Single-chain chimeric polypeptides and uses thereof

Abstract

Provided herein are single-chain chimeric polypeptides comprising: (i) a first target binding domain; (ii) a soluble tissue factor domain; and (iii) a second target binding domain. Also provided herein are methods of using these single-chain chimeric polypeptides and nucleic acids encoding these single-chain chimeric polypeptides.

Description

[0001] Cross-references to related applications

[0002] This application claims priority to the following U.S. patent applications: U.S. Patent Application Serial No. 62 / 725,038, filed August 30, 2018; U.S. Patent Application Serial No. 62 / 817,244, filed March 12, 2019; U.S. Patent Application Serial No. 62 / 746,832, filed October 17, 2018; U.S. Patent Application Serial No. 62 / 749,506, filed October 23, 2018; U.S. Patent Application Serial No. 62 / 817,241, filed March 12, 2019; U.S. Patent Application Serial No. 62 / 816,683, filed March 11, 2019; and U.S. Patent Application Serial No. 62 / 881,039, filed July 31, 2019; each of which is incorporated herein by reference in its entirety. Technical Field

[0003] This disclosure relates to the field of biotechnology, and more specifically, to antigen-binding molecules. Background Technology

[0004] Tissue factor (TF) is a monolithic membrane glycoprotein of 263 amino acids with a molecular weight of approximately 46 kDa. It is a trigger protein in the extrinsic coagulation pathway and a major initiator of coagulation in vivo. Tissue factor, which normally does not come into contact with circulating blood, triggers a coagulation cascade upon exposure to circulating coagulation serine protease factors. Vascular injury exposes subendothelial cells expressing TF, leading to the formation of a calcium-dependent high-affinity complex with pre-existing plasma factor VIIa (FVIIa). The binding of serine protease FVIIa to TF promotes the rapid cleavage of FX to FXa and FIX to FIXa. The resulting FXa and the proteolytic activity on the active membrane surface are thus unable to effectively convert small amounts of prothrombin into thrombin. The thrombin generated from FIXa triggers platelet activation and activates trace amounts of proto-cofactors: factor V (FV) and factor VIII (FVIII) into active cofactors: factor Va (FVa) and factor VIIIa (FVIIIa). FIXa and FVIIIa complex on the platelet surface to form an intrinsic liquefying enzyme (tenase) complex, thereby causing rapid production of FXa. FXa complexes with FVa to form a prothrombinase complex on the activated platelet surface, which causes rapid cleavage of prothrombin into thrombin.

[0005] In addition to the tissue factor-FVIIa complex, recent studies have shown that the tissue factor-FVIIa-FXa complex can also activate FVIII, thereby providing additional levels of FVIIIa in the initial phase. The extrinsic pathway primarily involves initiating coagulation through the activation of a limited amount of thrombin, while the intrinsic pathway maintains coagulation by significantly amplifying the initial signal.

[0006] Many tissue factors expressed on the cell surface are "encrypted" and must be "decrypted" to fully participate in coagulation. Currently, the mechanism of this "decryption" is unclear, but the exposure of anionic phospholipids plays a crucial role. Healthy cells actively isolate anionic phospholipids (such as phosphatidylserine (PS)) to the inner lobules of the plasma membrane. In cases of cell damage, activation, or cytoplasmic calcium... 2+ Upon increasing levels, this bilayer asymmetry disappears, leading to increased PS exposure on the outer leaflets, which in turn increases the specific activity of the cell surface tissue factor-FVIIa complex. PS exposure is known to reduce the apparent Km of tissue factor-FVIIa complex activation of FIX and FX, but other mechanisms may include conformational rearrangement of tissue factor or tissue factor-FVIIa and subsequent exposure to substrate binding sites. Summary of the Invention

[0007] This invention is based on the discovery that soluble tissue factor can be used as a scaffold for chimeric peptides including an antigen-binding domain. Based on this discovery, this document provides single-chain chimeric peptides comprising: (i) a first target-binding domain; (ii) a soluble tissue factor domain; and (iii) a second target-binding domain. This document also provides compositions comprising any of the single-chain chimeric peptides described herein, a nucleic acid encoding any of the single-chain chimeric peptides described herein, and cells comprising any of the nucleic acids encoding any of the single-chain chimeric peptides described herein. This document also provides methods for stimulating immune cells and methods for treating subjects in need (including using any of the single-chain chimeric peptides described herein). This document also provides methods for generating any of the single-chain chimeric peptides described herein. Other uses of any of the single-chain chimeric peptides are described herein.

[0008] Therefore, this article provides a single-chain chimeric polypeptide comprising:

[0009] (i) First target binding domain;

[0010] (ii) Soluble tissue factor domain; and

[0011] (iii) A second target-binding domain. In some embodiments, the first target-binding domain and the soluble tissue factor domain are directly adjacent to each other. In some embodiments, the single-chain chimeric polypeptide further includes a linker sequence between the first target-binding domain and the soluble tissue factor domain. In some embodiments, the soluble tissue factor domain and the second target-binding domain are directly adjacent to each other. In some embodiments, the single-chain chimeric polypeptide further includes a linker sequence between the soluble tissue factor domain and the second target-binding domain. In some embodiments, the first target-binding domain and the second target-binding domain are directly adjacent to each other. In some embodiments, the single-chain chimeric polypeptide further includes a linker sequence between the first target-binding domain and the second target-binding domain. In some embodiments, the second target-binding domain and the soluble tissue factor domain are directly adjacent to each other. In some embodiments, the single-chain chimeric polypeptide further includes a linker sequence between the second target-binding domain and the soluble tissue factor domain. In some embodiments, the first target-binding domain and the second target-binding domain specifically bind to the same antigen. In some embodiments, the first target-binding domain and the second target-binding domain specifically bind to the same epitope. In some embodiments, the first target-binding domain and the second target-binding domain contain the same amino acid sequence. In some embodiments, the first target-binding domain and the second target-binding domain specifically bind to different antigens. In some embodiments, one or both of the first target-binding domain and the second target-binding domain are antigen-binding domains. In some embodiments, the first target-binding domain and the second target-binding domain are each an antigen-binding domain. In some embodiments, the antigen-binding domain comprises scFv or a single-domain antibody.In some embodiments, one or both of the first and second target-binding domains bind to targets selected from the group consisting of: CD16a, CD28, CD3, 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, TNFα, CD26a, CD36, ULBP2, CD30, CD200, IGF-1R, MUC4AC, MUC5AC, Trop-2, CMET, EGFR, HER1, HER2, HER3, PSMA, CEA, B7H3, EPCAM, BCMA, P-cadherin, CEACAM5, aUL16 binding protein, HLA-DR, DLL4, TYRO3, AXL, MER Ligands of CD122, CD155, PDGF-DD, TGF-β receptor II (TGF-βRII), TGF-βRIII, DNAM-1, NKp46, NKp44, NKG2D, NKp30, scMHCI, scMHCII, scTCR, IL-1 receptor, IL-2 receptor, IL-3 receptor, and IL-7 receptor. Receptors for IL-8, IL-10, IL-12, IL-15, IL-17, IL-18, IL-21, PDGF-DD, stem cell factor (SCF), stem cell-like tyrosine kinase 3 ligand (FLT3L), MICA, MICB, ULP16 binding protein, CD155, CD122, and CD28. In some embodiments, one or both of the first and second target-binding domains are soluble interleukins, soluble cytokine proteins, or soluble cell surface proteins. In some embodiments, the soluble interleukin, soluble cytokine protein, or soluble cell surface protein is selected from the group consisting of: IL-1, IL-2, IL-3, IL-7, IL-8, IL-10, IL-12, IL-15, IL-17, IL-18, IL-21, PDGF-DD, SCF, FLT3L, MICA, MICB, and ULP16 binding proteins. In some embodiments, one or both of the first target-binding domain and the second target-binding domain are soluble interleukin receptors, soluble cytokine receptors, or soluble cell surface receptors.In some embodiments, the soluble interleukin receptor, soluble cytokine receptor, or soluble cell surface receptor is a soluble TGF-β receptor II (TGF-βRII), a soluble TGF-βRIII, a soluble NKG2D, a soluble NKp30, a soluble NKp44, a soluble NKp46, a soluble DNAM-1, scMHCI, scMHCII, scTCR, a soluble CD155, or a soluble CD28. In some embodiments, the soluble tissue factor domain is a human soluble tissue factor domain. In some embodiments, the human soluble tissue factor domain contains a sequence that is at least 80% identical to SEQ ID NO:9. In some embodiments, the human soluble tissue factor domain contains a sequence that is at least 90% identical to SEQ ID NO:9. In some embodiments, the human soluble tissue factor domain contains a sequence that is at least 95% identical to SEQ ID NO:9. In some embodiments, the single-chain chimeric polypeptide of the human soluble tissue factor domain does not contain one or more of the following: lysine at the amino acid position corresponding to amino acid position 20 of the human mature wild-type tissue factor protein; isoleucine at the amino acid position corresponding to amino acid position 22 of the human mature wild-type tissue factor protein; tryptophan at the amino acid position corresponding to amino acid position 45 of the human mature wild-type tissue factor protein; aspartic acid at the amino acid position corresponding to amino acid position 58 of the human mature wild-type tissue factor protein; tyrosine at the amino acid position corresponding to amino acid position 94 of the human mature wild-type tissue factor protein; arginine at the amino acid position corresponding to amino acid position 135 of the human mature wild-type tissue factor protein; and phenylalanine at the amino acid position corresponding to amino acid position 140 of the human mature wild-type tissue factor protein. In some embodiments, the human soluble tissue factor domain does not contain any of the following: lysine at the amino acid position corresponding to amino acid position 20 of the human mature wild-type tissue factor protein; isoleucine at the amino acid position corresponding to amino acid position 22 of the human mature wild-type tissue factor protein; tryptophan at the amino acid position corresponding to amino acid position 45 of the human mature wild-type tissue factor protein; aspartic acid at the amino acid position corresponding to amino acid position 58 of the human mature wild-type tissue factor protein; tyrosine at the amino acid position corresponding to amino acid position 94 of the human mature wild-type tissue factor protein; arginine at the amino acid position corresponding to amino acid position 135 of the human mature wild-type tissue factor protein; and phenylalanine at the amino acid position corresponding to amino acid position 140 of the human mature wild-type tissue factor protein. In some embodiments, the soluble tissue factor domain cannot bind factor VIIa. In some embodiments, the soluble tissue factor domain does not convert inactive factor X into factor Xa. In some embodiments, the single-chain chimeric polypeptide does not stimulate coagulation in mammals.In some embodiments, the single-chain chimeric polypeptide further includes one or more other target-binding domains at its N-terminus and / or C-terminus. In some embodiments, the single-chain chimeric polypeptide includes one or more other target-binding domains at its N-terminus. In some embodiments, the one or more other target-binding domains are directly adjacent to the first target-binding domain, the second target-binding domain, or the soluble tissue factor domain. In some embodiments, the single-chain chimeric polypeptide further includes a linker sequence between at least one of the other target-binding domains and the first target-binding domain, the second target-binding domain, or the soluble tissue factor domain. In some embodiments, the single-chain chimeric polypeptide includes one or more other target-binding domains at its C-terminus. In some embodiments, one or more other target-binding domains are directly adjacent to the first target-binding domain, the second target-binding domain, or the soluble tissue factor domain. In some embodiments, the single-chain chimeric polypeptide further includes a linker sequence between at least one of the other target-binding domains and the first target-binding domain, the second target-binding domain, or the soluble tissue factor domain. In some embodiments, the single-chain chimeric polypeptide includes one or more other target-binding domains at its N-terminus and C-terminus. In some embodiments, one of one or more other antigen-binding domains located at the N-terminus is directly adjacent to the first target-binding domain, the second target-binding domain, or the soluble tissue factor domain. In some embodiments, the single-chain chimeric polypeptide further includes a linker sequence between one of the one or more other antigen-binding domains at the N-terminus and the first target-binding domain, the second target-binding domain, or the soluble tissue factor domain. In some embodiments, one of one or more other antigen-binding domains at the C-terminus is directly adjacent to the first target-binding domain, the second target-binding domain, or the soluble tissue factor domain. In some embodiments, the single-chain chimeric polypeptide further includes a linker sequence between one of one or more other antigen-binding domains at the C-terminus and the first target-binding domain, the second target-binding domain, or the soluble tissue factor domain. In some embodiments, two or more of the first target-binding domain, the second target-binding domain, and one or more other target-binding domains specifically bind to the same antigen. In some embodiments, two or more of the first target-binding domain, the second target-binding domain, and one or more other target-binding domains specifically bind to the same epitope. In some embodiments, two or more of the first target-binding domain, the second target-binding domain, and one or more other target-binding domains comprise the same amino acid sequence. In some embodiments, the first target-binding domain, the second target-binding domain, and one or more other target-binding domains each specifically bind to the same antigen. In some embodiments, the first target-binding domain, the second target-binding domain, and one or more other target-binding domains each specifically bind to the same epitope. In some embodiments, the first target-binding domain, the second target-binding domain, and one or more other target-binding domains each comprise the same amino acid sequence. In some embodiments, the first target-binding domain, the second target-binding domain, and one or more other target-binding domains specifically bind to different antigens.In some embodiments, one or more of the first target-binding domain, the second target-binding domain, and one or more other target-binding domains are antigen-binding domains. In some embodiments, the first target-binding domain, the second target-binding domain, and one or more other target-binding domains are each an antigen-binding domain. In some embodiments, the antigen-binding domain comprises scFv or a single-domain antibody. In some embodiments, one or more of the first target-binding domain, the second target-binding domain, and one or more target-binding domains specifically bind to a target selected from the group consisting of: CD16a, CD28, CD3, 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, TNFα, CD26a, CD36, ULBP2, CD30, CD200, IGF-1R, MUC4AC, MUC5AC, Trop-2, CMET, EGFR, HER1, HER2, HER3, PSMA, CEA, B7H3, EPCAM, BCMA, P-cadherin, CEACAM5, UL16-binding protein, HLA-DR, DLL4, TYRO3. AXL, MER, CD122, CD155, PDGF-DD, ligands of TGF-β receptor II (TGF-βRII), ligands of TGF-βRIII, ligands of DNAM-1, ligands of NKp46, ligands of NKp44, ligands of NKG2D, ligands of NKp30, ligands of scMHCII, ligands of scTCR, receptors of IL-1, receptors of IL-2, receptors of IL-3, and IL-7. The receptors for IL-8, IL-10, IL-12, IL-15, IL-17, IL-18, IL-21, PDGF-D, stem cell factor (SCF), stem cell-like tyrosine kinase 3 ligand (FLT3L), MICA, MICB, ULP16 binding protein, CD155, CD122, and CD28 are included. In some embodiments, one or more of the first target-binding domain, the second target-binding domain, and one or more other target-binding domains are soluble interleukins or cytokine proteins. In some embodiments, the soluble interleukin or cytokine protein is selected from the group consisting of: IL-1, IL-2, IL-3, IL-7, IL-8, IL-10, IL-12, IL-15, IL-17, IL-18, IL-21, PDGF-DD, SCF, FLT3L, MICA, MICB, and ULP16 binding protein.In some embodiments, one or more of the first target-binding domain, the second target-binding domain, and one or more other target-binding domains are soluble interleukin or cytokine receptors. In some embodiments, the soluble receptor is a soluble TGF-β receptor II (TGF-βRII), a soluble TGF-βRIII, a soluble NKG2D, a soluble NKp30, a soluble NKp44, a soluble NKp46, a soluble DNAM-1, scMHCI, scMHCII, scTCR, a soluble CD155, a soluble CD122, a soluble CD3, or a soluble CD28. In some embodiments, the single-chain chimeric polypeptide further comprises a signal sequence located at its N-terminus. In some embodiments, the single-chain chimeric polypeptide further comprises a peptide tag located at the N-terminus or C-terminus of the single-chain chimeric polypeptide.

[0012] This document also provides compositions comprising any of the single-chain chimeric peptides described above. In some embodiments, the compositions are pharmaceutical compositions. In some embodiments, the single-chain chimeric peptide is contained in at least one dose of the composition within a kit.

[0013] This article also provides a method for stimulating immune cells, the method comprising: contacting the immune cells with an effective amount of any of the above-described single-chain chimeric polypeptides or compositions. In some embodiments, the method comprises in vitro immunization. In some embodiments, the method comprises obtaining immune cells from a subject. In some embodiments, the method further comprises obtaining immune cells from a subject prior to the contact step. In some embodiments, the method comprises in vivo immunization. In some embodiments, the immune cells are selected from the group consisting of: immature thymocytes, peripheral blood lymphocytes, naive T cells, pluripotent Th cell precursors, lymphoprogenitor cells, Treg cells, memory T cells, Th17 cells, Th22 cells, Th9 cells, Th2 cells, Th1 cells, Th3 cells, γδT cells, αβT cells, tumor-infiltrating T cells, CD8+ cells, etc. + T cells, CD4 +T cells, natural killer T cells, mast cells, macrophages, neutrophils, dendritic cells, basophils, eosinophils, and natural killer cells. In some embodiments, the immune cells have been previously genetically modified to express chimeric antigen receptors or recombinant T cell receptors. In some embodiments, the method further includes introducing nucleic acids encoding chimeric antigen receptors or recombinant T cell receptors into the immune cells after the contact step. In some embodiments, the method further includes administering the immune cells to a subject in need. In some embodiments, the method includes identifying or diagnosing whether the subject has an age-related disease or condition. In some embodiments, age-related diseases or symptoms are selected from the group consisting of: Alzheimer's disease, aneurysm, cystic fibrosis, pancreatic fibrosis, glaucoma, hypertension, idiopathic pulmonary fibrosis, inflammatory bowel disease, intervertebral disc degeneration, macular degeneration, osteoarthritis, type 2 diabetes, lipomatosis, lipid dystrophy, atherosclerosis, cataracts, COPD, idiopathic pulmonary fibrosis, failed kidney transplant, liver fibrosis, bone loss, myocardial infarction, sarcopenia, wound healing, alopecia, cardiomyocyte hypertrophy, osteoarthritis, Parkinson's disease, age-related loss of lung elasticity, macular degeneration, cachexia, glomerulosclerosis, cirrhosis, NAFLD, osteoporosis, amyotrophic lateral sclerosis, Huntington's disease, spinocerebellar ataxia, multiple sclerosis, and renal insufficiency. In some embodiments, the method includes identifying or diagnosing whether the subject has cancer. In some embodiments, the cancer is selected from the group consisting of: solid tumors, hematologic malignancies, sarcomas, osteosarcomas, glioblastoma, neuroblastoma, melanoma, rhabdomyosarcoma, Ewing 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 lymphoblastic leukemia (ALL), myelodysplastic syndrome (MDS), cutaneous T-cell lymphoma, retinoblastoma, gastric cancer, urothelial carcinoma, lung cancer, renal cell carcinoma, gastric and esophageal cancer, pancreatic cancer, prostate cancer, breast cancer, colorectal cancer, ovarian cancer, non-small cell lung cancer, head and neck squamous cell carcinoma, endometrial cancer, cervical cancer, liver cancer, and hepatocellular carcinoma. In some embodiments, the method includes diagnosing or identifying whether the subject has an infectious disease.In some embodiments, the infectious disease is infection with human immunodeficiency virus, cytomegalovirus, adenovirus, coronavirus, rhinovirus, rotavirus, smallpox, herpes simplex virus, hepatitis B virus, hepatitis A virus and hepatitis C virus, papillomavirus and influenza virus.

[0014] This document also provides a method for inducing or increasing the proliferation of immune cells, the method comprising: contacting immune cells with an effective amount of any of the single-chain chimeric polypeptides or compositions discussed herein. In some embodiments, the method comprises in vitro contact with immune cells. In some embodiments, the method comprises obtaining immune cells from a subject. In some embodiments, the method further comprises obtaining immune cells from a subject prior to the contact step. In some embodiments, the method comprises in vivo contact with immune cells. In some embodiments, the immune cells are selected from the group consisting of: immature thymocytes, peripheral blood lymphocytes, naive T cells, pluripotent Th cell precursors, lymphoprogenitor cells, Treg cells, memory T cells, Th17 cells, Th22 cells, Th9 cells, Th2 cells, Th1 cells, Th3 cells, γδ T cells, αβ T cells, tumor-infiltrating T cells, CD8+ cells, etc. + T cells, CD4 +T cells, natural killer T cells, mast cells, macrophages, neutrophils, dendritic cells, basophils, eosinophils, and natural killer cells. In some embodiments, the immune cells have been previously genetically modified to express a chimeric antigen receptor or a recombinant T cell receptor. In some embodiments, the method further includes introducing a nucleic acid encoding a chimeric antigen receptor or a recombinant T cell receptor into the immune cells after the contact step. In some embodiments, the method further includes administering the immune cells to a subject in need. In some embodiments, the method includes identifying or diagnosing whether the subject has an age-related disease or condition. In some embodiments, age-related diseases or symptoms are selected from the group consisting of: Alzheimer's disease, aneurysm, cystic fibrosis, pancreatic fibrosis, glaucoma, hypertension, idiopathic pulmonary fibrosis, inflammatory bowel disease, intervertebral disc degeneration, macular degeneration, osteoarthritis, type 2 diabetes, lipomatosis, lipid dystrophy, atherosclerosis, cataracts, COPD, idiopathic pulmonary fibrosis, failed kidney transplant, liver fibrosis, bone loss, myocardial infarction, sarcopenia, wound healing, alopecia, cardiomyocyte hypertrophy, osteoarthritis, Parkinson's disease, age-related loss of lung elasticity, macular degeneration, cachexia, glomerulosclerosis, cirrhosis, NAFLD, osteoporosis, amyotrophic lateral sclerosis, Huntington's disease, spinocerebellar ataxia, multiple sclerosis, and renal insufficiency. In some embodiments, the method includes identifying or diagnosing whether the subject has cancer. In some embodiments, the cancer is selected from the group consisting of: solid tumors, hematologic malignancies, sarcomas, osteosarcomas, 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 lymphoblastic leukemia (ALL), myelodysplastic syndrome (MDS), cutaneous T-cell lymphoma, retinoblastoma, gastric cancer, urothelial carcinoma, lung cancer, renal cell carcinoma, gastric and esophageal cancer, pancreatic cancer, prostate cancer, breast cancer, colorectal cancer, ovarian cancer, non-small cell lung cancer, head and neck squamous cell carcinoma, endometrial cancer, cervical cancer, liver cancer, and hepatocellular carcinoma. In some embodiments, the method includes diagnosing or identifying whether the subject has an infectious disease. In some embodiments, the infectious disease is infection with human immunodeficiency virus, cytomegalovirus, adenovirus, coronavirus, rhinovirus, rotavirus, smallpox, herpes simplex virus, hepatitis B virus, hepatitis A virus and hepatitis C virus, papillomavirus and influenza virus.

