprising SEQ ID NO:51, and a light chain variable domain comprising CDR1 comprising SEQ ID NO:52, CDR2 comprising SEQ ID NO:53, and CDR3 comprising SEQ ID NO:54. In some examples of any protein described herein, the heavy chain variable domain comprises a sequence that is at least 80% (e.g., at least 85%, at least 90%, at least 92%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) identical to SEQ ID NO:77. In some examples of any protein described herein, the light chain variable domain comprises a sequence that is at least 80% (e.g., at least 85%, at least 90%, at least 92%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) identical to SEQ ID NO:78.

[0078] In some examples of any of the proteins described herein, the antigen-binding domain comprises a heavy chain variable domain comprising a CDR1 comprising SEQ ID NO: 55, a CDR2 comprising SEQ ID NO: 56, and a CDR3 comprising SEQ ID NO: 57, and a light chain variable domain comprising a CDR1 comprising SEQ ID NO: 58, a CDR2 comprising SEQ ID NO: 59, and a CDR3 comprising SEQ ID NO: 60. In some examples of any of the proteins described herein, the heavy chain variable domain comprises a sequence that is at least 80% (e.g., at least 85%, at least 90%, at least 92%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) identical to SEQ ID NO: 79. In some examples of any of the proteins described herein, the light chain variable domain comprises a sequence that is at least 80% (e.g., at least 85%, at least 90%, at least 92%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) identical to SEQ ID NO: 80.

[0079] CD26 Ab-01D heavy chain variable domain CDR1 (SEQ ID NO: 1) TINDSYIH CD26 Ab-01D heavy chain variable domain CDR2 (SEQ ID NO: 2) WIWPYGGFTY CD26 Ab-01D heavy chain variable domain CDR3 (SEQ ID NO: 3) ARFLGSSSIMDY CD26 Ab-01D light chain variable domain CDR1 (SEQ ID NO: 4) RASQDVNSNVA CD26 Ab-01D light chain variable domain CDR2 (SEQ ID NO: 5) FGSGGLYS CD26 Ab-01D light chain variable domain CDR3 (SEQ ID NO: 6) QQYSSYPL CD26 Ab-04A Heavy Chain Variable Domain CDR1 (SEQ ID NO: 7) AINNYSIH CD26 Ab-04A heavy chain variable domain CDR2 (SEQ ID NO: 8) SIWPYGGFTS CD26 Ab-04A heavy chain variable domain CDR3 (SEQ ID NO: 9) ARFFSSYGDMDY CD26 Ab-04A Light Chain Variable Domain CDR1 (SEQ ID NO: 10) RASQDVSGGVA CD26 Ab-04A light chain variable domain CDR2 (SEQ ID NO: 11) YGTSGLYS CD26 Ab-04A Light Chain Variable Domain CDR3 (SEQ ID NO: 12) QQGGWPI CD26 Ab-10B heavy chain variable domain CDR1 (SEQ ID NO: 13) TISDYSIH CD26 Ab-10B heavy chain variable domain CDR2 (SEQ ID NO: 14) SIWPGGFTS CD26 Ab-10B heavy chain variable domain CDR3 (SEQ ID NO: 15) ARFHSSSGDMDY CD26 Ab-10B Light Chain Variable Domain CDR1 (SEQ ID NO: 16) RASQDVWGYVA CD26 Ab-10B light chain variable domain CDR2 (SEQ ID NO: 17) FASGALYS CD26 Ab-10B Light Chain Variable Domain CDR3 (SEQ ID NO: 18) QQYFNWPI CD26 Ab-12D Heavy Chain Variable Domain CDR1 (SEQ ID NO: 19) TINSSYIH CD26 Ab-12D heavy chain variable domain CDR2 (SEQ ID NO: 20) GIGPYWGFTS CD26 Ab-12D heavy chain variable domain CDR3 (SEQ ID NO: 21) ARFYSSYGFMDY CD26 Ab-12D Light Chain Variable Domain CDR1 (SEQ ID NO: 22) RASQDVYSWVA CD26 Ab-12D Light Chain Variable Domain CDR2 (SEQ ID NO: 23) YGPGSLYS CD26 Ab-12D Light Chain Variable Domain CDR3 (SEQ ID NO: 24) QQYYNYPL CD26 Ab-03B Heavy Chain Variable Domain CDR1 (SEQ ID NO: 25) TIGNSYIH CD26 Ab-03B heavy chain variable domain CDR2 (SEQ ID NO: 26) GIGPYWGFTS CD26 Ab-03B Heavy Chain Variable Domain CDR3 (SEQ ID NO: 27) ARFNGSSGFMDY CD26 Ab-03B Light Chain Variable Domain CDR1 (SEQ ID NO: 28) RASQDVYYFVA CD26 Ab-03B light chain variable domain CDR2 (SEQ ID NO: 29) SWPTGLYS CD26 Ab-03B Light Chain Variable Domain CDR3 (SEQ ID NO: 30) QQYFSYPI CD26 Ab-07H Heavy Chain Variable Domain CDR1 (SEQ ID NO: 31) KASGYTFARFGMY CD26 Ab-07H heavy chain variable domain CDR2 (SEQ ID NO: 32) FIAPNHGYTF CD26 Ab-07H heavy chain variable domain CDR3 (SEQ ID NO: 33) ARGHWYHGYMDY CD26 Ab-07H Light Chain Variable Domain CDR1 (SEQ ID NO: 34) KSNQNLLYSHGRTYLN CD26 Ab-07H light chain variable domain CDR2 (SEQ ID NO: 35) FGTSHLYS CD26 Ab-07H Light Chain Variable Domain CDR3 (SEQ ID NO: 36) YQGYHVPF CD26 Ab-01G heavy chain variable domain CDR1 (SEQ ID NO: 37) AASGFTIGNYGIH CD26 Ab-01G heavy chain variable domain CDR2 (SEQ ID NO: 38) WIGPSGGYTF CD26 Ab-01G heavy chain variable domain CDR3 (SEQ ID NO: 39) ARFDVHGFHGMDY CD26 Ab-01G light chain variable domain CDR1 (SEQ ID NO: 40) RASQDVNNSVA CD26 Ab-01G light chain variable domain CDR2 (SEQ ID NO: 41) FSPTGLYS CD26 Ab-01G light chain variable domain CDR3 (SEQ ID NO: 42) QQYFDFPL CD26 Ab-04E Heavy Chain Variable Domain CDR1 (SEQ ID NO: 43) AASGFTINDGFIH CD26 Ab-04E heavy chain variable domain CDR2 (SEQ ID NO: 44) GIWPFGGSTS CD26 Ab-04E Heavy Chain Variable Domain CDR3 (SEQ ID NO: 45) ARFDVVDWGVMDY CD26 Ab-04E Light Chain Variable Domain CDR1 (SEQ ID NO: 46) RASQDVNDGVA CD26 Ab-04E Light Chain Variable Domain CDR2 (SEQ ID NO: 47) YWASYLYS CD26 Ab-04E Light Chain Variable Domain CDR3 (SEQ ID NO: 48) QQSWNFPL CD26 Ab-03G heavy chain variable domain CDR1 (SEQ ID NO: 49) AASGFTIGNYGIH CD26 Ab-03G heavy chain variable domain CDR2 (SEQ ID NO: 50) WIGPYGGYTF CD26 Ab-03G heavy chain variable domain CDR3 (SEQ ID NO: 51) ARFNNLLWNGMDY CD26 Ab-03G light chain variable domain CDR1 (SEQ ID NO: 52) RASQDVSSSVA CD26 Ab-03G light chain variable domain CDR2 (SEQ ID NO: 53) SYPGWLYS CD26 Ab-03G light chain variable domain CDR3 (SEQ ID NO: 54) QQFGDFPM CD26 Ab-01F heavy chain variable domain CDR1 (SEQ ID NO: 55) AASGFTISDYSIH CD26 Ab-01F heavy chain variable domain CDR2 (SEQ ID NO: 56) SIWPYGGFTS CD26 Ab-01F heavy chain variable domain CDR3 (SEQ ID NO: 57) ARFHSSSGDMDY CD26 Ab-01F light chain variable domain CDR1 (SEQ ID NO: 58) RASQDVWGYVA CD26 Ab-01F light chain variable domain CDR2 (SEQ ID NO: 59) FSSRSLYS CD26 Ab-01F light chain variable domain CDR3 (SEQ ID NO: 60) QQYFNWPI

[0080] In some embodiments, any protein described herein may comprise an antigen-binding domain comprising a heavy chain variable domain comprising a sequence at least 80% identical (e.g., at least 85% identical, at least 90% identical, at least 92% identical, at least 94% identical, at least 96% identical, at least 98% identical, at least 99% identical, or 100% identical) to SEQ ID NO: 61, and a light chain variable domain comprising a sequence at least 80% identical (e.g., at least 85% identical, at least 90% identical, at least 92% identical, at least 94% identical, at least 96% identical, at least 98% identical, at least 99% identical, or 100% identical) to SEQ ID NO: 62.

[0081] CD26 Ab-01D heavy chain variable domain (SEQ ID NO: 61) EVQLVESGGGLVQPGGSLRLSCAASGFTINDSYIHWVRQAPGKGLEWVAWIWPYGGFTYYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARFLGSSSIMDYWGQG TLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDK THTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEK TISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK CD26 Ab-01D light chain variable domain (SEQ ID NO: 62) DIQMTQSPSSLSASVGDRVTITCRASQDVNSNVAWYQQKPGKAPKLLIFGSGGLYSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQYSSYPLTFGQGTKVETK RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC

[0082] In some embodiments, any protein described herein may comprise an antigen-binding domain comprising a heavy chain variable domain comprising a sequence at least 80% identical (e.g., at least 85% identical, at least 90% identical, at least 92% identical, at least 94% identical, at least 96% identical, at least 98% identical, at least 99% identical, or 100% identical) to SEQ ID NO: 63, and a light chain variable domain comprising a sequence at least 80% identical (e.g., at least 85% identical, at least 90% identical, at least 92% identical, at least 94% identical, at least 96% identical, at least 98% identical, at least 99% identical, or 100% identical) to SEQ ID NO: 64.

[0083] CD26 Ab-04A heavy chain variable domain (SEQ ID NO: 63) EVQLVESGGGLVQPGGSLRLSCAASGFAINNYSIHWVRQAPGKGLEWVASIWPYGGFTSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARFFSSYGDMDYWGQG TLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDK THTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEK TISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK CD26 Ab-04A light chain variable domain (SEQ ID NO: 64) DIQMTQSPSSLSASVGDRVTITCRASQDVSGGVAWYQQKPGKAPKLLIYGTSGLYSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQGGDWPITFGQGTKVEIK RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC

[0084] In some embodiments, any protein described herein may comprise an antigen-binding domain comprising a heavy chain variable domain comprising a sequence at least 80% identical (e.g., at least 85% identical, at least 90% identical, at least 92% identical, at least 94% identical, at least 96% identical, at least 98% identical, at least 99% identical, or 100% identical) to SEQ ID NO:65, and a light chain variable domain comprising a sequence at least 80% identical (e.g., at least 85% identical, at least 90% identical, at least 92% identical, at least 94% identical, at least 96% identical, at least 98% identical, at least 99% identical, or 100% identical) to SEQ ID NO:66.

[0085] CD26 Ab-10B heavy chain variable domain (SEQ ID NO: 65) EVQLVESGGGLVQPGGSLRLSCAASGFTISDYSIHWVRQAPGKGLEWVASIWPYGGFTSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARFHSSSGDMDYWGQG TLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDK THTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEK TISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK CD26 Ab-10B light chain variable domain (SEQ ID NO: 66) DIQMTQSPSSLSASVGDRVTITCRASQDVWGYVAWYQQKPGKAPKLLIFASGALYSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQYFNWPITFGQGTKVEIK RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC

[0086] In some embodiments, any protein described herein may comprise an antigen-binding domain comprising a heavy chain variable domain comprising a sequence at least 80% identical (e.g., at least 85% identical, at least 90% identical, at least 92% identical, at least 94% identical, at least 96% identical, at least 98% identical, at least 99% identical, or 100% identical) to SEQ ID NO: 67, and a light chain variable domain comprising a sequence at least 80% identical (e.g., at least 85% identical, at least 90% identical, at least 92% identical, at least 94% identical, at least 96% identical, at least 98% identical, at least 99% identical, or 100% identical) to SEQ ID NO: 68.

[0087] CD26 Ab-12D heavy chain variable domain (SEQ ID NO: 67) EVQLVESGGGLVQPGGSLRLSCAASGFTINSSYIHWVRQAPGKGLEWVAGIGPYWGFTSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARFYSSYGFMDYWGQG TLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDK THTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEK TISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK CD26 Ab-12D light chain variable domain (SEQ ID NO: 68) DIQMTQSPSSLSASVGDRVTITCRASQDVYSWVAWYQQKPGKAPKLLIYGPGSLYSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQYYNYPLTFGQGTKVEIK RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC

[0088] In some embodiments, any protein described herein may comprise an antigen-binding domain comprising a heavy chain variable domain comprising a sequence at least 80% identical (e.g., at least 85% identical, at least 90% identical, at least 92% identical, at least 94% identical, at least 96% identical, at least 98% identical, at least 99% identical, or 100% identical) to SEQ ID NO: 69, and a light chain variable domain comprising a sequence at least 80% identical (e.g., at least 85% identical, at least 90% identical, at least 92% identical, at least 94% identical, at least 96% identical, at least 98% identical, at least 99% identical, or 100% identical) to SEQ ID NO: 70.

[0089] CD26 Ab-03B heavy chain variable domain (SEQ ID NO: 69) EVQLVESGGGLVQPGGSLRLSCAASGFTIGNSYIHWVRQAPGKGLEWVAGIGPYWGFTSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARFNGSSGFMDYWGQG TLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDK THTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEK TISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK CD26 Ab-03B light chain variable domain (SEQ ID NO: 70) DIQMTQSPSSLSASVGDRVTITCRASQDVYYFVAWYQQKPGKAPKLLISWPTGLYSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQYFSYPITFGQGTKVEIK RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC

[0090] In some embodiments, any protein described herein may comprise an antigen-binding domain comprising a heavy chain variable domain comprising a sequence at least 80% identical (e.g., at least 85% identical, at least 90% identical, at least 92% identical, at least 94% identical, at least 96% identical, at least 98% identical, at least 99% identical, or 100% identical) to SEQ ID NO: 71, and a light chain variable domain comprising a sequence at least 80% identical (e.g., at least 85% identical, at least 90% identical, at least 92% identical, at least 94% identical, at least 96% identical, at least 98% identical, at least 99% identical, or 100% identical) to SEQ ID NO: 72.

[0091] CD26 Ab-07H heavy chain variable domain (SEQ ID NO: 71) EVQLVESGGGLVQPGGSLRLSCKASGYTFARFGMYWVRQAPGKGLEWVAFIAPNHGYTFYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARGHWYHGYMDYWGQGTLVTVSSAS CD26 Ab-07H light chain variable domain (SEQ ID NO: 72) DIQMTQSPSSLSASVGDRVTITCKSNQNLLYSHGRTYLNWYQQKPGKAPKLLIFGTSHLYSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCYQGYHVPFTFGQGTKVEIKR

[0092] In some embodiments, any protein described herein may comprise an antigen-binding domain comprising a heavy chain variable domain comprising a sequence at least 80% identical (e.g., at least 85% identical, at least 90% identical, at least 92% identical, at least 94% identical, at least 96% identical, at least 98% identical, at least 99% identical, or 100% identical) to SEQ ID NO: 73, and a light chain variable domain comprising a sequence at least 80% identical (e.g., at least 85% identical, at least 90% identical, at least 92% identical, at least 94% identical, at least 96% identical, at least 98% identical, at least 99% identical, or 100% identical) to SEQ ID NO: 74.

[0093] CD26 Ab-01G heavy chain variable domain (SEQ ID NO: 73) EVQLVESGGGLVQPGGSLRLSCAASGFTIGNYGIHWVRQAPGKGLEWVAWIGPSGGYTFYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARFDVHGFHGMDYWGQGTLVTVSSAS CD26 Ab-01G light chain variable domain (SEQ ID NO: 74) DIQMTQSPSSLSASVGDRVTITCRASQDVNNSVAWYQQKPGKAPKLLIFSPTGLYSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQYFDFPLTFGQGTKVEIKR

[0094] In some embodiments, any protein described herein may comprise an antigen-binding domain comprising a heavy chain variable domain comprising a sequence at least 80% identical (e.g., at least 85% identical, at least 90% identical, at least 92% identical, at least 94% identical, at least 96% identical, at least 98% identical, at least 99% identical, or 100% identical) to SEQ ID NO: 75, and a light chain variable domain comprising a sequence at least 80% identical (e.g., at least 85% identical, at least 90% identical, at least 92% identical, at least 94% identical, at least 96% identical, at least 98% identical, at least 99% identical, or 100% identical) to SEQ ID NO: 76.

[0095] CD26 Ab-04E heavy chain variable domain (SEQ ID NO: 75) EVQLVESGGGLVQPGGSLRLSCAASGFTINDGFIHWVRQAPGKGLEWVAGIWPFGGSTSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARFDVVDWGVMDYWGQGTLVTVSSAS CD26 Ab-04E light chain variable domain (SEQ ID NO: 76) DIQMTQSPSSLSASVGDRVTITCRASQDVNDGVAWYQQKPGKAPKLLIYWASYLYSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSWNFPLTFGQGTKVEIKR

[0096] In some embodiments, any protein described herein may comprise an antigen-binding domain comprising a heavy chain variable domain comprising a sequence at least 80% identical (e.g., at least 85% identical, at least 90% identical, at least 92% identical, at least 94% identical, at least 96% identical, at least 98% identical, at least 99% identical, or 100% identical) to SEQ ID NO: 77, and a light chain variable domain comprising a sequence at least 80% identical (e.g., at least 85% identical, at least 90% identical, at least 92% identical, at least 94% identical, at least 96% identical, at least 98% identical, at least 99% identical, or 100% identical) to SEQ ID NO: 78.

[0097] CD26 Ab-03G heavy chain variable domain (SEQ ID NO: 77) EVQLVESGGGLVQPGGSLRLSCAASGFTIGNYGIHWVRQAPGKGLEWVAWIGPYGGYTFYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARFNNLLWNGMDYWGQGTLVTVSSAS CD26 Ab-03G light chain variable domain (SEQ ID NO: 78) DIQMTQSPSSLSASVGDRVTITCRASQDVSSSVAWYQQKPGKAPKLLISYPGWLYSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQFGDFPMTFGQGTKVEIKR

[0098] In some embodiments, any protein described herein may comprise an antigen-binding domain comprising a heavy chain variable domain comprising a sequence at least 80% identical (e.g., at least 85% identical, at least 90% identical, at least 92% identical, at least 94% identical, at least 96% identical, at least 98% identical, at least 99% identical, or 100% identical) to SEQ ID NO: 79, and a light chain variable domain comprising a sequence at least 80% identical (e.g., at least 85% identical, at least 90% identical, at least 92% identical, at least 94% identical, at least 96% identical, at least 98% identical, at least 99% identical, or 100% identical) to SEQ ID NO: 80.

[0099] CD26 Ab-01F heavy chain variable domain (SEQ ID NO: 79) EVQLVESGGGLVQPGGSLRLSCAASGFTISDYSIHWVRQAPGKGLEWVASIWPYGGFTSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARFHSSSGDMDYWGQGTLVTVSSAS CD26 Ab-01F light chain variable domain (SEQ ID NO: 80) DIQMTQSPSSLSASVGDRVTITCRASQDVWGYVAWYQQKPGKAPKLLIFSSRSLYSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQYFNWPITFGQGTKVEIKR

[0100] In some embodiments, any protein described herein may comprise an antigen-binding domain comprising a heavy chain variable domain encoded by a nucleic acid comprising a sequence that is at least 80% identical (e.g., at least 85% identical, at least 90% identical, at least 92% identical, at least 94% identical, at least 96% identical, at least 98% identical, at least 99% identical, or 100% identical) to SEQ ID NO: 81, and a light chain variable domain encoded by a nucleic acid comprising a sequence that is at least 80% identical (e.g., at least 85% identical, at least 90% identical, at least 92% identical, at least 94% identical, at least 96% identical, at least 98% identical, at least 99% identical, or 100% identical) to SEQ ID NO: 82.

[0101] CD26 Ab-01D-DNA heavy chain variable domain (SEQ ID NO: 81) GAAGTGCAGCTGGTGGAATCGGGAGGCGGTCTGGTGCAACCTGGCGGCAGCCTTCGTCTGAGCTGTGCGGCGAGCGGGTTCACCATTAACGACTCTTATATTCATTGGGTGCGTCAAGCTCCCGGCAAGGGGCTGGAGTGGGTCGCGTGGATTTGGCCCTACGGGGTTTTACATATTATGCCG ACAGCTGAAAGGTCGCTTTACGATTAGTGCGGACACCAGCAAAAATACCGCGTACCTCAGATGAATAGCCTGCGTGCGGAAGACACAGCGGTGTATTATTGCGCGCGTTTCCTCGGTTCCTCCAGCATTATGGATTATTGGGGGCAGGGCACCCTTGTTACCGTGAGCTCGGCGTCAGCGCC CD26 Ab-01D-DNA light chain variable domain (SEQ ID NO: 82) GATATTCAATGACCCAGAGCCCGAGCAGCCTGAGCGCGAGCGTGGGAGATCGCGTGACCATTACCTGCCGTGCGAGCCAGGATGTTAATAGTAATGTCGCATGGTATCAGCAGAAACCAGGCAAAGCGCCGAAACTTCTGATATTTGGGTCTGGTGGGCTG TATAGCGGCGTGCCGTCGCGTTTTTCGGGCAGTGGCAGCGGCACGGACTTTACCCTGACGATATCTTCCTTACAACCGGAGGATTTTGCGACCTACTACTGTCAACAGTATTCTAGCTACCCGTTAACCTTCGGTCAAGGCACCAAAGTGGAAACCAAACGC

[0102] In some embodiments, any protein described herein may comprise an antigen-binding domain comprising a heavy chain variable domain encoded by a nucleic acid comprising a sequence at least 80% identical (e.g., at least 85% identical, at least 90% identical, at least 92% identical, at least 94% identical, at least 96% identical, at least 98% identical, at least 99% identical, or 100% identical) to SEQ ID NO: 83, and a light chain variable domain encoded by a nucleic acid comprising a sequence at least 80% identical (e.g., at least 85% identical, at least 90% identical, at least 92% identical, at least 94% identical, at least 96% identical, at least 98% identical, at least 99% identical, or 100% identical) to SEQ ID NO: 84.

[0103] CD26 Ab-04A-DNA heavy chain variable domain (SEQ ID NO: 83) GAAGTGCAGCTGGTGGAATCGGGAGGCGGTCTGGTGCAACCTGGCGGCAGCCTTCGTCTGAGCTGTGCGGCGAGCGGGTTCGCCATTAACAATTACTCCATTCATTGGGTGCGTCAAGCTCCCGGCAAGGGGCTGGAGTGGGTCGCGTCTATTTGGCCCTATGGGGGTTTTACATCTTATGCCGACAGCGTGAAAGGTCGCTTTACGATTAGTGCGGACACCAGCAAAAATACCGCGTACCTGCAGATGAATAGCCTGCGTGCGGAAGACACAGCGGTGTATTATTGCGCGCGTTTCTTTAGCTCCTATGGCGATATGGATTATTGGGGGCAGGGCACCCTTGTTACCGTGAGCTCGGCGTCAGCGGCC CD26 Ab-04A-DNA light chain variable domain (SEQ ID NO: 84) GATATTCAAATGACCCAGAGCCCGAGCAGCCTGAGCGCGAGCGTGGGAGATCGCGTGACCATTACCTGCCGTGCGAGCCAGGATGTTAGTGGCGGGGTCGCATGGTATCAGCAGAAACCAGGCAAAGCGCCGAAACTTCTGATATATGGGACAAGTGGGCTGTATAGCGGCGTGCCGTCGCGTTTTTCGGGCAGTGGCAGCGGCACGGACTTTACCCTGACGATATCTTCCTTACAACCGGAGGATTTTGCGACCTACTACTGTCAACAGGGGGGGGATTGGCCGATAACCTTCGGTCAAGGCACCAAAGTGGAAATCAAACGC

[0104] In some embodiments, any protein described herein may comprise an antigen-binding domain comprising a heavy chain variable domain encoded by a nucleic acid comprising a sequence that is at least 80% identical (e.g., at least 85% identical, at least 90% identical, at least 92% identical, at least 94% identical, at least 96% identical, at least 98% identical, at least 99% identical, or 100% identical) to SEQ ID NO: 85, and a light chain variable domain encoded by a nucleic acid comprising a sequence that is at least 80% identical (e.g., at least 85% identical, at least 90% identical, at least 92% identical, at least 94% identical, at least 96% identical, at least 98% identical, at least 99% identical, or 100% identical) to SEQ ID NO: 86.