[0015] This article also provides a method for inducing immune cells to differentiate into memory or memory-like immune cells, the method comprising: contacting the immune cells with an effective amount of any of the above-described single-chain chimeric polypeptides or compositions. In some embodiments, the method comprises in vitro contacting of the immune cells. In some embodiments, the method comprises obtaining immune cells from a subject. In some embodiments, the method further comprises obtaining immune cells from a subject prior to the contacting step. In some embodiments, the method comprises in vivo contacting of the immune cells. In some embodiments, the immune cells are selected from the group consisting of: immature thymocytes, peripheral blood lymphocytes, naive T cells, pluripotent Th cell precursors, lymphoprogenitor cells, Treg cells, memory T cells, Th17 cells, Th22 cells, Th9 cells, Th2 cells, Th1 cells, Th3 cells, γδ T cells, αβ T cells, tumor-infiltrating T cells, CD8+ cells, etc. + T cells, CD4 +T cells, natural killer T cells, mast cells, macrophages, neutrophils, dendritic cells, basophils, eosinophils, and natural killer cells. In some embodiments, the method includes genetically modifying immune cells to express a chimeric antigen receptor or a recombinant T cell receptor. In some embodiments, the method further includes introducing a nucleic acid encoding a chimeric antigen receptor or a recombinant T cell receptor into the immune cells after a contact step. In some embodiments, the method further includes administering the immune cells to a subject in need. In some embodiments, the method includes identifying or diagnosing whether a subject has an age-related disease or condition. In some embodiments, age-related diseases or symptoms are selected from the group consisting of: Alzheimer's disease, aneurysm, cystic fibrosis, pancreatic fibrosis, glaucoma, hypertension, idiopathic pulmonary fibrosis, inflammatory bowel disease, intervertebral disc degeneration, macular degeneration, osteoarthritis, type 2 diabetes, lipomatosis, lipid dystrophy, atherosclerosis, cataracts, COPD, idiopathic pulmonary fibrosis, failed kidney transplant, liver fibrosis, bone loss, myocardial infarction, sarcopenia, wound healing, alopecia, cardiomyocyte hypertrophy, osteoarthritis, Parkinson's disease, age-related loss of lung elasticity, macular degeneration, cachexia, glomerulosclerosis, cirrhosis, NAFLD, osteoporosis, amyotrophic lateral sclerosis, Huntington's disease, spinocerebellar ataxia, multiple sclerosis, and renal insufficiency. In some embodiments, the method includes identifying or diagnosing whether the subject has cancer. In some embodiments, the cancer is selected from the group consisting of: solid tumors, hematologic malignancies, sarcomas, osteosarcomas, 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 lymphoblastic leukemia (ALL), myelodysplastic syndrome (MDS), cutaneous T-cell lymphoma, retinoblastoma, gastric cancer, urothelial carcinoma, lung cancer, renal cell carcinoma, gastric and esophageal cancer, pancreatic cancer, prostate cancer, breast cancer, colorectal cancer, ovarian cancer, non-small cell lung cancer, head and neck squamous cell carcinoma, endometrial cancer, cervical cancer, liver cancer, and hepatocellular carcinoma. In some embodiments, the method includes diagnosing or identifying whether the subject has an infectious disease. In some embodiments, the infectious disease is infection with human immunodeficiency virus, cytomegalovirus, adenovirus, coronavirus, rhinovirus, rotavirus, smallpox, herpes simplex virus, hepatitis B virus, hepatitis A virus and hepatitis C virus, papillomavirus and influenza virus.

[0016] This article also provides a method for killing cancer cells, infected cells, or senescent cells in a subject in need, the method comprising administering to the subject a therapeutically effective amount of any of the above-described single-chain chimeric peptides or compositions. In some embodiments, the method includes identifying or diagnosing whether the subject has cancer. In some embodiments, the cancer is selected from the group consisting of: solid tumors, hematologic malignancies, sarcomas, osteosarcomas, 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 lymphoblastic leukemia (ALL), myelodysplastic syndrome (MDS), cutaneous T-cell lymphoma, retinoblastoma, gastric cancer, urothelial carcinoma, lung cancer, renal cell carcinoma, gastric and esophageal cancer, pancreatic cancer, prostate cancer, breast cancer, colorectal cancer, ovarian cancer, non-small cell lung cancer, head and neck squamous cell carcinoma, endometrial cancer, cervical cancer, liver cancer, and hepatocellular carcinoma. In some embodiments, the method identifies or diagnoses whether a subject has age-related diseases or conditions. In some embodiments, age-related diseases or conditions are selected from the group consisting of: Alzheimer's disease, aneurysm, cystic fibrosis, pancreatic fibrosis, glaucoma, hypertension, idiopathic pulmonary fibrosis, inflammatory bowel disease, intervertebral disc degeneration, macular degeneration, osteoarthritis, type 2 diabetes, lipomatosis, lipid dystrophy, atherosclerosis, cataracts, COPD, idiopathic pulmonary fibrosis, failed kidney transplant, liver fibrosis, bone loss, myocardial infarction, sarcopenia, wound healing, alopecia, cardiomyocyte hypertrophy, osteoarthritis, Parkinson's disease, age-related loss of elasticity of lung tissue, macular degeneration, cachexia, glomerulosclerosis, cirrhosis, NAFLD, osteoporosis, amyotrophic lateral sclerosis, Huntington's disease, spinocerebellar ataxia, multiple sclerosis, and renal insufficiency.

[0017] This article also provides a method for treating a subject in need, the method comprising administering to the subject a therapeutically effective amount of any of the above-described single-chain chimeric peptides or compositions. In some embodiments, the method includes identifying or diagnosing whether the subject has cancer. In some embodiments, the cancer is selected from the group consisting of: solid tumors, hematologic malignancies, sarcomas, osteosarcomas, 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 lymphoblastic leukemia (ALL), myelodysplastic syndrome (MDS), cutaneous T-cell lymphoma, retinoblastoma, gastric cancer, urothelial carcinoma, lung cancer, renal cell carcinoma, gastric and esophageal cancer, pancreatic cancer, prostate cancer, breast cancer, colorectal cancer, ovarian cancer, non-small cell lung cancer, head and neck squamous cell carcinoma, endometrial cancer, cervical cancer, liver cancer, and hepatocellular carcinoma. In some embodiments, the method includes identifying or diagnosing whether the subject has age-related diseases or conditions. In some embodiments, age-related diseases or conditions are selected from the group consisting of: Alzheimer's disease, aneurysm, cystic fibrosis, pancreatic fibrosis, glaucoma, hypertension, idiopathic pulmonary fibrosis, inflammatory bowel disease, intervertebral disc degeneration, macular degeneration, osteoarthritis, type 2 diabetes, lipomatosis, lipid dystrophy, atherosclerosis, cataracts, COPD, idiopathic pulmonary fibrosis, failed kidney transplant, liver fibrosis, bone loss, myocardial infarction, sarcopenia, wound healing, alopecia, cardiomyocyte hypertrophy, osteoarthritis, Parkinson's disease, age-related loss of lung elasticity, macular degeneration, cachexia, glomerulosclerosis, cirrhosis, NAFLD, osteoporosis, amyotrophic lateral sclerosis, Huntington's disease, spinocerebellar ataxia, multiple sclerosis, and renal insufficiency. In some embodiments, the method includes diagnosing or identifying whether the subject has an infectious disease. In some embodiments, the infectious disease is infection with human immunodeficiency virus, cytomegalovirus, adenovirus, coronavirus, rhinovirus, rotavirus, smallpox, herpes simplex virus, hepatitis B virus, hepatitis A virus and hepatitis C virus, papillomavirus and influenza virus.

[0018] This document also provides nucleic acids encoding any of the above-described single-stranded chimeric polypeptides. Some embodiments include vectors comprising the above-described nucleic acids. In some embodiments, the vector is an expression vector. Some embodiments include cells comprising the above-described nucleic acids or vectors.

[0019] This document also provides a method for generating single-chain chimeric peptides, the method comprising: culturing cells in a culture medium under conditions sufficient to induce the generation of single-chain chimeric peptides; and recovering the single-chain chimeric peptides from the cells and / or the culture medium. Some embodiments include single-chain chimeric peptides generated according to the above description. In some embodiments of any single-chain chimeric peptides provided herein, the human soluble tissue factor domain does not induce blood clotting. In some embodiments of any single-chain chimeric peptides provided herein, the soluble tissue factor domain comprises or consists of human soluble wild-type tissue factor.

[0020] In some embodiments, the mutant human soluble tissue factor domain comprises a sequence that is at least 80% identical to SEQ ID NO:96. In some embodiments, the human soluble tissue factor domain comprises a sequence that is at least 90% identical to SEQ ID NO:96. In some embodiments, the human soluble tissue factor domain comprises a sequence that is at least 95% identical to SEQ ID NO:96. In some embodiments, the human soluble tissue factor domain comprises a sequence that is at least 100% identical to SEQ ID NO:96. In some embodiments, the human soluble tissue factor domain comprises a sequence that is at least 80% identical to SEQ ID NO:97. In some embodiments, the human soluble tissue factor domain comprises a sequence that is at least 90% identical to SEQ ID NO:97. In some embodiments, the human soluble tissue factor domain comprises a sequence that is at least 95% identical to SEQ ID NO:97. In some embodiments, the human soluble tissue factor domain comprises a sequence that is at least 100% identical to SEQ ID NO:97.

[0021] As used herein, the term "chimerism" refers to a polypeptide comprising amino acid sequences (e.g., domains) originally derived from two different sources (e.g., two naturally occurring different proteins, such as from the same or different species). For example, a chimeric polypeptide may include domains derived from at least two naturally occurring different human proteins. In some instances, a chimeric polypeptide may include a domain that is a synthetic sequence (e.g., scFv) and a domain derived from a naturally occurring protein (e.g., a naturally occurring human protein). In some embodiments, a chimeric polypeptide may include at least two different domains that are synthetic sequences (e.g., two different scFvs).

[0022] An "antigen-binding domain" is one or more protein domains (e.g., formed from amino acids from a single polypeptide or from amino acids from two or more polypeptides, such as the same or different polypeptides) capable of specifically binding to one or more different antigens. In some instances, an antigen-binding domain is capable of binding to an antigen or epitope with similar specificity and affinity to naturally occurring antibodies. In some embodiments, the antigen-binding domain may be an antibody or a fragment thereof. In some embodiments, the antigen-binding domain may include an alternative scaffold. Non-limiting examples of antigen-binding domains are described herein. Other examples of antigen-binding domains are known in the art.

[0023] "Soluble tissue factor domain" refers to a polypeptide that has at least 70% identity (e.g., at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100% identity) with a segment of a wild-type mammalian tissue factor protein (e.g., wild-type human tissue factor protein) lacking a transmembrane domain and an intracellular domain. Non-limiting examples of soluble tissue factor domains are described herein.

[0024] The term "soluble interleukin protein" is used herein to refer to a mature and secreted interleukin protein or a biologically active fragment thereof. In some instances, a soluble interleukin protein may include a sequence that is at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100% identical to a mature and secreted mammalian wild-type interleukin protein (e.g., human wild-type interleukin protein) and retains its biological activity. Non-limiting examples of soluble interleukins are described herein.

[0025] The term "soluble cytokine protein" is used herein to refer to mature and secreted cytokine proteins or biologically active fragments thereof. In some instances, soluble cytokine proteins may include sequences that are at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100% identical to and retain the biological activity of mature and secreted mammalian wild-type interleukin proteins (e.g., human wild-type interleukin protein). Non-limiting examples of soluble cytokine proteins are described herein.

[0026] The term "soluble interleukin receptor" is used herein in its broadest sense to refer to a polypeptide that lacks a transmembrane domain (and optionally an intracellular domain) capable of binding one or more of its native ligands (e.g., under physiological conditions, such as at room temperature, in phosphate-buffered saline). For example, a soluble interleukin receptor may comprise a sequence that is at least 70% identical (e.g., at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100% identical) to the extracellular domain of a wild-type interleukin receptor and retains its ability to specifically bind to one or more of its native ligands, but lacks its transmembrane domain (and optionally, also its intracellular domain). Non-limiting examples of soluble interleukin receptors are described herein.

[0027] The term "soluble cytokine receptor" is used herein in its broadest sense to refer to a polypeptide that lacks a transmembrane domain (and optionally an intracellular domain) capable of binding one or more of its native ligands (e.g., under physiological conditions, such as at room temperature, in phosphate-buffered saline). For example, a soluble cytokine receptor may comprise a sequence that is at least 70% identical (e.g., at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100% identical) to the extracellular domain of a wild-type cytokine receptor and retains its ability to specifically bind to one or more of its native ligands, but lacks its transmembrane domain (and optionally, also its intracellular domain). Non-limiting examples of soluble cytokine receptors are described herein.

[0028] The term "antibody" is used herein in its broadest sense and includes certain types of immunoglobulin molecules that include 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), antibody fragments, and multispecific antibodies. One example of an antigen-binding domain is an antigen-binding domain formed by a VH-VL dimer. Other examples of antibodies are described herein. Other examples of antibodies are known in the art.

[0029] "Affinity" refers to the sum of non-covalent interactions between an antigen-binding site and its binding partner (e.g., an antigen or epitope). Unless otherwise stated, as used herein, "affinity" refers to 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 molecule X for its partner Y can be expressed using the dissociation equilibrium constant (K0). D The kinetic components that contribute to the dissociation equilibrium constant are described in more detail below. Affinity can be measured by conventional methods known in the art, including those described herein. Affinity can be measured, for example, using surface plasmon resonance (SPR) techniques (e.g., ) or biological layer interferometry (e.g. The affinity for the antigen-binding domain and its corresponding antigen or epitope is determined by means of other methods known in the art.

[0030] As used in this article, "single-chain polypeptide" refers to a single protein chain.

[0031] The term "a pair of affinity domains" refers to domains smaller than 1 × 10⁻⁶. -7 M (e.g., less than 1×10) -8 M, less than 1×10 -9 M, less than 1×10 -10 M or less than 1×10 -11 M) of K D A pair of affinity domains are two distinct protein domains that bind specifically to each other. In some instances, an affinity domain pair may be a pair of naturally occurring proteins. In some embodiments, an affinity domain pair may be a pair of synthetic proteins. Non-limiting examples of affinity domain pairs are described herein.

[0032] The term "epitope" refers to a portion of an antigen that specifically binds to an antigen-binding domain. An epitope may consist, for example, of surface-accessible amino acid residues and / or sugar side chains, and may possess specific three-dimensional structural features and specific charge characteristics. The difference between conformational epitopes and non-conformational epitopes is that binding to the former may disappear in the presence of denaturing solvents, while binding to the latter will not. An epitope may contain amino acid residues that directly participate in binding and other amino acid residues that do not directly participate in binding. Methods for identifying epitopes that bind to an antigen-binding domain are known in the art.

[0033] "Immune effector cells" are cells of the mammalian immune system that are capable of directly or indirectly recognizing pathogenic cells (e.g., cancer cells) in mammals and / or causing cell arrest or cell death in said cells. Non-limiting examples of immune effector cells include macrophages, T lymphocytes (e.g., cytotoxic T lymphocytes and T helper cells), natural killer cells, neutrophils, monocytes, and eosinophils. Other examples of immune effector cells are known in the art.

[0034] The term "treatment" refers to the improvement of at least one symptom of a disease. In some instances, the disease being treated is cancer, and improving at least one symptom of cancer includes reducing abnormal proliferation, gene expression, signal transduction, translation, and / or factor secretion. Generally, a treatment method involves administering a therapeutically effective amount of a composition to a subject who needs or has been identified as needing such treatment, said composition reducing at least one symptom of the disease.

[0035] 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 pertains. The methods and materials used in this invention are described herein; other suitable methods and materials known in the art may also be used. Materials, methods, and examples are illustrative only and not intended to be limiting. All publications, patent applications, patents, sequences, database entries, and other references mentioned herein are incorporated herein by reference in their entirety. In case of conflict, this specification (including definitions) shall prevail.

[0036] Other features and advantages of the invention will become apparent from the following detailed description and drawings, as well as from the claims. Attached Figure Description

[0037] Figure 1 This is a schematic diagram of an exemplary αCD3scFv / TF / αCD28scFv single-chain chimeric polypeptide.

[0038] Figure 2 It is a chromatogram showing the elution of an exemplary αCD3scFv / TF / αCD28scFv single-chain chimeric polypeptide from an anti-tissue factor antibody affinity column.

[0039] Figure 3 The chromatogram shows the elution of a Superdex 200 Increase 10 / 300GL gel filter column loaded with an exemplary αCD3scFv / TF / αCD28scFv single-chain chimeric polypeptide.

[0040] Figure 4 This is an electrophoresis image of an exemplary αCD3scFv / TF / αCD28scFv single-chain chimeric polypeptide purified using an anti-tissue factor antibody affinity column on a sodium dodecyl sulfate polyacrylamide gel (4-12% NuPage Bis-Tris gel).

[0041] Figure 5 This is a graph showing the ELISA quantification of an exemplary αCD3scFv / TF / αCD28scFv single-chain chimeric peptide performed using the method described in Example 1. Purified tissue factor was used as a control.

[0042] Figure 6 This demonstrates CD4 isolated from the blood of two donors stimulated by an exemplary αCD3scFv / TF / αCD28scFv single-chain chimeric peptide. + A graph showing the ability of T cells to express CD25. The experiment was conducted as described in Example 2.

[0043] Figure 7This demonstrates CD8 isolates from the blood of two donors stimulated by an exemplary αCD3scFv / TF / αCD28scFv single-chain chimeric peptide. + A graph showing the ability of T cells to express CD25. The experiment was conducted as described in Example 2.

[0044] Figure 8 This demonstrates CD4 isolated from the blood of two donors stimulated by an exemplary αCD3scFv / TF / αCD28scFv single-chain chimeric peptide. + A graph showing the ability of T cells to express CD69. The experiment was conducted as described in Example 2.

[0045] Figure 9 This is a schematic diagram of an exemplary IL-2 / TF / IL-2 single-chain chimeric polypeptide.

[0046] Figure 10 The study shows a comparison of IL-2 activity in IL-2 / TF IL-2 as measured by 32Dβ cell proliferation assay with recombinant IL-2.

[0047] Figure 11 The study shows a comparison of IL-2 activity in IL-2 / TF / IL-2 as measured by CTLL-2 cell proliferation assay with recombinant IL-2.

[0048] Figure 12 ApoE was shown to be fed a standard diet or a high-fat diet and treated with PBS control (untreated) or IL-2 / TF / IL-2. - / - Fasting blood glucose levels in mice.

[0049] Figure 13 This shows the CD4 count in blood lymphocytes of ApoE- / - mice fed a standard diet or a high-fat diet and treated with PBS control (untreated) or IL-2 / TF / IL-2. + CD25 + FoxP3 + T regulates cell ratio.

[0050] Figure 14 It is a graph showing the chromatographic characteristics of cell culture supernatant containing IL-2 / TF / IL-2 protein after binding and elution on anti-TF antibody resin.

[0051] Figure 15 The analytical SEC features of IL-2 / TF / IL-2 are shown.

[0052] Figure 16A and 16B The reduced SDS-PAGE analysis of IL-2 / TF / IL-2 before and after deglycosylation is shown. Figure 16A The reduced SDS-PAGE analysis of IL-2 / TF / IL-2 before deglycosylation is shown. Figure 16B The reduced SDS-PAGE analysis of IL-2 / TF / IL-2 after deglycosylation is shown.

[0053] Figures 17A and 17B show the results of immune stimulation of C57BL / 6 mice using IL-2 / TF / IL-2. Figure 17A The spleen weight after IL-2 / TF / IL-2 treatment is shown. Figure 17B The percentage of immune cell types after IL-2 / TF / IL-2 treatment is shown.

[0054] Figure 18 CD4 in mice treated with IL-2 / TF / IL-2 was shown. + CD25 expression is upregulated on T cells.

[0055] Figure 19 The pharmacokinetics of IL-2 / TF / IL-2 in C57BL / 6 mice were shown.

[0056] Figure 20A and 20B The study showed that IL-2 / TF / IL-2 alleviates ApoE. - / - The role of high-fat diet in the formation of atherosclerotic plaques in mice. Figure 20A ApoE was shown to be caused by feeding standard food or a high-fat diet and treatment with PBS control or IL-2 / TF / IL-2. - / - A representative view of atherosclerotic plaques in mice. Figure 20B The results of quantitative analysis of atherosclerotic plaques in each group are shown.

[0057] Figure 21 The fasting glucose levels of IL-2 / TF / IL-2 treated mice were shown compared to those of control mice.

[0058] Figure 22 The image shows CD4+ in the blood lymphocytes of mice treated with IL-2 / TF / IL-2 and mice treated with the control. + CD25 + FoxP3 + Tregs percentage.

[0059] Figure 23 This is a schematic diagram of an exemplary IL-15 / TF / IL-15 single-chain chimeric polypeptide.

[0060] Figure 24The study shows a comparison of IL-15 activity in IL-15 / TF / IL-15 as measured by 32Dβ cell proliferation analysis with that of recombinant IL-15.

[0061] Figure 25 It is a graph showing the chromatographic characteristics of cell culture supernatant containing IL-15 / TF / IL-15 protein after binding and elution on anti-TF antibody resin.

[0062] Figure 26A and 26B The reduced SDS-PAGE analysis of IL-15 / TF / IL-15 before and after deglycosylation is shown. Figure 26A The reduced SDS-PAGE analysis of IL-15 / TF / IL-15 before deglycosylation is shown. Figure 26B The reduced SDS-PAGE analysis of IL-15 / TF / IL-15 after deglycosylation is shown.

[0063] Figure 27 A-27C is a set of figures showing the immune stimulation of C57BL / 6 mice after 2t2 treatment.

[0064] Figure 28 A-28C is a set of figures showing ApoEs fed a Western diet and treated with 2t2. - / - Tregs, NK cells and CD8 in mice + In vivo stimulation of T cells.

[0065] Figure 29 Figures A and 29B are a set of figures showing how 2t2 induces spleen cell proliferation in C57BL / 6 mice.

[0066] Figure 30 A and 30B are a set of graphs showing ApoEs fed a Western diet and treated with 2t2. - / - NK cells and CD8 in mice + In vivo induction of T cell proliferation.

[0067] Figure 31 A-31C is a set of graphs that show the improvement of ApoE by 2t2. - / - Hyperglycemia induced in mice by a Western diet.

[0068] Figure 32 The study showed upregulation of CD44 memory T cells after treatment with 2t2.

[0069] Figure 33 The diagram shows the activation of factor X (FX) after treatment with single-chain or multi-chain chimeric peptides.

[0070] Figure 34 A-34C displays CD4 + CD25 hi T reg Cells, CD4 + CD25 - T con Cells or CD8 + T con Human blood lymphocytes in cells respond to pStat5a as a response to treatment with 2t2 or IL2. Figure 34 A shows CD4 + CD25 hi T reg pSTAT5 response in cells. Figure 34 B showed CD4 + CD25 - T con pSTAT5 response in cells. Figure 34 C displays CD8 + T con pSTAT5 response in cells. Detailed Implementation

[0071] This document provides a single-chain chimeric polypeptide comprising: (i) a first target-binding domain (e.g., any target-binding domain described herein or known in the art), (ii) a soluble tissue factor domain (e.g., any exemplary soluble tissue factor domain described herein or known in the art), and (iii) a second target-binding domain (e.g., any target-binding domain described herein or known in the art).

[0072] This document also provides compositions comprising any of the single-chain chimeric polypeptides described herein, nucleic acids encoding any of the single-chain chimeric polypeptides described herein, and cells comprising any of the nucleic acids encoding any of the single-chain chimeric polypeptides described herein. This document also provides methods for stimulating immune cells and methods for treating subjects in need (including using any of the single-chain chimeric polypeptides described herein). This document also provides methods for generating any of the single-chain chimeric polypeptides described herein.

[0073] In some examples of any of the single-chain chimeric polypeptides described herein, the single-chain chimeric polypeptide may have the following total lengths: about 50 amino acids to about 3000 amino acids, about 50 amino acids to about 2500 amino acids, about 50 amino acids to about 2000 amino acids, about 50 amino acids to about 1500 amino acids, about 50 amino acids to about 1000 amino acids, about 50 amino acids to about 950 amino acids, about 50 amino acids to about 900 amino acids, about 50 amino acids to about 850 amino acids, about 50 amino acids to about 800 amino acids, about 50 amino acids to about 750 amino acids, about 50 amino acids to about 700 amino acids, and about 50 amino acids to about 650 amino acids. Approximately 50 amino acids to approximately 600 amino acids, approximately 50 amino acids to approximately 550 amino acids, approximately 50 amino acids to approximately 500 amino acids, approximately 50 amino acids to approximately 480 amino acids, approximately 50 amino acids to approximately 460 amino acids, approximately 50 amino acids to approximately 440 amino acids, approximately 50 amino acids to approximately 420 amino acids, approximately 50 amino acids to approximately 400 amino acids, approximately 50 amino acids to approximately 380 amino acids, approximately 50 amino acids to approximately 360 amino acids, approximately 50 amino acids to approximately 340 amino acids, approximately 50 amino acids to approximately 320 amino acids, approximately 50 amino acids to approximately 300 amino acids, approximately 50 amino acids to approximately 280 amino acids, approximately 50 amino acids to approximately 260 amino acids, approximately 50 amino acids to approximately 240 amino acids, approximately 50 amino acids to approximately 220 amino acids, approximately 50 amino acids to approximately 200 amino acids, approximately 50 amino acids to approximately 150 amino acids, approximately 50 amino acids to approximately 100 amino acids, approximately 100 amino acids to approximately 3000 amino acids, approximately 100 amino acids to approximately 2500 amino acids, approximately 100 amino acids to approximately 2000 amino acids, approximately 100 amino acids to approximately 1500 amino acids, approximately 100 amino acids to approximately 1000 amino acids, approximately 100 amino acids to approximately 950 amino acids, approximately 100 amino acids to approximately 900 amino acids, approximately 100 amino acids to approximately 850 amino acids, approximately 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 650 amino acids, about 100 amino acids to about 600 amino acids, about 100 amino acids to about 550 amino acids, about 100 amino acids to about 500 amino acids, about 100 amino acids to about 480 amino acids, about 100 amino acids to about 460 amino acids, about 100 amino acids to about 440 amino acids, about 100 amino acids to about 420 amino acids, about 100 amino acids to about 400 amino acids, about 100 amino acids to about 380 amino acids, about 100 amino acids to about 360 amino acids.Approximately 100 amino acids to approximately 340 amino acids, approximately 100 amino acids to approximately 320 amino acids, approximately 100 amino acids to approximately 300 amino acids, approximately 100 amino acids to approximately 280 amino acids, approximately 100 amino acids to approximately 260 amino acids, approximately 100 amino acids to approximately 240 amino acids, approximately 100 amino acids to approximately 220 amino acids, approximately 100 amino acids to approximately 200 amino acids, approximately 100 amino acids to approximately 150 amino acids, approximately 150 amino acids to approximately 3000 amino acids, approximately 150 amino acids to approximately 2500 amino acids, approximately 150 amino acids to approximately 2000 amino acids, approximately 150 amino acids to approximately 1500 amino acids, approximately 150 amino acids to approximately 10 00 amino acids, approximately 150 amino acids to approximately 950 amino acids, approximately 150 amino acids to approximately 900 amino acids, approximately 150 amino acids to approximately 850 amino acids, approximately 150 amino acids to approximately 800 amino acids, approximately 150 amino acids to approximately 750 amino acids, approximately 150 amino acids to approximately 700 amino acids, approximately 150 amino acids to approximately 650 amino acids, approximately 150 amino acids to approximately 600 amino acids, approximately 150 amino acids to approximately 550 amino acids, approximately 150 amino acids to approximately 500 amino acids, approximately 150 amino acids to approximately 480 amino acids, approximately 150 amino acids to approximately 460 amino acids, approximately 150 amino acids to approximately 440 amino acids, approximately 150 amino acids to Approximately 420 amino acids, approximately 150 amino acids to approximately 400 amino acids, approximately 150 amino acids to approximately 380 amino acids, approximately 150 amino acids to approximately 360 amino acids, approximately 150 amino acids to approximately 340 amino acids, approximately 150 amino acids to approximately 320 amino acids, approximately 150 amino acids to approximately 300 amino acids, approximately 150 amino acids to approximately 280 amino acids, approximately 150 amino acids to approximately 260 amino acids, approximately 150 amino acids to approximately 240 amino acids, approximately 150 amino acids to approximately 220 amino acids, approximately 150 amino acids to approximately 200 amino acids, approximately 200 amino acids to approximately 3000 amino acids, approximately 200 amino acids to approximately 2500 amino acids, approximately 200 Amino acids to about 2000 amino acids, about 200 amino acids to about 1500 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 480 amino acids.Approximately 200 amino acids to approximately 460 amino acids, approximately 200 amino acids to approximately 440 amino acids, approximately 200 amino acids to approximately 420 amino acids, approximately 200 amino acids to approximately 400 amino acids, approximately 200 amino acids to approximately 380 amino acids, approximately 200 amino acids to approximately 360 amino acids, approximately 200 amino acids to approximately 340 amino acids, approximately 200 amino acids to approximately 320 amino acids, approximately 200 amino acids to approximately 300 amino acids, approximately 200 amino acids to approximately 280 amino acids, approximately 200 amino acids to approximately 260 amino acids, approximately 200 amino acids to approximately 240 amino acids, approximately 200 amino acids to approximately 220 amino acids, approximately 220 amino acids to approximately 3000 amino acids. Amino acids, approximately 220 amino acids to approximately 2500 amino acids, approximately 220 amino acids to approximately 2000 amino acids, approximately 220 amino acids to approximately 1500 amino acids, approximately 220 amino acids to approximately 1000 amino acids, approximately 220 amino acids to approximately 950 amino acids, approximately 220 amino acids to approximately 900 amino acids, approximately 220 amino acids to approximately 850 amino acids, approximately 220 amino acids to approximately 800 amino acids, approximately 220 amino acids to approximately 750 amino acids, approximately 220 amino acids to approximately 700 amino acids, approximately 220 amino acids to approximately 650 amino acids, approximately 220 amino acids to approximately 600 amino acids, approximately 220 amino acids to approximately 550 amino acids, approximately 220 amino acids to Approximately 500 amino acids, approximately 220 amino acids to approximately 480 amino acids, approximately 220 amino acids to approximately 460 amino acids, approximately 220 amino acids to approximately 440 amino acids, approximately 220 amino acids to approximately 420 amino acids, approximately 220 amino acids to approximately 400 amino acids, approximately 220 amino acids to approximately 380 amino acids, approximately 220 amino acids to approximately 360 amino acids, approximately 220 amino acids to approximately 340 amino acids, approximately 220 amino acids to approximately 320 amino acids, approximately 220 amino acids to approximately 300 amino acids, approximately 220 amino acids to approximately 280 amino acids, approximately 220 amino acids to approximately 260 amino acids, approximately 220 amino acids to approximately 240 amino acids, approximately 240 amino acids Acids up to 3000 amino acids, about 240 amino acids up to 2500 amino acids, about 240 amino acids up to 2000 amino acids, about 240 amino acids up to 1500 amino acids, about 240 amino acids up to 1000 amino acids, about 240 amino acids up to 950 amino acids, about 240 amino acids up to 900 amino acids, about 240 amino acids up to 850 amino acids, about 240 amino acids up to 800 amino acids, about 240 amino acids up to 750 amino acids, about 240 amino acids up to 700 amino acids, about 240 amino acids up to 650 amino acids, about 240 amino acids up to 600 amino acids, about 240 amino acids up to 550 amino acids.Approximately 240 amino acids to approximately 500 amino acids, approximately 240 amino acids to approximately 480 amino acids, approximately 240 amino acids to approximately 460 amino acids, approximately 240 amino acids to approximately 440 amino acids, approximately 240 amino acids to approximately 420 amino acids, approximately 240 amino acids to approximately 400 amino acids, approximately 240 amino acids to approximately 380 amino acids, approximately 240 amino acids to approximately 360 amino acids, approximately 240 amino acids to approximately 340 amino acids, approximately 240 amino acids to approximately 320 amino acids, approximately 240 amino acids to approximately 300 amino acids, approximately 240 amino acids to approximately 280 amino acids, approximately 240 amino acids to approximately 260 amino acids, approximately 260 amino acids to approximately 3000 amino acids. Amino acids, approximately 260 amino acids to approximately 2500 amino acids, approximately 260 amino acids to approximately 2000 amino acids, approximately 260 amino acids to approximately 1500 amino acids, approximately 260 amino acids to approximately 1000 amino acids, approximately 260 amino acids to approximately 950 amino acids, approximately 260 amino acids to approximately 900 amino acids, approximately 260 amino acids to approximately 850 amino acids, approximately 260 amino acids to approximately 800 amino acids, approximately 260 amino acids to approximately 750 amino acids, approximately 260 amino acids to approximately 700 amino acids, approximately 260 amino acids to approximately 650 amino acids, approximately 260 amino acids to approximately 600 amino acids, approximately 260 amino acids to approximately 550 amino acids, approximately 260 amino acids to Approximately 500 amino acids, approximately 260 amino acids to approximately 480 amino acids, approximately 260 amino acids to approximately 460 amino acids, approximately 260 amino acids to approximately 440 amino acids, approximately 260 amino acids to approximately 420 amino acids, approximately 260 amino acids to approximately 400 amino acids, approximately 260 amino acids to approximately 380 amino acids, approximately 260 amino acids to approximately 360 amino acids, approximately 260 amino acids to approximately 340 amino acids, approximately 260 amino acids to approximately 320 amino acids, approximately 260 amino acids to approximately 300 amino acids, approximately 260 amino acids to approximately 280 amino acids, approximately 280 amino acids to approximately 3000 amino acids, approximately 280 amino acids to approximately 2500 amino acids, approximately 280 Amino acids to about 2000 amino acids, about 280 amino acids to about 1500 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 480 amino acids.Approximately 280 amino acids to approximately 460 amino acids, approximately 280 amino acids to approximately 440 amino acids, approximately 280 amino acids to approximately 420 amino acids, approximately 280 amino acids to approximately 400 amino acids, approximately 280 amino acids to approximately 380 amino acids, approximately 280 amino acids to approximately 360 amino acids, approximately 280 amino acids to approximately 340 amino acids, approximately 280 amino acids to approximately 320 amino acids, approximately 280 amino acids to approximately 300 amino acids, approximately 300 amino acids to approximately 3000 amino acids, approximately 300 amino acids to approximately 2500 amino acids, approximately 300 amino acids to approximately 2000 amino acids, approximately 300 amino acids to approximately 1500 amino acids, approximately 300 amino acids to approximately 10 00 amino acids, approximately 300 amino acids to approximately 950 amino acids, approximately 300 amino acids to approximately 900 amino acids, approximately 300 amino acids to approximately 850 amino acids, approximately 300 amino acids to approximately 800 amino acids, approximately 300 amino acids to approximately 750 amino acids, approximately 300 amino acids to approximately 700 amino acids, approximately 300 amino acids to approximately 650 amino acids, approximately 300 amino acids to approximately 600 amino acids, approximately 300 amino acids to approximately 550 amino acids, approximately 300 amino acids to approximately 500 amino acids, approximately 300 amino acids to approximately 480 amino acids, approximately 300 amino acids to approximately 460 amino acids, approximately 300 amino acids to approximately 440 amino acids, approximately 300 amino acids to Approximately 420 amino acids, approximately 300 amino acids to approximately 400 amino acids, approximately 300 amino acids to approximately 380 amino acids, approximately 300 amino acids to approximately 360 amino acids, approximately 300 amino acids to approximately 340 amino acids, approximately 300 amino acids to approximately 320 amino acids, approximately 320 amino acids to approximately 3000 amino acids, approximately 320 amino acids to approximately 2500 amino acids, approximately 320 amino acids to approximately 2000 amino acids, approximately 320 amino acids to approximately 1500 amino acids, approximately 320 amino acids to approximately 1000 amino acids, approximately 320 amino acids to approximately 950 amino acids, approximately 320 amino acids to approximately 900 amino acids, approximately 320 amino acids to approximately 850 amino acids, approximately 3 20 amino acids to about 800 amino acids, about 320 amino acids to about 750 amino acids, about 320 amino acids to about 700 amino acids, about 320 amino acids to about 650 amino acids, about 320 amino acids to about 600 amino acids, about 320 amino acids to about 550 amino acids, about 320 amino acids to about 500 amino acids, about 320 amino acids to about 480 amino acids, about 320 amino acids to about 460 amino acids, about 320 amino acids to about 440 amino acids, about 320 amino acids to about 420 amino acids, about 320 amino acids to about 400 amino acids, about 320 amino acids to about 380 amino acids, about 320 amino acids to about 360 amino acids.Approximately 320 amino acids to approximately 340 amino acids, approximately 340 amino acids to approximately 3000 amino acids, approximately 340 amino acids to approximately 2500 amino acids, approximately 340 amino acids to approximately 2000 amino acids, approximately 340 amino acids to approximately 1500 amino acids, approximately 340 amino acids to approximately 1000 amino acids, approximately 340 amino acids to approximately 950 amino acids, approximately 340 amino acids to approximately 900 amino acids, approximately 340 amino acids to approximately 850 amino acids, approximately 340 amino acids to approximately 800 amino acids, approximately 340 amino acids to approximately 750 amino acids, approximately 340 amino acids to approximately 700 amino acids, approximately 340 amino acids to approximately 650 amino acids, approximately 340 amino acids Acids to about 600 amino acids, about 340 amino acids to about 550 amino acids, about 340 amino acids to about 500 amino acids, about 340 amino acids to about 480 amino acids, about 340 amino acids to about 460 amino acids, about 340 amino acids to about 440 amino acids, about 340 amino acids to about 420 amino acids, about 340 amino acids to about 400 amino acids, about 340 amino acids to about 380 amino acids, about 340 amino acids to about 360 amino acids, about 360 amino acids to about 3000 amino acids, about 360 amino acids to about 2500 amino acids, about 360 amino acids to about 2000 amino acids, about 360 amino acids to about 1500 amino acids. Acids, approximately 360 amino acids to approximately 1000 amino acids, approximately 360 amino acids to approximately 950 amino acids, approximately 360 amino acids to approximately 900 amino acids, approximately 360 amino acids to approximately 850 amino acids, approximately 360 amino acids to approximately 800 amino acids, approximately 360 amino acids to approximately 750 amino acids, approximately 360 amino acids to approximately 700 amino acids, approximately 360 amino acids to approximately 650 amino acids, approximately 360 amino acids to approximately 600 amino acids, approximately 360 amino acids to approximately 550 amino acids, approximately 360 amino acids to approximately 500 amino acids, approximately 360 amino acids to approximately 480 amino acids, approximately 360 amino acids to approximately 460 amino acids, approximately 360 amino acids to approximately 440 amino acids, approximately 360 amino acids to approximately 420 amino acids, approximately 360 amino acids to approximately 400 amino acids, approximately 360 amino acids to approximately 380 amino acids, approximately 380 amino acids to approximately 3000 amino acids, approximately 380 amino acids to approximately 2500 amino acids, approximately 380 amino acids to approximately 2000 amino acids, approximately 380 amino acids to approximately 1500 amino acids, approximately 380 amino acids to approximately 1000 amino acids, approximately 380 amino acids to approximately 950 amino acids, approximately 380 amino acids to approximately 900 amino acids, approximately 380 amino acids to approximately 850 amino acids, approximately 380 amino acids to approximately 800 amino acids, approximately 380 amino acids to approximately 750 amino acids.Approximately 380 amino acids to approximately 700 amino acids, approximately 380 amino acids to approximately 650 amino acids, approximately 380 amino acids to approximately 600 amino acids, approximately 380 amino acids to approximately 550 amino acids, approximately 380 amino acids to approximately 500 amino acids, approximately 380 amino acids to approximately 480 amino acids, approximately 380 amino acids to approximately 460 amino acids, approximately 380 amino acids to approximately 440 amino acids, approximately 380 amino acids to approximately 420 amino acids, approximately 380 amino acids to approximately 400 amino acids, approximately 400 amino acids to approximately 3000 amino acids, approximately 400 amino acids to approximately 2500 amino acids, approximately 400 amino acids to approximately 2000 amino acids, approximately 400 amino acids to... Approximately 1500 amino acids, approximately 400 amino acids to approximately 1000 amino acids, approximately 400 amino acids to approximately 950 amino acids, approximately 400 amino acids to approximately 900 amino acids, approximately 400 amino acids to approximately 850 amino acids, approximately 400 amino acids to approximately 800 amino acids, approximately 400 amino acids to approximately 750 amino acids, approximately 400 amino acids to approximately 700 amino acids, approximately 400 amino acids to approximately 650 amino acids, approximately 400 amino acids to approximately 600 amino acids, approximately 400 amino acids to approximately 550 amino acids, approximately 400 amino acids to approximately 500 amino acids, approximately 400 amino acids to approximately 480 amino acids, approximately 400 amino acids to approximately 460 amino acids, approximately 4 00 amino acids to about 440 amino acids, about 400 amino acids to about 420 amino acids, about 420 amino acids to about 3000 amino acids, about 420 amino acids to about 2500 amino acids, about 420 amino acids to about 2000 amino acids, about 420 amino acids to about 1500 amino acids, about 420 amino acids to about 1000 amino acids, about 420 amino acids to about 950 amino acids, about 420 amino acids to about 900 amino acids, about 420 amino acids to about 850 amino acids, about 420 amino acids to about 800 amino acids, about 420 amino acids to about 750 amino acids, about 420 amino acids to about 700 amino acids, about 420 amino acids to about 650 amino acids, approximately 420 amino acids to approximately 600 amino acids, approximately 420 amino acids to approximately 550 amino acids, approximately 420 amino acids to approximately 500 amino acids, approximately 420 amino acids to approximately 480 amino acids, approximately 420 amino acids to approximately 460 amino acids, approximately 420 amino acids to approximately 440 amino acids, approximately 440 amino acids to approximately 3000 amino acids, approximately 440 amino acids to approximately 2500 amino acids, approximately 440 amino acids to approximately 2000 amino acids, approximately 440 amino acids to approximately 1500 amino acids, approximately 440 amino acids to approximately 1000 amino acids, approximately 440 amino acids to approximately 950 amino acids, approximately 440 amino acids to approximately 900 amino acids.Approximately 440 amino acids to approximately 850 amino acids, approximately 440 amino acids to approximately 800 amino acids, approximately 440 amino acids to approximately 750 amino acids, approximately 440 amino acids to approximately 700 amino acids, approximately 440 amino acids to approximately 650 amino acids, approximately 440 amino acids to approximately 600 amino acids, approximately 440 amino acids to approximately 550 amino acids, approximately 440 amino acids to approximately 500 amino acids, approximately 440 amino acids to approximately 480 amino acids, approximately 440 amino acids to approximately 460 amino acids, approximately 460 amino acids to approximately 3000 amino acids, approximately 460 amino acids to approximately 2500 amino acids, approximately 460 amino acids to approximately 2000 amino acids, approximately 460 amino acids to Approximately 1500 amino acids, approximately 460 amino acids to approximately 1000 amino acids, approximately 460 amino acids to approximately 950 amino acids, approximately 460 amino acids to approximately 900 amino acids, approximately 460 amino acids to approximately 850 amino acids, approximately 460 amino acids to approximately 800 amino acids, approximately 460 amino acids to approximately 750 amino acids, approximately 460 amino acids to approximately 700 amino acids, approximately 460 amino acids to approximately 650 amino acids, approximately 460 amino acids to approximately 600 amino acids, approximately 460 amino acids to approximately 550 amino acids, approximately 460 amino acids to approximately 500 amino acids, approximately 460 amino acids to approximately 480 amino acids, approximately 480 amino acids to approximately 3000 amino acids, approximately 480 amino acids to about 2500 amino acids, about 480 amino acids to about 2000 amino acids, about 480 amino acids to about 1500 amino acids, about 480 amino acids to about 1000 amino acids, about 480 amino acids to about 950 amino acids, about 480 amino acids to about 900 amino acids, about 480 amino acids to about 850 amino acids, about 480 amino acids to about 800 amino acids, about 480 amino acids to about 750 amino acids, about 480 amino acids to about 700 amino acids, about 480 amino acids to about 650 amino acids, about 480 amino acids to about 600 amino acids, about 480 amino acids to about 550 amino acids, about 480 amino acids to about 500 amino acids, approximately 500 amino acids to approximately 3000 amino acids, approximately 500 amino acids to approximately 2500 amino acids, approximately 500 amino acids to approximately 2000 amino acids, approximately 500 amino acids to approximately 1500 amino acids, approximately 500 amino acids to approximately 1000 amino acids, approximately 500 amino acids to approximately 950 amino acids, approximately 500 amino acids to approximately 900 amino acids, approximately 500 amino acids to approximately 850 amino acids, approximately 500 amino acids to approximately 800 amino acids, approximately 500 amino acids to approximately 750 amino acids, approximately 500 amino acids to approximately 700 amino acids, approximately 500 amino acids to approximately 650 amino acids, approximately 500 amino acids to approximately 600 amino acids.Approximately 500 amino acids to approximately 550 amino acids, approximately 550 amino acids to approximately 3000 amino acids, approximately 550 amino acids to approximately 2500 amino acids, approximately 550 amino acids to approximately 2000 amino acids, approximately 550 amino acids to approximately 1500 amino acids, approximately 550 amino acids to approximately 1000 amino acids, approximately 550 amino acids to approximately 950 amino acids, approximately 550 amino acids to approximately 900 amino acids, approximately 550 amino acids to approximately 850 amino acids, approximately 550 amino acids to approximately 800 amino acids, approximately 550 amino acids to approximately 750 amino acids, approximately 550 amino acids to approximately 700 amino acids, approximately 550 amino acids to approximately 650 amino acids, approximately 550 amino acids to approximately 600 amino acids, approximately 600 amino acids to approximately 3000 amino acids, approximately 600 amino acids to approximately 2500 amino acids, approximately 600 amino acids to approximately 2000 amino acids, approximately 600 amino acids to approximately 1500 amino acids, approximately 600 amino acids to approximately 1000 amino acids, approximately 600 amino acids to approximately 950 amino acids, approximately 600 amino acids to approximately 900 amino acids, approximately 600 amino acids to approximately 850 amino acids, approximately 600 amino acids to approximately 800 amino acids, approximately 600 amino acids to approximately 750 amino acids, approximately 600 amino acids to approximately 700 amino acids, approximately 600 amino acids to approximately 650 amino acids, approximately 650 amino acids to approximately 3000 amino acids, approximately 650 amino acids to about 2500 amino acids, about 650 amino acids to about 2000 amino acids, about 650 amino acids to about 1500 amino acids, about 650 amino acids to about 1000 amino acids, about 650 amino acids to about 950 amino acids, about 650 amino acids to about 900 amino acids, about 650 amino acids to about 850 amino acids, about 650 amino acids to about 800 amino acids, about 650 amino acids to about 750 amino acids, about 650 amino acids to about 700 amino acids, about 700 amino acids to about 3000 amino acids, about 700 amino acids to about 2500 amino acids, about 700 amino acids to about 2000 amino acids, about 700 amino acids to Approximately 1500 amino acids, approximately 700 amino acids to approximately 1000 amino acids, approximately 700 amino acids to approximately 950 amino acids, approximately 700 amino acids to approximately 900 amino acids, approximately 700 amino acids to approximately 850 amino acids, approximately 700 amino acids to approximately 800 amino acids, approximately 700 amino acids to approximately 750 amino acids, approximately 750 amino acids to approximately 3000 amino acids, approximately 750 amino acids to approximately 2500 amino acids, approximately 750 amino acids to approximately 2000 amino acids, approximately 750 amino acids to approximately 1500 amino acids, approximately 750 amino acids to approximately 1000 amino acids, approximately 750 amino acids to approximately 950 amino acids, approximately 750 amino acids to approximately 900 amino acids.Approximately 750 amino acids to approximately 850 amino acids, approximately 750 amino acids to approximately 800 amino acids, approximately 800 amino acids to approximately 3000 amino acids, approximately 800 amino acids to approximately 2500 amino acids, approximately 800 amino acids to approximately 2000 amino acids, approximately 800 amino acids to approximately 1500 amino acids, approximately 800 amino acids to approximately 1000 amino acids, approximately 800 amino acids to approximately 950 amino acids, approximately 800 amino acids to approximately 900 amino acids, approximately 800 amino acids to approximately 850 amino acids. Amino acids, approximately 850 amino acids to approximately 3000 amino acids, approximately 850 amino acids to approximately 2500 amino acids, approximately 850 amino acids to approximately 2000 amino acids, approximately 850 amino acids to approximately 1500 amino acids, approximately 850 amino acids to approximately 1000 amino acids, approximately 850 amino acids to approximately 950 amino acids, approximately 850 amino acids to approximately 900 amino acids, approximately 900 amino acids to approximately 3000 amino acids, approximately 900 amino acids to approximately 2500 amino acids, approximately 900 amino acids From approximately 2000 amino acids, from approximately 900 amino acids to approximately 1500 amino acids, from approximately 900 amino acids to approximately 1000 amino acids, from approximately 900 amino acids to approximately 950 amino acids, from approximately 950 amino acids to approximately 3000 amino acids, from approximately 950 amino acids to approximately 2500 amino acids, from approximately 950 amino acids to approximately 2000 amino acids, from approximately 950 amino acids to approximately 1500 amino acids, from approximately 950 amino acids to approximately 1000 amino acids, from approximately 1000 amino acids to approximately 3000 amino acids Approximately 1000 amino acids to approximately 2500 amino acids, approximately 1000 amino acids to approximately 2000 amino acids, approximately 1000 amino acids to approximately 1500 amino acids, approximately 1500 amino acids to approximately 3000 amino acids, approximately 1500 amino acids to approximately 2500 amino acids, approximately 1500 amino acids to approximately 2000 amino acids, approximately 2000 amino acids to approximately 3000 amino acids, approximately 2000 amino acids to approximately 2500 amino acids, or approximately 2500 amino acids to approximately 3000 amino acids. Figure 1 The figure depicts an exemplary single-chain chimeric polypeptide provided herein.