[0105] CD26 Ab-10B-DNA heavy chain variable domain (SEQ ID NO: 85) GAAGTGCAGCTGGTGGAATCGGGAGGCGGTCTGGTGCAACCTGGCGGCAGCCTTCGTCTGAGCTGTGCGGCGAGCGGGTTCACCATTAGTGACTATTCTATTCATTGGGTGCGTCAAGCTCCCGGCAAGGGGCTGGAGTGGGTCGCGTCTATTTGGCCCTATGGGGGTTTTACATCTTATGCCG ACAGCTGAAAGGTCGCTTTACGATTAGTGCGGACACCAGCAAAAATACCGCGTACCTCAGATGAATAGCCTGCGTGCGGAAGACACAGCGGTGTATTATTGCGCGCGTTTCCACAGCTCCTCCGGCGACATGGATTATTGGGGGCAGGGCACCCTTGTTACCGTGAGCTCGGCGTCAGCGCC CD26 Ab-10B-DNA light chain variable domain (SEQ ID NO: 86) GATATTCAATGACCCAGAGCCCGAGCAGCCTGAGCGCGAGCGTGGGAGATCGCGTGACCATTACCTGCCGTGCGAGCCAGGATGTTTGGGGGTACGTCGCATGGTATCAGCAGAAACCAGGCAAAGCGCCGAAACTTCTGATATTTGCCTCCGGCGCCCTG TATAGCGGCGTGCCGTCGCGTTTTTCGGGCAGTGGCAGCGGCACGGACTTTACCCTGACGATATCTTCCTTACAACCGGAGGATTTTGCGACCTACTACTGTCAACAGTACTTTAATTGCCGATTACCTTCGGTCAAGGCACCAAAGTGGAAATCAAACGC

[0106] In some embodiments, any protein described herein may comprise an antigen-binding domain comprising a heavy chain variable domain encoded by a nucleic acid comprising a sequence at least 80% identical (e.g., at least 85% identical, at least 90% identical, at least 92% identical, at least 94% identical, at least 96% identical, at least 98% identical, at least 99% identical, or 100% identical) to SEQ ID NO: 87, and a light chain variable domain encoded by a nucleic acid comprising a sequence at least 80% identical (e.g., at least 85% identical, at least 90% identical, at least 92% identical, at least 94% identical, at least 96% identical, at least 98% identical, at least 99% identical, or 100% identical) to SEQ ID NO: 88.

[0107] CD26 Ab-12D-DNA heavy chain variable domain (SEQ ID NO: 87) GAAGTGCAGCTGGTGGAATCGGGAGGCGGTCTGGTGCAACCTGGCGGCAGCCTTCGTCTGAGCTGTGCGGCGAGCGGGTTCACCATTAACAGTTCCTACATTCATTGGGTGCGTCAAGCTCCCGGCAAGGGGCTGGAGTGGGTCGCGGGGATTGGGCCCTATTGGGGTTTCACATCTTATGCCGACAGCGTAAAAGGTCGCTTTACGATTAGTGCGGACACCAGCAAAAATACCGCGTACCTGCAGATGAATAGCCTGCGTGCGGAAGACACAGCGGTGTATTATTGCGCGCGTTTCTATAGCTCCTATGGCTTCATGGATTATTGGGGGCAGGGCACCCTTGTTACCGTGAGCTCGGCGTCAGCGGCC CD26 Ab-12D-DNA light chain variable domain (SEQ ID NO: 88) GATATTCAAATGACCCAGAGCCCGAGCAGCCTGAGCGCGAGCGTGGGAGATCGCGTGACCATTACCTGCCGTGCGAGCCAGGATGTTTACTCCTGGGTCGCATGGTATCAGCAGAAACCAGGCAAAGCGCCGAAACTTCTGATATACGGGCCCGGTTCCCTGTATAGCGGCGTGCCGTCGCGTTTTTCGGGCAGTGGCAGCGGCACGGACTTTACCCTGACGATATCTTCCTTACAACCGGAGGATTTTGCGACCTACTACTGTCAACAGTACTATAATTATCCGCTCACCTTCGGTCAAGGCACCAAAGTGGAAATCAAACGC

[0108] In some embodiments, any protein described herein may comprise an antigen-binding domain comprising a heavy chain variable domain encoded by a nucleic acid comprising a sequence that is at least 80% identical (e.g., at least 85% identical, at least 90% identical, at least 92% identical, at least 94% identical, at least 96% identical, at least 98% identical, at least 99% identical, or 100% identical) to SEQ ID NO: 89, and a light chain variable domain encoded by a nucleic acid comprising a sequence that is at least 80% identical (e.g., at least 85% identical, at least 90% identical, at least 92% identical, at least 94% identical, at least 96% identical, at least 98% identical, at least 99% identical, or 100% identical) to SEQ ID NO: 90.

[0109] CD26 Ab-03B-DNA heavy chain variable domain (SEQ ID NO: 89) GAAGTGCAGCTGGTGGAATCGGGAGGCGGTCTGGTGCAACCTGGCGGCAGCCTTCGTCTGAGCTGTGCGGCGAGCGGGTTCACCATTGGTAATTCTTATATTCATTGGGTGCGTCAAGCTCCCGGCAAGGGGCTGGAGTGGGTCGCGGGGATTGGGCCCTATTGGGGTTTCACATCTTATGCCG ACAGCTGAAAGGTCGCTTTACGATTAGTGCGGACACCAGCAAAAATACCGCGTACCTCAGATGAATAGCCTGCGTGCGGAAGACACAGCGGTGTATTATTGCGCGTTTCAACGGCTCTTCTGGTTTTATGGATTATTGGGGGCAGGGCACCCTTGTTACCGTGAGCTCGGCGTCAGCGCC CD26 Ab-03B-DNA light chain variable domain (SEQ ID NO: 90) GATATTCAAATGACCCAGAGCCCGAGCAGCCTGAGCGCGAGCGTGGGAGATCGCGTGACCATTACCTGCCGTGCGAGCCAGGATGTTTATTATTTTGTCGCATGGTATCAGCAGAAACCAGGCAAAGCGCCGAAACTTCTGATATCCTGGCCTACCGGGCTG TATAGCGGCGTGCCGTCGCGTTTTTCGGGCAGTGGCAGCGGCACGGACTTTACCCTGACGATATCTTCCTTACAACCGGAGGATTTTGCGACCTACTACTGTCAACAGTATTTTAGTATCCGATAACCTTCGGTCAAGGCACCAAAGTGGAAATCAAACGC

[0110] In some embodiments, any protein described herein may comprise an antigen-binding domain comprising a heavy chain variable domain encoded by a nucleic acid comprising a sequence that is at least 80% identical (e.g., at least 85% identical, at least 90% identical, at least 92% identical, at least 94% identical, at least 96% identical, at least 98% identical, at least 99% identical, or 100% identical) to SEQ ID NO:91, and a light chain variable domain encoded by a nucleic acid comprising a sequence that is at least 80% identical (e.g., at least 85% identical, at least 90% identical, at least 92% identical, at least 94% identical, at least 96% identical, at least 98% identical, at least 99% identical, or 100% identical) to SEQ ID NO:92.

[0111] CD26 Ab-07H-DNA heavy chain variable domain (SEQ ID NO: 91) GAAGTGCAGCTGGTGGAATCGGGAGGCGGTCTGGTGCAACCTGGCGGCAGCCTTCGTCTGAGCTGTAAGGCGTCTGGCTATACCTTCGCCCGCTTTGGGATGTATTGGGTGCGTCAAGCTCCCGGCAAGGGGCTGGAGTGGGTCGCGTTTATCGCTCCAAATCATGGCTATACATTTTATGCCGACAGCGTGAAAGGTCGCTTTACGATTAGTGCGGACACCAGCAAAAATACCGCGTACCTGCAGATGAATAGCCTGCGTGCGGAAGACACAGCGGTGTATTATTGCGCGCGTGGGCACTGGTACCATGGGTATATGGATTATTGGGGGCAGGGCACCCTTGTTACCGTGAGCTCGGCGTCAGCGGCC CD26 Ab-07H-DNA light chain variable domain (SEQ ID NO: 92) GATATTCAAATGACCCAGAGCCCGAGCAGCCTGAGCGCGAGCGTGGGAGATCGCGTGACCATTACCTGCAAAAGTAACCAGAACCTGCTGTACTCTCACGGCCGGACCTATCTGAATTGGTATCAGCAGAAACCAGGCAAAGCGCCGAAACTTCTGATATTTGGGACGTCTCATCTGTATAGTGGCGTGCCGTCGCGTTTTTCGGGCAGTGGCAGCGGCACGGACTTTACCCTGACGATATCTTCCTTACAACCGGAGGATTTTGCGACCTACTACTGTTATCAGGGCTATCATGTTCCCTTCACCTTCGGTCAAGGCACCAAAGTGGAAATCAAACGC

[0112] In some embodiments, any protein described herein may comprise an antigen-binding domain comprising a heavy chain variable domain encoded by a nucleic acid comprising a sequence that is at least 80% identical (e.g., at least 85% identical, at least 90% identical, at least 92% identical, at least 94% identical, at least 96% identical, at least 98% identical, at least 99% identical, or 100% identical) to SEQ ID NO:93, and a light chain variable domain encoded by a nucleic acid comprising a sequence that is at least 80% identical (e.g., at least 85% identical, at least 90% identical, at least 92% identical, at least 94% identical, at least 96% identical, at least 98% identical, at least 99% identical, or 100% identical) to SEQ ID NO:94.

[0113] CD26 Ab-01G-DNA heavy chain variable domain (SEQ ID NO: 93) GAAGTGCAGCTGGTGGAATCGGGAGGCGGTCTGGTGCAACCTGGCGGCAGCCTTCGTCTGAGCTGTGCGGCGAGCGGGTTCACCATTGGCAATTACGGGATTCATTGGGTGCGTCAAGCTCCCGGCAAGGGGCTGGAGTGGGTCGCGTGGATTGGGCCCTCGGGGTTATACATTCTATGCCGAC AGCGTGAAAGGTCGCTTTACGATTAGTGCGGACACCAGCAAAAATACCGCGTACCTGCAGATGAATAGCCTGCGTGCGGAAGACACAGCGGTGTATTATTGCGCGCGTTTCGACGTTCACGGGTTCCACGGGATGGATTATTGGGGGCAGGGCACCCTTGTTACCGTGAGCTCGGCGTCAGCGCC CD26 Ab-01G-DNA light chain variable domain (SEQ ID NO: 94) GATATTCAAATGACCCAGAGCCCGAGCAGCCTGAGCGCGAGCGTGGGAGATCGCGTGACCATTACCTGCCGTGCGAGCCAGGATGTTAACAACAGTGTCGCATGGTATCAGCAGAAACCAGGCAAAGCGCCGAAACTTCTGATATTTTCCCCTACTGGGCTG TATAGCGGCGTGCCGTCGCGTTTTTCGGGCAGTGGCAGCGGCACGGACTTTACCCTGACGATATCTTCCTTACAACCGGAGGATTTTGCGACCTACTACTGTCAACAGTACTTCGACTTCCCGTTAACCTTCGGTCAAGGCACCAAAGTGGAAATCAAACGT

[0114] In some embodiments, any protein described herein may comprise an antigen-binding domain comprising a heavy chain variable domain encoded by a nucleic acid comprising a sequence that is at least 80% identical (e.g., at least 85% identical, at least 90% identical, at least 92% identical, at least 94% identical, at least 96% identical, at least 98% identical, at least 99% identical, or 100% identical) to SEQ ID NO:95, and a light chain variable domain encoded by a nucleic acid comprising a sequence that is at least 80% identical (e.g., at least 85% identical, at least 90% identical, at least 92% identical, at least 94% identical, at least 96% identical, at least 98% identical, at least 99% identical, or 100% identical) to SEQ ID NO:96.

[0115] CD26 Ab-04E-DNA heavy chain variable domain (SEQ ID NO: 95) GAAGTGCAGCTGGTGGAATCGGGAGGCGGTCTGGTGCAACCTGGCGGCAGCCTTCGTCTGAGCTGTGCGGCGAGCGGGTTCACCATTAACGACGGGTTCATTCATTGGGTGCGTCAAGCTCCCGGCAAGGGGCTGGAGTGGGTCGCGGGGATTTGGCCCTTTGGGGGTTCCACATCCTATGCCGACAGCGTGAAAGGTCGCTTTACGATTAGTGCGGACACCAGCAAAAATACCGCGTACCTGCAGATGAATAGCCTGCGTGCGGAAGACACAGCGGTGTATTATTGCGCGCGTTTCGATGTCGTCGATTGGGGGGTCATGGATTATTGGGGGCAGGGCACCCTTGTTACCGTGAGCTCGGCGTCAGCGGCC CD26 Ab-04E-DNA light chain variable domain (SEQ ID NO: 96) GATATTCAAATGACCCAGAGCCCGAGCAGCCTGAGCGCGAGCGTGGGAGATCGCGTGACCATTACCTGCCGTGCGAGCCAGGATGTTAATGATGGCGTCGCATGGTATCAGCAGAAACCAGGCAAAGCGCCGAAACTTCTGATATATTGGGCGAGTTACCTGTATAGCGGCGTGCCGTCGCGTTTTTCGGGCAGTGGCAGCGGCACGGACTTTACCCTGACGATATCTTCCTTACAACCGGAGGATTTTGCGACCTACTACTGTCAACAGTCCTGGAATTTTCCGCTCACCTTCGGTCAAGGCACCAAAGTGGAAATCAAACGC

[0116] In some embodiments, any protein described herein may comprise an antigen-binding domain comprising a heavy chain variable domain encoded by a nucleic acid comprising a sequence at least 80% identical (e.g., at least 85% identical, at least 90% identical, at least 92% identical, at least 94% identical, at least 96% identical, at least 98% identical, at least 99% identical, or 100% identical) to SEQ ID NO:97, and a light chain variable domain encoded by a nucleic acid comprising a sequence at least 80% identical (e.g., at least 85% identical, at least 90% identical, at least 92% identical, at least 94% identical, at least 96% identical, at least 98% identical, at least 99% identical, or 100% identical) to SEQ ID NO:98.

[0117] CD26 Ab-03G-DNA heavy chain variable domain (SEQ ID NO: 97) GAAGTGCAGCTGGTGGAATCGGGAGGCGGTCTGGTGCAACCTGGCGGCAGCCTTCGTCTGAGCTGTGCGGCGAGCGGGTTCACCATTGGTAATTACGGGATTCATTGGGTGCGTCAAGCTCCCGGCAAGGGGCTGGAGTGGGTCGCGTGGATTGGGCCCTATGGGGGTTACACATTCTATGCCGAC AGCGTGAAAGGTCGCTTTACGATTAGTGCGGACACCAGCAAAAATACCGCGTACCTGCAGATGAATAGCCTGCGTGCGGAAGACACAGCGGTGTATTATTGCGCGCGTTTCAATAACCTTCTTTGGAATGGGATGGATTATTGGGGGCAGGGCACCCTTGTTACCGTGAGCTCGGCGTCAGCGCC CD26 Ab-03G-DNA light chain variable domain (SEQ ID NO: 98) GATATTCAAATGACCCAGAGCCCGAGCAGCCTGAGCGCGAGCGTGGGAGATCGCGTGACCATTACCTGCCGTGCGAGCCAGGATGTTTCCTCTTCCGTCGCATGGTATCAGCAGAAACCAGGCAAAGCGCCGAAACTTCTGATATCCTATCCTGGTTGGCTG TATAGCGGCGTGCCGTCGCGTTTTTCGGGCAGTGGCAGCGGCACGGACTTTACCCTGACGATATCTTCCTTACAACCGGAGGATTTTGCGACCTACTACTGTCAACAGTTTGGGGATTTTCCGATGACCTTCGGTCAAGGCACCAAAGTGGAAATCAAACGC

[0118] In some embodiments, any protein described herein may comprise an antigen-binding domain comprising a heavy chain variable domain encoded by a nucleic acid comprising a sequence that is at least 80% identical (e.g., at least 85% identical, at least 90% identical, at least 92% identical, at least 94% identical, at least 96% identical, at least 98% identical, at least 99% identical, or 100% identical) to SEQ ID NO:99, and a light chain variable domain encoded by a nucleic acid comprising a sequence that is at least 80% identical (e.g., at least 85% identical, at least 90% identical, at least 92% identical, at least 94% identical, at least 96% identical, at least 98% identical, at least 99% identical, or 100% identical) to SEQ ID NO:100.

[0119] CD26 Ab-01F-DNA heavy chain variable domain (SEQ ID NO: 99) GAAGTGCAGCTGGTGGAATCGGGAGGCGGTCTGGTGCAACCTGGCGGCAGCCTTCGTCTGAGCTGTGCGGCGAGCGGGTTCACCATTAGTGACTATTCTATTCATTGGGTGCGTCAAGCTCCCGGCAAGGGGCTGGAGTGGGTCGCGTCTATTTGGCCCTATGGGGGTTTTACATCTTATGCCG ACAGCTGAAAGGTCGCTTTACGATTAGTGCGGACACCAGCAAAAATACCGCGTACCTCAGATGAATAGCCTGCGTGCGGAAGACACAGCGGTGTATTATTGCGCGCGTTTCCACAGCTCCTCCGGCGACATGGATTATTGGGGGCAGGGCACCCTTGTTACCGTGAGCTCGGCGTCAGCGCC CD26 Ab-01F-DNA light chain variable domain (SEQ ID NO: 100) GATATTCAATGACCCAGAGCCCGAGCAGCCTGAGCGCGAGCGTGGGAGATCGCGTGACCATTACCTGCCGTGCGAGCCAGGATGTTTGGGGGTACGTCGCATGGTATCAGCAGAAACCAGGCAAAGCGCCGAAACTTCTGATATTTTCCTCCCGCTCCCTG TATAGCGGCGTGCCGTCGCGTTTTTCGGGCAGTGGCAGCGGCACGGACTTTACCCTGACGATATCTTCCTTACAACCGGAGGATTTTGCGACCTACTACTGTCAACAGTACTTTAATTGCCGATTACCTTCGGTCAAGGCACCAAAGTGGAAATCAAACGC

[0120] In some embodiments, the protein can be a single-chain polypeptide. In some embodiments, the protein can be a multi-chain polypeptide. In some examples, the proteins described herein can be antibodies, antigen-binding antibody fragments, or chimeric antigen receptors.

[0121] antigen-binding domain Each antigen-binding domain present in any of the proteins described herein (either the single-chain or multi-chain proteins described herein) is independently selected from the group consisting of a VHH domain, a VNAR domain, and an scFv. In some embodiments, any of the antigen-binding domains described herein is a BiTe, (scFv)2, a nanobody, a nanobody-HSA, a DART, a TandAb, a scDiabody, a scDiabody-CH3, a scFv-CH-CL-scFv, an HSAbody, a scDiabody-HAS, or a tandem-scFv. Additional examples of antigen-binding domains that can be used in any of the proteins described herein are known in the art.

[0122] VHH domains are single monomeric variable antibody domains that can be found in camelids. VNAR domains are single monomeric variable antibody domains that can be found in cartilaginous fish. Non-limiting embodiments of VHH domains and VNAR domains are described, for example, in Cromie et al., Curr. Top. Med. Chem. 15:2543-2557, 2016; De Genst et al., Dev. Comp. Immunol. 30:187-198, 2006; De Meyer et al., Trends Biotechnol. 32:263-270, 2014; Kijanka et al., Nanomedicine 10:161-174, 2015; Kovaleva et al., Expert. Opin. Biol. Ther. 14:1527-1539, 2014; Krah et al., Immunopharmacol. Immunotoxicol. 38:21-28, 2016; Mujic-Delic et al., Trends Pharmacol.Sci.35:247-255,2014, Muyldermans,J.Biotechnol.74:277-302,2001, Muyldermans et al.,Trends Biochem.Sci.26:230-235,2001, Muyldermans,Ann.Rev.Biochem.82:775-797,2013, Rahbarizadeh et al.,Immunol.Invest.40:299-338,2011, Van Audenhove et al.,EBioMedicine 8:40-48,2016, Van Bockstaele et al.,Curr.Opin.Investig.Drugs 10:1212-1224,2009, Vincke et al.,Methods Mol. Biol. 911:15-26, 2012, and Wesolowski et al., Med. Microbiol. Immunol. 198:157-174, 2009.

[0123] In some embodiments, two or more polypeptides present in a multi-chain protein are assembled (e.g., non-covalently assembled) to form any of the antigen-binding domains described herein, e.g., an antigen-binding fragment of an antibody (e.g., any of the antigen-binding fragments of an antibody described herein), VHH-scAb, VHH-Fab, double-scFab, F(ab'), diabody, crossMab, DAF (two-in-one), DAF (four-in-one), DutaMab, DT-IgG, knobs-in-hole common light chain, knobs-in-hole assembly, charge pair, Fab arm exchange, SEEDbody, LUZ-Y, Fcab, κλ-body, orthogonal Fab, DVD-IgG, IgG(H)-scFv, scFv-(H)IgG, IgG(L)-scFv, scFv-(L)IgG, IgG(L,H)-Fv, IgG(H)-V, V(H)-IgG, IgG(L)-V, V(L)-IgG, KIH IgG-scFab, 2scFv-IgG, IgG-2scFv, scFv4-Ig, Zybody, DVI-IgG, diabody-CH3, triplebody, minibody, minibody, TriBi minibody, scFv-CH3 KIH, Fab-scFv, F(ab')2-scFv2, scFv-KIH, Fab-scFv-Fc, tetravalent HCAb, sc diabody-Fc, diabody-Fc, tandem scFv-Fc, intrabody, dock-and-lock, lmmTAC, IgG-IgG conjugate, Cov-X-Body, and scFv1-PEG-scFv2 can be formed. For a description of these components, see, e.g., Spiess et al., Mol. Immunol. 67:95-106, 2015, which is incorporated herein in its entirety. Non-limiting examples of antigen-binding fragments of antibodies include Fv fragments, Fab fragments, F(ab')2 fragments, and Fab' fragments.Additional examples of antigen-binding fragments of antibodies are antigen-binding fragments of IgG (e.g., antigen-binding fragments of IgG1, IgG2, IgG3, or IgG4) (e.g., antigen-binding fragments of human or humanized IgG, e.g., human or humanized IgG1, IgG2, IgG3, or IgG4), antigen-binding fragments of IgA (e.g., antigen-binding fragments of IgA1 or IgA2) (e.g., antigen-binding fragments of human or humanized IgA, e.g., human or humanized IgA1 or IgA2), antigen-binding fragments of IgD (e.g., antigen-binding fragments of human or humanized IgD), antigen-binding fragments of IgE (e.g., antigen-binding fragments of human or humanized IgE), or antigen-binding fragments of IgM (e.g., antigen-binding fragments of human or humanized IgM).

[0124] An "Fv" fragment comprises a non-covalent dimer of one heavy-chain and one light-chain variable domain.

[0125] A "Fab" fragment contains the heavy and light chain variable domains of an Fv fragment as well as the constant domain of the light chain and the first constant domain of the heavy chain (C H1 ) is included.

[0126] An "F(ab')2" fragment contains two Fab fragments linked by a disulfide bond near the hinge region.

[0127] "Dual variable domain immunoglobulin" or "DVD-Ig" refers to the multivalent and multispecific binding proteins described, for example, in DiGiammarino et al., Methods Mol. Biol. 899:145-156, 2012; Jakob et al., MABs 5:358-363, 2013; and U.S. Pat. Nos. 7,612,181, 8,258,268, 8,586,714, 8,716,450, 8,722,855, 8,735,546, and 8,822,645, each of which is incorporated by reference in its entirety.

[0128] DART is described, for example, in Garber, Nature Reviews Drug Discovery 13:799-801, 2014.

[0129] antibody In some embodiments, a protein described herein can be an antibody (e.g., a human or humanized antibody). In some embodiments, the antibody can be a human or humanized IgG, e.g., a human or humanized IgG1, IgG2, IgG3, or IgG4. In some embodiments, the antibody can be a human or humanized IgA (e.g., a human or humanized IgA1 or IgA2). In some embodiments, the antibody can be a human or humanized IgD, a human or humanized IgE, or a human or humanized IgM.

[0130] In some embodiments, any of the proteins described herein can include an Fc receptor (eg, an Fc receptor comprising three substitutions: S239D, A330L, and I332E).

[0131] Chimeric Antigen Receptor In some embodiments, the proteins described herein may be chimeric antigen receptors. A chimeric antigen receptor comprises an extracellular antigen-binding domain (e.g., any of the anti-CD26 antigen-binding regions described herein), a transmembrane domain, a costimulatory domain (e.g., an intracellular CD28 domain), and a CD3 zeta signaling domain. In some examples, a chimeric antigen receptor may comprise an extracellular antigen-binding domain (e.g., any of the anti-CD26 antigen-binding regions described herein), a transmembrane domain (e.g., a CD8 alpha transmembrane domain), a CD28 intracellular signaling domain, and a CD3 zeta intracellular signaling domain. In some embodiments, a chimeric antigen receptor may comprise a hinge region (e.g., a CD8 alpha hinge region) disposed between the extracellular antigen-binding domain and the transmembrane domain.

[0132] In some embodiments, the hinge region can comprise a sequence that is at least 80% identical (e.g., at least 85%, at least 90%, at least 92%, at least 94%, at least 96%, at least 98%, at least 99%, or 100% identical) to SEQ ID NO: 254. In some embodiments, the hinge region can be encoded by a sequence that is at least 80% identical (e.g., at least 85%, at least 90%, at least 92%, at least 94%, at least 96%, at least 98%, at least 99%, or 100% identical) to SEQ ID NO: 255.

[0133] Human CD8 alpha hinge (SEQ ID NO: 254) ALSNSIMYFSHFVPVFLPAKPTTTPAPRPPTPAPTIASQPLSLRPEASRPAAGGAVHTRGLD Human CD8 alpha hinge (SEQ ID NO: 255) GCTTTAAGCAACTCCATCATGTACTTCTCCCACTTCGTGCCCGTTTTTTTACCCGCTAAGCCCACCACAACCCCCGCTCCCAGACCCCCTACCCCCGCTCCTACCATCGCCTCCCAGCCTTTATCTTTAAGACCCGAAGCCTCTCGTCCCGCTGCCGGCGGCGCCGTGCACACAAGGGGTTTAGAC

[0134] For example, the transmembrane domain can comprise a sequence at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100% identical to SEQ ID NO: 101 (FWVLVVVGGVLACYSLLVTVAFIIFWV). For example, the transmembrane domain can be a transmembrane domain derived from CD28 (e.g., human CD28). In some embodiments, the chimeric antigen receptor can comprise a transmembrane and cytoplasmic signaling domain derived from CD28 comprising a sequence at least 80% identical (e.g., at least 85%, at least 90%, at least 92%, at least 94%, at least 96%, at least 98%, at least 99%, or 100% identical) to SEQ ID NO: 256. In some embodiments, the transmembrane and cytoplasmic signaling domains from CD28 can be encoded by a sequence that is at least 80% identical (e.g., at least 85%, at least 90%, at least 92%, at least 94%, at least 96%, at least 98%, at least 99%, or 100% identical) to SEQ ID NO:257.

[0135] Human CD28 transmembrane domain (SEQ ID NO: 256) KPFWVLVVVGGVLACYSLLVTVAFIIFWVRSKRSRLLHSDYMNMTPRRPGPTRKHYQPYAPPRDFAAYRS Human CD28 transmembrane domain (SEQ ID NO: 257) AAGCCTTTTTGGGTTTTAGTGGTGGTGGGCGGCGTGCTGGCTTGTTACTCTTTACTGGTGACCGTGGCCTTCATCATCTTCTGGGTTCGTTCCAAGAGGTCTCGTCTGCTGCACTCCGACTATATGAACATGACCCCTAGGAGGCCCGGCCCTACCAGAAAACACTATCAGCCCTATGCCCCTCCTCGTGACTTTGCCGCTTATCGTTCT

[0136] For example, the costimulatory domain can comprise a sequence that is at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100% identical to SEQ ID NO: 102 (RSKRSRLLHSDYMNMTPRRPGPTRKHYQPYAPPRDFAAYRS).

[0137] For example, the CD3 zeta signaling domain can comprise a sequence that is at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100% identical to SEQ ID NO: 103 (RVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPQRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALPPR).

[0138] For example, the CD3 zeta signaling domain can comprise a sequence that is at least 80% (e.g., at least 85%, at least 90%, at least 92%, at least 94%, at least 96%, at least 98%, at least 99%, or 100% identical) to SEQ ID NO: 258. For example, the CD3 zeta signaling domain can be encoded by a sequence that is at least 80% identical (e.g., at least 85%, at least 90%, at least 92%, at least 94%, at least 96%, at least 98%, at least 99%, or 100% identical) to SEQ ID NO: 259.

[0139] Human CD3 zeta signaling domain (SEQ ID NO: 258) RVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPQRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALPPR Human CD3 zeta signaling domain (SEQ ID NO: 259) CGTGTGAAGTTCTCCAGATCCGCCGATGCCCCCGCTTACCAGCAAGGTCAAAACCAGCTCTATAACGAGCTGAATTTAGGTCGTAGAGAGGAGTACGACGTGCTGGATAAAAGAAGGGGCAGAGACCCCGAAATGGGAGGCAAACCCCAGAGAAGGAAGAACCCCCAAG AAGGACTGTACAACGAACTGCAGAAGGATAAGATGGCCGAGGCCTACTCCGAGATTGGCATGAAAGGCGAGAGGAGGAGGGGCAAGGGCCATGATGGTTTATACCAAGGTTTATCCACAGCTACAAAGGACACCTACGACGCTTTACACATGCAAGCTTTACCTCCTAGA

[0140] In some embodiments, the chimeric antigen receptor may comprise a sequence that is at least 80% identical (e.g., at least 85%, at least 90%, at least 92%, at least 94%, at least 96%, at least 98%, at least 99%, or 100% identical) to SEQ ID NO: 253. In some embodiments, the chimeric antigen receptor may be encoded by a sequence that is at least 80% identical (e.g., at least 85%, at least 90%, at least 92%, at least 94%, at least 96%, at least 98%, at least 99%, or 100% identical) to SEQ ID NO: 252.

[0141] Exemplary anti-CD26 chimeric antigen receptor (with signal sequence) (SEQ ID NO: 253) MDRLTSSFLLLIVPAYVLSDIQMTQSPSSLSASVGDRVTITCRASQDVNSNVAWYQQKPGKAPKLLIFGSGGLYSGVPSRFSGSGSGTFTLTISSLQPEDFATYYCQQYSSYPLTFGQGTKVETKGGG GSGGGGSGGGGSEVQLVESGGGLVQPGGSLRLSCAASGFTINDSYIHWVRQAPGKGLEWVAWIWPYGGFTYYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARFLGSSSIMDYWGQGTLVTV SSAEQKLISEEDLALSNSIMYFSHFVPVFLPAKPTTTPAPRPPTPAPTIASQPLSLRPEASRPAAGGAVHTRGLDKPFWVLVVVGGVLACYSLLVTVAFIIFWVRSKRSRLLHSDYMNMTPRRPGPTRK HYQPYAPPRDFAAYRSRVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPQRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALPPR Exemplary anti-CD26 chimeric antigen receptor (with signal sequence) (SEQ ID NO: 252)

[0142] In some embodiments, the chimeric antigen receptor may comprise a sequence that is at least 80% identical (e.g., at least 85%, at least 90%, at least 92%, at least 94%, at least 96%, at least 98%, at least 99%, or 100% identical) to SEQ ID NO: 260. In some embodiments, the chimeric antigen receptor may be encoded by a sequence that is at least 80% identical (e.g., at least 85%, at least 90%, at least 92%, at least 94%, at least 96%, at least 98%, at least 99%, or 100% identical) to SEQ ID NO: 261.

[0143] Exemplary anti-CD26 chimeric antigen receptor (without signal sequence) (SEQ ID NO: 260) DIQMTQSPSSLSASVGDRVTITCRASQDVNSNVAWYQQKPGKAPKLLIFGSGGLYSGVPSRFSGSGSGTTDFTLTISSLQPEDFATYYCQQYSSYPLTFGQGTKVETKGGGGSGGGGSGGGSEV QLVESGGGLVQPGGSLRLSCAASGFTINDSYIHWVRQAPGKGLEWVAWIWPYGGFTYYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARFLGSSSIMDYWGQGTLVTVSSAEQKLIS EEDLALSNSIMYFSHFVPVFLPAKPTTTPAPRPPTPAPTIASQPLSLRPEASRPAAGGAVHTRGLDKPFWVLVVVGGVLACYSLLVTVAFIIFWVRSKRSRLLHSDYMNMTPRRPGPTRKHYQP YAPPRDFAAYRSRVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPQRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALPPR Exemplary anti-CD26 chimeric antigen receptor (without signal sequence) (SEQ ID NO: 261)

[0144] nucleic acid Also provided herein are nucleic acids comprising sequences encoding any of the proteins described herein. Also provided herein are sets of nucleic acids which together comprise sequences encoding any of the multi-chain proteins described herein.

[0145] Also provided herein are vectors containing sequences encoding any of the proteins described herein. Also provided herein are sets of vectors that together contain sequences encoding any of the multi-chain proteins described herein. Non-limiting examples of vectors include expression vectors. Examples of expression vectors include viral vectors (e.g., lentiviral vectors, adeno-associated viral vectors, or retroviral vectors).

[0146] Some embodiments of any of the vectors or nucleic acids described herein can further comprise a promoter operably linked to the protein-encoding sequence or sequences.

[0147] cell Also provided herein are cells comprising a nucleic acid encoding any of the proteins described herein or a vector comprising any of the nucleic acids described herein. In some examples of any of the cells described herein, the cell is an immune cell. Alternatively, the cell is a production cell line, including, but not limited to, Chinese hamster ovary (CHO) cells (e.g., CHO.K1, CD-CHO, CHO-S, GS CHO, CHO-DG44, etc.), HEK293, Cos, NS0, Sp2 / 0, and PerC6 cells.

[0148] As used herein, "immune cell" refers to a cell of the immune system that can be classified as a lymphocyte (e.g., a T cell, a B cell, and a NK cell), a neutrophil, and a monocyte / macrophage. In some examples of any cell described herein, the immune cell is a T cell, a B cell, or a natural killer (NK) cell. In some examples of any cell described herein, the immune cell can be a T cell, e.g., a population of T cells including CD4+ T cells, CD8+ T cells, Treg cells, Th1 T cells, Th2 T cells, Th17 T cells, nonspecific T cells, or a combination thereof.

[0149] composition Also provided herein are compositions (e.g., pharmaceutical compositions) comprising at least one of any of the proteins described herein or any of the cells described herein. Also provided herein are compositions (e.g., pharmaceutical compositions) comprising at least one of any of the nucleic acids described herein or any of the vectors described herein. In some embodiments, the compositions (e.g., pharmaceutical compositions) can be placed in sterile vials or pre-loaded syringes.

[0150] In some embodiments, the compositions (e.g., pharmaceutical compositions) are formulated for different routes of administration (e.g., intravenous, subcutaneous, intramuscular, or intratumor). In some embodiments, the compositions (e.g., pharmaceutical compositions) may include a pharmaceutically acceptable carrier (e.g., phosphate-buffered saline). Single or multiple administrations of any of the pharmaceutical compositions described herein may be given to a subject, depending, for example, on the dosage and frequency required and tolerated by the patient. The dose of the pharmaceutical composition should provide a sufficient quantity of protein, nucleic acid, vector, or cell to effectively treat or ameliorate a condition, disease, or symptom.

[0151] Also provided herein are methods of treating a subject having cancer (e.g., any of the cancers described herein), comprising administering a therapeutically effective amount of at least one of any of the compositions or pharmaceutical compositions provided herein.

[0152] kit Also provided herein is a kit comprising any of the pharmaceutical compositions described herein. In some embodiments, the kit may include instructions for performing any of the methods described herein. In some embodiments, the kit may include at least one dose of any of the compositions (e.g., pharmaceutical compositions) described herein. In some embodiments, the kit may provide a syringe for administering any of the pharmaceutical compositions described herein.

[0153] Methods of treating age-related and inflammatory diseases in a subject Provided herein are methods of treating an age-related or inflammatory disease in a subject, comprising administering a therapeutically effective amount of any of the proteins described herein or any of the pharmaceutical compositions described herein. Also provided herein are methods of treating an age-related or inflammatory disease in a subject, comprising administering a therapeutically effective amount of any of the nucleic acids described herein or any of the pharmaceutical compositions described herein. Also provided herein are methods of treating an age-related or inflammatory disease in a subject, comprising administering a therapeutically effective amount of any of the cells described herein or any of the pharmaceutical compositions described herein.

[0154] In some embodiments, the method further comprises administering (i) a therapeutically effective amount of an NK cell activator and / or NK cells and / or a monoclonal antibody, and (ii) a therapeutically effective amount of a Treg cell activator and / or Treg cells and / or a monoclonal antibody and / or an advanced glycation end product (AGE) inhibitor. In some embodiments, the age-related disease is inflammation-aging associated.

[0155] Also provided herein is a method for treating an age-related disease or an inflammatory disease, comprising administering (i) a therapeutically effective amount of an NK cell activator and / or NK cells and / or a monoclonal antibody, and (ii) a therapeutically effective amount of a Treg cell activator and / or Treg cells and / or a monoclonal antibody and / or an advanced glycation end product (AGE) inhibitor.

[0156] In some embodiments of any of the methods described herein, (i) is administered to the subject substantially simultaneously with (ii). In some embodiments of any of the methods described herein, (i) is administered to the subject before (ii) is administered to the subject. In some embodiments of any of the methods described herein, (ii) is administered to the subject before (i) is administered to the subject. In some embodiments, the subject is administered the protein, cells, or nucleic acid substantially simultaneously with (i) and (ii). In some embodiments, the subject is administered the protein, cells, or nucleic acid before administration of (i) and (ii). In some embodiments, the subject is administered the protein, cells, or nucleic acid after administration of (i) and (ii).

[0157] In some embodiments of any of the methods described herein, the method comprises administering to a subject a therapeutically effective amount of NK cells. In some embodiments, the NK cells are autologous NK cells. In some embodiments, the method can further comprise isolating NK cells from the subject and culturing the isolated NK cells in a liquid culture medium under conditions sufficient to induce or increase the proliferation of the NK cells, wherein after the isolating and culturing steps, the NK cells are administered to the subject. In some embodiments, the liquid culture medium comprises one or more multi-chain chimeric polypeptides (e.g., any of the exemplary multi-chain chimeric polypeptides described herein).

[0158] In some embodiments, the NK cells comprise a chimeric antigen receptor (e.g., the chimeric antigen receptor comprises an extracellular domain that specifically binds tissue factor or to CD26) (e.g., any of the chimeric antigen receptors described herein that comprise any of the anti-CD26 antigen binding domains described herein).

[0159] In some embodiments, the method may include administering to the subject a therapeutically effective amount of an NK cell activator. In some embodiments, the NK cell activator is one or more multi-chain chimeric polypeptides (e.g., one or more of any of the multi-chain chimeric polypeptides described herein). In some embodiments, the NK cell activator is one or more of an anti-tissue factor antibody, an anti-CD26 antibody (e.g., any of the anti-CD26 antibodies described herein), and / or an anti-CD36 antibody. In some embodiments, the NK cell activator comprises one or more multi-chain chimeric polypeptides and one or more of an anti-tissue factor antibody, an anti-CD26 antibody (e.g., any of the anti-CD26 antibodies described herein), and / or an anti-CD36 antibody.

[0160] In some embodiments, the method comprises administering to the subject a therapeutically effective amount of Treg cells. In some embodiments, the Treg cells are autologous Treg cells. In some embodiments, the method further comprises culturing the isolated Treg cells in a liquid culture medium under conditions sufficient to induce or increase the proliferation of Treg cells, wherein after the isolating and culturing steps, the Treg cells are administered to the subject. In some embodiments, the liquid culture medium comprises one or more single-chain chimeric polypeptides.

[0161] In some embodiments, the Treg cells comprise a chimeric antigen receptor (e.g., a chimeric antigen receptor comprising an extracellular domain that specifically binds to tissue factor, to CD26 (e.g., any of the anti-CD26 antibodies described herein), and / or to CD36).

[0162] In some embodiments, the method includes administering to the subject a therapeutically effective amount of a Treg cell activator. In some embodiments, the Treg cell activator is one or more single-chain chimeric polypeptides (e.g., one or more of any of the single-chain chimeric polypeptides described herein). In some embodiments, the Treg cell activator is one or both of an anti-tissue factor antibody, an anti-CD26 antibody (e.g., any of the anti-CD26 antibodies described herein), and / or an anti-CD36 antibody. In some embodiments, the Treg cell activator is a soluble RAGE trap.

[0163] In some embodiments, the Treg cell activator comprises one or more single-chain chimeric polypeptides and one or more of an anti-tissue factor antibody, an anti-CD26 antibody (e.g., any of the anti-CD26 antibodies described herein), an anti-CD36 antibody, and a soluble RAGE trap.

[0164] In some embodiments, the methods include administering to the subject a therapeutically effective amount of a monoclonal antibody, hi some embodiments, the monoclonal antibody comprises one or more of an anti-tissue factor antibody, an anti-CD26 antibody (e.g., any of the anti-CD26 antibodies described herein), and / or an anti-CD36 antibody that can directly or indirectly reduce inflammasome or senescent cell activity.

[0165] In some embodiments, the methods include administering to the subject a therapeutically effective amount of an advanced glycation end product (AGE) inhibitor, in some embodiments, the AGE inhibitor comprises one or more soluble RAGE traps that can directly or indirectly reduce the activity of inflammasomes or senescent cells.

[0166] In some embodiments of any of the methods described herein, the age-related disease is inflammation-aging associated. Non-limiting examples of age-related diseases are selected from the group consisting of Alzheimer's disease, aneurysms, cystic fibrosis, fibrosis in pancreatitis, glaucoma, hypertension, idiopathic pulmonary fibrosis, inflammatory bowel disease, intervertebral disc degeneration, macular degeneration, osteoarthritis, type 2 diabetes mellitus, lipoatrophy, lipodystrophy, atherosclerosis, cataracts, COPD, idiopathic pulmonary fibrosis, renal transplant failure, liver fibrosis, bone loss, myocardial infarction, sarcopenia, wound healing, alopecia, cardiomyocyte hypertrophy, osteoarthritis, Parkinson's disease, age-related loss of lung tissue elasticity, macular degeneration, cachexia, glomerulosclerosis, liver cirrhosis, NAFLD, osteoporosis, amyotrophic lateral sclerosis, Huntington's disease, spinocerebellar ataxia, multiple sclerosis, neurodegeneration, stroke, cancer, dementia, vascular disease, susceptibility to infection, chronic inflammation, and renal dysfunction.

[0167] Non-limiting examples of inflammatory diseases include rheumatoid arthritis, inflammatory bowel disease, lupus erythematosus, lupus nephritis, amyotrophic lateral sclerosis, diabetic nephropathy, CNS injury, Alzheimer's disease, Parkinson's disease, Crohn's disease, multiple sclerosis, Guillain-Barré syndrome, psoriasis, Graves' disease, ulcerative colitis, non-alcoholic steatohepatitis, and mood disorders.

[0168] In some embodiments, the age-related disease is cancer. Non-limiting examples of cancer are selected from the group consisting of solid tumors, hematological tumors, sarcoma, osteosarcoma, glioblastoma, neuroblastoma, melanoma, rhabdomyosarcoma, Ewing's sarcoma, osteosarcoma, B-cell neoplasms, multiple myeloma, B-cell lymphoma, B-cell non-Hodgkin's lymphoma, Hodgkin's lymphoma, chronic lymphocytic leukemia (CLL), acute myeloid leukemia (AML), chronic myeloid leukemia (CML), acute lymphocytic leukemia (ALL), myelodysplastic syndrome (MDS), cutaneous T-cell lymphoma, retinoblastoma, gastric cancer, urothelial cancer, lung cancer, renal cell carcinoma, gastroesophageal cancer, pancreatic cancer, prostate cancer, breast cancer, colorectal cancer, ovarian cancer, non-small cell lung cancer, squamous cell head and neck cancer, endometrial cancer, cervical cancer, liver cancer, and hepatocellular carcinoma.

[0169] In some examples, the subject may be a subject who has been identified or diagnosed as having an age-related disease or chronic inflammation.

[0170] Also provided herein is a method for treating cancer in a subject, comprising administering to the subject a therapeutically effective amount of any one of the proteins described herein. Also provided herein is a method for treating an infectious disease in a subject, comprising administering to the subject a therapeutically effective amount of any one of the proteins described herein. Also provided herein is a method for treating an infectious disease in a subject, comprising administering to the subject a therapeutically effective amount of any one of the proteins described herein or any one of the pharmaceutical compositions described herein.

[0171] Non-limiting examples of infectious diseases include anthrax, arboviral diseases, babesiosis, botulism, brucellosis, campylobacteriosis, cholera, congenital syphilis, covid-19, dengue virus infection, diphtheria, ehrlichiosis and anaplasmosis, gonorrhea, leprosy, hantavirus infection, hepatitis, HIV infection, invasive pneumococcal disease, legionellosis, listeriosis, Lyme disease, malaria, measles, meningococcal disease, whooping cough, rubella, salmonellosis, smallpox, tetanus, tuberculosis, viral hemorrhagic fever, and Zika virus disease.

[0172] In some embodiments, these methods may result in a decrease in the number, severity, or frequency of one or more symptoms of an age-related disease in a subject (e.g., compared to the number, severity, or frequency of one or more symptoms of cancer in the subject before treatment).