[0074] In some embodiments of any single-chain chimeric polypeptide described herein, a first target-binding domain (e.g., any exemplary target-binding domain described herein or known in the art) and a soluble tissue factor domain (e.g., any exemplary soluble tissue factor domain described herein) are directly adjacent to each other. In some embodiments of any single-chain chimeric polypeptide described herein, the single-chain chimeric polypeptide further includes a linker sequence (e.g., any exemplary linker sequence described herein or known in the art) between the first target-binding domain (e.g., any exemplary first target-binding domain described herein or known in the art) and the soluble tissue factor domain (e.g., any exemplary soluble tissue factor domain described herein). In some embodiments of any single-chain chimeric polypeptide described herein, a soluble tissue factor domain (e.g., any exemplary soluble tissue factor domain described herein) and a second target-binding domain (e.g., any exemplary target-binding domain described herein or known in the art) are directly adjacent to each other. In some embodiments of any single-chain chimeric polypeptide described herein, the single-chain chimeric polypeptide further comprises a linker sequence (e.g., any exemplary soluble tissue factor domain described herein) between a soluble tissue factor domain (e.g., any exemplary soluble tissue factor domain described herein) and a second target-binding domain (e.g., any exemplary target-binding domain described herein or known in the art).

[0075] In some embodiments of any single-chain chimeric polypeptide described herein, a first target-binding domain (e.g., any exemplary target-binding domain described herein or known in the art) and a second target-binding domain (e.g., any exemplary target-binding domain described herein or known in the art) are directly adjacent to each other. In some embodiments of any single-chain chimeric polypeptide described herein, the single-chain chimeric polypeptide further includes a linker sequence (e.g., any exemplary linker sequence described herein or known in the art) between the first target-binding domain (e.g., any exemplary target-binding domain described herein or known in the art) and the second target-binding domain (e.g., any exemplary target-binding domain described herein or known in the art). In some embodiments of any single-chain chimeric polypeptide described herein…

[0076] The second target-binding domain (e.g., any exemplary target-binding domain described herein or known in the art) and the soluble tissue factor domain (e.g., any exemplary soluble tissue factor domain described herein) are directly adjacent to each other. In some embodiments of any single-chain chimeric polypeptide described herein, the single-chain chimeric polypeptide further includes a linker sequence (e.g., any exemplary linker sequence described herein or known in the art) between the second target-binding domain (e.g., any exemplary target-binding domain described herein or known in the art) and the soluble tissue factor domain (e.g., any exemplary soluble tissue factor domain described herein or known in the art).

[0077] In some embodiments, the single-chain chimeric polypeptide may include a sequence that is at least 70% identical to (e.g., at least 75% identical, at least 80% identical, at least 85% identical, at least 90% identical, at least 95% identical, at least 99% identical, or 100% identical).

[0078] QIVLTQSPAIMSASPGEKVTMTCSASSSVSYMNWYQQKSGTSPKRWIYDTSKLASGVPAHFRGSGSGTSYSLTISGMEAEDAATYYCQQWSSNPFTFGSGTKLEINRGGGGSGGGGSGGGGSQVQLQQSGAELARPGASVKMSCKASGYTFTRYTMHWVKQRPGQGLEWIGYINPSRGYTNYNQKFKDKATLTTDKSSSTAYMQLSSLTSEDSAVYYCARYYDDHYCLDYWGQGTTLTVSSSGTTNTVAAYNLTWKSTNFKTILEWEPKPVNQVYTVQISTKSGDWKSKCFYTTDTECDLTDEIVKDVKQTYLARVFSYPAGNVESTGSAGEPLYENSPEFTPYLETNLGQPTIQSFEQVGTKVNVTVEDERTLVRRNNTFLSLRDVFGKDLIYTLYYWKSSSSGKKTAKTNTNEFLIDVDKGENYCFSVQAVIPSRTVNRKSTDSPVECMGQEKGEFREVQLQQSGPELVKPGASVKMSCKASGYTFTSYVIQWVKQKPGQGLEWIGSINPYNDYTKYNEKFKGKATLTSDKSSITAYMEFSSLTSEDSALYYCARWGDGNYWGRGTTLTVSSGGGGSGGGGSGGGGSDIEMTQSPAIMSASLGERVTMTCTASSSVSSSYFHWYQQKPGSSPKLCIYSTSNLASGVPPRFSGSGSTSYSLTISSMEAEDAATYFCHQYHRSPTFGGGTKLETKR(SEQ ID NO:1)。

[0079]

[0080] In some embodiments, the single-chain chimeric polypeptide may include a sequence that is at least 70% identical to (e.g., at least 75% identical, at least 80% identical, at least 85% identical, at least 90% identical, at least 95% identical, at least 99% identical, or 100% identical).

[0081] (SEQ ID NO:3).

[0082] In some embodiments, the single-chain chimeric polypeptide is encoded by a nucleic acid comprising a sequence that is at least 70% identical to (e.g., at least 75% identical, at least 80% identical, at least 85% identical, at least 90% identical, at least 95% identical, at least 99% identical, or 100% identical).

[0083]

[0084] In some embodiments, the single-chain chimeric polypeptide may include a sequence that is at least 70% identical to (e.g., at least 75% identical, at least 80% identical, at least 85% identical, at least 90% identical, at least 95% identical, at least 99% identical, or 100% identical).

[0085] (SEQ ID NO:5).

[0086] In some embodiments, the single-chain chimeric polypeptide is encoded by a nucleic acid comprising a sequence that is at least 70% identical to (e.g., at least 75% identical, at least 80% identical, at least 85% identical, at least 90% identical, at least 95% identical, at least 99% identical, or 100% identical).

[0087]

[0088] In some embodiments, the single-chain chimeric polypeptide may include a sequence that is at least 70% identical to (e.g., at least 75% identical, at least 80% identical, at least 85% identical, at least 90% identical, at least 95% identical, at least 99% identical, or 100% identical).

[0089] (SEQ ID NO:7).

[0090] In some embodiments, the single-chain chimeric polypeptide is encoded by a nucleic acid comprising a sequence that is at least 70% identical to (e.g., at least 75% identical, at least 80% identical, at least 85% identical, at least 90% identical, at least 95% identical, at least 99% identical, or 100% identical).

[0091]

[0092] Non-limiting aspects of these chimeric peptides, nucleic acids, vectors, cells, and methods are described below and can be used in any combination without limitation. Further aspects of these chimeric peptides, nucleic acids, vectors, cells, and methods are known in the art.

[0093] Organization Factor

[0094] Human tissue factor (TGF) 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 C-terminal tail (residues 242-263) (UniProtKB identifier: P13726). The cytoplasmic tail contains two phosphorylation sites at Ser253 and Ser258 and one S-palmitoylation site at Cys245. No deletions or mutations in the cytoplasmic domains were found to affect TGF's coagulation activity. TGF has an S-palmitoylation site at Cys245 in its intracellular domain. Cys245 is located at the amino acid terminus of the intracellular domain and is close to the membrane surface. The TGF transmembrane domain consists of a single-span α-helix.

[0095] The extracellular domain of tissue factor, composed of two fibronectin type III domains, is linked to the transmembrane domain via a six-amino acid linker. This linker provides conformational flexibility to uncouple the extracellular domain of tissue factor from its transmembrane and cytoplasmic domains. Each tissue factor fibronectin type III module consists of two overlapping β-sheets, with the top fold containing three antiparallel β-chains and the bottom fold containing four β-chains. β-chains βA and βB, βC and βD, and βE and βF are linked by β-loops, all of which have a conserved conformation in both modules. Three short α-helical segments connect the β-chains. A unique feature of tissue factor is the 17-amino acid β-hairpin between β10 and β11, which is not a common component of the fibronectin superfamily. The N-terminal domain also contains a 12-amino acid loop between β6F and β7G, a loop not present in the C-terminal domain and specific to tissue factor. This fibronectin type III domain structure is characteristic of immunoglobulin-like family protein folding and is conserved in a wide variety of extracellular proteins.

[0096] Once proenzyme FVII binds to tissue to form an active tissue factor-FVIIa complex, it can be rapidly converted to FVIIa through limited proteolysis. FVIIa, circulating at a concentration of approximately 0.1 nM (1% of plasma FVII), can also directly bind to tissue factor. The allosteric interaction between tissue factor and FVIIa on the tissue factor-FVIIa complex significantly enhances the enzymatic activity of FVIIa: the hydrolysis rate of small chromophore substrates increases by approximately 20 to 100 times, and the activation rate of natural macromolecular substrates FIX and FX increases by nearly one million times. Consistent 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., when phosphatidyl-L-serine is exposed on the membrane surface) via Ca2+... 2+ The tissue factor-FVIIa-phospholipid complex increases the rate of FIX or FX activation by 1000-fold in a dependent manner. The approximately million-fold overall increase in FX activation by the tissue factor-FVIIa-phospholipid complex relative to free FVIIa is a key regulator of the coagulation cascade.

[0097] FVII is a single-chain polypeptide of approximately 50 kDa, composed of 406 amino acid residues, featuring an N-terminal γ-carboxyglutamate-rich domain, two epidermal growth factor-like domains (EGF1 and EFG2), and a C-terminal serine protease domain. FVII communicates with the EGF2 protease domain via an Ile- 154 -Arg 152 The specific protein hydrolysis of the bond activates it to FVIIa. This cleavage causes the light and heavy chains to pass through Cys... 135 and Cys 262 The individual disulfide bonds hold them together. FVIIa is held together by its N-terminal GLA domain via Ca 2+ It binds to the phospholipid membrane in a dependent manner. The C-terminus of the GLA domain is adjacent to the aromatic stack and two EGF domains. The aromatic stack connects the GLA to the binding of a single Ca2+ domain. 2+ The EGF1 domain of the ion. Occupying this Ca 2+ The binding site increases the amide degradation activity of FVIIa and tissue factor association. The catalytic triad is composed of His... 193 Asp 242 and Ser 344 Composition, and a single Ca within the FVIIa protease domain 2+ The binding of ions is crucial to their catalytic activity. FVII activates FVIIa through proteolytic activity, releasing Ile. 153 The newly formed amino terminus is folded back and inserted into the activation bag, along with Asp 343The carboxylate forms a salt bridge to create an oxyanion pore. This salt bridge formation is crucial for FVIIa activity. However, after proteolytic activation, the formation of oxyanion pores does not occur in free FVIIa. As a result, FVIIa cycles in its enzyme-as-an enzyme state, which plasma protease inhibitors do not readily recognize, allowing it to cycle with a half-life of approximately 90 minutes.

[0098] The tissue factor-mediated localization of the FVIIa active site above the membrane surface is important for FVIIa orientation toward homologous substrates. When free FVIIa binds to the membrane, its active site is located approximately above the membrane surface. It adopts a stable extended structure. After FVIIa binds to tissue factors, the FVa active site relocates to a position closer to the membrane. This regulation helps the FVIIa catalytic triad to properly align with the target substrate cleavage site. Using FVIIa without the GLA domain, the active site has been shown to remain at a similar distance above the membrane, demonstrating that tissue factor can fully support the localization of the FVIIa active site even in the absence of FVIIa-membrane interactions. Further data show that tissue factor supports full FVIIa proteolytic activity as long as its extracellular domain is somehow tethered to the membrane surface. However, elevating the FVIIa active site above the membrane surface requires more than [a certain amount of time / effort]. It will greatly reduce the ability of the tissue factor-FVIIa complex to activate FX, but will not reduce the amide hydrolysis activity of tissue factor-FVIIa.

[0099] Alanine scanning mutagenesis has been used to assess the role of specific amino acid side chains in the extracellular domain of tissue factor in their interaction with FVIIa (Gibbs et al., Biochemistry 33(47):14003-14010, 1994; Schullek et al., Journal of Biochemistry 269(30):19399-19403, 1994). Alanine substitution identified a limited number of residue sites where alanine substitution resulted in a 5- to 10-fold decrease in affinity for FVIIa binding. Most of these residue side chains were found to be well exposed to the solvent in the crystal structure and consistent with interactions with macromolecular ligands. The FVIIa ligand binding sites were located over a broad region at the boundary between the two modules. In module C, the Arg residue located on the prominent BC ring was... 135 and Phe 140 Provides independent contact with FVIIa. Leu 133 Located at the base of the finger-like structure, and squeezed into the crack between the two modules. This is provided by Lys 20 Thr 60 Asp 58and Ile 22 The continuity of the main clusters of important binding residues. Thr 60 It is only partially exposed to the solvent and may play a localized structural role rather than having significant contact with the ligand. The binding site extends to the concave surface of the intermodal corner, which involves Glu. 24 and Gln 110 And may involve more distant residues Val 207 The binding region from Asp 58 Extended to Lys 48 Lys 46 、Gln 37 Asp 44 and Trp 45 The resulting convex surface region. Trp 45 and Asp 44 It does not interact independently with FVIIa, indicating that Trp 45 The mutation effect at this location may reflect the effect of this side chain on neighboring Asp. 44 and Gln 37 The structural importance of localized side chain stacking. The interaction region further includes two surface-exposed aromatic residues, Phe 76 and Tyr 78 It forms part of the hydrophobic cluster in the N module.

[0100] The known physiological substrates of tissue factor-FVIIa are FVII, FIX, and FX, as well as receptors activated by certain proteases. Mutation analysis has identified numerous residues that, when mutated, support full FVIIa amideolytic activity against small peptide substrates but lack the ability to support activation against macromolecular substrates (i.e., FVII, FIX, and FX) (Ruf et al., Journal of Biochemistry 267(31):22206-22210, 1992; Ruf et al., Journal of Biochemistry 267(9):6375-6381, 1992; Huang et al., Journal of Biochemistry 271(36):21752-21757, 1996; Kirchhofer et al., Biochemistry 39(25):7380-7387, 2000). The tissue factor loop region at residues 159-165 and residues in or near this flexible loop have been shown to be crucial for the proteolytic activity of the tissue factor-FVIIa complex. This defines the extra-substrate binding site region of the proposed tissue factor, which is located far from the active site of FVIIa. Substitution of glycine residues with the slightly larger alanine residue significantly impairs the proteolytic activity of tissue factor-FVIIa. This suggests that the flexibility provided by glycine is crucial for the loop at residues 159-165 used for macromolecular substrate recognition by tissue factor.

[0101] It also proved the residue Lys 165 and Lys 166 This is important for substrate recognition and binding. Mutations of any of these residues to alanine result in a significant reduction in the function of tissue factor cofactors. In most tissue factor-FVIIa structures, Lys 165 and Lys 166 Back to back, Lys 165 Pointing to FVIIa, while Lys 166 Pointing to the region outside the substrate binding site in the crystal structure. Lys of FVIIa 165 and Gla 35 The proposed salt bridge formation between them would support the idea that the interaction between tissue factor and the GLA domain of FVIIa regulates substrate recognition. These results suggest that the C-terminal portion of the extracellular domain of tissue factor interacts directly with the GLA domains of FIX and FX, and possibly the neighboring EGF1 domain, and that the presence of the FVIIa GLA domain can directly or indirectly modulate these interactions.

[0102] Soluble tissue factor domain

[0103] In some embodiments of any of the single-chain chimeric peptides, compositions, or methods described herein, the soluble tissue factor domain may be a wild-type tissue factor peptide lacking a signal sequence, transmembrane domain, and intracellular domain. In some instances, the soluble tissue factor domain may be a tissue factor mutant, wherein the wild-type tissue factor peptide lacks a signal sequence, transmembrane domain, and intracellular domain, and has been further modified at selected amino acids. In some instances, the soluble tissue factor domain may be a human soluble tissue factor domain. In some instances, the soluble tissue factor domain may be a mouse soluble tissue factor domain. In some instances, the soluble tissue factor domain may be a rat soluble tissue factor domain. Non-limiting examples of human soluble tissue factor domains, mouse soluble tissue factor domains, rat soluble tissue factor domains, and mutant soluble tissue factor domains are shown below. Exemplary human soluble tissue factor domain (SEQ ID NO: 9)

[0104] SGTTNTVAAYNLTWKSTNFKTILEWEPKPVNQVYTVQISTKSGDWKSKCFYTTDTECDLTDEIVKDVKQTYLARVFSYPAGNVESTGSAGEPLYENSPEFTPYLETNLG QPTIQSFEQVGTKVNVTVEDERTLVRRNNTFLSLRDVFGKDLIYTLYYWKSSSSGKKTAKTNTNEFLIDVDKGENYCFSVQAVIPSRTVNRKSTDSPVECMGQEKGEFRE

[0105] Exemplary nucleic acid encoding the human soluble tissue factor domain (SEQ ID NO:10)

[0106] AGCGGCACAACCAACACAGTCGCTGCCTATAACCTCACTTGGAAGAGCACCAACTTCAAAACCATCCTCGAATGGGAACCCAAACCCGTTAACCAAGTTTACACCGTGCAGATCAGCACCAAGTCCGGCGACTGGAAGTCCAAATGTTTCTATACCACCGACACCGAGTGCGATCTCACCGATGAGATCGTGAAAGATGTGAAACAGACCTACCTCGCCCGGGTGTTTAGCTACCCCGCCGGCAATGTGGAGAGCACTGGTTCCGCTGGCGAGCCTTTATACGAGAACAGCCCCGAATTTACCCCTTACCTCGAGACCAATTTAGGACAGCCCACCATCCAAAGCTTTGAGCAAGTTGGCACAAAGGTGAATGTGACAGTGGAGGACGAGCGGACTTTAGTGCGGCGGAACAACACCTTTCTCAGCCTCCGGGATGTGTTCGGCAAAGATTTAATCTACACACTGTATTACTGGAAGTCCTCTTCCTCCGGCAAGAAGACAGCTAAAACCAACACAAACGAGTTTTTAATCGACGTGGATAAAGGCGAAAACTACTGTTTCAGCGTGCAAGCTGTGATCCCCTCCCGGACCGTGAATAGGAAAAGCACCGATAGCCCCGTTGAGTGCATGGGCCAAGAAAAGGGCGAGTTCCGGGAG

[0107] Exemplary mouse soluble tissue factor domain (SEQ ID NO:11)

[0108] agipekafnltwistdfktilewqpkptnytytvqisdrsrnwknkcfsttdtecdltdeivkdvtwayeakvlsvprrnsvhgdgdqlvihgeeppftnapkflpyrdtn lgqpviqqfeqdgrklnvvvkdsltlvrkngtfltlrqvfgkdlgyiityrkgsstgkktnitntnefsidveegvsycffvqamifsrktnqnspgsstvcteqwksflge

[0109] Exemplary rat soluble tissue factor domain (SEQ ID NO:12)

[0110] Agtppgkafnltwistdfktilewqpkptnytytvqisdrsrnwkykctgttdtecdltdeivkdvnwtyearvlsvpwrnsthgketlfgthgeeppftnarkflpyrdtk igqpviqkyeqggtklkvtvkdsftlvrkngtfltlrqvfgndlgyiltyrkdsstgrktntthtneflidvekgvsycffaqavifsrktnhkspesitkcteqwksvlge

[0111] Exemplary mutant human soluble tissue factor domain (SEQ ID NO:96)

[0112] SGTTNTVAAYNLTWKSTNFATALEWEPKPVNQVYTVQISTKSGDWKSKCFYTTDTECALTDEIVKDVKQTYLARVFSYPAGNVESTGSAGEPLYENSPEFTPYLETNLG QPTIQSFEQVGTKVNVTVEDERTLVARNNTALSLRDVFGKDLIYTLYYWKSSSSGKKTAKTNTNEFLIDVDKGENYCFSVQAVIPSRTVNRKSTDSPVECMGQEKGEFRE

[0113] Exemplary mutant human soluble tissue factor domain (SEQ ID NO:97)

[0114] SGTTNTVAAYNLTWKSTNFATALEWEPKPVNQVYTVQISTKSGDAKSKCFYTTDTECALTDEIVKDVKQTYLARVFSYPAGNVESTGSAGEPLAENSPEFTPYLETNLG QPTIQSFEQVGTKVNVTVEDERTLVARNNTALSLRDVFGKDLIYTLYYWKSSSSGKKTAKTNTNEFLIDVDKGENYCFSVQAVIPSRTVNRKSTDSPVECMGQEKGEFRE

[0115] In some embodiments, the soluble tissue factor domain may include a sequence that is at least 70% consistent with, at least 72% consistent with, at least 74% consistent with, at least 76% consistent with, at least 78% consistent with, at least 80% consistent with, at least 82% consistent with, at least 84% consistent with, at least 86% consistent with, at least 88% consistent with, at least 90% consistent with, at least 92% consistent with, at least 94% consistent with, at least 96% consistent with, at least 98% consistent with, at least 99% consistent with, or 100% consistent with SEQ ID NO: 9, 11, 12, 96 or 97. In some embodiments, the soluble tissue factor domain may include the sequence SEQ ID NO: 9, 11, 12, 96 or 97, wherein one to twenty 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) have been removed from its N-terminus and / or one to twenty 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) have been removed from its C-terminus.