[0173] In some examples, the methods result in a reduction in the number of senescent cells in a subject (e.g., a reduction in the number of senescent cells in one or more specific tissues involved and / or associated with an age-related disease or disorder in a subject), e.g., compared to the number of senescent cells in the subject before treatment (e.g., between about a 1% reduction and about a 99% reduction, between about a 1% reduction and about a 95% reduction, between about a 1% reduction and about a 90% reduction, between about a 1% reduction and about a 85% reduction, between about a 1% reduction and about a 80% reduction, between about a 1% reduction and about a 75% reduction, between about a 1% reduction and about a 70% reduction, between about a 1% reduction and about a 65% reduction, between about a 1% reduction and about a 60% reduction). , approximately 1% decrease to approximately 55% decrease, approximately 1% decrease to approximately 50% decrease, approximately 1% decrease to approximately 45% decrease, approximately 1% decrease to approximately 40% decrease, approximately 1% decrease to approximately 35% decrease, approximately 1% decrease to approximately 30% decrease, approximately 1% decrease to approximately 25% decrease, approximately 1% decrease to approximately 20% decrease, approximately 1% decrease to approximately 15% decrease, approximately 1% decrease to approximately 10% decrease, approximately 1% decrease to approximately 5% decrease, approximately 5% decrease to approximately 99% decrease, approximately 5% decrease to approximately 95% decrease, approximately 5% decrease to approximately 90% decrease, approximately 5% decrease to approximately 85% decrease, approximately 5% decrease to approximately 80% decrease, approximately 5% decrease to approximately 7 5% decrease, approximately 5% to approximately 70% decrease, approximately 5% to approximately 65% ​​decrease, approximately 5% to approximately 60% decrease, approximately 5% to approximately 55% decrease, approximately 5% to approximately 50% decrease, approximately 5% to approximately 45% decrease, approximately 5% to approximately 40% decrease, approximately 5% to approximately 35% decrease, approximately 5% to approximately 30% decrease, approximately 5% to approximately 25% decrease, approximately 5% to approximately 20% decrease, approximately 5% to approximately 15% decrease, approximately 5% to approximately 10% decrease, approximately 10% to approximately 99% decrease, approximately 10% to approximately 95% decrease, approximately 10% to approximately 90% decrease, approximately 10 % reduction to approximately 85% reduction, approximately 10% reduction to approximately 80% reduction, approximately 10% reduction to approximately 75% reduction, approximately 10% reduction to approximately 70% reduction, approximately 10% reduction to approximately 65% ​​reduction, approximately 10% reduction to approximately 60% reduction, approximately 10% reduction to approximately 55% reduction, approximately 10% reduction to approximately 50% reduction, approximately 10% reduction to approximately 45% reduction, approximately 10% reduction to approximately 40% reduction, approximately 10% reduction to approximately 35% reduction, approximately 10% reduction to approximately 30% reduction, approximately 10% reduction to approximately 25% reduction, approximately 10% reduction to approximately 20% reduction, approximately 10% reduction to approximately 15% reduction, approximately 15% reduction to approximately 99% reduction,Approximately 15% decrease to approximately 95% decrease, approximately 15% decrease to approximately 90% decrease, approximately 15% decrease to approximately 85% decrease, approximately 15% decrease to approximately 80% decrease, approximately 15% decrease to approximately 75% decrease, approximately 15% decrease to approximately 70% decrease, approximately 15% decrease to approximately 65% ​​decrease, approximately 15% decrease to approximately 60% decrease, approximately 15% decrease to approximately 55% decrease, approximately 15% decrease to approximately 50% decrease, approximately 15% decrease to approximately 45% decrease, approximately 15% decrease to approximately 40% decrease, approximately 15% decrease to approximately 35% decrease, approximately 15% decrease to approximately 30% decrease, approximately 15% decrease to approximately 25% decrease, approximately 15% decrease to approximately 20% reduction, approximately 20% reduction to approximately 99% reduction, approximately 20% reduction to approximately 95% reduction, approximately 20% reduction to approximately 90% reduction, approximately 20% reduction to approximately 85% reduction, approximately 20% reduction to approximately 80% reduction, approximately 20% reduction to approximately 75% reduction, approximately 20% reduction to approximately 70% reduction, approximately 20% reduction to approximately 65% ​​reduction, approximately 20% reduction to approximately 60% reduction, approximately 20% reduction to approximately 55% reduction, approximately 20% reduction to approximately 50% reduction, approximately 20% reduction to approximately 45% reduction, approximately 20% reduction to approximately 40% reduction, approximately 20% reduction to approximately 35% reduction, approximately 20% reduction to approximately 30% reduction, approximately 20% reduction to Approximately 25% reduction, approximately 25% reduction to approximately 99% reduction, approximately 25% reduction to approximately 95% reduction, approximately 25% reduction to approximately 90% reduction, approximately 25% reduction to approximately 85% reduction, approximately 25% reduction to approximately 80% reduction, approximately 25% reduction to approximately 75% reduction, approximately 25% reduction to approximately 70% reduction, approximately 25% reduction to approximately 65% ​​reduction, approximately 25% reduction to approximately 60% reduction, approximately 25% reduction to approximately 55% reduction, approximately 25% reduction to approximately 50% reduction, approximately 25% reduction to approximately 45% reduction, approximately 25% reduction to approximately 40% reduction, approximately 25% reduction to approximately 35% reduction, approximately 25% reduction to approximately 30% reduction, approximately 30% Reduction of about 99% reduction, Reduction of about 30% reduction to about 95% reduction, Reduction of about 30% reduction to about 90% reduction, Reduction of about 30% reduction to about 85% reduction, Reduction of about 30% reduction to about 80% reduction, Reduction of about 30% reduction to about 75% reduction, Reduction of about 30% reduction to about 70% reduction, Reduction of about 30% reduction to about 65% reduction, Reduction of about 30% reduction to about 60% reduction, Reduction of about 30% reduction to about 55% reduction, Reduction of about 30% reduction to about 50% reduction, Reduction of about 30% reduction to about 45% reduction, Reduction of about 30% reduction to about 40% reduction, Reduction of about 30% reduction to about 35% reduction, Reduction of about 35% reduction to about 99% reduction, Reduction of about 35% reduction to about 95% reduction,Approximately 35% reduction to approximately 90% reduction, approximately 35% reduction to approximately 85% reduction, approximately 35% reduction to approximately 80% reduction, approximately 35% reduction to approximately 75% reduction, approximately 35% reduction to approximately 70% reduction, approximately 35% reduction to approximately 65% ​​reduction, approximately 35% reduction to approximately 60% reduction, approximately 35% reduction to approximately 55% reduction, approximately 35% reduction to approximately 50% reduction, approximately 35% reduction to approximately 45% reduction, approximately 35% reduction to approximately 40% reduction, approximately 40% reduction to approximately 99% reduction, approximately 40% reduction to approximately 95% reduction, approximately 40% reduction to approximately 90% reduction, approximately 40% reduction to approximately 85% reduction, approximately 40% reduction to approximately 8 0% reduction, approximately 40% reduction to approximately 75% reduction, approximately 40% to approximately 70% reduction, approximately 40% reduction to approximately 65% ​​reduction, approximately 40% reduction to approximately 60% reduction, approximately 40% reduction to approximately 55% reduction, approximately 40% reduction to approximately 50% reduction, approximately 40% reduction to approximately 45% reduction, approximately 45% reduction to approximately 99% reduction, approximately 45% reduction to approximately 95% reduction, approximately 45% reduction to approximately 90% reduction, approximately 45% reduction to approximately 85% reduction, approximately 45% reduction to approximately 80% reduction, approximately 45% reduction to approximately 75% reduction, approximately 45% reduction to approximately 70% reduction, approximately 45% reduction to approximately 65% ​​reduction, approximately 45% reduction Approximately 60% reduction, approximately 45% reduction to approximately 55% reduction, approximately 45% reduction to approximately 50% reduction, approximately 50% reduction to approximately 99% reduction, approximately 50% reduction to approximately 95% reduction, approximately 50% reduction to approximately 90% reduction, approximately 50% reduction to approximately 85% reduction, approximately 50% reduction to approximately 80% reduction, approximately 50% reduction to approximately 75% reduction, approximately 50% reduction to approximately 70% reduction, approximately 50% reduction to approximately 65% ​​reduction, approximately 50% reduction to approximately 60% reduction, approximately 50% reduction to approximately 55% reduction, approximately 55% reduction to approximately 99% reduction, approximately 55% reduction to approximately 95% reduction, approximately 55% reduction to approximately 90% reduction, approximately 5 5% decrease to approximately 85% decrease, approximately 55% decrease to approximately 80% decrease, approximately 55% decrease to approximately 75% decrease, approximately 55% decrease to approximately 70% decrease, approximately 55% decrease to approximately 65% ​​decrease, approximately 55% decrease to approximately 60% decrease, approximately 60% decrease to approximately 99% decrease, approximately 60% decrease to approximately 95% decrease, approximately 60% decrease to approximately 90% decrease, approximately 60% decrease to approximately 85% decrease, approximately 60% decrease to approximately 80% decrease, approximately 60% decrease to approximately 75% decrease, approximately 60% decrease to approximately 70% decrease, approximately 60% decrease to approximately 65% ​​decrease, approximately 65% ​​decrease to approximately 99% decrease, approximately 65% ​​decrease to approximately 95% decrease,Approximately 65% ​​decrease to approximately 90% decrease, approximately 65% ​​decrease to approximately 85% decrease, approximately 65% ​​decrease to approximately 80% decrease, approximately 65% ​​decrease to approximately 75% decrease, approximately 65% ​​decrease to approximately 70% decrease, approximately 70% decrease to approximately 99% decrease, approximately 70% decrease to approximately 95% decrease, approximately 70% decrease to approximately 90% decrease, approximately 70% decrease to approximately 85% decrease, approximately 70% decrease to approximately 80% decrease, approximately 70% decrease to approximately 75% decrease, approximately 75% decrease to approximately 99% decrease, approximately 75% decrease to approximately 95% decrease, approximately 75% decrease a reduction of about 80% to about 99%, a reduction of about 80% to about 95%, a reduction of about 80% to about 90%, a reduction of about 80% to about 95%, a reduction of about 80% to about 90%, a reduction of about 80% to about 85%, a reduction of about 85% to about 99%, a reduction of about 85% to about 95%, a reduction of about 85% to about 90%, a reduction of about 90% to about 99%, a reduction of about 90% to about 95%, or a reduction of about 95% to about 99%.

[0174] The term "subject" refers to any mammal. In some embodiments, the subject or "subject in need of treatment" can be a canine (e.g., dog), feline (e.g., cat), equine (e.g., horse), sheep, cow, pig, goat, primate, such as a simian (e.g., monkey (e.g., marmoset, baboon), or ape (e.g., gorilla, chimpanzee, orangutan, or gibbon), or human, or a rodent (e.g., mouse, guinea pig, hamster, or rat). In some embodiments, the subject or "subject in need of treatment" can be a non-human mammal, particularly a mammal (e.g., mouse, lapwing, pig, dog, or primate) that is traditionally used as a model to demonstrate therapeutic efficacy in humans.

[0175] Treg cells In some embodiments, Treg cells may be administered to a subject. In some embodiments, the Treg cells administered to a subject may be autologous, haploidentical, or allogeneic Treg cells isolated from peripheral blood or umbilical cord blood. In some embodiments, the methods described herein may further comprise isolating Treg cells from the subject, culturing the isolated Treg cells in a liquid culture medium, and administering the Treg cells back to the subject. In some embodiments, Treg cells may be prepared from commercially available kits (e.g., EasySep™ Human CD4 + CD127 low CD25 + Regulatory T Cell Isolation Kit or Dynabeads CD4 + CD25 + Regulatory T cells can be isolated using a variety of methods, including the use of a regulatory T cell kit (see regulatory T cell kit). In some embodiments, the liquid culture medium can include one or more of the single-chain chimeric polypeptides (e.g., any of the exemplary single-chain chimeric polypeptides described herein, e.g., 2t2 or 3t28). In some embodiments, the liquid culture medium can include the use of beads having CD3 and CD28 and recombinant IL-2 or 2t2 on their surface.

[0176] In some embodiments, Treg cells may comprise a chimeric antigen receptor (e.g., a chimeric antigen receptor comprising an extracellular domain that specifically binds to tissue factor, to CD26 (e.g., any of the anti-CD26 antigen binding domains described herein), or to CD36). Non-limiting examples of extracellular domains that can bind to tissue factor, CD26, or CD36 are scFvs. Non-limiting examples of anti-CD36 antibodies are those commercially available from Invitrogen, Abcam, GeneTex, Novus Biologicals, Proteintech, and EMD Millipore. Non-limiting examples of anti-tissue factor heavy chain variable domains and light chain variable domains are described in U.S. Pat. No. 7,968,094 and U.S. Pat. No. 8,007,795.

[0177] Treg cell activator In some embodiments, one or more Treg cell activators may be administered to a subject. In some embodiments, the Treg cell activator may be a single-chain chimeric polypeptide (e.g., any of the exemplary single-chain chimeric polypeptides described herein), an anti-tissue factor antibody (e.g., the anti-tissue factor antibodies described in U.S. Pat. Nos. 7,968,094 and 8,007,795), a soluble RAGE protein, an anti-CD26 antibody (e.g., any of the anti-CD26 antibodies described herein), or an anti-CD36 antibody.

[0178] The soluble RAGE protein may have a sequence that is at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100% identical to SEQ ID NO:104 or SEQ ID NO:105.

[0179] Soluble human RAGE variant 1 (SEQ ID NO: 104) TIFF0007734693000001.tif15138 Soluble human RAGE variant 2 (SEQ ID NO: 105) TIFF0007734693000002.tif17152

[0180] In some examples, the soluble RAGE protein is encoded by a nucleic acid having a sequence that is at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100% identical to SEQ ID NO:106 or SEQ ID NO:107.

[0181] Soluble human RAGE variant 1 cDNA (SEQ ID NO: 106) TIFF0007734693000003.tif45138 Mouse RAGE cDNA (SEQ ID NO: 107) TIFF0007734693000004.tif39151

[0182] As can be understood by one of skill in the art, substitutions / mutations made at positions that are not conserved between different species are unlikely to adversely affect the activity of the protein / nucleic acid, while substitutions / mutations made at positions that are conserved between species are likely to adversely affect the activity of the protein / nucleic acid.

[0183] NK cells In some embodiments, NK cells may be administered to a subject. In some embodiments, the NK cells administered to a subject may be autologous, haploidentical, or allogeneic NK cells isolated from peripheral blood, isolated from umbilical cord blood, or isolated and differentiated from iPSCs. In some embodiments, the methods described herein may further include isolating NK cells from the subject, culturing the isolated NK cells in a liquid culture medium, and administering the NK cells back to the subject. In some embodiments, NK cells may be isolated using commercially available kits (see, e.g., EasySep™ Human NK Cell Isolation Kit, MojoSort Human NK Cell Isolation Kit, and Novus Biologicals Human NK Cell Isolation Kit). In some embodiments, the liquid culture medium may include one or more of the multi-chain chimeric polypeptides (e.g., any of the exemplary multi-chain chimeric polypeptides described herein, e.g., 18t15-12s and / or 7t15-21s).

[0184] In some embodiments, the NK cells may comprise a chimeric antigen receptor (e.g., a chimeric antigen receptor comprising an extracellular domain that specifically binds to tissue factor or to CD26). A non-limiting example of an extracellular domain capable of binding to tissue factor or CD26 is an scFv. Non-limiting examples of anti-CD26 antibodies are commercially available from Abcam, Invitrogen, and GeneTex. Further examples of anti-CD26 antibodies are the anti-CD26 antibodies described herein. Non-limiting examples of anti-tissue factor heavy chain variable domains and light chain variable domains are described in U.S. Pat. Nos. 7,968,094 and 8,007,795. The chimeric antigen receptor comprises a transmembrane domain, a costimulatory domain (e.g., an intracellular CD28 domain), and a CD3 zeta signaling domain. For example, the transmembrane domain may comprise a sequence at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100% identical to SEQ ID NO: 101. For example, the costimulatory domain can comprise a sequence that is at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100% identical to SEQ ID NO: 102. For example, the CD3 zeta signaling domain can comprise a sequence that is at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100% identical to SEQ ID NO: 103.

[0185] NK cell activator In some embodiments, one or more NK cell activators may be administered to a subject. In some embodiments, the NK cell activators may be one or more multi-chain chimeric polypeptides (e.g., any of the exemplary multi-chain chimeric polypeptides described herein), anti-tissue factor antibodies (e.g., the anti-tissue factor antibodies described in U.S. Pat. Nos. 7,968,094 and 8,007,795), anti-CD36 antibodies (e.g., anti-CD36 antibodies commercially available from Invitrogen, Abcam, GeneTex, Novus Biologicals, Proteintech, and EMD Millipore), or anti-CD26 antibodies (e.g., anti-CD26 antibodies commercially available from Abcam, Invitrogen, and GeneTex). NK cell activators, such as cytokine-based agents, can act by directing NK cell activation, or NK cell activators, such as antibodies that mediate antibody-dependent cellular cytotoxicity (ADCC) of NK cells, can enhance NK cell activity.

[0186] Multi-chain chimeric polypeptides Provided herein is a multi-chain chimeric polypeptide comprising: (a) a first chimeric polypeptide comprising (i) a first target binding domain, (ii) a soluble tissue factor domain, and (iii) a first domain of a pair of affinity domains; and (b) a second chimeric polypeptide comprising (i) a second domain of the pair of affinity domains and (ii) a second target binding domain, wherein the first chimeric polypeptide and the second chimeric polypeptide are associated via binding between the first domain and the second domain of the pair of affinity domains.

[0187] In some embodiments of any of the multi-chain chimeric polypeptides described herein, the first target binding domain (e.g., any of the first target binding domains described herein) and the soluble tissue factor domain (e.g., any of the exemplary soluble tissue factor domains described herein) are immediately adjacent to each other within the first chimeric polypeptide. In some embodiments of any of the multi-chain chimeric polypeptides described herein, the first chimeric polypeptide further comprises a linker sequence (e.g., any of the exemplary linker sequences described herein or known in the art) between the first target binding domain (e.g., any of the exemplary first target binding domains described herein) and the soluble tissue factor domain (e.g., any of the exemplary soluble tissue factor domains described herein) within the first chimeric polypeptide.

[0188] In some embodiments of any of the multi-chain chimeric polypeptides described herein, the soluble tissue factor domain (e.g., any of the exemplary soluble tissue factor domains described herein) and the first domain of the pair of affinity domains (e.g., any of the exemplary first domains of any of the exemplary affinity domain pairs described herein) are immediately adjacent to each other within the first chimeric polypeptide. In some embodiments of any of the multi-chain chimeric polypeptides described herein, the first chimeric polypeptide further comprises a linker sequence (e.g., any of the exemplary linker sequences described herein or known in the art) between the soluble tissue factor domain (e.g., any of the exemplary soluble tissue factor domains described herein) and the first domain of the pair of affinity domains (e.g., any of the exemplary first domains of any of the exemplary affinity domain pairs described herein) within the first chimeric polypeptide.

[0189] In some embodiments of any of the multi-chain chimeric polypeptides described herein, the second domain of the pair of affinity domains (e.g., any of the exemplary second domains of any of the exemplary affinity domain pairs described herein) and the second target binding domain (e.g., any of the exemplary second target binding domains described herein) are immediately adjacent to each other in the second chimeric polypeptide. In some embodiments of any of the multi-chain chimeric polypeptides described herein, the second chimeric polypeptide further comprises a linker sequence (e.g., any of the exemplary linker sequences described herein or known in the art) between the second domain of the pair of affinity domains (e.g., any of the exemplary second domains of any of the exemplary affinity domain pairs described herein) and the second target binding domain (e.g., any of the exemplary second target binding domains described herein) in the second chimeric polypeptide.

[0190] tissue factor Human tissue factor is a 263-amino acid transmembrane protein containing three domains: (1) a 219-amino acid N-terminal extracellular domain (residues 1–219), (2) a 22-amino acid transmembrane domain (residues 220–242), and (3) a 21-amino acid cytoplasmic C-terminal tail (residues 242–263) (UniProtKB Identifier Number: P13726). The cytoplasmic tail contains two phosphorylation sites at Ser253 and Ser258 and one S-palmitoylation site at Cys245. Deletions or mutations in the cytoplasmic domain have not been found to affect tissue factor clotting activity. Tissue factor has one S-palmitoylation site within the intracellular domain of the protein at Cys245, which is located at the amino acid terminus of the intracellular domain and is close to the membrane surface. The tissue factor transmembrane domain consists of a single transmembrane α-helix.

[0191] The extracellular domain of tissue factor, consisting of two fibronectin type III domains, is connected to the transmembrane domain via a six-amino acid linker. This linker provides conformational flexibility to separate the tissue factor extracellular domain from its transmembrane and cytoplasmic domains. Each tissue factor fibronectin type III module is composed of two overlapping β-sheets, with the upper sheet domain containing three antiparallel β-strands and the lower sheet containing four β-strands. The β-strands are connected by β-loops between strands βA and βB, βC and βD, and βE and βF, all of which conformations are conserved within the two modules. Three short α-helical segments connect the β-strands. A unique feature of tissue factor is the 17-amino acid β-hairpin between strands β10 and β11, which is not a common element of the fibronectin superfamily. The N-terminal domain also contains a 12-amino acid loop between β6F and β7G, which is unique to tissue factor and absent in the C-terminal domain. Such a fibronectin type III domain structure is characteristic of the immunoglobulin-like protein family fold and is conserved among a wide variety of extracellular proteins.

[0192] Zymogen FVII is rapidly converted to FVIIa by limited proteolysis upon binding to tissues to form the active tissue factor-FVIIa complex. FVIIa, which circulates as an enzyme at a concentration of approximately 0.1 nM (1% of plasma FVII), can also bind directly to tissue factor. The allosteric interaction between tissue factor and FVIIa on the tissue factor-FVIIa complex significantly increases the enzymatic activity of FVIIa, increasing the hydrolysis rate of small chromogenic peptidyl substrates by approximately 20- to 100-fold and the activation rate of the natural polymer substrates FIX and FX by nearly one million-fold. In concert with the allosteric activation of the FVIIa active site upon binding to tissue factor, the formation of the tissue factor-FVIIa complex on the phospholipid bilayer (i.e., upon exposure of phosphatidyl-L-serine on the membrane surface) induces Ca + ... 2+It further increases the activation rate of FIX or FX by 1,000-fold in a coagulation-dependent manner. The approximately 1,000,000-fold overall increase in FX activation by tissue factor-FVIIa-phospholipid complexes compared to free FVIIa is a critical regulatory point in the coagulation cascade.

[0193] FVII is a single polypeptide chain of approximately 50 kDa, consisting of 406 amino acid residues, with an N-terminal γ-carboxyglutamic acid-rich (GLA) domain, two epidermal growth factor-like domains (EGF1 and EGFR2), and a C-terminal serine protease domain. FVII is characterized by an Ile-Ile linker in the short linker region between the EGF2 and protease domains. 154 -Arg 152 Activation to FVIIa occurs by specific proteolytic cleavage of the bond between the light and heavy chains. 135 and Cys 262 FVIIa binds to Ca via its N-terminal GLA domain. 2+ Immediately C-terminal to the GLA domain is an aromatic stack and two EGF domains. The aromatic stack binds the GLA domain to a single Ca 2+ This Ca 2+ Occupation of the binding site increases FVIIa amidolytic activity and tissue factor association. The catalytic triad consists of His 193 , Asp 242 , and Ser 344 and a single Ca domain in the FVIIa protease domain. 2+ Ion binding is essential for its catalytic activity. The proteolytic activity of FVII to FVIIa is mediated by Ile 153 releases the newly formed amino terminus, folds back, and inserts into the activation pocket, forming Asp 343It forms a salt bridge with the carboxylate of FVIIa to generate an oxyanion hole. This salt bridge formation is essential for FVIIa activity. However, the formation of the oxyanion hole does not occur in free FVIIa upon proteolytic activation. As a result, FVIIa circulates in a zymogen-like state that is poorly recognized by plasma protease inhibitors, allowing it to circulate with a half-life of approximately 90 minutes.

[0194] Tissue factor-mediated positioning of the FVIIa active site on the membrane surface is important for FVIIa to its cognate substrate. Free FVIIa adopts a stable extended conformation upon membrane binding, with its active site positioned approximately 80 Å above the membrane surface. Upon FVIIa binding to tissue factor, the FVIIa active site repositions approximately 6 Å closer to the membrane. This adjustment may aid in the proper alignment of the FVIIa catalytic triad with the target substrate cleavage site. Using GLA domain-less FVIIa, it was shown that the active site was still positioned at a similar distance from the membrane, indicating that tissue factor can fully support FVIIa active site positioning even in the absence of FVIIa-membrane interactions. Additional data showed that tissue factor supports full FVIIa proteolytic activity as long as the tissue factor extracellular domain is somehow tethered to the membrane surface. However, elevating the active site of FVIIa more than 80 Å from the membrane surface significantly reduced the ability of the tissue factor-FVIIa complex to activate FX, but did not reduce tissue factor-FVIIa amidolytic activity.

[0195] Alanine-scanning mutagenesis has been used to evaluate the role of specific amino acid side chains in the tissue factor extracellular domain for interaction with FVIIa (Gibbs et al., Biochemistry 33(47):14003-14010, 1994; Schullek et al., J Biol Chem 269(30):19399-19403, 1994). Alanine substitutions identified a limited number of residue positions where alanine substitutions caused a 5- to 10-fold lower affinity for FVIIa binding. Most of these residue side chains were found to be well exposed to solvent in the crystal structure, consistent with macromolecular ligand interactions. The FVIIa ligand-binding site spans an extensive region of the interface between the two modules. In the C-module, residues Arg, located on the protruding BC loop, 135 and Phe 140 provides independent contact with FVIIa. 133 is located at the base of the finger-like structure and is packed into the gap between the two modules. 20 , Thr 60 , Asp 58 , and Ile 22 Continuity is provided for the major cluster of important binding residues consisting of Thr 60 The Glu is only partially solvent exposed and may play a local structural role rather than making significant contacts with the ligand. 24 and Gln 110 , as well as potentially more distal residue Val 207 The binding region extends from Asp58 to Lys 48 , Lys 46 , Gln 37 , Asp 44 , and Trp 45 The convex area formed by the Trp 45 and Asp 44 does not interact independently with FVIIa, and Trp 45 The effect of mutation at position 1 is similar to that of the adjacent Asp 44 and Gln 37The interaction region is located between two surface-exposed aromatic residues, Phe, which form part of a hydrophobic cluster within the N-module. 76 and Tyr 78 Further includes:

[0196] The known physiological substrates of tissue factor-FVIIa are FVII, FIX, and FX, as well as certain proteinase-activated receptors. Mutational analysis has identified several residues that, when mutated, support full FVIIa amidolytic activity toward small peptidyl substrates but lack the ability to support activation of macromolecular substrates (i.e., FVII, FIX, and FX) (Ruf et al., J Biol Chem 267(31):22206-22210, 1992; Ruf et al., J Biol Chem 267(9):6375-6381, 1992; Huang et al., J Biol Chem 271(36):21752-21757, 1996; Kirchhofer et al., Biochemistry 39(25):7380-7387, 2000). The tissue factor loop region from residues 159 to 165, and residues within or adjacent to this flexible loop, have been shown to be essential for the proteolytic activity of the tissue factor-FVIIa complex. This defines a proposed substrate-binding exosite region of tissue factor, located far from the FVIIa active site. Substitution of the glycine residue with a slightly more bulky alanine residue significantly impairs the proteolytic activity of tissue factor-FVIIa. This suggests that the flexibility conferred by glycine is essential for the loop from residues 159 to 165 for tissue factor macromolecular substrate recognition.