[0116] As is understood in the art, those skilled in the art will appreciate that mutations in conserved amino acids across different mammalian species are more likely to reduce protein activity and / or structural stability, while mutations in non-conserved amino acids across different mammalian species are less likely to reduce protein activity and / or structural stability.

[0117] In some examples of any of the single-chain chimeric peptides described herein, the soluble tissue factor domain cannot bind to factor VIIa. In some examples of any of the single-chain chimeric peptides described herein, the soluble tissue factor domain does not convert inactive factor X into factor Xa. In some embodiments of any of the single-chain chimeric peptides described herein, the single-chain chimeric peptide does not stimulate blood clotting in mammals.

[0118] In some instances, the soluble tissue factor domain may be the human soluble tissue factor domain. In some embodiments, the soluble tissue factor domain may be the mouse soluble tissue factor domain. In some embodiments, the soluble tissue factor domain may be the rat soluble tissue factor domain.

[0119] In some instances, the soluble tissue factor domain does not include one or more of the following (e.g., two, three, four, five, six, or seven): lysine at the amino acid position corresponding to amino acid position 20 of the human mature wild-type tissue factor protein; isoleucine at the amino acid position corresponding to amino acid position 22 of the human mature wild-type tissue factor protein; tryptophan at the amino acid position corresponding to amino acid position 45 of the human mature wild-type tissue factor protein; aspartic acid at the amino acid position corresponding to amino acid position 58 of the human mature wild-type tissue factor protein; tyrosine at the amino acid position corresponding to amino acid position 94 of the human mature wild-type tissue factor protein; arginine at the amino acid position corresponding to amino acid position 135 of the human mature wild-type tissue factor protein; and phenylalanine at the amino acid position corresponding to amino acid position 140 of the human mature wild-type tissue factor protein. In some embodiments, the mutant soluble tissue factor has the amino acid sequence of SEQ ID NO:96 or SEQ ID NO:97.

[0120] In some instances, the soluble tissue factor domain may be encoded by nucleic acids comprising sequences that are 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:10.

[0121] In some embodiments, the soluble tissue factor domain may have the following total lengths: about 20 amino acids to about 220 amino acids, about 20 amino acids to about 215 amino acids, about 20 amino acids to about 210 amino acids, about 20 amino acids to about 205 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... 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 2 0 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 60 amino acids, about 20 amino acids to about 50 amino acids, about 20 amino acids to about 40 amino acids, about 20 amino acids to about 30 amino acids, about 30 amino acids to about 220 amino acids, about 30 amino acids to about 215 amino acids, about 30 amino acids to about 210 amino acids, about 30 amino acids to about 205 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 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.Approximately 30 amino acids to approximately 105 amino acids, approximately 30 amino acids to approximately 100 amino acids, approximately 30 amino acids to approximately 95 amino acids, approximately 30 amino acids to approximately 90 amino acids, approximately 30 amino acids to approximately 85 amino acids, approximately 30 amino acids to approximately 80 amino acids, approximately 30 amino acids to approximately 75 amino acids, approximately 30 amino acids to approximately 70 amino acids, approximately 30 amino acids to approximately 60 amino acids, approximately 30 amino acids to approximately 50 amino acids, approximately 30 amino acids to approximately 40 amino acids, approximately 40 amino acids to approximately 220 amino acids, approximately 40 amino acids to approximately 215 amino acids, approximately 40 amino acids to approximately 210 amino acids, approximately 40 amino acids to approximately 205 amino acids. Approximately 40 amino acids to approximately 200 amino acids, approximately 40 amino acids to approximately 195 amino acids, approximately 40 amino acids to approximately 190 amino acids, approximately 40 amino acids to approximately 185 amino acids, approximately 40 amino acids to approximately 180 amino acids, approximately 40 amino acids to approximately 175 amino acids, approximately 40 amino acids to approximately 170 amino acids, approximately 40 amino acids to approximately 165 amino acids, approximately 40 amino acids to approximately 160 amino acids, approximately 40 amino acids to approximately 155 amino acids, approximately 40 amino acids to approximately 150 amino acids, approximately 40 amino acids to approximately 145 amino acids, approximately 40 amino acids to approximately 140 amino acids, approximately 40 amino acids to approximately 135 amino acids, approximately 40 amino acids to approximately 130 amino acids, approximately 40 amino acids to approximately 125 amino acids, approximately 40 amino acids to approximately 120 amino acids, approximately 40 amino acids to approximately 115 amino acids, approximately 40 amino acids to approximately 110 amino acids, approximately 40 amino acids to approximately 105 amino acids, approximately 40 amino acids to approximately 100 amino acids, approximately 40 amino acids to approximately 95 amino acids, approximately 40 amino acids to approximately 90 amino acids, approximately 40 amino acids to approximately 85 amino acids, approximately 40 amino acids to approximately 80 amino acids, approximately 40 amino acids to approximately 75 amino acids, approximately 40 amino acids to approximately 70 amino acids, approximately 40 amino acids to approximately 60 amino acids, approximately 40 amino acids to approximately 50 amino acids, approximately 50 amino acids to Approximately 220 amino acids, approximately 50 amino acids to approximately 215 amino acids, approximately 50 amino acids to approximately 210 amino acids, approximately 50 amino acids to approximately 205 amino acids, approximately 50 amino acids to approximately 200 amino acids, approximately 50 amino acids to approximately 195 amino acids, approximately 50 amino acids to approximately 190 amino acids, approximately 50 amino acids to approximately 185 amino acids, approximately 50 amino acids to approximately 180 amino acids, approximately 50 amino acids to approximately 175 amino acids, approximately 50 amino acids to approximately 170 amino acids, approximately 50 amino acids to approximately 165 amino acids, approximately 50 amino acids to approximately 160 amino acids, approximately 50 amino acids to approximately 155 amino acids, approximately 50 amino acids to approximately 150 amino acids.Approximately 50 amino acids to approximately 145 amino acids, approximately 50 amino acids to approximately 140 amino acids, approximately 50 amino acids to approximately 135 amino acids, approximately 50 amino acids to approximately 130 amino acids, approximately 50 amino acids to approximately 125 amino acids, approximately 50 amino acids to approximately 120 amino acids, approximately 50 amino acids to approximately 115 amino acids, approximately 50 amino acids to approximately 110 amino acids, approximately 50 amino acids to approximately 105 amino acids, approximately 50 amino acids to approximately 100 amino acids, approximately 50 amino acids to approximately 95 amino acids, approximately 50 amino acids to approximately 90 amino acids, approximately 50 amino acids to approximately 85 amino acids, approximately 50 amino acids to approximately 80 amino acids, approximately 50 amino acids to approximately 75 amino acids. Amino acids, approximately 50 amino acids to approximately 70 amino acids, approximately 50 amino acids to approximately 60 amino acids, approximately 60 amino acids to approximately 220 amino acids, approximately 60 amino acids to approximately 215 amino acids, approximately 60 amino acids to approximately 210 amino acids, approximately 60 amino acids to approximately 205 amino acids, approximately 60 amino acids to approximately 200 amino acids, approximately 60 amino acids to approximately 195 amino acids, approximately 60 amino acids to approximately 190 amino acids, approximately 60 amino acids to approximately 185 amino acids, approximately 60 amino acids to approximately 180 amino acids, approximately 60 amino acids to approximately 175 amino acids, approximately 60 amino acids to approximately 170 amino acids, approximately 60 amino acids to approximately 165 amino acids, approximately 60 amino acids to approximately 160 amino acids, approximately 60 amino acids to approximately 155 amino acids, approximately 60 amino acids to approximately 150 amino acids, approximately 60 amino acids to approximately 145 amino acids, approximately 60 amino acids to approximately 140 amino acids, approximately 60 amino acids to approximately 135 amino acids, approximately 60 amino acids to approximately 130 amino acids, approximately 60 amino acids to approximately 125 amino acids, approximately 60 amino acids to approximately 120 amino acids, approximately 60 amino acids to approximately 115 amino acids, approximately 60 amino acids to approximately 110 amino acids, approximately 60 amino acids to approximately 105 amino acids, approximately 60 amino acids to approximately 100 amino acids, approximately 60 amino acids to approximately 95 amino acids, approximately 60 amino acids to approximately 90 amino acids, approximately 60 From approximately 85 amino acids, approximately 60 amino acids to approximately 80 amino acids, approximately 60 amino acids to approximately 75 amino acids, approximately 60 amino acids to approximately 70 amino acids, approximately 70 amino acids to approximately 220 amino acids, approximately 70 amino acids to approximately 215 amino acids, approximately 70 amino acids to approximately 210 amino acids, approximately 70 amino acids to approximately 205 amino acids, approximately 70 amino acids to approximately 200 amino acids, approximately 70 amino acids to approximately 195 amino acids, approximately 70 amino acids to approximately 190 amino acids, approximately 70 amino acids to approximately 185 amino acids, approximately 70 amino acids to approximately 180 amino acids, approximately 70 amino acids to approximately 175 amino acids, approximately 70 amino acids to approximately 170 amino acids.Approximately 70 amino acids to approximately 165 amino acids, approximately 70 amino acids to approximately 160 amino acids, approximately 70 amino acids to approximately 155 amino acids, approximately 70 amino acids to approximately 150 amino acids, approximately 70 amino acids to approximately 145 amino acids, approximately 70 amino acids to approximately 140 amino acids, approximately 70 amino acids to approximately 135 amino acids, approximately 70 amino acids to approximately 130 amino acids, approximately 70 amino acids to approximately 125 amino acids, approximately 70 amino acids to approximately 120 amino acids, approximately 70 amino acids to approximately 115 amino acids, approximately 70 amino acids to approximately 110 amino acids, approximately 70 amino acids to approximately 105 amino acids, approximately 70 amino acids to approximately 100 amino acids, approximately 70 amino acids to approximately 95 amino acids. 1 amino acid, approximately 70 amino acids to approximately 90 amino acids, approximately 70 amino acids to approximately 85 amino acids, approximately 70 amino acids to approximately 80 amino acids, approximately 80 amino acids to approximately 220 amino acids, approximately 80 amino acids to approximately 215 amino acids, approximately 80 amino acids to approximately 210 amino acids, approximately 80 amino acids to approximately 205 amino acids, approximately 80 amino acids to approximately 200 amino acids, approximately 80 amino acids to approximately 195 amino acids, approximately 80 amino acids to approximately 190 amino acids, approximately 80 amino acids to approximately 185 amino acids, approximately 80 amino acids to approximately 180 amino acids, approximately 80 amino acids to approximately 175 amino acids, approximately 80 amino acids to approximately 170 amino acids, approximately 80 amino acids to approximately 165 amino acids, approximately 80 amino acids to approximately 160 amino acids, approximately 80 amino acids to approximately 155 amino acids, approximately 80 amino acids to approximately 150 amino acids, approximately 80 amino acids to approximately 145 amino acids, approximately 80 amino acids to approximately 140 amino acids, approximately 80 amino acids to approximately 135 amino acids, approximately 80 amino acids to approximately 130 amino acids, approximately 80 amino acids to approximately 125 amino acids, approximately 80 amino acids to approximately 120 amino acids, approximately 80 amino acids to approximately 115 amino acids, approximately 80 amino acids to approximately 110 amino acids, approximately 80 amino acids to approximately 105 amino acids, approximately 80 amino acids to approximately 100 amino acids, approximately 80 amino acids to approximately 95 amino acids, approximately 80 Amino acids to about 90 amino acids, about 90 amino acids to about 220 amino acids, about 90 amino acids to about 215 amino acids, about 90 amino acids to about 210 amino acids, about 90 amino acids to about 205 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.Approximately 90 amino acids to approximately 150 amino acids, approximately 90 amino acids to approximately 145 amino acids, approximately 90 amino acids to approximately 140 amino acids, approximately 90 amino acids to approximately 135 amino acids, approximately 90 amino acids to approximately 130 amino acids, approximately 90 amino acids to approximately 125 amino acids, approximately 90 amino acids to approximately 120 amino acids, approximately 90 amino acids to approximately 115 amino acids, approximately 90 amino acids to approximately 110 amino acids, approximately 90 amino acids to approximately 105 amino acids, approximately 90 amino acids to approximately 100 amino acids, approximately 100 amino acids to approximately 220 amino acids, approximately 100 amino acids to approximately 215 amino acids, approximately 100 amino acids to approximately 210 amino acids, approximately 100 amino acids From approximately 205 amino acids, from approximately 100 amino acids to approximately 200 amino acids, from approximately 100 amino acids to approximately 195 amino acids, from approximately 100 amino acids to approximately 190 amino acids, from approximately 100 amino acids to approximately 185 amino acids, from approximately 100 amino acids to approximately 180 amino acids, from approximately 100 amino acids to approximately 175 amino acids, from approximately 100 amino acids to approximately 170 amino acids, from approximately 100 amino acids to approximately 165 amino acids, from approximately 100 amino acids to approximately 160 amino acids, from approximately 100 amino acids to approximately 155 amino acids, from approximately 100 amino acids to approximately 150 amino acids, from approximately 100 amino acids to approximately 145 amino acids, from approximately 100 amino acids to approximately 140 amino acids, from approximately 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 110 amino acids to about 220 amino acids, about 110 amino acids to about 215 amino acids, about 110 amino acids to about 210 amino acids, about 110 amino acids to about 205 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 1 From 10 amino acids to about 180 amino acids, from about 110 amino acids to about 175 amino acids, from about 110 amino acids to about 170 amino acids, from about 110 amino acids to about 165 amino acids, from about 110 amino acids to about 160 amino acids, from about 110 amino acids to about 155 amino acids, from about 110 amino acids to about 150 amino acids, from about 110 amino acids to about 145 amino acids, from about 110 amino acids to about 140 amino acids, from about 110 amino acids to about 135 amino acids, from about 110 amino acids to about 130 amino acids, from about 110 amino acids to about 125 amino acids, from about 110 amino acids to about 120 amino acids, from about 110 amino acids to about 115 amino acids.Approximately 115 amino acids to approximately 220 amino acids, approximately 115 amino acids to approximately 215 amino acids, approximately 115 amino acids to approximately 210 amino acids, approximately 115 amino acids to approximately 205 amino acids, approximately 115 amino acids to approximately 200 amino acids, approximately 115 amino acids to approximately 195 amino acids, approximately 115 amino acids to approximately 190 amino acids, approximately 115 amino acids to approximately 185 amino acids, approximately 115 amino acids to approximately 180 amino acids, approximately 115 amino acids to approximately 175 amino acids, approximately 115 amino acids to approximately 170 amino acids, approximately 115 amino acids to approximately 165 amino acids, approximately 115 amino acids to approximately 160 amino acids, approximately 115 amino acids to approximately 155 amino acids. 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 220 amino acids, about 120 amino acids to about 215 amino acids, about 120 amino acids to about 210 amino acids, about 120 amino acids to about 205 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 Approximately 190 amino acids, approximately 120 amino acids to approximately 185 amino acids, approximately 120 amino acids to approximately 180 amino acids, approximately 120 amino acids to approximately 175 amino acids, approximately 120 amino acids to approximately 170 amino acids, approximately 120 amino acids to approximately 165 amino acids, approximately 120 amino acids to approximately 160 amino acids, approximately 120 amino acids to approximately 155 amino acids, approximately 120 amino acids to approximately 150 amino acids, approximately 120 amino acids to approximately 145 amino acids, approximately 120 amino acids to approximately 140 amino acids, approximately 120 amino acids to approximately 135 amino acids, approximately 120 amino acids to approximately 130 amino acids, approximately 120 amino acids to approximately 125 amino acids, approximately 125 From approximately 125 amino acids to approximately 215 amino acids, from approximately 125 amino acids to approximately 210 amino acids, from approximately 125 amino acids to approximately 205 amino acids, from approximately 125 amino acids to approximately 200 amino acids, from approximately 125 amino acids to approximately 195 amino acids, from approximately 125 amino acids to approximately 190 amino acids, from approximately 125 amino acids to approximately 185 amino acids, from approximately 125 amino acids to approximately 180 amino acids, from approximately 125 amino acids to approximately 175 amino acids, from approximately 125 amino acids to approximately 170 amino acids, from approximately 125 amino acids to approximately 165 amino acids, from approximately 125 amino acids to approximately 160 amino acids, from approximately 125 amino acids to approximately 155 amino acids.Approximately 125 amino acids to approximately 150 amino acids, approximately 125 amino acids to approximately 145 amino acids, approximately 125 amino acids to approximately 140 amino acids, approximately 125 amino acids to approximately 135 amino acids, approximately 125 amino acids to approximately 130 amino acids, approximately 130 amino acids to approximately 220 amino acids, approximately 130 amino acids to approximately 215 amino acids, approximately 130 amino acids to approximately 210 amino acids, approximately 130 amino acids to approximately 205 amino acids, approximately 130 amino acids to approximately 200 amino acids, approximately 130 amino acids to approximately 195 amino acids, approximately 130 amino acids to approximately 190 amino acids, approximately 130 amino acids to approximately 185 amino acids, approximately 130 amino acids to approximately 180 130 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 220 amino acids, about 135 amino acids to about 215 amino acids, about 135 amino acids to about 210 amino acids, about 135 amino acids to about 205 amino acids, about 135 amino acids to Approximately 200 amino acids, approximately 135 amino acids to approximately 195 amino acids, approximately 135 amino acids to approximately 190 amino acids, approximately 135 amino acids to approximately 185 amino acids, approximately 135 amino acids to approximately 180 amino acids, approximately 135 amino acids to approximately 175 amino acids, approximately 135 amino acids to approximately 170 amino acids, approximately 135 amino acids to approximately 165 amino acids, approximately 135 amino acids to approximately 160 amino acids, approximately 135 amino acids to approximately 155 amino acids, approximately 135 amino acids to approximately 150 amino acids, approximately 135 amino acids to approximately 145 amino acids, approximately 135 amino acids to approximately 140 amino acids, approximately 140 amino acids to approximately 220 amino acids, approximately 140 From approximately 140 amino acids to approximately 215 amino acids, from approximately 140 amino acids to approximately 210 amino acids, from approximately 140 amino acids to approximately 205 amino acids, from approximately 140 amino acids to approximately 200 amino acids, from approximately 140 amino acids to approximately 195 amino acids, from approximately 140 amino acids to approximately 190 amino acids, from approximately 140 amino acids to approximately 185 amino acids, from approximately 140 amino acids to approximately 180 amino acids, from approximately 140 amino acids to approximately 175 amino acids, from approximately 140 amino acids to approximately 170 amino acids, from approximately 140 amino acids to approximately 165 amino acids, from approximately 140 amino acids to approximately 160 amino acids, from approximately 140 amino acids to approximately 155 amino acids, from approximately 140 amino acids to approximately 150 amino acids.Approximately 140 amino acids to approximately 145 amino acids, approximately 145 amino acids to approximately 220 amino acids, approximately 145 amino acids to approximately 215 amino acids, approximately 145 amino acids to approximately 210 amino acids, approximately 145 amino acids to approximately 205 amino acids, approximately 145 amino acids to approximately 200 amino acids, approximately 145 amino acids to approximately 195 amino acids, approximately 145 amino acids to approximately 190 amino acids, approximately 145 amino acids to approximately 185 amino acids, approximately 145 amino acids to approximately 180 amino acids, approximately 145 amino acids to approximately 175 amino acids, approximately 145 amino acids to approximately 170 amino acids, approximately 145 amino acids to approximately 165 amino acids, approximately 145 amino acids to approximately 160 145 amino acids to about 155 amino acids, about 145 amino acids to about 150 amino acids, about 150 amino acids to about 220 amino acids, about 150 amino acids to about 215 amino acids, about 150 amino acids to about 210 amino acids, about 150 amino acids to about 205 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 Approximately 165 amino acids, approximately 150 amino acids to approximately 160 amino acids, approximately 150 amino acids to approximately 155 amino acids, approximately 155 amino acids to approximately 220 amino acids, approximately 155 amino acids to approximately 215 amino acids, approximately 155 amino acids to approximately 210 amino acids, approximately 155 amino acids to approximately 205 amino acids, approximately 155 amino acids to approximately 200 amino acids, approximately 155 amino acids to approximately 195 amino acids, approximately 155 amino acids to approximately 190 amino acids, approximately 155 amino acids to approximately 185 amino acids, approximately 155 amino acids to approximately 180 amino acids, approximately 155 amino acids to approximately 175 amino acids, approximately 155 amino acids to approximately 170 amino acids, approximately 155 From approximately 165 amino acids, from approximately 155 amino acids to approximately 160 amino acids, from approximately 160 amino acids to approximately 220 amino acids, from approximately 160 amino acids to approximately 215 amino acids, from approximately 160 amino acids to approximately 210 amino acids, from approximately 160 amino acids to approximately 205 amino acids, from approximately 160 amino acids to approximately 200 amino acids, from approximately 160 amino acids to approximately 195 amino acids, from approximately 160 amino acids to approximately 190 amino acids, from approximately 160 amino acids to approximately 185 amino acids, from approximately 160 amino acids to approximately 180 amino acids, from approximately 160 amino acids to approximately 175 amino acids, from approximately 160 amino acids to approximately 170 amino acids, from approximately 160 amino acids to approximately 165 amino acids.Approximately 165 amino acids to approximately 220 amino acids, approximately 165 amino acids to approximately 215 amino acids, approximately 165 amino acids to approximately 210 amino acids, approximately 165 amino acids to approximately 205 amino acids, approximately 165 amino acids to approximately 200 amino acids, approximately 165 amino acids to approximately 195 amino acids, approximately 165 amino acids to approximately 190 amino acids, approximately 165 amino acids to approximately 185 amino acids, approximately 165 amino acids to approximately 180 amino acids, approximately 165 amino acids to approximately 175 amino acids, approximately 165 amino acids to approximately 170 amino acids, approximately 170 amino acids to approximately 220 amino acids, approximately 170 amino acids to approximately 215 amino acids, approximately 170 amino acids to approximately 210 170 amino acids to about 205 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 220 amino acids, about 175 amino acids to about 215 amino acids, about 175 amino acids to about 210 amino acids, about 175 amino acids to about 205 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 Approximately 190 amino acids, approximately 175 amino acids to approximately 185 amino acids, approximately 175 amino acids to approximately 180 amino acids, approximately 180 amino acids to approximately 220 amino acids, approximately 180 amino acids to approximately 215 amino acids, approximately 180 amino acids to approximately 210 amino acids, approximately 180 amino acids to approximately 205 amino acids, approximately 180 amino acids to approximately 200 amino acids, approximately 180 amino acids to approximately 195 amino acids, approximately 180 amino acids to approximately 190 amino acids, approximately 180 amino acids to approximately 185 amino acids, approximately 185 amino acids to approximately 220 amino acids, approximately 185 amino acids to approximately 215 amino acids, approximately 185 amino acids to approximately 210 amino acids, approximately 185 From approximately 185 amino acids to approximately 200 amino acids, from approximately 185 amino acids to approximately 195 amino acids, from approximately 185 amino acids to approximately 190 amino acids, from approximately 190 amino acids to approximately 220 amino acids, from approximately 190 amino acids to approximately 215 amino acids, from approximately 190 amino acids to approximately 210 amino acids, from approximately 190 amino acids to approximately 205 amino acids, from approximately 190 amino acids to approximately 200 amino acids, from approximately 190 amino acids to approximately 195 amino acids, from approximately 195 amino acids to approximately 220 amino acids, from approximately 195 amino acids to approximately 215 amino acids, from approximately 195 amino acids to approximately 210 amino acids, from approximately 195 amino acids to approximately 205 amino acids.Approximately 195 amino acids to approximately 200 amino acids, approximately 200 amino acids to approximately 220 amino acids, approximately 200 amino acids to approximately 215 amino acids, approximately 200 amino acids to approximately 210 amino acids, approximately 200 amino acids to approximately 205 amino acids, approximately 205 amino acids to approximately 220 amino acids, approximately 205 amino acids to approximately 215 amino acids, approximately 205 amino acids to approximately 210 amino acids, approximately 210 amino acids to approximately 220 amino acids, approximately 210 amino acids to approximately 215 amino acids, or approximately 215 amino acids to approximately 220 amino acids.

[0122] In some embodiments, the soluble tissue factor domain may include or consist of human soluble wild-type tissue factor (or any sequence thereof).

[0123] Connector sequence

[0124] In some embodiments, the adapter sequence may be a flexible adapter sequence. Non-limiting examples of adapter sequences that can be used are described in Klein et al., Protein Engineering, Design & Selection, Vol. 27, No. 10, pp. 325-330, 2014; Priyanka et al., Protein Science, Feb. 2013; 22(2):153-167. In some instances, the adapter sequence is a synthetic adapter sequence.

[0125] In some embodiments, any single-chain chimeric polypeptide described herein may include one, two, three, four, five, six, seven, eight, nine, or ten adapter sequences (e.g., the same or different adapter sequences, such as any exemplary adapter sequence described herein or known in the art).

[0126] In some embodiments, the linker sequence may have the following total lengths: 1 amino acid to about 100 amino acids, 1 amino acid to about 90 amino acids, 1 amino acid to about 80 amino acids, 1 amino acid to about 70 amino acids, 1 amino acid to about 60 amino acids, 1 amino acid to about 50 amino acids, 1 amino acid to about 45 amino acids, 1 amino acid to about 40 amino acids, 1 amino acid to about 35 amino acids, 1 amino acid to about 30 amino acids, 1 amino acid to about 25 amino acids, 1 amino acid to about 24 amino acids, 1 amino acid to about 22 amino acids, 1 amino acid to about 20 amino acids, 1 amino acid to about 18 amino acids, 1 amino acid to about 16 amino acids, 1 amino acid... The amino acids consist of approximately 14 amino acids; 1 amino acid consists of approximately 12 amino acids; 1 amino acid consists of approximately 10 amino acids; 1 amino acid consists of approximately 8 amino acids; 1 amino acid consists of approximately 6 amino acids; 1 amino acid consists of approximately 4 amino acids; approximately 2 amino acids consist of approximately 100 amino acids; approximately 2 amino acids consist of approximately 90 amino acids; approximately 2 amino acids consist of approximately 80 amino acids; approximately 2 amino acids consist of approximately 70 amino acids; approximately 2 amino acids consist of approximately 60 amino acids; approximately 2 amino acids consist of approximately 50 amino acids; approximately 2 amino acids consist of approximately 45 amino acids; approximately 2 amino acids consist of approximately 40 amino acids; approximately 2 amino acids consist of approximately 35 amino acids; approximately 2 amino acids consist of approximately 30 amino acids; approximately 2 amino acids consist of approximately 25 amino acids; approximately 2 amino acids consist of approximately 2 amino acids. From about 24 amino acids, from about 2 amino acids to about 22 amino acids, from about 2 amino acids to about 20 amino acids, from about 2 amino acids to about 18 amino acids, from about 2 amino acids to about 16 amino acids, from about 2 amino acids to about 14 amino acids, from about 2 amino acids to about 12 amino acids, from about 2 amino acids to about 10 amino acids, from about 2 amino acids to about 8 amino acids, from about 2 amino acids to about 6 amino acids, from about 2 amino acids to about 4 amino acids, from about 4 amino acids to about 100 amino acids, from about 4 amino acids to about 90 amino acids, from about 4 amino acids to about 80 amino acids, from about 4 amino acids to about 70 amino acids, from about 4 amino acids to about 60 amino acids, from about 4 amino acids to about 50 amino acids, from about 4 Amino acids to about 45 amino acids, about 4 amino acids to about 40 amino acids, about 4 amino acids to about 35 amino acids, about 4 amino acids to about 30 amino acids, about 4 amino acids to about 25 amino acids, about 4 amino acids to about 24 amino acids, about 4 amino acids to about 22 amino acids, about 4 amino acids to about 20 amino acids, about 4 amino acids to about 18 amino acids, about 4 amino acids to about 16 amino acids, about 4 amino acids to about 14 amino acids, about 4 amino acids to about 12 amino acids, about 4 amino acids to about 10 amino acids, about 4 amino acids to about 8 amino acids, about 4 amino acids to about 6 amino acids, about 6 amino acids to about 100 amino acids, about 6 amino acids 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 Approximately 10 amino acids, approximately 6 amino acids to approximately 8 amino acids, approximately 8 amino acids to approximately 100 amino acids, approximately 8 amino acids to approximately 90 amino acids, approximately 8 amino acids to approximately 80 amino acids, approximately 8 amino acids to approximately 70 amino acids, approximately 8 amino acids to approximately 60 amino acids, approximately 8 amino acids to approximately 50 amino acids, approximately 8 amino acids to approximately 45 amino acids, approximately 8 amino acids to approximately 40 amino acids, approximately 8 amino acids to approximately 35 amino acids, approximately 8 amino acids to approximately 30 amino acids, approximately 8 amino acids to approximately 25 amino acids, approximately 8 amino acids to approximately 24 amino acids, approximately 8 amino acids to approximately 22 amino acids, approximately 8 amino acids to approximately 20 amino acids, approximately 8 amino acids to approximately 18 amino acids. Approximately 8 amino acids to approximately 16 amino acids, approximately 8 amino acids to approximately 14 amino acids, approximately 8 amino acids to approximately 12 amino acids, approximately 8 amino acids to approximately 10 amino acids, approximately 10 amino acids to approximately 100 amino acids, approximately 10 amino acids to approximately 90 amino acids, approximately 10 amino acids to approximately 80 amino acids, approximately 10 amino acids to approximately 70 amino acids, approximately 10 amino acids to approximately 60 amino acids, approximately 10 amino acids to approximately 50 amino acids, approximately 10 amino acids to approximately 45 amino acids, approximately 10 amino acids to approximately 40 amino acids, approximately 10 amino acids to approximately 35 amino acids, approximately 10 amino acids to approximately 30 amino acids, approximately 10 amino acids to approximately 25 amino acids, approximately 10 amino acids to approximately 24 amino acids, about 10 amino acids to about 22 amino acids, about 10 amino acids to about 20 amino acids, about 10 amino acids to about 18 amino acids, about 10 amino acids to about 16 amino acids, about 10 amino acids to about 14 amino acids, about 10 amino acids to about 12 amino acids, about 12 amino acids to about 100 amino acids, about 12 amino acids to about 90 amino acids, about 12 amino acids to about 80 amino acids, about 12 amino acids to about 70 amino acids, about 12 amino acids to about 60 amino acids, about 12 amino acids to about 50 amino acids, about 12 amino acids to about 45 amino acids, about 12 amino acids to about 40 amino acids, about 12 amino acids to about 35 amino acids.Approximately 12 amino acids to approximately 30 amino acids, approximately 12 amino acids to approximately 25 amino acids, approximately 12 amino acids to approximately 24 amino acids, approximately 12 amino acids to approximately 22 amino acids, approximately 12 amino acids to approximately 20 amino acids, approximately 12 amino acids to approximately 18 amino acids, approximately 12 amino acids to approximately 16 amino acids, approximately 12 amino acids to approximately 14 amino acids, approximately 14 amino acids to approximately 100 amino acids, approximately 14 amino acids to approximately 90 amino acids, approximately 14 amino acids to approximately 80 amino acids, approximately 14 amino acids to approximately 70 amino acids, approximately 14 amino acids to approximately 60 amino acids, approximately 14 amino acids to approximately 50 amino acids, approximately 14 amino acids to approximately 45 amino acids, approximately 14 amino acids to Approximately 40 amino acids, approximately 14 amino acids to approximately 35 amino acids, approximately 14 amino acids to approximately 30 amino acids, approximately 14 amino acids to approximately 25 amino acids, approximately 14 amino acids to approximately 24 amino acids, approximately 14 amino acids to approximately 22 amino acids, approximately 14 amino acids to approximately 20 amino acids, approximately 14 amino acids to approximately 18 amino acids, approximately 14 amino acids to approximately 16 amino acids, approximately 16 amino acids to approximately 100 amino acids, approximately 16 amino acids to approximately 90 amino acids, approximately 16 amino acids to approximately 80 amino acids, approximately 16 amino acids to approximately 70 amino acids, approximately 16 amino acids to approximately 60 amino acids, approximately 16 amino acids to approximately 50 amino acids, approximately 16 amino acids to approximately 45 amino acids. Approximately 16 amino acids to approximately 40 amino acids, approximately 16 amino acids to approximately 35 amino acids, approximately 16 amino acids to approximately 30 amino acids, approximately 16 amino acids to approximately 25 amino acids, approximately 16 amino acids to approximately 24 amino acids, approximately 16 amino acids to approximately 22 amino acids, approximately 16 amino acids to approximately 20 amino acids, approximately 16 amino acids to approximately 18 amino acids, approximately 18 amino acids to approximately 100 amino acids, approximately 18 amino acids to approximately 90 amino acids, approximately 18 amino acids to approximately 80 amino acids, approximately 18 amino acids to approximately 70 amino acids, approximately 18 amino acids to approximately 60 amino acids, approximately 18 amino acids to approximately 50 amino acids, approximately 18 amino acids to approximately 45 amino acids, approximately 18 amino acids to Approximately 40 amino acids, approximately 18 amino acids to approximately 35 amino acids, approximately 18 amino acids to approximately 30 amino acids, approximately 18 amino acids to approximately 25 amino acids, approximately 18 amino acids to approximately 24 amino acids, approximately 18 amino acids to approximately 22 amino acids, approximately 18 amino acids to approximately 20 amino acids, approximately 20 amino acids to approximately 100 amino acids, approximately 20 amino acids to approximately 90 amino acids, approximately 20 amino acids to approximately 80 amino acids, approximately 20 amino acids to approximately 70 amino acids, approximately 20 amino acids to approximately 60 amino acids, approximately 20 amino acids to approximately 50 amino acids, approximately 20 amino acids to approximately 45 amino acids, approximately 20 amino acids to approximately 40 amino acids, approximately 20 amino acids to approximately 35 amino acids.Approximately 20 amino acids to approximately 30 amino acids, approximately 20 amino acids to approximately 25 amino acids, approximately 20 amino acids to approximately 24 amino acids, approximately 20 amino acids to approximately 22 amino acids, approximately 22 amino acids to approximately 100 amino acids, approximately 22 amino acids to approximately 90 amino acids, approximately 22 amino acids to approximately 80 amino acids, approximately 22 amino acids to approximately 70 amino acids, approximately 22 amino acids to approximately 60 amino acids, approximately 22 amino acids to approximately 50 amino acids, approximately 22 amino acids to approximately 45 amino acids, approximately 22 amino acids to approximately 40 amino acids, approximately 22 amino acids to approximately 35 amino acids, approximately 22 amino acids to approximately 30 amino acids, approximately 22 amino acids to approximately 25 amino acids, approximately 22 amino acids to approximately 24 amino acids, approximately 25 amino acids to approximately 100 amino acids, approximately 25 amino acids to approximately 90 amino acids, approximately 25 amino acids to approximately 80 amino acids, approximately 25 amino acids to approximately 70 amino acids, approximately 25 amino acids to approximately 60 amino acids, approximately 25 amino acids to approximately 50 amino acids, approximately 25 amino acids to approximately 45 amino acids, approximately 25 amino acids to approximately 40 amino acids, approximately 25 amino acids to approximately 35 amino acids, approximately 25 amino acids to approximately 30 amino acids, approximately 30 amino acids to approximately 100 amino acids, approximately 30 amino acids to approximately 90 amino acids, approximately 30 amino acids to approximately 80 amino acids, approximately 30 amino acids to approximately 70 amino acids, approximately 30 amino acids to approximately 60 amino acids, approximately 30 amino acids to about 50 amino acids, about 30 amino acids to about 45 amino acids, about 30 amino acids to about 40 amino acids, about 30 amino acids to about 35 amino acids, about 35 amino acids to about 100 amino acids, about 35 amino acids to about 90 amino acids, about 35 amino acids to about 80 amino acids, about 35 amino acids to about 70 amino acids, about 35 amino acids to about 60 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 100 amino acids, about 40 amino acids to about 90 amino acids, about 40 amino acids to about 80 amino acids, about 40 amino acids to about 70 amino acids, approximately 40 amino acids to approximately 60 amino acids, approximately 40 amino acids to approximately 50 amino acids, approximately 40 amino acids to approximately 45 amino acids, approximately 45 amino acids to approximately 100 amino acids, approximately 45 amino acids to approximately 90 amino acids, approximately 45 amino acids to approximately 80 amino acids, approximately 45 amino acids to approximately 70 amino acids, approximately 45 amino acids to approximately 60 amino acids, approximately 45 amino acids to approximately 50 amino acids, approximately 50 amino acids to approximately 100 amino acids, approximately 50 amino acids to approximately 90 amino acids, approximately 50 amino acids to approximately 80 amino acids, approximately 50 amino acids to approximately 70 amino acids, approximately 50 amino acids to approximately 60 amino acids, approximately 60 amino acids to approximately 100 amino acids.Approximately 60 amino acids to approximately 90 amino acids, approximately 60 amino acids to approximately 80 amino acids, approximately 60 amino acids to approximately 70 amino acids, approximately 70 amino acids to approximately 100 amino acids, approximately 70 amino acids to approximately 90 amino acids, approximately 70 amino acids to approximately 80 amino acids, approximately 80 amino acids to approximately 100 amino acids, approximately 80 amino acids to approximately 90 amino acids, or approximately 90 amino acids to approximately 100 amino acids.

[0127] In some embodiments, the linker is enriched with glycine (Gly or G) residues. In some embodiments, the linker is enriched with serine (Ser or S) residues. In some embodiments, the linker is enriched with both glycine and serine residues. In some embodiments, the linker has one or more glycine-serine residue pairs (GS), such as 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 or more GS pairs. In some embodiments, the linker has one or more Gly-Gly-Gly-Ser (GGGS) sequences, such as 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 or more GGGS sequences. In some embodiments, the linker has one or more Gly-Gly-Ser-Gly (GGSG) sequences, such as 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 or more GGGGS sequences. In some embodiments, the linker has one or more Gly-Gly-Ser-Gly (GGSG) sequences, such as 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 or more GGSG sequences.

[0128] In some embodiments, the adapter sequence may comprise or consist of the following: GGGGSGGGGSGGGGS (SEQ ID NO: 13). In some embodiments, the adapter sequence may be encoded by a nucleic acid comprising or consisting of the following: GGCGGTGGAGGATCCGGAGGAGGTGGCTCCGGCGGCGGAGGATCT (SEQ ID NO: 14). In some embodiments, the adapter sequence may comprise or consist of GGGSGGGS (SEQ ID NO: 15).

[0129] Target binding domain

[0130] In some embodiments of any single-chain chimeric polypeptide described herein, the first target-binding domain, the second target-binding domain, and / or one or more other target-binding domains may be an antigen-binding domain (e.g., any exemplary antigen-binding domain described herein or known in the art), a soluble interleukin or cytokine protein (e.g., any exemplary soluble interleukin protein or soluble cytokine protein described herein), and a soluble interleukin or cytokine receptor (e.g., any exemplary soluble interleukin receptor or soluble cytokine receptor described herein).

[0131] In some embodiments of any single-chain chimeric polypeptide described herein, one or more of a first target-binding domain (e.g., any exemplary first target-binding domain described herein or known in the art), a second target-binding domain (e.g., any exemplary second target-binding domain described herein or known in the art), and one or more other target-binding domains are capable of independently and specifically binding to targets selected from the group consisting of: CD16a, CD28, CD3 (e.g., one or more of CD3α, CD3β, CD3δ, CD3ε, and CD3γ), CD33, CD20, C D19, 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, TNFα, CD2 6a, CD36, ULBP2, CD30, CD200, IGF-1R, MUC4AC, MUC5AC, Trop-2, CMET, EGFR, HER1, HER2, HER3, PSMA, CEA, B7H3, EPCAM, BCMA, P - Ligands of cadherin, CEACAM5, 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), TGF-βRIII, DNAM1, NKp46, NKp44, NKG2D, NKp30, scMHCI, and scMHCII. , 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.