[0197] Residue Lys 165 and Lys 166 It has also been demonstrated that Lys is important for substrate recognition and binding. Mutation of either of these residues to alanine significantly reduces tissue factor cofactor function. 165 and Lys 166 are facing outwards, and Lys 165refers to FVIIa in most tissue factor-FVIIa structures, and Lys 166 refers to the substrate-binding exosite region in the crystal structure. Lys of FVIIa 165 and FVIIa Gla 35 The putative salt bridge formation between these domains would support the notion that tissue factor interaction with the GLA domain of FVIIa regulates substrate recognition. These results suggest that the C-terminal portion of the tissue factor ectodomain interacts directly with the GLA domains of FIX and FX, and with the possible adjacent EGF1 domain, and that the presence of the FVIIa GLA domain may regulate these interactions either directly or indirectly.

[0198] Soluble tissue factor domain In some embodiments of any of the polypeptides described herein, the soluble tissue factor domain can be a wild-type tissue factor polypeptide lacking the signal sequence, transmembrane domain, and intracellular domain. In some examples, the soluble tissue factor domain can be a tissue factor variant, where the wild-type tissue factor polypeptide lacks the signal sequence, transmembrane domain, and intracellular domain and is further modified with selected amino acids. In some examples, the soluble tissue factor domain can be a soluble human tissue factor domain. In some examples, the soluble tissue factor domain can be a soluble mouse tissue factor domain. In some examples, the soluble tissue factor domain can be a soluble rat tissue factor domain. Non-limiting examples of soluble human tissue factor domains, soluble mouse tissue factor domains, soluble rat tissue factor domains, and mutant soluble tissue factor domains are provided below.

[0199] Exemplary soluble human tissue factor domain (SEQ ID NO: 120) SGTTNTVAAYNLTWKSTNFKTILEWEPKPVNQVYTVQISTKSGDWKSKCFYTTDTECDLTDEIVKDVKQTYLARVFSYPAGNVESTGSAGEPLYENSPEFTPYLETNLG QPTIQSFEQVGTKVNVTVEDERTLVRRNNTFLSLRDVFGKDLIYTLYYWKSSSSGKKTAKTNTNEFLIDVDKGENYCFSVQAVIPSRTVNRKSTDSPVECMGQEKGEFRE Exemplary nucleic acid encoding a soluble human tissue factor domain (SEQ ID NO: 121) AGCGGCACAACCAACACAGTCGCTGCCTATAACCTCACTTGGAAGAGCACCAACTTCAAAACCATCCTCGAATGGGAACCCAAACCCGTTAACCAAGTTTACACCGTGCAGATCAGCACCAAGTCCGGCGACTGGAAGTCCAAATGTTTCTATACCACCGACAC CGAGTGCGATCTCACCGATGAGATCGTGAAAGATGTGAAACAGACCTACCTCGCCCGGGTGTTTAGCTACCCCGCCGGCAATGTGGAGAGCACTGGTTCCGCTGGCGAGCCTTTATACGAGAACAGCCCCGAATTTACCCCTTACCTCGAGACCAATTTAGGAC AGCCCACCATCCAAAGCTTTGAGCAAGTTGGCACAAAGGTGAATGTGACAGTGGAGGACGAGCGGACTTTAGTGCGGCGGAACAACACCTTTCTCAGCCTCCGGGATGTGTTCGGCAAAGATTTAATCTACACACTGTATTACTGGAAGTCCTCTTCCTCCGGC AAGAAGACAGCTAAAACCAACACAAACGAGTTTTTAATCGACGTGGATAAAGGCGAAAACTACTGTTTCAGCGTGCAAGCTGTGATCCCCTCCCGGACCGTGAATAGGAAAAGCACCGATAGCCCCGTTGAGTGCATGGGCCAAGAAAAGGGCGAGTTCCGGGAG Exemplary mutant soluble human tissue factor domain (SEQ ID NO: 122) SGTTNTVAAYNLTWKSTNFATALEWEPKPVNQVYTVQISTKSGDWKSKCFYTTDTECALTDEIVKDVKQTYLARVFSYPAGNVESTGSAGEPLYENSPEFTPYLETNLG QPTIQSFEQVGTKVNVTVEDERTLVARNNTALSLRDVFGKDLIYTLYYWKSSSSGKKTAKTNTNEFLIDVDKGENYCFSVQAVIPSRTVNRKSTDSPVECMGQEKGEFRE Exemplary mutant soluble human tissue factor domain (SEQ ID NO: 123) SGTTNTVAAYNLTWKSTNFATALEWEPKPVNQVYTVQISTKSGDAKSKCFYTTDTECALTDEIVKDVKQTYLARVFSYPAGNVESTGSAGEPLAENSPEFTPYLETNLG QPTIQSFEQVGTKVNVTVEDERTLVARNNTALSLRDVFGKDLIYTLYYWKSSSSGKKTAKTNTNEFLIDVDKGENYCFSVQAVIPSRTVNRKSTDSPVECMGQEKGEFRE Exemplary soluble mouse tissue factor domain (SEQ ID NO: 124) AGIPEKAFNLTWISTDFKTILEWQPKPTNYTYTVQISDRSRNWKNKCFSTTDTECDLTDEIVKDVTWAYEAKVLSVPRRNSVHGDGDQLVIHGEEPPFTNAPKFLPYRDTN LGQPVIQQFEQDGRKLNVVVKDSLTLVRKNGTFLTLRQVFGKDLGYIITYRKGSSTGKKTNITNTNEFSIDVEEGVSYCFFVQAMIFSRKTNQNSPGSSTVCTEQWKSFLGE Exemplary soluble rat tissue factor domain (SEQ ID NO: 125) AGTPPGKAFNLTWISTDFKTILEWQPKPTNYTYTVQISDRSRNWKYKCTGTTDTECDLTDEIVKDVNWTYEARVLSVPWRNSTHGKETLFGTHGEEPPFTNARKFLPYRDTK IGQPVIQKYEQGGTKLKVTVKDSFTLVRKNGTFLTLRQVFGNDLGYILTYRKDSSTGRKTNTTHTNEFLIDVEKGVSYCFFAQAVIFSRKTNHKSPESITKCTEQWKSVLGE

[0200] In some embodiments, the soluble tissue factor domain can comprise a sequence that is at least 70% identical, at least 72% identical, at least 74% identical, at least 76% identical, at least 78% identical, at least 80% identical, at least 82% identical, at least 84% identical, at least 86% identical, at least 88% identical, at least 90% identical, at least 92% identical, at least 94% identical, at least 96% identical, at least 98% identical, at least 99% identical, or 100% identical to SEQ ID NO: 120, 122, 123, 124, or 125. In some embodiments, the soluble tissue factor domain may comprise the sequence of SEQ ID NO: 120, 122, 123, 124, or 125, with 1 to 20 amino acids (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 amino acids) removed from its N-terminus and / or 1 to 20 amino acids (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 amino acids) removed from its C-terminus.

[0201] As can be understood in the art, a person skilled in the art will understand that mutations in amino acids that are conserved among different mammalian species are likely to reduce the activity and / or structural stability of a protein, while mutations in amino acids that are not conserved among different mammalian species are unlikely to reduce the activity and / or structural stability of a protein.

[0202] In some examples of any of the single-chain or multi-chain chimeric polypeptides described herein, the soluble tissue factor domain is unable to bind to factor VIIa. In some examples of any of the single-chain or multi-chain chimeric polypeptides described herein, the soluble tissue factor domain does not convert inactive factor X to factor Xa. In some embodiments of any of the single-chain or multi-chain chimeric polypeptides described herein, the single-chain or multi-chain chimeric polypeptide does not stimulate blood clotting in a mammal.

[0203] In some examples, the soluble tissue factor domain can be a soluble human tissue factor domain. In some embodiments, the soluble tissue factor domain can be a soluble mouse tissue factor domain. In some embodiments, the soluble tissue factor domain can be a soluble rat tissue factor domain.

[0204] In some examples, the soluble tissue factor domain does not include one or more (e.g., 2, 3, 4, 5, 6, or 7) of: a lysine at the amino acid position corresponding to amino acid position 20 of the mature wild-type human tissue factor protein, an isoleucine at the amino acid position corresponding to amino acid position 22 of the mature wild-type human tissue factor protein, a tryptophan at the amino acid position corresponding to amino acid position 45 of the mature wild-type human tissue factor protein, an aspartic acid at the amino acid position corresponding to amino acid position 58 of the mature wild-type human tissue factor protein, a tyrosine at the amino acid position corresponding to amino acid position 94 of the mature wild-type human tissue factor protein, an arginine at the amino acid position corresponding to amino acid position 135 of the mature wild-type human tissue factor protein, and a phenylalanine at the amino acid position corresponding to amino acid position 140 of the mature wild-type human tissue factor protein. In some embodiments, the mutant soluble tissue factor has the amino acid sequence of SEQ ID NO: 122 or SEQ ID NO: 123.

[0205] In some examples, the soluble tissue factor domain can be encoded by a nucleic acid comprising a sequence that is at least 70% identical, at least 72% identical, at least 74% identical, at least 76% identical, at least 78% identical, at least 80% identical, at least 82% identical, at least 84% identical, at least 86% identical, at least 88% identical, at least 90% identical, at least 92% identical, at least 94% identical, at least 96% identical, at least 98% identical, at least 99% identical, or 100% identical to SEQ ID NO:121.

[0206] Linker sequence In some embodiments, the linker sequence may be a flexible linker sequence. Non-limiting examples of linker sequences that can be used are described in Klein et al., Protein Engineering, Design & Selection 27(10):325-330, 2014, and Priyanka et al., Protein Sci. 22(2):153-167, 2013. In some examples, the linker sequence is a synthetic linker sequence.

[0207] In some embodiments of any of the multi-chain chimeric polypeptides described herein, the first chimeric polypeptide can include 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 linker sequences (e.g., the same or different linker sequences, e.g., any of the exemplary linker sequences described herein or known in the art). In some embodiments of any of the multi-chain chimeric polypeptides described herein, the second chimeric polypeptide can include 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 linker sequences (e.g., the same or different linker sequences, e.g., any of the exemplary linker sequences described herein or known in the art).

[0208] In some embodiments, the linker sequence may be from 1 to about 100 amino acids, from 1 to about 90 amino acids, from 1 to about 80 amino acids, from 1 to about 70 amino acids, from 1 to about 60 amino acids, from 1 to about 50 amino acids, from 1 to about 45 amino acids, from 1 to about 40 amino acids, from 1 to about 35 amino acids, from 1 to about 30 amino acids, from 1 to about 25 amino acids, from 1 to about 24 amino acids, from 1 to about 22 amino acids, from 1 to about 20 amino acids, from 1 to about 18 amino acids, from 1 to about 16 amino acids, or from 1 to about 2 to about 14 amino acids, 1 to about 12 amino acids, 1 to about 10 amino acids, 1 to about 8 amino acids, 1 to about 6 amino acids, 1 to about 4 amino acids, about 2 to about 100 amino acids, about 2 to about 90 amino acids, about 2 to about 80 amino acids, about 2 to about 70 amino acids, about 2 to about 60 amino acids, about 2 to about 50 amino acids, about 2 to about 45 amino acids, about 2 to about 40 amino acids, about 2 to about 35 amino acids, about 2 to about 30 amino acids, about 2 to about 25 amino acids, about 2 to about 3 About 24 amino acids, about 2 to about 22 amino acids, about 2 to about 20 amino acids, about 2 to about 18 amino acids, about 2 to about 16 amino acids, about 2 to about 14 amino acids, about 2 to about 12 amino acids, about 2 to about 10 amino acids, about 2 to about 8 amino acids, about 2 to about 6 amino acids, about 2 to about 4 amino acids, about 4 to about 100 amino acids, about 4 to about 90 amino acids, about 4 to about 80 amino acids, about 4 to about 70 amino acids, about 4 to about 60 amino acids, about 4 to about 50 amino acids, about 4 amino acids about 4 to about 45 amino acids, about 4 to about 40 amino acids, about 4 to about 35 amino acids, about 4 to about 30 amino acids, about 4 to about 25 amino acids, about 4 to about 24 amino acids, about 4 to about 22 amino acids, about 4 to about 20 amino acids, about 4 to about 18 amino acids, about 4 to about 16 amino acids, about 4 to about 14 amino acids, about 4 to about 12 amino acids, about 4 to about 10 amino acids, about 4 to about 8 amino acids, about 4 to about 6 amino acids, about 6 to about 100 amino acids, about 6 to about 90 amino acids,About 6 amino acids to about 80 amino acids, about 6 amino acids to about 70 amino acids, about 6 amino acids to about 60 amino acids, about 6 amino acids to about 50 amino acids, about 6 amino acids to about 45 amino acids, about 6 amino acids to about 40 amino acids, about 6 amino acids to about 35 amino acids, about 6 amino acids to about 30 amino acids, about 6 amino acids to about 25 amino acids, about 6 amino acids to about 24 amino acids, about 6 amino acids to about 22 amino acids, about 6 amino acids to about 20 amino acids, about 6 amino acids to about 18 amino acids, about 6 amino acids to about 16 amino acids, about 6 amino acids to about 14 amino acids, about 6 amino acids to about 12 amino acids, about 6 amino acids to about 1 About 10 amino acids, about 6 to about 8 amino acids, about 8 to about 100 amino acids, about 8 to about 90 amino acids, about 8 to about 80 amino acids, about 8 to about 70 amino acids, about 8 to about 60 amino acids, about 8 to about 50 amino acids, about 8 to about 45 amino acids, about 8 to about 40 amino acids, about 8 to about 35 amino acids, about 8 to about 30 amino acids, about 8 to about 25 amino acids, about 8 to about 24 amino acids, about 8 to about 22 amino acids, about 8 to about 20 amino acids, about 8 to about 18 amino acids , about 8 amino acids to about 16 amino acids, about 8 amino acids to about 14 amino acids, about 8 amino acids to about 12 amino acids, about 8 amino acids to about 10 amino acids, about 10 amino acids to about 100 amino acids, about 10 amino acids to about 90 amino acids, about 10 amino acids to about 80 amino acids, about 10 amino acids to about 70 amino acids, about 10 amino acids to about 60 amino acids, about 10 amino acids to about 50 amino acids, about 10 amino acids to about 45 amino acids, about 10 amino acids to about 40 amino acids, about 10 amino acids to about 35 amino acids, about 10 amino acids to about 30 amino acids, about 10 amino acids to about 25 amino acids, about 10 amino acids to about 24 amino acids, about 10 to about 22 amino acids, about 10 to about 20 amino acids, about 10 to about 18 amino acids, about 10 to about 16 amino acids, about 10 to about 14 amino acids, about 10 to about 12 amino acids, about 12 to about 100 amino acids, about 12 to about 90 amino acids, about 12 to about 80 amino acids, about 12 to about 70 amino acids, about 12 to about 60 amino acids, about 12 to about 50 amino acids, about 12 to about 45 amino acids, about 12 to about 40 amino acids, about 12 to about 35 amino acids,About 12 amino acids to about 30 amino acids, about 12 amino acids to about 25 amino acids, about 12 amino acids to about 24 amino acids, about 12 amino acids to about 22 amino acids, about 12 amino acids to about 20 amino acids, about 12 amino acids to about 18 amino acids, about 12 amino acids to about 16 amino acids, about 12 amino acids to about 14 amino acids, about 14 amino acids to about 100 amino acids, about 14 amino acids to about 90 amino acids, about 14 amino acids to about 80 amino acids, about 14 amino acids to about 70 amino acids, about 14 amino acids to about 60 amino acids, about 14 amino acids to about 50 amino acids, about 14 amino acids to about 45 amino acids, about 14 amino acids to about 5 about 40 amino acids, about 14 amino acids to about 35 amino acids, about 14 amino acids to about 30 amino acids, about 14 amino acids to about 25 amino acids, about 14 amino acids to about 24 amino acids, about 14 amino acids to about 22 amino acids, about 14 amino acids to about 20 amino acids, about 14 amino acids to about 18 amino acids, about 14 amino acids to about 16 amino acids, about 16 amino acids to about 100 amino acids, about 16 amino acids to about 90 amino acids, about 16 amino acids to about 80 amino acids, about 16 amino acids to about 70 amino acids, about 16 amino acids to about 60 amino acids, about 16 amino acids to about 50 amino acids, about 16 amino acids to about 45 amino acids, About 16 amino acids to about 40 amino acids, about 16 amino acids to about 35 amino acids, about 16 amino acids to about 30 amino acids, about 16 amino acids to about 25 amino acids, about 16 amino acids to about 24 amino acids, about 16 amino acids to about 22 amino acids, about 16 amino acids to about 20 amino acids, about 16 amino acids to about 18 amino acids, about 18 amino acids to about 100 amino acids, about 18 amino acids to about 90 amino acids, about 18 amino acids to about 80 amino acids, about 18 amino acids to about 70 amino acids, about 18 amino acids to about 60 amino acids, about 18 amino acids to about 50 amino acids, about 18 amino acids to about 45 amino acids, about 18 amino acids to about about 40 amino acids, about 18 amino acids to about 35 amino acids, about 18 amino acids to about 30 amino acids, about 18 amino acids to about 25 amino acids, about 18 amino acids to about 24 amino acids, about 18 amino acids to about 22 amino acids, about 18 amino acids to about 20 amino acids, about 20 amino acids to about 100 amino acids, about 20 amino acids to about 90 amino acids, about 20 amino acids to about 80 amino acids, about 20 amino acids to about 70 amino acids, about 20 amino acids to about 60 amino acids, about 20 amino acids to about 50 amino acids, about 20 amino acids to about 45 amino acids, about 20 amino acids to about 40 amino acids, about 20 amino acids to about 35 amino acids,About 20 to about 30 amino acids, about 20 to about 25 amino acids, about 20 to about 24 amino acids, about 20 to about 22 amino acids, about 22 to about 100 amino acids, about 22 to about 90 amino acids, about 22 to about 80 amino acids, about 22 to about 70 amino acids, about 22 to about 60 amino acids, about 22 to about 50 amino acids, about 22 to about 45 amino acids, about 22 to about 40 amino acids, about 22 to about 35 amino acids, about 22 to about 30 amino acids, about 22 to about 25 amino acids, about 22 to about 24 amino acids, about 25 to about 100 amino acids, about 25 to about 90 amino acids, about 25 to about 80 amino acids, about 25 to about 70 amino acids, about 25 to about 60 amino acids, about 25 to about 50 amino acids, about 25 to about 45 amino acids, about 25 to about 40 amino acids, about 25 to about 35 amino acids, about 25 to about 30 amino acids, about 30 to about 100 amino acids, about 30 to about 90 amino acids, about 30 to about 80 amino acids, about 30 to about 70 amino acids, about 30 to about 60 amino acids, 30 to about 50 amino acids, about 30 to about 45 amino acids, about 30 to about 40 amino acids, about 30 to about 35 amino acids, about 35 to about 100 amino acids, about 35 to about 90 amino acids, about 35 to about 80 amino acids, about 35 to about 70 amino acids, about 35 to about 60 amino acids, about 35 to about 50 amino acids, about 35 to about 45 amino acids, about 35 to about 40 amino acids, about 40 to about 100 amino acids, about 40 to about 90 amino acids, about 40 to about 80 amino acids, about 40 to about 70 amino acids, about 40 amino acids to about 60 amino acids, about 40 amino acids to about 50 amino acids, about 40 amino acids to about 45 amino acids, about 45 amino acids to about 100 amino acids, about 45 amino acids to about 90 amino acids, about 45 amino acids to about 80 amino acids, about 45 amino acids to about 70 amino acids, about 45 amino acids to about 60 amino acids, about 45 amino acids to about 50 amino acids, about 50 amino acids to about 100 amino acids, about 50 amino acids to about 90 amino acids, about 50 amino acids to about 80 amino acids, about 50 amino acids to about 70 amino acids, about 50 amino acids to about 60 amino acids, about 60 amino acids to about 100 amino acids,The total length may be about 60 to about 90 amino acids, about 60 to about 80 amino acids, about 60 to about 70 amino acids, about 70 to about 100 amino acids, about 70 to about 90 amino acids, about 70 to about 80 amino acids, about 80 to about 100 amino acids, about 80 to about 90 amino acids, or about 90 to about 100 amino acids.

[0209] In some embodiments, the linker is rich in glycine (Gly or G) residues. In some embodiments, the linker is rich in serine (Ser or S) residues. In some embodiments, the linker is rich in glycine and serine residues. In some embodiments, the linker has one or more glycine-serine residue pairs (GS), for example, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more GS pairs. In some embodiments, the linker has one or more Gly-Gly-Gly-Ser (GGGS) sequences, for example, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more GGGS sequences. In some embodiments, the linker has one or more Gly-Gly-Gly-Gly-Ser (GGGGS) sequences, for example, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more GGGGS sequences. In some embodiments, the linker has one or more Gly-Gly-Ser-Gly (GGSG) sequences, for example, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more GGSG sequences.

[0210] In some embodiments, the linker sequence may comprise or consist of GGGGSGGGGSGGGGS (SEQ ID NO: 126). In some embodiments, the linker sequence may be encoded by a nucleic acid comprising or consisting of GGCGGTGGAGGATCCGGAGGAGGTGGCTCCGGCGGCGGAGGATCT (SEQ ID NO: 127). In some embodiments, the linker sequence may comprise or consist of GGGSGGGS (SEQ ID NO: 128).

[0211] Target Binding Domain In some embodiments of any of the single-chain or multi-chain chimeric polypeptides described herein, the first target binding domain, the second target binding domain, and / or the additional target binding domain or domains can be an antigen binding domain (e.g., any of the exemplary antigen binding domains described herein or known in the art), a soluble interleukin or cytokine protein (e.g., any of the exemplary soluble interleukin or soluble cytokine proteins described herein), and a soluble interleukin or cytokine receptor (e.g., any of the exemplary soluble interleukin or soluble cytokine receptors described herein).

[0212] In some embodiments of any of the single-chain or multi-chain chimeric polypeptides described herein, one or more of the first target binding domain (e.g., any of the exemplary first target binding domains described herein or known in the art), the second target binding domain (e.g., any of the exemplary second target binding domains described herein or known in the art), and the one or more additional target binding domains are each independently selected from the group consisting of CD16a, CD28, CD3 (e.g., CD3α, CD3β, CD3δ, CD3ε, and CD3β). one or more of D3γ), CD33, CD20, CD19, CD22, CD123, IL-1R, IL-1, VEGF, IL-6R, IL-4, IL-10, PDL-1, TIGIT, PD-1, TIM3, CTLA4, MICA, MICB, IL-6, IL-8, T NFα, CD26, CD36, ULBP2, CD30, CD200, IGF-1R, MUC4AC, MUC5AC, Trop-2, CMET, EGFR, HER1, HER2, HER3, PSMA, CEA, B7H3, EPCAM, BCMA, P-cadherin, CEACA M5, UL16 binding proteins (e.g., ULBP1, ULBP2, ULBP3, ULBP4, ULBP5, and ULBP6), HLA-DR, DLL4, TYRO3, AXL, MER, CD122, CD155, PDGF-DD, TGF-β receptor II (TGF-βRII) ligand, TGF-βRIII ligand, DNAM-1 ligand, NKp46 ligand, NKp44 ligand, NKG2D ligand, NKp30 ligand, scMHC1 ligand, scMHCII ligand, scTCR ligand, IL-1 receptor, IL-2 receptor , IL-3 receptor, IL-7 receptor, IL-8 receptor, IL-10 receptor, IL-12 receptor, IL-15 receptor, IL-17 receptor, IL-18 receptor, IL-21 receptor, PDGF-DD receptor, stem cell factor (SCF) receptor, stem cell-like tyrosine kinase 3 ligand (FLT3L) receptor, MICA receptor, MICB receptor, ULP16 binding protein receptor, CD155 receptor, CD122 receptor, and CD28 receptor.