[0132] In some embodiments of any single-chain chimeric polypeptide described herein, the first target-binding domain, the second target-binding domain, and / or one or more other target-binding domains may independently each have the following total amino acid counts: about 5 amino acids to about 1000 amino acids, about 5 amino acids to about 950 amino acids, about 5 amino acids to about 900 amino acids, about 5 amino acids to about 850 amino acids, about 5 amino acids to about 800 amino acids, about 5 amino acids to about 750 amino acids, about 5 amino acids to about 700 amino acids, about 5 amino acids to about 650 amino acids, about 5 amino acids to about 600 amino acids, about 5 amino acids to about 550 amino acids, about 5 amino acids to about 500 amino acids, about 5 From approximately 450 amino acids, from approximately 5 amino acids to approximately 400 amino acids, from approximately 5 amino acids to approximately 350 amino acids, from approximately 5 amino acids to approximately 300 amino acids, from approximately 5 amino acids to approximately 280 amino acids, from approximately 5 amino acids to approximately 260 amino acids, from approximately 5 amino acids to approximately 240 amino acids, from approximately 5 amino acids to approximately 220 amino acids, from approximately 5 amino acids to approximately 200 amino acids, from approximately 5 amino acids to approximately 195 amino acids, from approximately 5 amino acids to approximately 190 amino acids, from approximately 5 amino acids to approximately 185 amino acids, from approximately 5 amino acids to approximately 180 amino acids, from approximately 5 amino acids to approximately 175 amino acids, from approximately 5 amino acids to approximately 170 amino acids, from approximately 5 amino acids to approximately 165 amino acids. Amino acids, approximately 5 amino acids to approximately 160 amino acids, approximately 5 amino acids to approximately 155 amino acids, approximately 5 amino acids to approximately 150 amino acids, approximately 5 amino acids to approximately 145 amino acids, approximately 5 amino acids to approximately 140 amino acids, approximately 5 amino acids to approximately 135 amino acids, approximately 5 amino acids to approximately 130 amino acids, approximately 5 amino acids to approximately 125 amino acids, approximately 5 amino acids to approximately 120 amino acids, approximately 5 amino acids to approximately 115 amino acids, approximately 5 amino acids to approximately 110 amino acids, approximately 5 amino acids to approximately 105 amino acids, approximately 5 amino acids to approximately 100 amino acids, approximately 5 amino acids to approximately 95 amino acids, approximately 5 amino acids to approximately 90 amino acids, approximately 5 amino acids to approximately 85 amino acids, approximately 5 amino acids to approximately 80 amino acids, approximately 5 amino acids to approximately 75 amino acids, approximately 5 amino acids to approximately 70 amino acids, approximately 5 amino acids to approximately 65 amino acids, approximately 5 amino acids to approximately 60 amino acids, approximately 5 amino acids to approximately 55 amino acids, approximately 5 amino acids to approximately 50 amino acids, approximately 5 amino acids to approximately 45 amino acids, approximately 5 amino acids to approximately 40 amino acids, approximately 5 amino acids to approximately 35 amino acids, approximately 5 amino acids to approximately 30 amino acids, approximately 5 amino acids to approximately 25 amino acids, approximately 5 amino acids to approximately 20 amino acids, approximately 5 amino acids to approximately 15 amino acids, approximately 5 amino acids to approximately 10 amino acids, approximately 10 amino acids to approximately 1000 amino acids.Approximately 10 amino acids to approximately 950 amino acids, approximately 10 amino acids to approximately 900 amino acids, approximately 10 amino acids to approximately 850 amino acids, approximately 10 amino acids to approximately 800 amino acids, approximately 10 amino acids to approximately 750 amino acids, approximately 10 amino acids to approximately 700 amino acids, approximately 10 amino acids to approximately 650 amino acids, approximately 10 amino acids to approximately 600 amino acids, approximately 10 amino acids to approximately 550 amino acids, approximately 10 amino acids to approximately 500 amino acids, approximately 10 amino acids to approximately 450 amino acids, approximately 10 amino acids to approximately 400 amino acids, approximately 10 amino acids to approximately 350 amino acids, approximately 10 amino acids to approximately 300 amino acids, approximately 10 amino acids to Approximately 280 amino acids, approximately 10 amino acids to approximately 260 amino acids, approximately 10 amino acids to approximately 240 amino acids, approximately 10 amino acids to approximately 220 amino acids, approximately 10 amino acids to approximately 200 amino acids, approximately 10 amino acids to approximately 195 amino acids, approximately 10 amino acids to approximately 190 amino acids, approximately 10 amino acids to approximately 185 amino acids, approximately 10 amino acids to approximately 180 amino acids, approximately 10 amino acids to approximately 175 amino acids, approximately 10 amino acids to approximately 170 amino acids, approximately 10 amino acids to approximately 165 amino acids, approximately 10 amino acids to approximately 160 amino acids, approximately 10 amino acids to approximately 155 amino acids, approximately 10 amino acids to approximately 150 amino acids. Approximately 10 amino acids to approximately 145 amino acids, approximately 10 amino acids to approximately 140 amino acids, approximately 10 amino acids to approximately 135 amino acids, approximately 10 amino acids to approximately 130 amino acids, approximately 10 amino acids to approximately 125 amino acids, approximately 10 amino acids to approximately 120 amino acids, approximately 10 amino acids to approximately 115 amino acids, approximately 10 amino acids to approximately 110 amino acids, approximately 10 amino acids to approximately 105 amino acids, approximately 10 amino acids to approximately 100 amino acids, approximately 10 amino acids to approximately 95 amino acids, approximately 10 amino acids to approximately 90 amino acids, approximately 10 amino acids to approximately 85 amino acids, approximately 10 amino acids to approximately 80 amino acids, approximately 10 amino acids to approximately 75 amino acids. Amino acids, approximately 10 amino acids to approximately 70 amino acids, approximately 10 amino acids to approximately 65 amino acids, approximately 10 amino acids to approximately 60 amino acids, approximately 10 amino acids to approximately 55 amino acids, approximately 10 amino acids to approximately 50 amino acids, approximately 10 amino acids to approximately 45 amino acids, approximately 10 amino acids to approximately 40 amino acids, approximately 10 amino acids to approximately 35 amino acids, approximately 10 amino acids to approximately 30 amino acids, approximately 10 amino acids to approximately 25 amino acids, approximately 10 amino acids to approximately 20 amino acids, approximately 10 amino acids to approximately 15 amino acids, approximately 15 amino acids to approximately 1000 amino acids, approximately 15 amino acids to approximately 950 amino acids, approximately 15 amino acids to approximately 900 amino acids.Approximately 15 amino acids to approximately 850 amino acids, approximately 15 amino acids to approximately 800 amino acids, approximately 15 amino acids to approximately 750 amino acids, approximately 15 amino acids to approximately 700 amino acids, approximately 15 amino acids to approximately 650 amino acids, approximately 15 amino acids to approximately 600 amino acids, approximately 15 amino acids to approximately 550 amino acids, approximately 15 amino acids to approximately 500 amino acids, approximately 15 amino acids to approximately 450 amino acids, approximately 15 amino acids to approximately 400 amino acids, approximately 15 amino acids to approximately 350 amino acids, approximately 15 amino acids to approximately 300 amino acids, approximately 15 amino acids to approximately 280 amino acids, approximately 15 amino acids to approximately 260 amino acids, approximately 15 amino acids to approximately 240 amino acids, approximately 15 amino acids to approximately 220 amino acids, approximately 15 amino acids to approximately 200 amino acids, approximately 15 amino acids to approximately 195 amino acids, approximately 15 amino acids to approximately 190 amino acids, approximately 15 amino acids to approximately 185 amino acids, approximately 15 amino acids to approximately 180 amino acids, approximately 15 amino acids to approximately 175 amino acids, approximately 15 amino acids to approximately 170 amino acids, approximately 15 amino acids to approximately 165 amino acids, approximately 15 amino acids to approximately 160 amino acids, approximately 15 amino acids to approximately 155 amino acids, approximately 15 amino acids to approximately 150 amino acids, approximately 15 amino acids to approximately 145 amino acids, approximately 15 amino acids to approximately 140 amino acids, approximately 15 amino acids to about 135 amino acids, about 15 amino acids to about 130 amino acids, about 15 amino acids to about 125 amino acids, about 15 amino acids to about 120 amino acids, about 15 amino acids to about 115 amino acids, about 15 amino acids to about 110 amino acids, about 15 amino acids to about 105 amino acids, about 15 amino acids to about 100 amino acids, about 15 amino acids to about 95 amino acids, about 15 amino acids to about 90 amino acids, about 15 amino acids to about 85 amino acids, about 15 amino acids to about 80 amino acids, about 15 amino acids to about 75 amino acids, about 15 amino acids to about 70 amino acids, about 15 amino acids to about 65 amino acids. Approximately 15 amino acids to approximately 60 amino acids, approximately 15 amino acids to approximately 55 amino acids, approximately 15 amino acids to approximately 50 amino acids, approximately 15 amino acids to approximately 45 amino acids, approximately 15 amino acids to approximately 40 amino acids, approximately 15 amino acids to approximately 35 amino acids, approximately 15 amino acids to approximately 30 amino acids, approximately 15 amino acids to approximately 25 amino acids, approximately 15 amino acids to approximately 20 amino acids, approximately 20 amino acids to approximately 1000 amino acids, approximately 20 amino acids to approximately 950 amino acids, approximately 20 amino acids to approximately 900 amino acids, approximately 20 amino acids to approximately 850 amino acids, approximately 20 amino acids to approximately 800 amino acids, approximately 20 amino acids to approximately 750 amino acids.Approximately 20 amino acids to approximately 700 amino acids, approximately 20 amino acids to approximately 650 amino acids, approximately 20 amino acids to approximately 600 amino acids, approximately 20 amino acids to approximately 550 amino acids, approximately 20 amino acids to approximately 500 amino acids, approximately 20 amino acids to approximately 450 amino acids, approximately 20 amino acids to approximately 400 amino acids, approximately 20 amino acids to approximately 350 amino acids, approximately 20 amino acids to approximately 300 amino acids, approximately 20 amino acids to approximately 280 amino acids, approximately 20 amino acids to approximately 260 amino acids, approximately 20 amino acids to approximately 240 amino acids, approximately 20 amino acids to approximately 220 amino acids, approximately 20 amino acids to approximately 200 amino acids, approximately 20 amino acids to approximately... 195 amino acids, approximately 20 amino acids to approximately 190 amino acids, approximately 20 amino acids to approximately 185 amino acids, approximately 20 amino acids to approximately 180 amino acids, approximately 20 amino acids to approximately 175 amino acids, approximately 20 amino acids to approximately 170 amino acids, approximately 20 amino acids to approximately 165 amino acids, approximately 20 amino acids to approximately 160 amino acids, approximately 20 amino acids to approximately 155 amino acids, approximately 20 amino acids to approximately 150 amino acids, approximately 20 amino acids to approximately 145 amino acids, approximately 20 amino acids to approximately 140 amino acids, approximately 20 amino acids to approximately 135 amino acids, approximately 20 amino acids to approximately 130 amino acids, approximately 20 amino acids to approximately 125 amino acids, approximately 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 The numbers appear to be a series of amino acids and symbols, possibly related to a list of amino acids and their composition. A direct translation wouldn't be meaningful without further context.Approximately 25 amino acids to approximately 500 amino acids, approximately 25 amino acids to approximately 450 amino acids, approximately 25 amino acids to approximately 400 amino acids, approximately 25 amino acids to approximately 350 amino acids, approximately 25 amino acids to approximately 300 amino acids, approximately 25 amino acids to approximately 280 amino acids, approximately 25 amino acids to approximately 260 amino acids, approximately 25 amino acids to approximately 240 amino acids, approximately 25 amino acids to approximately 220 amino acids, approximately 25 amino acids to approximately 200 amino acids, approximately 25 amino acids to approximately 195 amino acids, approximately 25 amino acids to approximately 190 amino acids, approximately 25 amino acids to approximately 185 amino acids, approximately 25 amino acids to approximately 180 amino acids, approximately 25 amino acids to approximately 175 amino acids, approximately 25 amino acids to approximately 170 amino acids, approximately 25 amino acids to approximately 165 amino acids, approximately 25 amino acids to approximately 160 amino acids, approximately 25 amino acids to approximately 155 amino acids, approximately 25 amino acids to approximately 150 amino acids, approximately 25 amino acids to approximately 145 amino acids, approximately 25 amino acids to approximately 140 amino acids, approximately 25 amino acids to approximately 135 amino acids, approximately 25 amino acids to approximately 130 amino acids, approximately 25 amino acids to approximately 125 amino acids, approximately 25 amino acids to approximately 120 amino acids, approximately 25 amino acids to approximately 115 amino acids, approximately 25 amino acids to approximately 110 amino acids, approximately 25 amino acids to approximately 105 amino acids, approximately 2 5 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 35 amino acids, about 25 amino acids to about 30 amino acids, about 30 amino acids to Approximately 1000 amino acids, approximately 30 amino acids to approximately 950 amino acids, approximately 30 amino acids to approximately 900 amino acids, approximately 30 amino acids to approximately 850 amino acids, approximately 30 amino acids to approximately 800 amino acids, approximately 30 amino acids to approximately 750 amino acids, approximately 30 amino acids to approximately 700 amino acids, approximately 30 amino acids to approximately 650 amino acids, approximately 30 amino acids to approximately 600 amino acids, approximately 30 amino acids to approximately 550 amino acids, approximately 30 amino acids to approximately 500 amino acids, approximately 30 amino acids to approximately 450 amino acids, approximately 30 amino acids to approximately 400 amino acids, approximately 30 amino acids to approximately 350 amino acids, approximately 30 amino acids to approximately 300 amino acids.Approximately 30 amino acids to approximately 280 amino acids, approximately 30 amino acids to approximately 260 amino acids, approximately 30 amino acids to approximately 240 amino acids, approximately 30 amino acids to approximately 220 amino acids, approximately 30 amino acids to approximately 200 amino acids, approximately 30 amino acids to approximately 195 amino acids, approximately 30 amino acids to approximately 190 amino acids, approximately 30 amino acids to approximately 185 amino acids, approximately 30 amino acids to approximately 180 amino acids, approximately 30 amino acids to approximately 175 amino acids, approximately 30 amino acids to approximately 170 amino acids, approximately 30 amino acids to approximately 165 amino acids, approximately 30 amino acids to approximately 160 amino acids, approximately 30 amino acids to approximately 155 amino acids, approximately 30 amino acids to approximately 150 amino acids, approximately 30 amino acids to approximately 145 amino acids, approximately 30 amino acids to approximately 140 amino acids, approximately 30 amino acids to approximately 135 amino acids, approximately 30 amino acids to approximately 130 amino acids, approximately 30 amino acids to approximately 125 amino acids, approximately 30 amino acids to approximately 120 amino acids, approximately 30 amino acids to approximately 115 amino acids, approximately 30 amino acids to approximately 110 amino acids, approximately 30 amino acids to approximately 105 amino acids, approximately 30 amino acids to approximately 100 amino acids, approximately 30 amino acids to approximately 95 amino acids, approximately 30 amino acids to approximately 90 amino acids, approximately 30 amino acids to approximately 85 amino acids, approximately 30 amino acids to approximately 80 amino acids, approximately 30 amino acids Acid to about 75 amino acids, about 30 amino acids to about 70 amino acids, about 30 amino acids to about 65 amino acids, about 30 amino acids to about 60 amino acids, about 30 amino acids to about 55 amino acids, about 30 amino acids to about 50 amino acids, about 30 amino acids to about 45 amino acids, about 30 amino acids to about 40 amino acids, about 30 amino acids 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 From approximately 700 amino acids, from approximately 35 amino acids to approximately 650 amino acids, from approximately 35 amino acids to approximately 600 amino acids, from approximately 35 amino acids to approximately 550 amino acids, from approximately 35 amino acids to approximately 500 amino acids, from approximately 35 amino acids to approximately 450 amino acids, from approximately 35 amino acids to approximately 400 amino acids, from approximately 35 amino acids to approximately 350 amino acids, from approximately 35 amino acids to approximately 300 amino acids, from approximately 35 amino acids to approximately 280 amino acids, from approximately 35 amino acids to approximately 260 amino acids, from approximately 35 amino acids to approximately 240 amino acids, from approximately 35 amino acids to approximately 220 amino acids, from approximately 35 amino acids to approximately 200 amino acids, from approximately 35 amino acids to approximately 195 amino acids.Approximately 35 amino acids to approximately 190 amino acids, approximately 35 amino acids to approximately 185 amino acids, approximately 35 amino acids to approximately 180 amino acids, approximately 35 amino acids to approximately 175 amino acids, approximately 35 amino acids to approximately 170 amino acids, approximately 35 amino acids to approximately 165 amino acids, approximately 35 amino acids to approximately 160 amino acids, approximately 35 amino acids to approximately 155 amino acids, approximately 35 amino acids to approximately 150 amino acids, approximately 35 amino acids to approximately 145 amino acids, approximately 35 amino acids to approximately 140 amino acids, approximately 35 amino acids to approximately 135 amino acids, approximately 35 amino acids to approximately 130 amino acids, approximately 35 amino acids to approximately 125 amino acids, approximately 35 amino acids to approximately 1 20 amino acids, approximately 35 amino acids to approximately 115 amino acids, approximately 35 amino acids to approximately 110 amino acids, approximately 35 amino acids to approximately 105 amino acids, approximately 35 amino acids to approximately 100 amino acids, approximately 35 amino acids to approximately 95 amino acids, approximately 35 amino acids to approximately 90 amino acids, approximately 35 amino acids to approximately 85 amino acids, approximately 35 amino acids to approximately 80 amino acids, approximately 35 amino acids to approximately 75 amino acids, approximately 35 amino acids to approximately 70 amino acids, approximately 35 amino acids to approximately 65 amino acids, approximately 35 amino acids to approximately 60 amino acids, approximately 35 amino acids to approximately 55 amino acids, approximately 35 amino acids to approximately 50 amino acids, approximately 35 amino acids to approximately 45 amino acids Amino acids, approximately 35 amino acids to approximately 40 amino acids, approximately 40 amino acids to approximately 1000 amino acids, approximately 40 amino acids to approximately 950 amino acids, approximately 40 amino acids to approximately 900 amino acids, approximately 40 amino acids to approximately 850 amino acids, approximately 40 amino acids to approximately 800 amino acids, approximately 40 amino acids to approximately 750 amino acids, approximately 40 amino acids to approximately 700 amino acids, approximately 40 amino acids to approximately 650 amino acids, approximately 40 amino acids to approximately 600 amino acids, approximately 40 amino acids to approximately 550 amino acids, approximately 40 amino acids to approximately 500 amino acids, approximately 40 amino acids to approximately 450 amino acids, approximately 40 amino acids to approximately 400 amino acids, approximately 40 amino acids From approximately 350 amino acids, from approximately 40 amino acids to approximately 300 amino acids, from approximately 40 amino acids to approximately 280 amino acids, from approximately 40 amino acids to approximately 260 amino acids, from approximately 40 amino acids to approximately 240 amino acids, from approximately 40 amino acids to approximately 220 amino acids, from approximately 40 amino acids to approximately 200 amino acids, from approximately 40 amino acids to approximately 195 amino acids, from approximately 40 amino acids to approximately 190 amino acids, from approximately 40 amino acids to approximately 185 amino acids, from approximately 40 amino acids to approximately 180 amino acids, from approximately 40 amino acids to approximately 175 amino acids, from approximately 40 amino acids to approximately 170 amino acids, from approximately 40 amino acids to approximately 165 amino acids, from approximately 40 amino acids to approximately 160 amino acids.Approximately 40 amino acids to approximately 155 amino acids, approximately 40 amino acids to approximately 150 amino acids, approximately 40 amino acids to approximately 145 amino acids, approximately 40 amino acids to approximately 140 amino acids, approximately 40 amino acids to approximately 135 amino acids, approximately 40 amino acids to approximately 130 amino acids, approximately 40 amino acids to approximately 125 amino acids, approximately 40 amino acids to approximately 120 amino acids, approximately 40 amino acids to approximately 115 amino acids, approximately 40 amino acids to approximately 110 amino acids, approximately 40 amino acids to approximately 105 amino acids, approximately 40 amino acids to approximately 100 amino acids, approximately 40 amino acids to approximately 95 amino acids, approximately 40 amino acids to approximately 90 amino acids, approximately 40 amino acids to approximately 85 amino acids. Amino acids, approximately 40 amino acids to approximately 80 amino acids, approximately 40 amino acids to approximately 75 amino acids, approximately 40 amino acids to approximately 70 amino acids, approximately 40 amino acids to approximately 65 amino acids, approximately 40 amino acids to approximately 60 amino acids, approximately 40 amino acids to approximately 55 amino acids, approximately 40 amino acids to approximately 50 amino acids, approximately 40 amino acids to approximately 45 amino acids, approximately 45 amino acids to approximately 1000 amino acids, approximately 45 amino acids to approximately 950 amino acids, approximately 45 amino acids to approximately 900 amino acids, approximately 45 amino acids to approximately 850 amino acids, approximately 45 amino acids to approximately 800 amino acids, approximately 45 amino acids to approximately 750 amino acids, approximately 45 amino acids to approximately 700 amino acids. Amino acids, approximately 45 amino acids to approximately 650 amino acids, approximately 45 amino acids to approximately 600 amino acids, approximately 45 amino acids to approximately 550 amino acids, approximately 45 amino acids to approximately 500 amino acids, approximately 45 amino acids to approximately 450 amino acids, approximately 45 amino acids to approximately 400 amino acids, approximately 45 amino acids to approximately 350 amino acids, approximately 45 amino acids to approximately 300 amino acids, approximately 45 amino acids to approximately 280 amino acids, approximately 45 amino acids to approximately 260 amino acids, approximately 45 amino acids to approximately 240 amino acids, approximately 45 amino acids to approximately 220 amino acids, approximately 45 amino acids to approximately 200 amino acids, approximately 45 amino acids to approximately 195 amino acids, approximately 45 amino groups. Acids up to 190 amino acids, about 45 amino acids up to 185 amino acids, about 45 amino acids up to 180 amino acids, about 45 amino acids up to 175 amino acids, about 45 amino acids up to 170 amino acids, about 45 amino acids up to 165 amino acids, about 45 amino acids up to 160 amino acids, about 45 amino acids up to 155 amino acids, about 45 amino acids up to 150 amino acids, about 45 amino acids up to 145 amino acids, about 45 amino acids up to 140 amino acids, about 45 amino acids up to 135 amino acids, about 45 amino acids up to 130 amino acids, about 45 amino acids up to 125 amino acids, about 45 amino acids up to 120 amino acids.Approximately 45 amino acids to approximately 115 amino acids, approximately 45 amino acids to approximately 110 amino acids, approximately 45 amino acids to approximately 105 amino acids, approximately 45 amino acids to approximately 100 amino acids, approximately 45 amino acids to approximately 95 amino acids, approximately 45 amino acids to approximately 90 amino acids, approximately 45 amino acids to approximately 85 amino acids, approximately 45 amino acids to approximately 80 amino acids, approximately 45 amino acids to approximately 75 amino acids, approximately 45 amino acids to approximately 70 amino acids, approximately 45 amino acids to approximately 65 amino acids, approximately 45 amino acids to approximately 60 amino acids, approximately 45 amino acids to approximately 55 amino acids, approximately 45 amino acids to approximately 50 amino acids, approximately 50 amino acids to approximately 1000 amino acids, approximately 50 amino acids to about 950 amino acids, about 50 amino acids to about 900 amino acids, about 50 amino acids to about 850 amino acids, about 50 amino acids to about 800 amino acids, about 50 amino acids to about 750 amino acids, about 50 amino acids to about 700 amino acids, about 50 amino acids to about 650 amino acids, about 50 amino acids to about 600 amino acids, about 50 amino acids to about 550 amino acids, about 50 amino acids to about 500 amino acids, about 50 amino acids to about 450 amino acids, about 50 amino acids to about 400 amino acids, about 50 amino acids to about 350 amino acids, about 50 amino acids to about 300 amino acids, about 50 amino acids to about 2 80 amino acids, approximately 50 amino acids to approximately 260 amino acids, approximately 50 amino acids to approximately 240 amino acids, approximately 50 amino acids to approximately 220 amino acids, approximately 50 amino acids to approximately 200 amino acids, approximately 50 amino acids to approximately 195 amino acids, approximately 50 amino acids to approximately 190 amino acids, approximately 50 amino acids to approximately 185 amino acids, approximately 50 amino acids to approximately 180 amino acids, approximately 50 amino acids to approximately 175 amino acids, approximately 50 amino acids to approximately 170 amino acids, approximately 50 amino acids to approximately 165 amino acids, approximately 50 amino acids to approximately 160 amino acids, approximately 50 amino acids to approximately 155 amino acids, approximately 50 amino acids to approximately 150 amino acids, approximately 5 0 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.Approximately 50 amino acids to approximately 70 amino acids, approximately 50 amino acids to approximately 65 amino acids, approximately 50 amino acids to