[0213] In some embodiments of any of the single-chain or multi-chain chimeric polypeptides described herein, the first target binding domain, the second target binding domain, and / or the one or more additional target binding domains are each independently from about 5 amino acids to about 1000 amino acids, from about 5 amino acids to about 950 amino acids, from about 5 amino acids to about 900 amino acids, from about 5 amino acids to about 850 amino acids, from about 5 amino acids to about 800 amino acids, from about 5 amino acids to about 750 amino acids, from about 5 amino acids to about 700 amino acids, from about 5 amino acids to about 650 amino acids, from about 5 amino acids to about 600 amino acids, from about 5 amino acids to about 700 amino acids, from about 5 amino acids to about 850 amino acids, from about 5 amino acids to about 800 amino acids, to about 550 amino acids, about 5 amino acids to about 500 amino acids, about 5 amino acids to about 450 amino acids, about 5 amino acids to about 400 amino acids, about 5 amino acids to about 350 amino acids, about 5 amino acids to about 300 amino acids, about 5 amino acids to about 280 amino acids, about 5 amino acids to about 260 amino acids, about 5 amino acids to about 240 amino acids, about 5 amino acids to about 220 amino acids, about 5 amino acids to about 200 amino acids, about 5 amino acids to about 195 amino acids, about 5 amino acids to about 190 amino acids, about 5 amino acids to about 185 amino acids, about 5 amino acids to about 180 amino acids, about 5 amino acids to about 175 amino acids acid, about 5 amino acids to about 170 amino acids, about 5 amino acids to about 165 amino acids, about 5 amino acids to about 160 amino acids, about 5 amino acids to about 155 amino acids, about 5 amino acids to about 150 amino acids, about 5 amino acids to about 145 amino acids, about 5 amino acids to about 140 amino acids, about 5 amino acids to about 135 amino acids, about 5 amino acids to about 130 amino acids, about 5 amino acids to about 125 amino acids, about 5 amino acids to about 120 amino acids, about 5 amino acids to about 115 amino acids, about 5 amino acids to about 110 amino acids, about 5 amino acids to about 105 amino acids, about 5 amino acids to about 100 amino acids, about 5 amino acids about 95 amino acids, about 5 amino acids to about 90 amino acids, about 5 amino acids to about 85 amino acids, about 5 amino acids to about 80 amino acids, about 5 amino acids to about 75 amino acids, about 5 amino acids to about 70 amino acids, about 5 amino acids to about 65 amino acids, about 5 amino acids to about 60 amino acids, about 5 amino acids to about 55 amino acids, about 5 amino acids to about 50 amino acids, about 5 amino acids to about 45 amino acids, about 5 amino acids to about 40 amino acids, about 5 amino acids to about 35 amino acids, about 5 amino acids to about 30 amino acids, about 5 amino acids to about 25 amino acids, about 5 amino acids to about 20 amino acids, about 5 amino acids to about 15 amino acids,About 5 amino acids to about 10 amino acids, about 10 amino acids to about 1000 amino acids, about 10 amino acids to about 950 amino acids, about 10 amino acids to about 900 amino acids, about 10 amino acids to about 850 amino acids, about 10 amino acids to about 800 amino acids, about 10 amino acids to about 750 amino acids, about 10 amino acids to about 700 amino acids, about 10 amino acids to about 650 amino acids, about 10 amino acids to about 600 amino acids, about 10 amino acids to about 550 amino acids, about 10 amino acids to about 500 amino acids, about 10 amino acids to about 450 amino acids, about 10 amino acids to about 400 amino acids, about 10 amino acids to about 350 amino acids, about 10 amino acids to about 300 amino acids, about 10 amino acids to about 280 amino acids, about 10 amino acids to about 260 amino acids, about 10 amino acids to about 240 amino acids, about 10 amino acids to about 220 amino acids, about 10 amino acids to about 200 amino acids, about 10 amino acids to about 195 amino acids, about 10 amino acids to about 190 amino acids, about 10 amino acids to about 185 amino acids, about 10 amino acids to about 180 amino acids, about 10 amino acids to about 175 amino acids, about 10 amino acids to about 170 amino acids, about 10 amino acids to about 165 amino acids, about 10 amino acids to about 160 amino acids, about 10 amino acids to about 155 amino acids, about 10 amino acids to about 150 amino acids, about 10 amino acids to about 145 amino acids, about 10 amino acids to about 140 amino acids, about 10 amino acids to about 135 amino acids, about 10 amino acids to about 130 amino acids, about 10 amino acids to about 125 amino acids, about 10 amino acids to about 120 amino acids, about 10 amino acids to about 115 amino acids, about 10 amino acids to about 110 amino acids, about 10 amino acids to about 105 amino acids, about 10 amino acids to about 100 amino acids, about 10 amino acids to about 95 amino acids, about 10 amino acids to about 90 amino acids, about 10 amino acids to about 85 Amino acids, about 10 to about 80 amino acids, about 10 to about 75 amino acids, about 10 to about 70 amino acids, about 10 to about 65 amino acids, about 10 to about 60 amino acids, about 10 to about 55 amino acids, about 10 to about 50 amino acids, about 10 to about 45 amino acids, about 10 to about 40 amino acids, about 10 to about 35 amino acids, about 10 to about 30 amino acids, about 10 to about 25 amino acids, about 10 to about 20 amino acids, about 10 to about 15 amino acids, about 15 to about 1000 amino acids,About 15 amino acids to about 950 amino acids, about 15 amino acids to about 900 amino acids, about 15 amino acids to about 850 amino acids, about 15 amino acids to about 800 amino acids, about 15 amino acids to about 750 amino acids, about 15 amino acids to about 700 amino acids, about 15 amino acids to about 650 amino acids, about 15 amino acids to about 600 amino acids, about 15 amino acids to about 550 amino acids, about 15 amino acids to about 500 amino acids, about 15 amino acids to about 450 amino acids, about 15 amino acids to about 400 amino acids, about 15 amino acids to about 350 amino acids, about 15 amino acids to about 300 amino acids, about 15 amino acids to about 280 amino acids, about 15 amino acids to about 260 amino acids, about 15 amino acids to about 240 amino acids, about 15 amino acids to about 220 amino acids, about 15 amino acids to about 200 amino acids, about 15 amino acids to about 195 amino acids, about 15 amino acids to about 190 amino acids, about 15 amino acids to about 185 amino acids, about 15 amino acids to about 180 amino acids, about 15 amino acids to about 175 amino acids, about 15 amino acids to about 170 amino acids, about 15 amino acids to about 165 amino acids, about 15 amino acids to about 160 amino acids, about 15 amino acids to about 155 amino acids, about 15 amino acids to about 150 amino acids, about 15 to about 145 amino acids, about 15 to about 140 amino acids, about 15 to about 135 amino acids, about 15 to about 130 amino acids, about 15 to about 125 amino acids, about 15 to about 120 amino acids, about 15 to about 115 amino acids, about 15 to about 110 amino acids, about 15 to about 105 amino acids, about 15 to about 100 amino acids, about 15 to about 95 amino acids, about 15 to about 90 amino acids, about 15 to about 85 amino acids, about 15 to about 80 amino acids, about 15 to about 75 amino acids amino acids, about 15 to about 70 amino acids, about 15 to about 65 amino acids, about 15 to about 60 amino acids, about 15 to about 55 amino acids, about 15 to about 50 amino acids, about 15 to about 45 amino acids, about 15 to about 40 amino acids, about 15 to about 35 amino acids, about 15 to about 30 amino acids, about 15 to about 25 amino acids, about 15 to about 20 amino acids, about 20 to about 1000 amino acids, about 20 to about 950 amino acids, about 20 to about 900 amino acids, about 20 to about 850 amino acids,About 20 amino acids to about 800 amino acids, about 20 amino acids to about 750 amino acids, about 20 amino acids to about 700 amino acids, about 20 amino acids to about 650 amino acids, about 20 amino acids to about 600 amino acids, about 20 amino acids to about 550 amino acids, about 20 amino acids to about 500 amino acids, about 20 amino acids to about 450 amino acids, about 20 amino acids to about 400 amino acids, about 20 amino acids to about 350 amino acids, about 20 amino acids to about 300 amino acids, about 20 amino acids to about 280 amino acids, about 20 amino acids to about 260 amino acids, about 20 amino acids to about 240 amino acids, about 20 amino acids to about 220 amino acids, about 20 amino acids to about 200 amino acids, about 20 amino acids to about 195 amino acids, about 20 amino acids to about 190 amino acids, about 20 amino acids to about 185 amino acids, about 20 amino acids to about 180 amino acids, about 20 amino acids to about 175 amino acids, about 20 amino acids to about 170 amino acids, about 20 amino acids to about 165 amino acids, about 20 amino acids to about 160 amino acids, about 20 amino acids to about 155 amino acids, about 20 amino acids to about 150 amino acids, about 20 amino acids to about 145 amino acids, about 20 amino acids to about 140 amino acids, about 20 amino acids to about 135 amino acids, about 20 amino acids to about 130 amino acids, about 20 amino acids to about 125 amino acids, about 20 amino acids to about 120 amino acids, about 20 amino acids to about 115 amino acids, about 20 amino acids to about 110 amino acids, about 20 amino acids to about 105 amino acids, about 20 amino acids to about 100 amino acids, about 20 amino acids to about 95 amino acids, about 20 amino acids to about 90 amino acids, about 20 amino acids to about 85 amino acids, about 20 amino acids to about 80 amino acids, about 20 amino acids to about 75 amino acids, about 20 amino acids to about 70 amino acids, about 20 amino acids to about 65 amino acids, about 20 amino acids to about 60 amino acids, about 20 amino acids to about 55 amino acids, about 20 amino acids to about 50 amino acids, about 20 amino acids to about 45 amino acids, about 20 amino acids to about 40 amino acids, about 20 amino acids to about 35 amino acids, about 20 amino acids to about 30 amino acids, about 20 amino acids to about 25 amino acids, about 25 amino acids to about 1000 amino acids, about 25 amino acids to about 950 amino acids, about 25 amino acids to about 900 amino acids, about 25 amino acids to about 850 amino acids, about 25 amino acids to about 800 amino acids, about 25 amino acids to about 750 amino acids, about 25 amino acids to about 700 amino acids, about 25 amino acids to about 650 amino acids,About 25 amino acids to about 600 amino acids, about 25 amino acids to about 550 amino acids, about 25 amino acids to about 500 amino acids, about 25 amino acids to about 450 amino acids, about 25 amino acids to about 400 amino acids, about 25 amino acids to about 350 amino acids, about 25 amino acids to about 300 amino acids, about 25 amino acids to about 280 amino acids, about 25 amino acids to about 260 amino acids, about 25 amino acids to about 240 amino acids, about 25 amino acids to about 220 amino acids, about 25 amino acids to about 200 amino acids, about 25 amino acids to about 195 amino acids, about 25 amino acids to about 190 amino acids, about 25 amino acids to about 185 amino acids, about 25 amino acids to about 180 amino acids, about 25 amino acids to about 175 amino acids, about 25 amino acids to about 170 amino acids, about 25 amino acids to about 165 amino acids, about 25 amino acids to about 160 amino acids, about 25 amino acids to about 155 amino acids, about 25 amino acids to about 150 amino acids, about 25 amino acids to about 145 amino acids, about 25 amino acids to about 140 amino acids, about 25 amino acids to about 135 amino acids, about 25 amino acids to about 130 amino acids, about 25 amino acids to about 125 amino acids, about 25 amino acids to about 120 amino acids, about 25 amino acids to about 115 amino acids, about 2 5 amino acids to about 110 amino acids, about 25 amino acids to about 105 amino acids, about 25 amino acids to about 100 amino acids, about 25 amino acids to about 95 amino acids, about 25 amino acids to about 90 amino acids, about 25 amino acids to about 85 amino acids, about 25 amino acids to about 80 amino acids, about 25 amino acids to about 75 amino acids, about 25 amino acids to about 70 amino acids, about 25 amino acids to about 65 amino acids, about 25 amino acids to about 60 amino acids, about 25 amino acids to about 55 amino acids, about 25 amino acids to about 50 amino acids, about 25 amino acids to about 45 amino acids, about 25 amino acids to about 40 amino acids, about 25 amino acids to about 5 amino acids to about 35 amino acids, about 25 amino acids to about 30 amino acids, about 30 amino acids to about 1000 amino acids, about 30 amino acids to about 950 amino acids, about 30 amino acids to about 900 amino acids, about 30 amino acids to about 850 amino acids, about 30 amino acids to about 800 amino acids, about 30 amino acids to about 750 amino acids, about 30 amino acids to about 700 amino acids, about 30 amino acids to about 650 amino acids, about 30 amino acids to about 600 amino acids, about 30 amino acids to about 550 amino acids, about 30 amino acids to about 500 amino acids, about 30 amino acids to about 450 amino acids, about 30 amino acids to about 400 amino acids,About 30 amino acids to about 350 amino acids, about 30 amino acids to about 300 amino acids, about 30 amino acids to about 280 amino acids, about 30 amino acids to about 260 amino acids, about 30 amino acids to about 240 amino acids, about 30 amino acids to about 220 amino acids, about 30 amino acids to about 200 amino acids, about 30 amino acids to about 195 amino acids, about 30 amino acids to about 190 amino acids, about 30 amino acids to about 185 amino acids, about 30, Amino acids up to about 180 amino acids, about 30 amino acids to about 175 amino acids, about 30 amino acids to about 170 amino acids, about 30 amino acids to about 165 amino acids, about 30 amino acids to about 160 amino acids, about 30 amino acids to about 155 amino acids, about 30 amino acids to about 150 amino acids, about 30 amino acids to about 145 amino acids, about 30 amino acids to about 140 amino acids, about 30 amino acids to about 135 amino acids, about 30 amino acids to about 130 amino acids, about 30 amino acids to about 125 amino acids, about 30 amino acids to about 120 amino acids, about 30 amino acids to about 115 amino acids, about 30 amino acids to about 110 amino acids, about 30 to about 105 amino acids, about 30 to about 100 amino acids, about 30 to about 95 amino acids, about 30 to about 90 amino acids, about 30 to about 85 amino acids, about 30 to about 80 amino acids, about 30 to about 75 amino acids, about 30 to about 70 amino acids, about 30 to about 65 amino acids, about 30 to about 60 amino acids, about 30 to about 55 amino acids, about 30 to about 50 amino acids, about 30 to about 45 amino acids, about 30 to about 40 amino acids, about 30 to about 35 amino acids, About 35 amino acids to about 1000 amino acids, about 35 amino acids to about 950 amino acids, about 35 amino acids to about 900 amino acids, about 35 amino acids to about 850 amino acids, about 35 amino acids to about 800 amino acids, about 35 amino acids to about 750 amino acids, about 35 amino acids to about 700 amino acids, about 35 amino acids to about 650 amino acids, about 35 amino acids to about 600 amino acids, about 35 amino acids to about 550 amino acids, about 35 amino acids to about 500 amino acids, about 35 amino acids to about 450 amino acids, about 35 amino acids to about 400 amino acids, about 35 amino acids to about 350 amino acids, about 35 amino acids to about 5 about 300 amino acids, about 35 amino acids to about 280 amino acids, about 35 amino acids to about 260 amino acids, about 35 amino acids to about 240 amino acids, about 35 amino acids to about 220 amino acids, about 35 amino acids to about 200 amino acids, about 35 amino acids to about 195 amino acids, about 35 amino acids to about 190 amino acids, about 35 amino acids to about 185 amino acids, about 35 amino acids to about 180 amino acids, about 35 amino acids to about 175 amino acids, about 35 amino acids to about 170 amino acids, about 35 amino acids to about 165 amino acids, about 35 amino acids to about 160 amino acids, about 35 amino acids to about 155 amino acids,About 35 amino acids to about 150 amino acids, about 35 amino acids to about 145 amino acids, about 35 amino acids to about 140 amino acids, about 35 amino acids to about 135 amino acids, about 35 amino acids to about 130 amino acids, about 35 amino acids to about 125 amino acids, about 35 amino acids to about 120 amino acids, about 35 amino acids to about 115 amino acids, about 35 amino acids to about 110 amino acids, about 35 amino acids to about 105 amino acids, about 35 amino acids to about 100 amino acids, about 35 amino acids to about 95 amino acids, about 35 amino acids to about 90 amino acids, about 35 amino acids to about 85 amino acids, about 35 amino acids to about 80 amino acids amino acids, about 35 amino acids to about 75 amino acids, about 35 amino acids to about 70 amino acids, about 35 amino acids to about 65 amino acids, about 35 amino acids to about 60 amino acids, about 35 amino acids to about 55 amino acids, about 35 amino acids to about 50 amino acids, about 35 amino acids to about 45 amino acids, about 35 amino acids to about 40 amino acids, about 40 amino acids to about 1000 amino acids, about 40 amino acids to about 950 amino acids, about 40 amino acids to about 900 amino acids, about 40 amino acids to about 850 amino acids, about 40 amino acids to about 800 amino acids, about 40 amino acids to about 750 amino acids, about 40 amino acids to about 700 amino acids amino acids, about 40 amino acids to about 650 amino acids, about 40 amino acids to about 600 amino acids, about 40 amino acids to about 550 amino acids, about 40 amino acids to about 500 amino acids, about 40 amino acids to about 450 amino acids, about 40 amino acids to about 400 amino acids, about 40 amino acids to about 350 amino acids, about 40 amino acids to about 300 amino acids, about 40 amino acids to about 280 amino acids, about 40 amino acids to about 260 amino acids, about 40 amino acids to about 240 amino acids, about 40 amino acids to about 220 amino acids, about 40 amino acids to about 200 amino acids, about 40 amino acids to about 195 amino acids, about 40 amino acids to about 190 amino acids, about 40 amino acids to about 185 amino acids, about 40 amino acids to about 180 amino acids, about 40 amino acids to about 175 amino acids, about 40 amino acids to about 170 amino acids, about 40 amino acids to about 165 amino acids, about 40 amino acids to about 160 amino acids, about 40 amino acids to about 155 amino acids, about 40 amino acids to about 150 amino acids, about 40 amino acids to about 145 amino acids, about 40 amino acids to about 140 amino acids, about 40 amino acids to about 135 amino acids, about 40 amino acids to about 130 amino acids, about 40 amino acids to about 125 amino acids, about 40 amino acids to about 120 amino acids,About 40 amino acids to about 115 amino acids, about 40 amino acids to about 110 amino acids, about 40 amino acids to about 105 amino acids, about 40 amino acids to about 100 amino acids, about 40 amino acids to about 95 amino acids, about 40 amino acids to about 90 amino acids, about 40 amino acids to about 85 amino acids, about 40 amino acids to about 80 amino acids, about 40 amino acids to about 75 amino acids, about 40 amino acids to about 70 amino acids, about 40 amino acids to about 65 amino acids, about 40 amino acids to about 60 amino acids, about 40 amino acids to about 55 amino acids, about 40 amino acids to about 50 amino acids, about 40 amino acids to about 45 amino acids, about 45 Amino acids up to about 1000 amino acids, about 45 amino acids to about 950 amino acids, about 45 amino acids to about 900 amino acids, about 45 amino acids to about 850 amino acids, about 45 amino acids to about 800 amino acids, about 45 amino acids to about 750 amino acids, about 45 amino acids to about 700 amino acids, about 45 amino acids to about 650 amino acids, about 45 amino acids to about 600 amino acids, about 45 amino acids to about 550 amino acids, about 45 amino acids to about 500 amino acids, about 45 amino acids to about 450 amino acids, about 45 amino acids to about 400 amino acids, about 45 amino acids to about 350 amino acids, about 45 amino acids to about 30 0 amino acids, about 45 amino acids to about 280 amino acids, about 45 amino acids to about 260 amino acids, about 45 amino acids to about 240 amino acids, about 45 amino acids to about 220 amino acids, about 45 amino acids to about 200 amino acids, about 45 amino acids to about 195 amino acids, about 45 amino acids to about 190 amino acids, about 45 amino acids to about 185 amino acids, about 45 amino acids to about 180 amino acids, about 45 amino acids to about 175 amino acids, about 45 amino acids to about 170 amino acids, about 45 amino acids to about 165 amino acids, about 45 amino acids to about 160 amino acids, about 45 amino acids to about 155 amino acids, about 45 Amino acids up to about 150 amino acids, about 45 amino acids to about 145 amino acids, about 45 amino acids to about 140 amino acids, about 45 amino acids to about 135 amino acids, about 45 amino acids to about 130 amino acids, about 45 amino acids to about 125 amino acids, about 45 amino acids to about 120 amino acids, about 45 amino acids to about 115 amino acids, about 45 amino acids to about 110 amino acids, about 45 amino acids to about 105 amino acids, about 45 amino acids to about 100 amino acids, about 45 amino acids to about 95 amino acids, about 45 amino acids to about 90 amino acids, about 45 amino acids to about 85 amino acids, about 45 amino acids to about 80 amino acids,About 45 to about 75 amino acids, about 45 to about 70 amino acids, about 45 to about 65 amino acids, about 45 to about 60 amino acids, about 45 to about 55 amino acids, about 45 to about 50 amino acids, about 50 to about 1000 amino acids, about 50 to about 950 amino acids, about 50 to about 900 amino acids, about 50 to about 850 amino acids, about 50 to about 800 amino acids, about 50 to about 750 amino acids, about 50 to about 700 amino acids, about 50 to about 650 amino acids, about 50 to about 600 amino acids amino acids, about 50 to about 550 amino acids, about 50 to about 500 amino acids, about 50 to about 450 amino acids, about 50 to about 400 amino acids, about 50 to about 350 amino acids, about 50 to about 300 amino acids, about 50 to about 280 amino acids, about 50 to about 260 amino acids, about 50 to about 240 amino acids, about 50 to about 220 amino acids, about 50 to about 200 amino acids, about 50 to about 195 amino acids, about 50 to about 190 amino acids, about 50 to about 185 amino acids, about 50 amino acids to about 180 amino acids, about 50 amino acids to about 175 amino acids, about 50 amino acids to about 170 amino acids, about 50 amino acids to about 165 amino acids, about 50 amino acids to about 160 amino acids, about 50 amino acids to about 155 amino acids, about 50 amino acids to about 150 amino acids, about 50 amino acids to about 145 amino acids, about 50 amino acids to about 140 amino acids, about 50 amino acids to about 135 amino acids, about 50 amino acids to about 130 amino acids, about 50 amino acids to about 125 amino acids, about 50 amino acids to about 120 amino acids, about 50 amino acids to about 115 amino acids, about 50 amino acids to about 110 amino acids , about 50 amino acids to about 105 amino acids, about 50 amino acids to about 100 amino acids, about 50 amino acids to about 95 amino acids, about 50 amino acids to about 90 amino acids, about 50 amino acids to about 85 amino acids, about 50 amino acids to about 80 amino acids, about 50 amino acids to about 75 amino acids, about 50 amino acids to about 70 amino acids, about 50 amino acids to about 65 amino acids, about 50 amino acids to about 60 amino acids, about 50 amino acids to about 55 amino acids, about 55 amino acids to about 1000 amino acids, about 55 amino acids to about 950 amino acids, about 55 amino acids to about 900 amino acids, about 55 amino acids to about 850 amino acids,About 55 amino acids to about 800 amino acids, about 55 amino acids to about 750 amino acids, about 55 amino acids to about 700 amino acids, about 55 amino acids to about 650 amino acids, about 55 amino acids to about 600 amino acids, about 55 amino acids to about 550 amino acids, about 55 amino acids to about 500 amino acids, about 55 amino acids to about 450 amino acids, about 55 amino acids to about 400 amino acids, about 55 amino acids to about 350 amino acids, about 55 amino acids to about 300 amino acids, about 55 amino acids to about 280 amino acids, about 55 amino acids to about 260 amino acids, about 55 amino acids to about 240 amino acids, about 55 amino acids to about 22 0 amino acids, about 55 amino acids to about 200 amino acids, about 55 amino acids to about 195 amino acids, about 55 amino acids to about 190 amino acids, about 55 amino acids to about 185 amino acids, about 55 amino acids to about 180 amino acids, about 55 amino acids to about 175 amino acids, about 55 amino acids to about 170 amino acids, about 55 amino acids to about 165 amino acids, about 55 amino acids to about 160 amino acids, about 55 amino acids to about 155 amino acids, about 55 amino acids to about 150 amino acids, about 55 amino acids to about 145 amino acids, about 55 amino acids to about 140 amino acids, about 55 amino acids to about 135 amino acids, about 55 amino acids to about 130 amino acids, about 55 amino acids to about 125 amino acids, about 55 amino acids to about 120 amino acids, about 55 amino acids to about 115 amino acids, about 55 amino acids to about 110 amino acids, about 55 amino acids to about 105 amino acids, about 55 amino acids to about 100 amino acids, about 55 amino acids to about 95 amino acids, about 55 amino acids to about 90 amino acids, about 55 amino acids to about 85 amino acids, about 55 amino acids to about 80 amino acids, about 55 amino acids to about 75 amino acids, about 55 amino acids to about 70 amino acids, about 55 amino acids to about 65 amino acids, about 55 amino acids to about 60 amino acids, about 60 amino acids to about 1000 amino acids, about 60 amino acids to about 950 amino acids, about 60 amino acids to about 900 amino acids, about 60 amino acids to about 850 amino acids, about 60 amino acids to about 800 amino acids, about 60 amino acids to about 750 amino acids, about 60 amino acids to about 700 amino acids, about 60 amino acids to about 650 amino acids, about 60 amino acids to about 600 amino acids, about 60 amino acids to about 550 amino acids, about 60 amino acids to about 500 amino acids, about 60 amino acids to about 450 amino acids, about 60 amino acids to about 400 amino acids, about 60 amino acids to about 350 amino acids, about 60 amino acids to about 300 amino acids,About 60 to about 280 amino acids, about 60 to about 260 amino acids, about 60 to about 240 amino acids, about 60 to about 220 amino acids, about 60 to about 200 amino acids, about 60 to about 195 amino acids, about 60 to about 190 amino acids, about 60 to about 185 amino acids, about 60 to about 180 amino acids, about 60 to about 175 amino acids, Amino acids, about 60 amino acids to about 170 amino acids, about 60 amino acids to about 165 amino acids, about 60 amino acids to about 160 amino acids, about 60 amino acids to about 155 amino acids, about 60 amino acids to about 150 amino acids, about 60 amino acids to about 145 amino acids, about 60 amino acids to about 140 amino acids, about 60 amino acids to about 135 amino acids, about 60 amino acids to about 130 amino acids, about 60 amino acids to about 125 amino acids, about 60 amino acids to about 120 amino acids, about 60 amino acids to about 115 amino acids, about 60 amino acids to about 110 amino acids, about 60 amino acids to about 105 amino acids, about 60 Amino acids to about 100 amino acids, about 60 amino acids to about 95 amino acids, about 60 amino acids to about 90 amino acids, about 60 amino acids to about 85 amino acids, about 60 amino acids to about 80 amino acids, about 60 amino acids to about 75 amino acids, about 60 amino acids to about 70 amino acids, about 60 amino acids to about 65 amino acids, about 65 amino acids to about 1000 amino acids, about 65 amino acids to about 950 amino acids, about 65 amino acids to about 900 amino acids, about 65 amino acids to about 850 amino acids, about 65 amino acids to about 800 amino acids, about 65 amino acids to about 750 amino acids, about 65 amino acids to about 700 amino acids, About 65 amino acids to about 650 amino acids, about 65 amino acids to about 600 amino acids, about 65 amino acids to about 550 amino acids, about 65 amino acids to about 500 amino acids, about 65 amino acids to about 450 amino acids, about 65 amino acids to about 400 amino acids, about 65 amino acids to about 350 amino acids, about 65 amino acids to about 300 amino acids, about 65 amino acids to about 280 amino acids, about 65 amino acids to about 260 amino acids, about 65 amino acids to about 240 amino acids, about 65 amino acids to about 220 amino acids, about 65 amino acids to about 200 amino acids, about 65 amino acids to about 195 ... about 190 amino acids, about 65 amino acids to about 185 amino acids, about 65 amino acids to about 180 amino acids, about 65 amino acids to about 175 amino acids, about 65 amino acids to about 170 amino acids, about 65 amino acids to about 165 amino acids, about 65 amino acids to about 160 amino acids, about 65 amino acids to about 155 amino acids, about 65 amino acids to about 150 amino acids, about 65 amino acids to about 145 amino acids, about 65 amino acids to about 140 amino acids, about 65 amino acids to about 135 amino acids, about 65 amino acids to about 130 amino acids, about 65 amino acids to about 125 amino acids, about 65 amino acids to about 120 amino acids,About 65 amino acids to about 115 amino acids, about 65 amino acids to about 110 amino acids, about 65 amino acids to about 105 amino acids, about 65 amino acids to about 100 amino acids, about 65 amino acids to about 95 amino acids, about 65 amino acids to about 90 amino acids, about 65 amino acids to about 85 amino acids, about 65 amino acids to about 80 amino acids, about 65 amino acids to about 75 amino acids, about 65 amino acids to about 70 amino acids, about 70 amino acids to about 1000 amino acids, about 70 amino acids to about 950 amino acids, about 70 amino acids to about 900 amino acids, about 70 amino acids to about 850 amino acids, about 70 amino acids to about 800 amino acids amino acids, about 70 to about 750 amino acids, about 70 to about 700 amino acids, about 70 to about 650 amino acids, about 70 to about 600 amino acids, about 70 to about 550 amino acids, about 70 to about 500 amino acids, about 70 to about 450 amino acids, about 70 to about 400 amino acids, about 70 to about 350 amino acids, about 70 to about 300 amino acids, about 70 to about 280 amino acids, about 70 to about 260 amino acids, about 70 to about 240 amino acids, about 70 to about 220 amino acids, about 70 to about 200 amino acids, about 70 amino acids to about 195 amino acids, about 70 amino acids to about 190 amino acids, about 70 amino acids to about 185 amino acids, about 70 amino acids to about 180 amino acids, about 70 amino acids to about 175 amino acids, about 70 amino acids to about 170 amino acids, about 70 amino acids to about 165 amino acids, about 70 amino acids to about 160 amino acids, about 70 amino acids to about 155 amino acids, about 70 amino acids to about 150 amino acids, about 70 amino acids to about 145 amino acids, about 70 amino acids to about 140 amino acids, about 70 amino acids to about 135 amino acids, about 70 amino acids to about 130 amino acids, about 7 0 amino acids to about 125 amino acids, about 70 amino acids to about 120 amino acids, about 70 amino acids to about 115 amino acids, about 70 amino acids to about 110 amino acids, about 70 amino acids to about 105 amino acids, about 70 amino acids to about 100 amino acids, about 70 amino acids to about 95 amino acids, about 70 amino acids to about 90 amino acids, about 70 amino acids to about 85 amino acids, about 70 amino acids to about 80 amino acids, about 70 amino acids to about 75 amino acids, about 75 amino acids to about 1000 amino acids, about 75 amino acids to about 950 amino acids, about 75 amino acids to about 900 amino acids, about 75 amino acids to about 850 amino acids,About 75 amino acids to about 800 amino acids, about 75 amino acids to about 750 amino acids, about 75 amino acids to about 700 amino acids, about 75 amino acids to about 650 amino acids, about 75 amino acids to about 600 amino acids, about 75 amino acids to about 550 amino acids, about 75 amino acids to about 500 amino acids, about 75 amino acids to about 450 amino acids, about 75 amino acids to about 400 amino acids, about 75 amino acids to about 350 amino acids, about 75 amino acids to about 300 amino acids, about 75 amino acids to about 280 amino acids, about 75 amino acids to about 260 amino acids, about 75 amino acids to about 240 amino acids, about 75 amino acids to about 220 Amino acids, about 75 to about 200 amino acids, about 75 to about 195 amino acids, about 75 to about 190 amino acids, about 75 to about 185 amino acids, about 75 to about 180 amino acids, about 75 to about 175 amino acids, about 75 to about 170 amino acids, about 75 to about 165 amino acids, about 75 to about 160 amino acids, about 75 to about 155 amino acids, about 75 to about 150 amino acids, about 75 to about 145 amino acids, about 75 to about 140 amino acids, about 75 to about 135 amino acids, about 75 amino acids about 130 amino acids, about 75 amino acids to about 125 amino acids, about 75 amino acids to about 120 amino acids, about 75 amino acids to about 115 amino acids, about 75 amino acids to about 110 amino acids, about 75 amino acids to about 105 amino acids, about 75 amino acids to about 100 amino acids, about 75 amino acids to about 95 amino acids, about 75 amino acids to about 90 amino acids, about 75 amino acids to about 85 amino acids, about 75 amino acids to about 80 amino acids, about 80 amino acids to about 1000 amino acids, about 80 amino acids to about 950 amino acids, about 80 amino acids to about 900 amino acids, about 80 amino acids to about 850 amino acids, about 80 amino acids from about 800 amino acids, from about 80 amino acids to about 750 amino acids, from about 80 amino acids to about 700 amino acids, from about 80 amino acids to about 650 amino acids, from about 80 amino acids to about 600 amino acids, from about 80 amino acids to about 550 amino acids, from about 80 amino acids to about 500 amino acids, from about 80 amino acids to about 450 amino acids, from about 80 amino acids to about 400 amino acids, from about 80 amino acids to about 350 amino acids, from about 80 amino acids to about 300 amino acids, from about 80 amino acids to about 280 amino acids, from about 80 amino acids to about 260 amino acids, from about 80 amino acids to about 240 amino acids, from about 80 amino acids to about 220 amino acids,About 80 amino acids to about 200 amino acids, about 80 amino acids to about 195 amino acids, about 80 amino acids to about 190 amino acids, about 80 amino acids to about 185 amino acids, about 80 amino acids to about 180 amino acids, about 80 amino acids to about 175 amino acids, about 80 amino acids to about 170 amino acids, about 80 amino acids to about 165 amino acids, about 80 amino acids to about 160 amino acids, about 80 amino acids to about 155 amino acids, about 80 amino acids to about 150 amino acids, about 80 amino acids to about 145 amino acids, about 80 amino acids to about 140 amino acids, about 80 amino acids to about 135 amino acids, about 80 amino acids to about 130 Amino acids, about 80 to about 125 amino acids, about 80 to about 120 amino acids, about 80 to about 115 amino acids, about 80 to about 110 amino acids, about 80 to about 105 amino acids, about 80 to about 100 amino acids, about 80 to about 95 amino acids, about 80 to about 90 amino acids, about 80 to about 85 amino acids, about 85 to about 1000 amino acids, about 85 to about 950 amino acids, about 85 to about 900 amino acids, about 85 to about 850 amino acids, about 85 to about 800 amino acids, about 85 to about 750 amino acids, about 85 amino acids to about 700 amino acids, about 85 amino acids to about 650 amino acids, about 85 amino acids to about 600 amino acids, about 85 amino acids to about 550 amino acids, about 85 amino acids to about 500 amino acids, about 85 amino acids to about 450 amino acids, about 85 amino acids to about 400 amino acids, about 85 amino acids to about 350 amino acids, about 85 amino acids to about 300 amino acids, about 85 amino acids to about 280 amino acids, about 85 amino acids to about 260 amino acids, about 85 amino acids to about 240 amino acids, about 85 amino acids to about 220 amino acids, about 85 amino acids to about 200 amino acids, about 85 amino acids amino acids to about 195 amino acids, about 85 amino acids to about 190 amino acids, about 85 amino acids to about 185 amino acids, about 85 amino acids to about 180 amino acids, about 85 amino acids to about 175 amino acids, about 85 amino acids to about 170 amino acids, about 85 amino acids to about 165 amino acids, about 85 amino acids to about 160 amino acids, about 85 amino acids to about 155 amino acids, about 85 amino acids to about 150 amino acids, about 85 amino acids to about 145 amino acids, about 85 amino acids to about 140 amino acids, about 85 amino acids to about 135 amino acids, about 85 amino acids to about 130 amino acids, about 85 amino acids to about 125 amino acids,About 85 amino acids to about 120 amino acids, about 85 amino acids to about 115 amino acids, about 85 amino acids to about 110 amino acids, about 85 amino acids to about 105 amino acids, about 85 amino acids to about 100 amino acids, about 85 amino acids to about 95 amino acids, about 85 amino acids to about 90 amino acids, about 90 amino acids to about 1000 amino acids, about 90 amino acids to about 950 amino acids, about 90 amino acids to about 900 amino acids, about 90 amino acids to about 850 amino acids, about 90 amino acids to about 800 amino acids, about 90 amino acids to about 750 amino acids, about 90 amino acids to about 700 amino acids, about 90 amino acids to about 650 amino acids, about 90 amino acids to about 600 amino acids, about 90 amino acids to about 550 amino acids, about 90 amino acids to about 500 amino acids, about 90 amino acids to about 450 amino acids, about 90 amino acids to about 400 amino acids, about 90 amino acids to about 350 amino acids, about 90 amino acids to about 300 amino acids, about 90 amino acids to about 280 amino acids, about 90 amino acids to about 260 amino acids, about 90 amino acids to about 240 amino acids, about 90 amino acids to about 220 amino acids, about 90 amino acids to about 200 amino acids, about 90 amino acids to about 195 amino acids, about 90 amino acids to about 190 amino acids, About 90 amino acids to about 185 amino acids, about 90 amino acids to about 180 amino acids, about 90 amino acids to about 175 amino acids, about 90 amino acids to about 170 amino acids, about 90 amino acids to about 165 amino acids, about 90 amino acids to about 160 amino acids, about 90 amino acids to about 155 amino acids, about 90 amino acids to about 150 amino acids, about 90 amino acids to about 145 amino acids, about 90 amino acids to about 140 amino acids, about 90 amino acids to about 135 amino acids, about 90 amino acids to about 130 amino acids, about 90 amino acids to about 125 amino acids, about 90 amino acids to about 12 ... about 115 amino acids, about 90 amino acids to about 110 amino acids, about 90 amino acids to about 105 amino acids, about 90 amino acids to about 100 amino acids, about 90 amino acids to about 95 amino acids, about 95 amino acids to about 1000 amino acids, about 95 amino acids to about 950 amino acids, about 95 amino acids to about 900 amino acids, about 95 amino acids to about 850 amino acids, about 95 amino acids to about 800 amino acids, about 95 amino acids to about 750 amino acids, about 95 amino acids to about 700 amino acids, about 95 amino acids to about 650 amino acids, about 95 amino acids to about 600 amino acids, about 95 amino acids to about 550 amino acids,About 95 amino acids to about 500 amino acids, about 95 amino acids to about 450 amino acids, about 95 amino acids to about 400 amino acids, about 95 amino acids to about 350 amino acids, about 95 amino acids to about 300 amino acids, about 95 amino acids to about 280 amino acids, about 95 amino acids to about 260 amino acids, about 95 amino acids to about 240 amino acids, about 95 amino acids to about 220 amino acids, about 9, 5 amino acids to about 200 amino acids, about 95 amino acids to about 195 amino acids, about 95 amino acids to about 190 amino acids, about 95 amino acids to about 185 amino acids, about 95 amino acids to about 180 amino acids, about 95 amino acids to about 175 amino acids, about 95 amino acids to about 170 amino acids, about 95 amino acids to about 165 amino acids, about 95 amino acids to about 160 amino acids, about 95 amino acids to about 155 amino acids, about 95 amino acids to about 150 amino acids, about 95 amino acids to about 145 amino acids, about 95 amino acids to about 140 amino acids, about 95 amino acids to about 135 amino acids, about 95 amino acids to about 1 30 amino acids, about 95 amino acids to about 125 amino acids, about 95 amino acids to about 120 amino acids, about 95 amino acids to about 115 amino acids, about 95 amino acids to about 110 amino acids, about 95 amino acids to about 105 amino acids, about 95 amino acids to about 100 amino acids, about 100 amino acids to about 1000 amino acids, about 100 amino acids to about 950 amino acids, about 100 amino acids to about 900 amino acids, about 100 amino acids to about 850 amino acids, about 100 amino acids to about 800 amino acids, about 100 amino acids to about 750 amino acids, about 100 amino acids to about 700 amino acids, about 100 amino acids to about 6 50 amino acids, about 100 to about 600 amino acids, about 100 to about 550 amino acids, about 100 to about 500 amino acids, about 100 to about 450 amino acids, about 100 to about 400 amino acids, about 100 to about 350 amino acids, about 100 to about 300 amino acids, about 100 to about 280 amino acids, about 100 to about 260 amino acids, about 100 to about 240 amino acids, about 100 to about 220 amino acids, about 100 to about 200 amino acids, about 100 to about 195 amino acids, about 100 amino acids amino acids to about 190 amino acids, about 100 amino acids to about 185 amino acids, about 100 amino acids to about 180 amino acids, about 100 amino acids to about 175 amino acids, about 100 amino acids to about 170 amino acids, about 100 amino acids to about 165 amino acids, about 100 amino acids to about 160 amino acids, about 100 amino acids to about 155 amino acids, about 100 amino acids to about 150 amino acids, about 100 amino acids to about 145 amino acids, about 100 amino acids to about 140 amino acids, about 100 amino acids to about 135 amino acids, about 100 amino acids to about 130 amino acids, about 100 amino acids to about 125 amino acids,About 100 amino acids to about 120 amino acids, about 100 amino acids to about 115 amino acids, about 100 amino acids to about 110 amino acids, about 100 amino acids to about 105 amino acids, about 105 amino acids to about 1000 amino acids, about 105 amino acids to about 950 amino acids, about 105 amino acids to about 900 amino acids, about 105 amino acids to about 850 amino acids, about 105 amino acids to about 800 amino acids, about 105 amino acids to about 750 amino acids, about 105 amino acids to about 700 amino acids, about 105 amino acids to about 650 amino acids, about 105 amino acids to about 600 amino acids, about 105 amino acids to about 550 Amino acids, about 105 amino acids to about 500 amino acids, about 105 amino acids to about 450 amino acids, about 105 amino acids to about 400 amino acids, about 105 amino acids to about 350 amino acids, about 105 amino acids to about 300 amino acids, about 105 amino acids to about 280 amino acids, about 105 amino acids to about 260 amino acids, about 105 amino acids to about 240 amino acids, about 105 amino acids to about 220 amino acids, about 105 amino acids to about 200 amino acids, about 105 amino acids to about 195 amino acids, about 105 amino acids to about 190 amino acids, about 105 amino acids to about 185 ... about 180 amino acids, about 105 amino acids to about 175 amino acids, about 105 amino acids to about 170 amino acids, about 105 amino acids to about 165 amino acids, about 105 amino acids to about 160 amino acids, about 105 amino acids to about 155 amino acids, about 105 amino acids to about 150 amino acids, about 105 amino acids to about 145 amino acids, about 105 amino acids to about 140 amino acids, about 105 amino acids to about 135 amino acids, about 105 amino acids to about 130 amino acids, about 105 amino acids to about 125 amino acids, about 105 amino acids to about 120 amino acids, about 105 amino acids to about 115 ... amino acids to about 110 amino acids, about 110 amino acids to about 1000 amino acids, about 110 amino acids to about 950 amino acids, about 110 amino acids to about 900 amino acids, about 110 amino acids to about 850 amino acids, about 110 amino acids to about 800 amino acids, about 110 amino acids to about 750 amino acids, about 110 amino acids to about 700 amino acids, about 110 amino acids to about 650 amino acids, about 110 amino acids to about 600 amino acids, about 110 amino acids to about 550 amino acids, about 110 amino acids to about 500 amino acids, about 110 amino acids to about 450 amino acids, about 110 amino acids to about 400 amino acids,About 110 amino acids to about 350 amino acids, about 110 amino acids to about 300 amino acids, about 110 amino acids to about 280 amino acids, about 110 amino acids to about 260 amino acids, about 110 amino acids to about 240 amino acids, about 110 amino acids to about 220 amino acids, about 110 amino acids to about 200 amino acids, about 110 amino acids to about 195 amino acids, about 110 amino acids to about 190 amino acids, about 110 amino acids to about 185 amino acids, about 110 amino acids to about 180 amino acids, about 110 amino acids to about 175 amino acids, about 110 amino acids to about 170 amino acids, about 110 amino acids to about 165 Amino acids, about 110 amino acids to about 160 amino acids, about 110 amino acids to about 155 amino acids, about 110 amino acids to about 150 amino acids, about 110 amino acids to about 145 amino acids, about 110 amino acids to about 140 amino acids, about 110 amino acids to about 135 amino acids, about 110 amino acids to about 130 amino acids, about 110 amino acids to about 125 amino acids, about 110 amino acids to about 120 amino acids, about 110 amino acids to about 115 amino acids, about 115 amino acids to about 1000 amino acids, about 115 amino acids to about 950 amino acids, about 115 amino acids to about 900 amino acids, about 115 amino acids to about 850 amino acids, about 115 amino acids to about 800 amino acids, about 115 amino acids to about 750 amino acids, about 115 amino acids to about 700 amino acids, about 115 amino acids to about 650 amino acids, about 115 amino acids to about 600 amino acids, about 115 amino acids to about 550 amino acids, about 115 amino acids to about 500 amino acids, about 115 amino acids to about 450 amino acids, about 115 amino acids to about 400 amino acids, about 115 amino acids to about 350 amino acids, about 115 amino acids to about 300 amino acids, about 115 amino acids to about 280 amino acids, about 115 amino acids to about 260 amino acids, about 115 Amino acids to about 240 amino acids, about 115 amino acids to about 220 amino acids, about 115 amino acids to about 200 amino acids, about 115 amino acids to about 195 amino acids, about 115 amino acids to about 190 amino acids, about 115 amino acids to about 185 amino acids, about 115 amino acids to about 180 amino acids, about 115 amino acids to about 175 amino acids, about 115 amino acids to about 170 amino acids, about 115 amino acids to about 165 amino acids, about 115 amino acids to about 160 amino acids, about 115 amino acids to about 155 amino acids, about 115 amino acids to about 150 amino acids, about 115 amino acids to about 145 amino acids,About 115 amino acids to about 140 amino acids, about 115 amino acids to about 135 amino acids, about 115 amino acids to about 130 amino acids, about 115 amino acids to about 125 amino acids, about 115 amino acids to about 120 amino acids, about 120 amino acids to about 1000 amino acids, about 120 amino acids to about 950 amino acids, about 120 amino acids to about 900 amino acids, about 120 amino acids to about 850 amino acids, about 120 amino acids to about 800 amino acids, about 120 amino acids to about 750 amino acids, about 120 amino acids to about 700 amino acids, about 120 amino acids to about 650 amino acids, about 120 amino acids to about 600 Amino acids, about 120 amino acids to about 550 amino acids, about 120 amino acids to about 500 amino acids, about 120 amino acids to about 450 amino acids, about 120 amino acids to about 400 amino acids, about 120 amino acids to about 350 amino acids, about 120 amino acids to about 300 amino acids, about 120 amino acids to about 280 amino acids, about 120 amino acids to about 260 amino acids, about 120 amino acids to about 240 amino acids, about 120 amino acids to about 220 amino acids, about 120 amino acids to about 200 amino acids, about 120 amino acids to about 195 amino acids, about 120 amino acids to about 190 amino acids, about 120 amino acids to about 2 About 185 amino acids, about 120 amino acids to about 180 amino acids, about 120 amino acids to about 175 amino acids, about 120 amino acids to about 170 amino acids, about 120 amino acids to about 165 amino acids, about 120 amino acids to about 160 amino acids, about 120 amino acids to about 155 amino acids, about 120 amino acids to about 150 amino acids, about 120 amino acids to about 145 amino acids, about 120 amino acids to about 140 amino acids, about 120 amino acids to about 135 amino acids, about 120 amino acids to about 130 amino acids, about 120 amino acids to about 125 amino acids, about 125 amino acids to about 1000 amino acids, about 125 Amino acids to about 950 amino acids, about 125 amino acids to about 900 amino acids, about 125 amino acids to about 850 amino acids, about 125 amino acids to about 800 amino acids, about 125 amino acids to about 750 amino acids, about 125 amino acids to about 700 amino acids, about 125 amino acids to about 650 amino acids, about 125 amino acids to about 600 amino acids, about 125 amino acids to about 550 amino acids, about 125 amino acids to about 500 amino acids, about 125 amino acids to about 450 amino acids, about 125 amino acids to about 400 amino acids, about 125 amino acids to about 350 amino acids, about 125 amino acids to about 300 amino acids,About 125 amino acids to about 280 amino acids, about 125 amino acids to about 260 amino acids, about 125 amino acids to about 240 amino acids, about 125 amino acids to about 220 amino acids, about 125 amino acids to about 200 amino acids, about 125 amino acids to about 195 amino acids, about 125 amino acids to about 190 amino acids, about 125 amino acids to about 185 amino acids, about 125 amino acids to about 180 amino acids, about 125 amino acids to about 175 amino acids, about 125 amino acids to about 170 amino acids, about 125 amino acids to about 165 amino acids, about 125 amino acids to about 160 amino acids, about 125 amino acids to about 155 amino acids amino acids, about 125 amino acids to about 150 amino acids, about 125 amino acids to about 145 amino acids, about 125 amino acids to about 140 amino acids, about 125 amino acids to about 135 amino acids, about 125 amino acids to about 130 amino acids, about 130 amino acids to about 1000 amino acids, about 130 amino acids to about 950 amino acids, about 130 amino acids to about 900 amino acids, about 130 amino acids to about 850 amino acids, about 130 amino acids to about 800 amino acids, about 130 amino acids to about 750 amino acids, about 130 amino acids to about 700 amino acids, about 130 amino acids to about 650 amino acids, about 130 amino acids to about 1000 amino acids About 600 amino acids, about 130 amino acids to about 550 amino acids, about 130 amino acids to about 500 amino acids, about 130 amino acids to about 450 amino acids, about 130 amino acids to about 400 amino acids, about 130 amino acids to about 350 amino acids, about 130 amino acids to about 300 amino acids, about 130 amino acids to about 280 amino acids, about 130 amino acids to about 260 amino acids, about 130 amino acids to about 240 amino acids, about 130 amino acids to about 220 amino acids, about 130 amino acids to about 200 amino acids, about 130 amino acids to about 195 amino acids, about 130 amino acids to about 190 amino acids, about 130 amino acids amino acids to about 185 amino acids, about 130 amino acids to about 180 amino acids, about 130 amino acids to about 175 amino acids, about 130 amino acids to about 170 amino acids, about 130 amino acids to about 165 amino acids, about 130 amino acids to about 160 amino acids, about 130 amino acids to about 155 amino acids, about 130 amino acids to about 150 amino acids, about 130 amino acids to about 145 amino acids, about 130 amino acids to about 140 amino acids, about 130 amino acids to about 135 amino acids, about 135 amino acids to about 1000 amino acids, about 135 amino acids to about 950 amino acids, about 135 amino acids to about 900 amino acids,About 135 amino acids to about 850 amino acids, about 135 amino acids to about 800 amino acids, about 135 amino acids to about 750 amino acids, about 135 amino acids to about 700 amino acids, about 135 amino acids to about 650 amino acids, about 135 amino acids to about 600 amino acids, about 135 amino acids to about 550 amino acids, about 135 amino acids to about 500 amino acids, about 135 amino acids to about 450 amino acids, about 135 amino acids, amino acids to about 400 amino acids, about 135 amino acids to about 350 amino acids, about 135 amino acids to about 300 amino acids, about 135 amino acids to about 280 amino acids, about 135 amino acids to about 260 amino acids, about 135 amino acids to about 240 amino acids, about 135 amino acids to about 220 amino acids, about 135 amino acids to about 200 amino acids, about 135 amino acids to about 195 amino acids, about 135 amino acids to about 190 amino acids, about 135 amino acids to about 185 amino acids, about 135 amino acids to about 180 amino acids, about 135 amino acids to about 175 amino acids, about 135 amino acids to about 170 amino acids , about 135 amino acids to about 165 amino acids, about 135 amino acids to about 160 amino acids, about 135 amino acids to about 155 amino acids, about 135 amino acids to about 150 amino acids, about 135 amino acids to about 145 amino acids, about 135 amino acids to about 140 amino acids, about 140 amino acids to about 1000 amino acids, about 140 amino acids to about 950 amino acids, about 140 amino acids to about 900 amino acids, about 140 amino acids to about 850 amino acids, about 140 amino acids to about 800 amino acids, about 140 amino acids to about 750 amino acids, about 140 amino acids to about 700 amino acids, about 140 amino acids to about 6 50 amino acids, about 140 amino acids to about 600 amino acids, about 140 amino acids to about 550 amino acids, about 140 amino acids to about 500 amino acids, about 140 amino acids to about 450 amino acids, about 140 amino acids to about 400 amino acids, about 140 amino acids to about 350 amino acids, about 140 amino acids to about 300 amino acids, about 140 amino acids to about 280 amino acids, about 140 amino acids to about 260 amino acids, about 140 amino acids to about 240 amino acids, about 140 amino acids to about 220 amino acids, about 140 amino acids to about 200 amino acids, about 140 amino acids to about 195 amino acids, about 140 amino acids to about 190 amino acids, about 140 amino acids to about 185 amino acids, about 140 amino acids to about 180 amino acids, about 140 amino acids to about 175 amino acids, about 140 amino acids to about 170 amino acids, about 140 amino acids to about 165 amino acids, about 140 amino acids to about 160 amino acids, about 140 amino acids to about 155 amino acids, about 140 amino acids to about 150 amino acids, about 140 amino acids to about 145 amino acids, about 145 amino acids to about 1000 amino acids, about 145 amino acids to about 950 amino acids, about 145 amino acids to about 900 amino acids, about 145 amino acids to about 850 amino acids,About 145 amino acids to about 800 amino acids, about 145 amino acids to about 750 amino acids, about 145 amino acids to about 700 amino acids, about 145 amino acids to about 650 amino acids, about 145 amino acids to about 600 amino acids, about 145 amino acids to about 550 amino acids, about 145 amino acids to about 500 amino acids, about 145 amino acids to about 450 amino acids, about 145 amino acids to about 400 amino acids, about 145 amino acids to about 350 amino acids, about 145 amino acids to about 300 amino acids, about 145 amino acids to about 280 amino acids, about 145 amino acids to about 260 amino acids, about 145 amino acids to about 240 Amino acids, about 145 to about 220 amino acids, about 145 to about 200 amino acids, about 145 to about 195 amino acids, about 145 to about 190 amino acids, about 145 to about 185 amino acids, about 145 to about 180 amino acids, about 145 to about 175 amino acids, about 145 to about 170 amino acids, about 145 to about 165 amino acids, about 145 to about 160 amino acids, about 145 to about 155 amino acids, about 145 to about 150 amino acids, about 150 to about 1000 amino acids, about 150 amino acids to about 950 amino acids, about 150 amino acids to about 900 amino acids, about 150 amino acids to about 850 amino acids, about 150 amino acids to about 800 amino acids, about 150 amino acids to about 750 amino acids, about 150 amino acids to about 700 amino acids, about 150 amino acids to about 650 amino acids, about 150 amino acids to about 600 amino acids, about 150 amino acids to about 550 amino acids, about 150 amino acids to about 500 amino acids, about 150 amino acids to about 450 amino acids, about 150 amino acids to about 400 amino acids, about 150 amino acids to about 350 amino acids, about 150 amino acids to about 300 amino acids, about 150 Amino acids to about 280 amino acids, about 150 amino acids to about 260 amino acids, about 150 amino acids to about 240 amino acids, about 150 amino acids to about 220 amino acids, about 150 amino acids to about 200 amino acids, about 150 amino acids to about 195 amino acids, about 150 amino acids to about 190 amino acids, about 150 amino acids to about 185 amino acids, about 150 amino acids to about 180 amino acids, about 150 amino acids to about 175 amino acids, about 150 amino acids to about 170 amino acids, about 150 amino acids to about 165 amino acids, about 150 amino acids to about 160 amino acids, about 150 amino acids to about 155 amino acids,About 155 amino acids to about 1000 amino acids, about 155 amino acids to about 950 amino acids, about 155 amino acids to about 900 amino acids, about 155 amino acids to about 850 amino acids, about 155 amino acids to about 800 amino acids, about 155 amino acids to about 750 amino acids, about 155 amino acids to about 700 amino acids, about 155 amino acids to about 650 amino acids, about 155 amino acids to about 600 amino acids, about 155 amino acids to about 550 amino acids, about 155 amino acids to about 500 amino acids, about 155 amino acids to about 450 amino acids, about 155 amino acids to about 400 amino acids, about 155 amino acids to about 350 Amino acids, about 155 amino acids to about 300 amino acids, about 155 amino acids to about 280 amino acids, about 155 amino acids to about 260 amino acids, about 155 amino acids to about 240 amino acids, about 155 amino acids to about 220 amino acids, about 155 amino acids to about 200 amino acids, about 155 amino acids to about 195 amino acids, about 155 amino acids to about 190 amino acids, about 155 amino acids to about 185 amino acids, about 155 amino acids to about 180 amino acids, about 155 amino acids to about 175 amino acids, about 155 amino acids to about 170 amino acids, about 155 amino acids to about 165 ...65 amino acids, about 155 amino acids to about 195 amino acids, about 155 amino acids to about 190 amino acids, about 155 amino acids to about 185 amino acids, about 155 amino acids to about 1 About 160 amino acids, about 160 amino acids to about 1000 amino acids, about 160 amino acids to about 950 amino acids, about 160 amino acids to about 900 amino acids, about 160 amino acids to about 850 amino acids, about 160 amino acids to about 800 amino acids, about 160 amino acids to about 750 amino acids, about 160 amino acids to about 700 amino acids, about 160 amino acids to about 650 amino acids, about 160 amino acids to about 600 amino acids, about 160 amino acids to about 550 amino acids, about 160 amino acids to about 500 amino acids, about 160 amino acids to about 450 amino acids, about 160 amino acids to about 400 amino acids, about 160 Amino acids up to about 350 amino acids, about 160 amino acids to about 300 amino acids, about 160 amino acids to about 280 amino acids, about 160 amino acids to about 260 amino acids, about 160 amino acids to about 240 amino acids, about 160 amino acids to about 220 amino acids, about 160 amino acids to about 200 amino acids, about 160 amino acids to about 195 amino acids, about 160 amino acids to about 190 amino acids, about 160 amino acids to about 185 amino acids, about 160 amino acids to about 180 amino acids, about 160 amino acids to about 175 amino acids, about 160 amino acids to about 170 amino acids, about 160 amino acids to about 165 amino acids,About 165 amino acids to about 1000 amino acids, about 165 amino acids to about 950 amino acids, about 165 amino acids to about 900 amino acids, about 165 amino acids to about 850 amino acids, about 165 amino acids to about 800 amino acids, about 165 amino acids to about 750 amino acids, about 165 amino acids to about 700 amino acids, about 165 amino acids to about 650 amino acids, about 165 amino acids to about 600 amino acids, about 165 amino acids to about 550 amino acids, about 165 amino acids to about 500 amino acids, about 165 amino acids to about 450 amino acids, about 165 amino acids to about 400 amino acids, about 165 amino acids to about 350 Amino acids, about 165 amino acids to about 300 amino acids, about 165 amino acids to about 280 amino acids, about 165 amino acids to about 260 amino acids, about 165 amino acids to about 240 amino acids, about 165 amino acids to about 220 amino acids, about 165 amino acids to about 200 amino acids, about 165 amino acids to about 195 amino acids, about 165 amino acids to about 190 amino acids, about 165 amino acids to about 185 amino acids, about 165 amino acids to about 180 amino acids, about 165 amino acids to about 175 amino acids, about 165 amino acids to about 170 amino acids, about 170 amino acids to about 1000 amino acids, about 170 amino acids to about 950 amino acids, about 170 amino acids to about 900 amino acids, about 170 amino acids to about 850 amino acids, about 170 amino acids to about 800 amino acids, about 170 amino acids to about 750 amino acids, about 170 amino acids to about 700 amino acids, about 170 amino acids to about 650 amino acids, about 170 amino acids to about 600 amino acids, about 170 amino acids to about 550 amino acids, about 170 amino acids to about 500 amino acids, about 170 amino acids to about 450 amino acids, about 170 amino acids to about 400 amino acids, about 170 amino acids to about 350 amino acids, about 170 amino acids to about 300 amino acids, about 170 Amino acids to about 280 amino acids, about 170 amino acids to about 260 amino acids, about 170 amino acids to about 240 amino acids, about 170 amino acids to about 220 amino acids, about 170 amino acids to about 200 amino acids, about 170 amino acids to about 195 amino acids, about 170 amino acids to about 190 amino acids, about 170 amino acids to about 185 amino acids, about 170 amino acids to about 180 amino acids, about 170 amino acids to about 175 amino acids, about 175 amino acids to about 1000 amino acids, about 175 amino acids to about 950 amino acids, about 175 amino acids to about 900 amino acids, about 175 amino acids to about 850 amino acids,About 175 amino acids to about 800 amino acids, about 175 amino acids to about 750 amino acids, about 175 amino acids to about 700 amino acids, about 175 amino acids to about 650 amino acids, about 175 amino acids to about 600 amino acids, about 175 amino acids to about 550 amino acids, about 175 amino acids to about 500 amino acids, about 175 amino acids to about 450 amino acids, about 175 amino acids to about 400 amino acids, about 175 amino acids to about 350 amino acids, about 175 amino acids to about 300 amino acids, about 175 amino acids to about 280 amino acids, about 175 amino acids to about 260 amino acids, about 175 amino acids to about 240 amino acids amino acids, about 175 amino acids to about 220 amino acids, about 175 amino acids to about 200 amino acids, about 175 amino acids to about 195 amino acids, about 175 amino acids to about 190 amino acids, about 175 amino acids to about 185 amino acids, about 175 amino acids to about 180 amino acids, about 180 amino acids to about 1000 amino acids, about 180 amino acids to about 950 amino acids, about 180 amino acids to about 900 amino acids, about 180 amino acids to about 850 amino acids, about 180 amino acids to about 800 amino acids, about 180 amino acids to about 750 amino acids, about 180 amino acids to about 700 amino acids, about 180 amino acids to about 1000 amino acids About 650 amino acids, about 180 amino acids to about 600 amino acids, about 180 amino acids to about 550 amino acids, about 180 amino acids to about 500 amino acids, about 180 amino acids to about 450 amino acids, about 180 amino acids to about 400 amino acids, about 180 amino acids to about 350 amino acids, about 180 amino acids to about 300 amino acids, about 180 amino acids to about 280 amino acids, about 180 amino acids to about 260 amino acids, about 180 amino acids to about 240 amino acids, about 180 amino acids to about 220 amino acids, about 180 amino acids to about 200 amino acids, about 180 amino acids to about 195 amino acids, about 180 amino acids amino acids to about 190 amino acids, about 180 amino acids to about 185 amino acids, about 185 amino acids to about 1000 amino acids, about 185 amino acids to about 950 amino acids, about 185 amino acids to about 900 amino acids, about 185 amino acids to about 850 amino acids, about 185 amino acids to about 800 amino acids, about 185 amino acids to about 750 amino acids, about 185 amino acids to about 700 amino acids, about 185 amino acids to about 650 amino acids, about 185 amino acids to about 600 amino acids, about 185 amino acids to about 550 amino acids, about 185 amino acids to about 500 amino acids, about 185 amino acids to about 450 amino acids,About 185 amino acids to about 400 amino acids, about 185 amino acids to about 350 amino acids, about 185 amino acids to about 300 amino acids, about 185 amino acids to about 280 amino acids, about 185 amino acids to about 260 amino acids, about 185 amino acids to about 240 amino acids, about 185 amino acids to about 220 amino acids, about 185 amino acids to about 200 amino acids, about 185 amino acids to about 195 amino acids, about 18, 5 amino acids to about 190 amino acids, about 190 amino acids to about 1000 amino acids, about 190 amino acids to about 950 amino acids, about 190 amino acids to about 900 amino acids, about 190 amino acids to about 850 amino acids, about 190 amino acids to about 800 amino acids, about 190 amino acids to about 750 amino acids, about 190 amino acids to about 700 amino acids, about 190 amino acids to about 650 amino acids, about 190 amino acids to about 600 amino acids, about 190 amino acids to about 550 amino acids, about 190 amino acids to about 500 amino acids, about 190 amino acids to about 450 amino acids, about 190 amino acids to about 400 amino acids amino acids, about 190 amino acids to about 350 amino acids, about 190 amino acids to about 300 amino acids, about 190 amino acids to about 280 amino acids, about 190 amino acids to about 260 amino acids, about 190 amino acids to about 240 amino acids, about 190 amino acids to about 220 amino acids, about 190 amino acids to about 200 amino acids, about 190 amino acids to about 195 amino acids, about 195 amino acids to about 1000 amino acids, about 195 amino acids to about 950 amino acids, about 195 amino acids to about 900 amino acids, about 195 amino acids to about 850 amino acids, about 195 amino acids to about 800 amino acids, about 195 amino acids to about 750 amino acids, about 195 amino acids to about 700 amino acids, about 195 amino acids to about 650 amino acids, about 195 amino acids to about 600 amino acids, about 195 amino acids to about 550 amino acids, about 195 amino acids to about 500 amino acids, about 195 amino acids to about 450 amino acids, about 195 amino acids to about 400 amino acids, about 195 amino acids to about 350 amino acids, about 195 amino acids to about 300 amino acids, about 195 amino acids to about 280 amino acids, about 195 amino acids to about 260 amino acids, about 195 amino acids to about 240 amino acids, about 195 amino acids to about 2 ... amino acids to about 200 amino acids, about 200 amino acids to about 1000 amino acids, about 200 amino acids to about 950 amino acids, about 200 amino acids to about 900 amino acids, about 200 amino acids to about 850 amino acids, about 200 amino acids to about 800 amino acids, about 200 amino acids to about 750 amino acids, about 200 amino acids to about 700 amino acids, about 200 amino acids to about 650 amino acids, about 200 amino acids to about 600 amino acids, about 200 amino acids to about 550 amino acids, about 200 amino acids to about 500 amino acids, about 200 amino acids to about 450 amino acids, about 200 amino acids to about 400 amino acids,About 200 amino acids to about 350 amino acids, about 200 amino acids to about 300 amino acids, about 200 amino acids to about 280 amino acids, about 200 amino acids to about 260 amino acids, about 200 amino acids to about 240 amino acids, about 200 amino acids to about 220 amino acids, about 220 amino acids to about 1000 amino acids, about 220 amino acids to about 950 amino acids, about 220 amino acids to about 900 amino acids, about 220 amino acids to about 850 amino acids, about 220 amino acids to about 800 amino acids, about 220 amino acids to about 750 amino acids, about 220 amino acids to about 700 amino acids, about 220 amino acids to about 650 Amino acids, about 220 amino acids to about 600 amino acids, about 220 amino acids to about 550 amino acids, about 220 amino acids to about 500 amino acids, about 220 amino acids to about 450 amino acids, about 220 amino acids to about 400 amino acids, about 220 amino acids to about 350 amino acids, about 220 amino acids to about 300 amino acids, about 220 amino acids to about 280 amino acids, about 220 amino acids to about 260 amino acids, about 220 amino acids to about 240 amino acids, about 240 amino acids to about 1000 amino acids, about 240 amino acids to about 950 amino acids, about 240 amino acids to about 900 amino acids, about 240 amino acids about 850 amino acids, about 240 amino acids to about 800 amino acids, about 240 amino acids to about 750 amino acids, about 240 amino acids to about 700 amino acids, about 240 amino acids to about 650 amino acids, about 240 amino acids to about 600 amino acids, about 240 amino acids to about 550 amino acids, about 240 amino acids to about 500 amino acids, about 240 amino acids to about 450 amino acids, about 240 amino acids to about 400 amino acids, about 240 amino acids to about 350 amino acids, about 240 amino acids to about 300 amino acids, about 240 amino acids to about 280 amino acids, about 240 amino acids to about 260 amino acids, about 260 Amino acids to about 1000 amino acids, about 260 amino acids to about 950 amino acids, about 260 amino acids to about 900 amino acids, about 260 amino acids to about 850 amino acids, about 260 amino acids to about 800 amino acids, about 260 amino acids to about 750 amino acids, about 260 amino acids to about 700 amino acids, about 260 amino acids to about 650 amino acids, about 260 amino acids to about 600 amino acids, about 260 amino acids to about 550 amino acids, about 260 amino acids to about 500 amino acids, about 260 amino acids to about 450 amino acids, about 260 amino acids to about 400 amino acids, about 260 amino acids to about 350 amino acids,About 260 amino acids to about 300 amino acids, about 260 amino acids to about 280 amino acids, about 280 amino acids to about 1000 amino acids, about 280 amino acids to about 950 amino acids, about 280 amino acids to about 900 amino acids, about 280 amino acids to about 850 amino acids, about 280 amino acids to about 800 amino acids, about 280 amino acids to about 750 amino acids, about 280 amino acids to about 700 amino acids, about 280 amino acids to about 650 amino acids, about 280 amino acids to about 600 amino acids, about 280 amino acids to about 550 amino acids, about 280 amino acids to about 500 amino acids, about 280 amino acids to about 450 Amino acids, about 280 amino acids to about 400 amino acids, about 280 amino acids to about 350 amino acids, about 280 amino acids to about 300 amino acids, about 300 amino acids to about 1000 amino acids, about 300 amino acids to about 950 amino acids, about 300 amino acids to about 900 amino acids, about 300 amino acids to about 850 amino acids, about 300 amino acids to about 800 amino acids, about 300 amino acids to about 750 amino acids, about 300 amino acids to about 700 amino acids, about 300 amino acids to about 650 amino acids, about 300 amino acids to about 600 amino acids, about 300 amino acids to about 550 amino acids, about 300 amino acids to about About 500 amino acids, about 300 to about 450 amino acids, about 300 to about 400 amino acids, about 300 to about 350 amino acids, about 350 to about 1000 amino acids, about 350 to about 950 amino acids, about 350 to about 900 amino acids, about 350 to about 850 amino acids, about 350 to about 800 amino acids, about 350 to about 750 amino acids, about 350 to about 700 amino acids, about 350 to about 650 amino acids, about 350 to about 600 amino acids, about 350 to about 550 amino acids, about 350 Amino acids to about 500 amino acids, about 350 amino acids to about 450 amino acids, about 350 amino acids ...

Claims

1. A protein comprising an anti-CD26 antigen-binding domain, the anti-CD26 antigen-binding domain a heavy chain variable domain comprising a CDR1 comprising SEQ ID NO: 1, a CDR2 comprising SEQ ID NO: 2, and a CDR3 comprising SEQ ID NO: 3; and a light chain variable domain comprising a CDR1 comprising SEQ ID NO:4, a CDR2 comprising SEQ ID NO:5, and a CDR3 comprising SEQ ID NO:

6. The protein comprising:

2. 2. The protein of claim 1, wherein the heavy chain variable domain comprises a sequence that is at least 80% identical to SEQ ID NO: 61 and the light chain variable domain comprises a sequence that is at least 80% identical to SEQ ID NO:

62.

3. 3. The protein of claim 1 or 2, which is a multi-chain protein.

4. 3. The protein of claim 1 or 2, which is a single-chain protein.

5. 3. The protein of claim 1 or 2, which is an antibody or an antigen-binding antibody fragment.

6. The protein of claim 1 or 2, which is a chimeric antigen receptor (CAR).

7. A pharmaceutical composition comprising the protein of any one of claims 1 to 6 and a pharmaceutically acceptable carrier.

8. A kit comprising the pharmaceutical composition of claim 7.

9. A nucleic acid encoding the protein according to any one of claims 1 to 6.

10. A vector comprising the nucleic acid of claim 9.

11. A cell comprising the nucleic acid of claim 9 or the vector of claim 10.

12. The cell of claim 11, which is an immune cell.

13. The cell of claim 12 , wherein the immune cell is a T cell, a B cell, or a natural killer (NK) cell.

14. A pharmaceutical composition comprising the cells of any one of claims 11 to 13 and a pharmaceutically acceptable carrier.

15. A pharmaceutical composition for treating an age-related disease or an inflammatory disease in a subject, comprising the protein according to any one of claims 1 to 6 or the pharmaceutical composition according to claim 7.

16. A pharmaceutical composition for treating an age-related or inflammatory disease in a subject, comprising the nucleic acid of claim 9 or the vector of claim 10.

17. A pharmaceutical composition for treating an age-related or inflammatory disease in a subject, comprising the cell of any one of claims 11 to 13 or the pharmaceutical composition of claim 14.

18. (i) NK cell activators and / or NK cells and / or monoclonal antibodies, and / or (ii) a Treg cell activator and / or a Treg cell and / or a monoclonal antibody and / or an advanced glycation end product (AGE) inhibitor The pharmaceutical composition of any one of claims 15 to 17, further comprising:

19. 19. The pharmaceutical composition of claim 18, comprising NK cells.

20. A pharmaceutical composition for treating cancer in a subject, comprising the protein according to any one of claims 1 to 6 or the pharmaceutical composition according to claim 7.

21. A pharmaceutical composition for treating an infectious disease in a subject, comprising a protein according to any one of claims 1 to 6 or a pharmaceutical composition according to claim 7.

Images