approximately 60 amino acids, approximately 50 amino acids to approximately 55 amino acids, approximately 55 amino acids to approximately 1000 amino acids, approximately 55 amino acids to approximately 950 amino acids, approximately 55 amino acids to approximately 900 amino acids, approximately 55 amino acids to approximately 850 amino acids, approximately 55 amino acids to approximately 800 amino acids, approximately 55 amino acids to approximately 750 amino acids, approximately 55 amino acids to approximately 700 amino acids, approximately 55 amino acids to approximately 650 amino acids, approximately 55 amino acids to approximately 600 amino acids, approximately 55 amino acids to approximately 550 amino acids, approximately 55 amino acids to approximately 500 amino acids. Amino acids, approximately 55 amino acids to approximately 450 amino acids, approximately 55 amino acids to approximately 400 amino acids, approximately 55 amino acids to approximately 350 amino acids, approximately 55 amino acids to approximately 300 amino acids, approximately 55 amino acids to approximately 280 amino acids, approximately 55 amino acids to approximately 260 amino acids, approximately 55 amino acids to approximately 240 amino acids, approximately 55 amino acids to approximately 220 amino acids, approximately 55 amino acids to approximately 200 amino acids, approximately 55 amino acids to approximately 195 amino acids, approximately 55 amino acids to approximately 190 amino acids, approximately 55 amino acids to approximately 185 amino acids, approximately 55 amino acids to approximately 180 amino acids, approximately 55 amino acids to approximately 175 amino acids, approximately 55 amino groups. Acids up to 170 amino acids, about 55 amino acids up to 165 amino acids, about 55 amino acids up to 160 amino acids, about 55 amino acids up to 155 amino acids, about 55 amino acids up to 150 amino acids, about 55 amino acids up to 145 amino acids, about 55 amino acids up to 140 amino acids, about 55 amino acids up to 135 amino acids, about 55 amino acids up to 130 amino acids, about 55 amino acids up to 125 amino acids, about 55 amino acids up to 120 amino acids, about 55 amino acids up to 115 amino acids, about 55 amino acids up to 110 amino acids, about 55 amino acids up to 105 amino acids, about 55 amino acids up to 100 amino acids. Approximately 55 amino acids to approximately 95 amino acids, approximately 55 amino acids to approximately 90 amino acids, approximately 55 amino acids to approximately 85 amino acids, approximately 55 amino acids to approximately 80 amino acids, approximately 55 amino acids to approximately 75 amino acids, approximately 55 amino acids to approximately 70 amino acids, approximately 55 amino acids to approximately 65 amino acids, approximately 55 amino acids to approximately 60 amino acids, approximately 60 amino acids to approximately 1000 amino acids, approximately 60 amino acids to approximately 950 amino acids, approximately 60 amino acids to approximately 900 amino acids, approximately 60 amino acids to approximately 850 amino acids, approximately 60 amino acids to approximately 800 amino acids, approximately 60 amino acids to approximately 750 amino acids, approximately 60 amino acids to approximately 700 amino acids.Approximately 60 amino acids to approximately 650 amino acids, approximately 60 amino acids to approximately 600 amino acids, approximately 60 amino acids to approximately 550 amino acids, approximately 60 amino acids to approximately 500 amino acids, approximately 60 amino acids to approximately 450 amino acids, approximately 60 amino acids to approximately 400 amino acids, approximately 60 amino acids to approximately 350 amino acids, approximately 60 amino acids to approximately 300 amino acids, approximately 60 amino acids to approximately 280 amino acids, approximately 60 amino acids to approximately 260 amino acids, approximately 60 amino acids to approximately 240 amino acids, approximately 60 amino acids to approximately 220 amino acids, approximately 60 amino acids to approximately 200 amino acids, approximately 60 amino acids to approximately 195 amino acids, approximately 60 amino acids to approximately 19 0 amino acids, approximately 60 amino acids to approximately 185 amino acids, approximately 60 amino acids to approximately 180 amino acids, approximately 60 amino acids to approximately 175 amino acids, approximately 60 amino acids to approximately 170 amino acids, approximately 60 amino acids to approximately 165 amino acids, approximately 60 amino acids to approximately 160 amino acids, approximately 60 amino acids to approximately 155 amino acids, approximately 60 amino acids to approximately 150 amino acids, approximately 60 amino acids to approximately 145 amino acids, approximately 60 amino acids to approximately 140 amino acids, approximately 60 amino acids to approximately 135 amino acids, approximately 60 amino acids to approximately 130 amino acids, approximately 60 amino acids to approximately 125 amino acids, approximately 60 amino acids to approximately 120 amino acids, approximately 60 amino acids The sequence of amino acids is approximately 115 amino acids, approximately 60 amino acids to approximately 110 amino acids, approximately 60 amino acids to approximately 105 amino acids, approximately 60 amino acids to approximately 100 amino acids, approximately 60 amino acids to approximately 95 amino acids, approximately 60 amino acids to approximately 90 amino acids, approximately 60 amino acids to approximately 85 amino acids, approximately 60 amino acids to approximately 80 amino acids, approximately 60 amino acids to approximately 75 amino acids, approximately 60 amino acids to approximately 70 amino acids, approximately 60 amino acids to approximately 65 amino acids, approximately 65 amino acids to approximately 1000 amino acids, approximately 65 amino acids to approximately 950 amino acids, approximately 65 amino acids to approximately 900 amino acids, approximately 65 amino acids to approximately 850 amino acids, approximately 65 amino acids... The list includes approximately 800 amino acids, approximately 65 amino acids, approximately 750 amino acids, approximately 65 amino acids, approximately 700 amino acids, approximately 65 amino acids, approximately 65 amino acids, approximately 600 amino acids, approximately 65 amino acids, approximately 550 amino acids, approximately 65 amino acids, approximately 450 amino acids, approximately 400 amino acids, approximately 65 amino acids, approximately 350 amino acids, approximately 300 amino acids, approximately 65 amino acids, approximately 280 amino acids, approximately 260 amino acids, approximately 240 amino acids, and approximately 220 amino acids.Approximately 65 amino acids to approximately 200 amino acids, approximately 65 amino acids to approximately 195 amino acids, approximately 65 amino acids to approximately 190 amino acids, approximately 65 amino acids to approximately 185 amino acids, approximately 65 amino acids to approximately 180 amino acids, approximately 65 amino acids to approximately 175 amino acids, approximately 65 amino acids to approximately 170 amino acids, approximately 65 amino acids to approximately 165 amino acids, approximately 65 amino acids to approximately 160 amino acids, approximately 65 amino acids to approximately 155 amino acids, approximately 65 amino acids to approximately 150 amino acids, approximately 65 amino acids to approximately 145 amino acids, approximately 65 amino acids to approximately 140 amino acids, approximately 65 amino acids to approximately 135 amino acids, approximately 65 amino acids to approximately 13 0 amino acids, approximately 65 amino acids to approximately 125 amino acids, approximately 65 amino acids to approximately 120 amino acids, approximately 65 amino acids to approximately 115 amino acids, approximately 65 amino acids to approximately 110 amino acids, approximately 65 amino acids to approximately 105 amino acids, approximately 65 amino acids to approximately 100 amino acids, approximately 65 amino acids to approximately 95 amino acids, approximately 65 amino acids to approximately 90 amino acids, approximately 65 amino acids to approximately 85 amino acids, approximately 65 amino acids to approximately 80 amino acids, approximately 65 amino acids to approximately 75 amino acids, approximately 65 amino acids to approximately 70 amino acids, approximately 70 amino acids to approximately 1000 amino acids, approximately 70 amino acids to approximately 950 amino acids, approximately 70 amino acids to approximately 90 0 amino acids, approximately 70 amino acids to approximately 850 amino acids, approximately 70 amino acids to approximately 800 amino acids, approximately 70 amino acids to approximately 750 amino acids, approximately 70 amino acids to approximately 700 amino acids, approximately 70 amino acids to approximately 650 amino acids, approximately 70 amino acids to approximately 600 amino acids, approximately 70 amino acids to approximately 550 amino acids, approximately 70 amino acids to approximately 500 amino acids, approximately 70 amino acids to approximately 450 amino acids, approximately 70 amino acids to approximately 400 amino acids, approximately 70 amino acids to approximately 350 amino acids, approximately 70 amino acids to approximately 300 amino acids, approximately 70 amino acids to approximately 280 amino acids, approximately 70 amino acids to approximately 260 amino acids, approximately 70 amino acids The range is approximately 240 amino acids, approximately 70 amino acids to approximately 220 amino acids, approximately 70 amino acids to approximately 200 amino acids, approximately 70 amino acids to approximately 195 amino acids, approximately 70 amino acids to approximately 190 amino acids, approximately 70 amino acids to approximately 185 amino acids, approximately 70 amino acids to approximately 180 amino acids, approximately 70 amino acids to approximately 175 amino acids, approximately 70 amino acids to approximately 170 amino acids, approximately 70 amino acids to approximately 165 amino acids, approximately 70 amino acids to approximately 160 amino acids, approximately 70 amino acids to approximately 155 amino acids, approximately 70 amino acids to approximately 150 amino acids, approximately 70 amino acids to approximately 145 amino acids, and approximately 70 amino acids to approximately 140 amino acids.Approximately 70 amino acids to approximately 135 amino acids, approximately 70 amino acids to approximately 130 amino acids, approximately 70 amino acids to approximately 125 amino acids, approximately 70 amino acids to approximately 120 amino acids, approximately 70 amino acids to approximately 115 amino acids, approximately 70 amino acids to approximately 110 amino acids, approximately 70 amino acids to approximately 105 amino acids, approximately 70 amino acids to approximately 100 amino acids, approximately 70 amino acids to approximately 95 amino acids, approximately 70 amino acids to approximately 90 amino acids, approximately 70 amino acids to approximately 85 amino acids, approximately 70 amino acids to approximately 80 amino acids, approximately 70 amino acids to approximately 75 amino acids, approximately 75 amino acids to approximately 1000 amino acids, approximately 75 amino acids to approximately 950 amino acids. Acid, approximately 75 amino acids to approximately 900 amino acids, approximately 75 amino acids to approximately 850 amino acids, approximately 75 amino acids to approximately 800 amino acids, approximately 75 amino acids to approximately 750 amino acids, approximately 75 amino acids to approximately 700 amino acids, approximately 75 amino acids to approximately 650 amino acids, approximately 75 amino acids to approximately 600 amino acids, approximately 75 amino acids to approximately 550 amino acids, approximately 75 amino acids to approximately 500 amino acids, approximately 75 amino acids to approximately 450 amino acids, approximately 75 amino acids to approximately 400 amino acids, approximately 75 amino acids to approximately 350 amino acids, approximately 75 amino acids to approximately 300 amino acids, approximately 75 amino acids to approximately 280 amino acids, approximately 75 amino acids to approximately 260 amino acids, approximately 75 amino acids to approximately 240 amino acids, approximately 75 amino acids to approximately 220 amino acids, approximately 75 amino acids to approximately 200 amino acids, approximately 75 amino acids to approximately 195 amino acids, approximately 75 amino acids to approximately 190 amino acids, approximately 75 amino acids to approximately 185 amino acids, approximately 75 amino acids to approximately 180 amino acids, approximately 75 amino acids to approximately 175 amino acids, approximately 75 amino acids to approximately 170 amino acids, approximately 75 amino acids to approximately 165 amino acids, approximately 75 amino acids to approximately 160 amino acids, approximately 75 amino acids to approximately 155 amino acids, approximately 75 amino acids to approximately 150 amino acids, approximately 75 amino acids to approximately 145 amino acids, approximately 75 From approximately 140 amino acids, from approximately 75 amino acids to approximately 135 amino acids, from approximately 75 amino acids to approximately 130 amino acids, from approximately 75 amino acids to approximately 125 amino acids, from approximately 75 amino acids to approximately 120 amino acids, from approximately 75 amino acids to approximately 115 amino acids, from approximately 75 amino acids to approximately 110 amino acids, from approximately 75 amino acids to approximately 105 amino acids, from approximately 75 amino acids to approximately 100 amino acids, from approximately 75 amino acids to approximately 95 amino acids, from approximately 75 amino acids to approximately 90 amino acids, from approximately 75 amino acids to approximately 85 amino acids, from approximately 75 amino acids to approximately 80 amino acids, from approximately 80 amino acids to approximately 1000 amino acids, from approximately 80 amino acids to approximately 950 amino acids.Approximately 80 amino acids to approximately 900 amino acids, approximately 80 amino acids to approximately 850 amino acids, approximately 80 amino acids to approximately 800 amino acids, approximately 80 amino acids to approximately 750 amino acids, approximately 80 amino acids to approximately 700 amino acids, approximately 80 amino acids to approximately 650 amino acids, approximately 80 amino acids to approximately 600 amino acids, approximately 80 amino acids to approximately 550 amino acids, approximately 80 amino acids to approximately 500 amino acids, approximately 80 amino acids to approximately 450 amino acids, approximately 80 amino acids to approximately 400 amino acids, approximately 80 amino acids to approximately 350 amino acids, approximately 80 amino acids to approximately 300 amino acids, approximately 80 amino acids to approximately 280 amino acids, approximately 80 amino acids to approximately 260 amino acids. 1 amino acid, approximately 80 amino acids to approximately 240 amino acids, approximately 80 amino acids to approximately 220 amino acids, approximately 80 amino acids to approximately 200 amino acids, approximately 80 amino acids to approximately 195 amino acids, approximately 80 amino acids to approximately 190 amino acids, approximately 80 amino acids to approximately 185 amino acids, approximately 80 amino acids to approximately 180 amino acids, approximately 80 amino acids to approximately 175 amino acids, approximately 80 amino acids to approximately 170 amino acids, approximately 80 amino acids to approximately 165 amino acids, approximately 80 amino acids to approximately 160 amino acids, approximately 80 amino acids to approximately 155 amino acids, approximately 80 amino acids to approximately 150 amino acids, approximately 80 amino acids to approximately 145 amino acids, approximately 80 amino acids From approximately 140 amino acids, from approximately 80 amino acids to approximately 135 amino acids, from approximately 80 amino acids to approximately 130 amino acids, from approximately 80 amino acids to approximately 125 amino acids, from approximately 80 amino acids to approximately 120 amino acids, from approximately 80 amino acids to approximately 115 amino acids, from approximately 80 amino acids to approximately 110 amino acids, from approximately 80 amino acids to approximately 105 amino acids, from approximately 80 amino acids to approximately 100 amino acids, from approximately 80 amino acids to approximately 95 amino acids, from approximately 80 amino acids to approximately 90 amino acids, from approximately 80 amino acids to approximately 85 amino acids, from approximately 85 amino acids to approximately 1000 amino acids, from approximately 85 amino acids to approximately 950 amino acids, from approximately 85 amino acids to approximately 900 amino acids, from approximately 85 Amino acids to about 850 amino acids, about 85 amino acids to about 800 amino acids, about 85 amino acids 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.Approximately 85 amino acids to approximately 220 amino acids, approximately 85 amino acids to approximately 200 amino acids, approximately 85 amino acids to approximately 195 amino acids, approximately 85 amino acids to approximately 190 amino acids, approximately 85 amino acids to approximately 185 amino acids, approximately 85 amino acids to approximately 180 amino acids, approximately 85 amino acids to approximately 175 amino acids, approximately 85 amino acids to approximately 170 amino acids, approximately 85 amino acids to approximately 165 amino acids, approximately 85 amino acids to approximately 160 amino acids, approximately 85 amino acids to approximately 155 amino acids, approximately 85 amino acids to approximately 150 amino acids, approximately 85 amino acids to approximately 145 amino acids, approximately 85 amino acids to approximately 140 amino acids, approximately 85 amino acids to Approximately 135 amino acids, approximately 85 amino acids to approximately 130 amino acids, approximately 85 amino acids to approximately 125 amino acids, approximately 85 amino acids to approximately 120 amino acids, approximately 85 amino acids to approximately 115 amino acids, approximately 85 amino acids to approximately 110 amino acids, approximately 85 amino acids to approximately 105 amino acids, approximately 85 amino acids to approximately 100 amino acids, approximately 85 amino acids to approximately 95 amino acids, approximately 85 amino acids to approximately 90 amino acids, approximately 90 amino acids to approximately 1000 amino acids, approximately 90 amino acids to approximately 950 amino acids, approximately 90 amino acids to approximately 900 amino acids, approximately 90 amino acids to approximately 850 amino acids, approximately 90 amino acids to approximately 800 amino acids. Approximately 90 amino acids to approximately 750 amino acids, approximately 90 amino acids to approximately 700 amino acids, approximately 90 amino acids to approximately 650 amino acids, approximately 90 amino acids to approximately 600 amino acids, approximately 90 amino acids to approximately 550 amino acids, approximately 90 amino acids to approximately 500 amino acids, approximately 90 amino acids to approximately 450 amino acids, approximately 90 amino acids to approximately 400 amino acids, approximately 90 amino acids to approximately 350 amino acids, approximately 90 amino acids to approximately 300 amino acids, approximately 90 amino acids to approximately 280 amino acids, approximately 90 amino acids to approximately 260 amino acids, approximately 90 amino acids to approximately 240 amino acids, approximately 90 amino acids to approximately 220 amino acids, approximately 90 amino acids to Approximately 200 amino acids, approximately 90 amino acids to approximately 195 amino acids, approximately 90 amino acids to approximately 190 amino acids, approximately 90 amino acids to approximately 185 amino acids, approximately 90 amino acids to approximately 180 amino acids, approximately 90 amino acids to approximately 175 amino acids, approximately 90 amino acids to approximately 170 amino acids, approximately 90 amino acids to approximately 165 amino acids, approximately 90 amino acids to approximately 160 amino acids, approximately 90 amino acids to approximately 155 amino acids, approximately 90 amino acids to approximately 150 amino acids, approximately 90 amino acids to approximately 145 amino acids, approximately 90 amino acids to approximately 140 amino acids, approximately 90 amino acids to approximately 135 amino acids, approximately 90 amino acids to approximately 130 amino acids.Approximately 90 amino acids to approximately 125 amino acids, approximately 90 amino acids to approximately 120 amino acids, approximately 90 amino acids to approximately 115 amino acids, approximately 90 amino acids to approximately 110 amino acids, approximately 90 amino acids to approximately 105 amino acids, approximately 90 amino acids to approximately 100 amino acids, approximately 90 amino acids to approximately 95 amino acids, approximately 95 amino acids to approximately 1000 amino acids, approximately 95 amino acids to approximately 950 amino acids, approximately 95 amino acids to approximately 900 amino acids, approximately 95 amino acids to approximately 850 amino acids, approximately 95 amino acids to approximately 800 amino acids, approximately 95 amino acids to approximately 750 amino acids, approximately 95 amino acids to approximately 700 amino acids, approximately 95 amino acids to approximately 65 0 amino acids, approximately 95 amino acids to approximately 600 amino acids, approximately 95 amino acids to approximately 550 amino acids, approximately 95 amino acids to approximately 500 amino acids, approximately 95 amino acids to approximately 450 amino acids, approximately 95 amino acids to approximately 400 amino acids, approximately 95 amino acids to approximately 350 amino acids, approximately 95 amino acids to approximately 300 amino acids, approximately 95 amino acids to approximately 280 amino acids, approximately 95 amino acids to approximately 260 amino acids, approximately 95 amino acids to approximately 240 amino acids, approximately 95 amino acids to approximately 220 amino acids, approximately 95 amino acids to approximately 200 amino acids, approximately 95 amino acids to approximately 195 amino acids, approximately 95 amino acids to approximately 190 amino acids, approximately 95 amino acids Acids up to 185 amino acids, about 95 amino acids up to 180 amino acids, about 95 amino acids up to 175 amino acids, about 95 amino acids up to 170 amino acids, about 95 amino acids up to 165 amino acids, about 95 amino acids up to 160 amino acids, about 95 amino acids up to 155 amino acids, about 95 amino acids up to 150 amino acids, about 95 amino acids up to 145 amino acids, about 95 amino acids up to 140 amino acids, about 95 amino acids up to 135 amino acids, about 95 amino acids up to 130 amino acids, about 95 amino acids up to 125 amino acids, about 95 amino acids up to 120 amino acids, about 95 amino acids up to 115 amino acids. Approximately 95 amino acids to approximately 110 amino acids, approximately 95 amino acids to approximately 105 amino acids, approximately 95 amino acids to approximately 100 amino acids, approximately 100 amino acids to approximately 1000 amino acids, approximately 100 amino acids to approximately 950 amino acids, approximately 100 amino acids to approximately 900 amino acids, approximately 100 amino acids to approximately 850 amino acids, approximately 100 amino acids to approximately 800 amino acids, approximately 100 amino acids to approximately 750 amino acids, approximately 100 amino acids to approximately 700 amino acids, approximately 100 amino acids to approximately 650 amino acids, approximately 100 amino acids to approximately 600 amino acids, approximately 100 amino acids to approximately 550 amino acids, approximately 100 amino acids to approximately 500 amino acids.Approximately 100 amino acids to approximately 450 amino acids, approximately 100 amino acids to approximately 400 amino acids, approximately 100 amino acids to approximately 350 amino acids, approximately 100 amino acids to approximately 300 amino acids, approximately 100 amino acids to approximately 280 amino acids, approximately 100 amino acids to approximately 260 amino acids, approximately 100 amino acids to approximately 240 amino acids, approximately 100 amino acids to approximately 220 amino acids, approximately 100 amino acids to approximately 200 amino acids, approximately 100 amino acids to approximately 195 amino acids, approximately 100 amino acids to approximately 190 amino acids, approximately 100 amino acids to approximately 185 amino acids, approximately 100 amino acids to approximately 180 amino acids, approximately 100 amino acids to approximately 175 amino acids. 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 Approximately 105 amino acids, approximately 105 amino acids to approximately 1000 amino acids, approximately 105 amino acids to approximately 950 amino acids, approximately 105 amino acids to approximately 900 amino acids, approximately 105 amino acids to approximately 850 amino acids, approximately 105 amino acids to approximately 800 amino acids, approximately 105 amino acids to approximately 750 amino acids, approximately 105 amino acids to approximately 700 amino acids, approximately 105 amino acids to approximately 650 amino acids, approximately 105 amino acids to approximately 600 amino acids, approximately 105 amino acids to approximately 550 amino acids, approximately 105 amino acids to approximately 500 amino acids, approximately 105 amino acids to approximately 450 amino acids, approximately 105 amino acids to approximately 400 amino acids, approximately 105 The numbers appear to be a series of amino acids and their components, possibly related to a list of amino acids and their composition. A direct translation wouldn't be meaningful without further context.Approximately 105 amino acids to approximately 160 amino acids, approximately 105 amino acids to approximately 155 amino acids, approximately 105 amino acids to approximately 150 amino acids, approximately 105 amino acids to approximately 145 amino acids, approximately 105 amino acids to approximately 140 amino acids, approximately 105 amino acids to approximately 135 amino acids, approximately 105 amino acids to approximately 130 amino acids, approximately 105 amino acids to approximately 125 amino acids, approximately 105 amino acids to approximately 120 amino acids, approximately 105 amino acids to approximately 115 amino acids, approximately 105 amino acids to approximately 110 amino acids, approximately 110 amino acids to approximately 1000 amino acids, approximately 110 amino acids to approximately 950 amino acids, approximately 110 amino acids to approximately 900 amino acids. 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 Approximately 260 amino acids, approximately 110 amino acids to approximately 240 amino acids, approximately 110 amino acids to approximately 220 amino acids, approximately 110 amino acids to approximately 200 amino acids, approximately 110 amino acids to approximately 195 amino acids, approximately 110 amino acids to approximately 190 amino acids, approximately 110 amino acids to approximately 185 amino acids, approximately 110 amino acids to approximately 180 amino acids, approximately 110 amino acids to approximately 175 amino acids, approximately 110 amino acids to approximately 170 amino acids, approximately 110 amino acids to approximately 165 amino acids, approximately 110 amino acids to approximately 160 amino acids, approximately 110 amino acids to approximately 155 amino acids, approximately 110 amino acids to approximately 150 amino acids, approximately 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.Approximately 115 amino acids to approximately 650 amino acids, approximately 115 amino acids to approximately 600 amino acids, approximately 115 amino acids to approximately 550 amino acids, approximately 115 amino acids to approximately 500 amino acids, approximately 115 amino acids to approximately 450 amino acids, approximately 115 amino acids to approximately 400 amino acids, approximately 115 amino acids to approximately 350 amino acids, approximately 115 amino acids to approximately 300 amino acids, approximately 115 amino acids to approximately 280 amino acids, approximately 115 amino acids to approximately 260 amino acids, approximately 115 amino acids to approximately 240 amino acids, approximately 115 amino acids to approximately 220 amino acids, approximately 115 amino acids to approximately 200 amino acids, approximately 115 amino acids to approximately 195 amino acids. 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 Approximately 125 amino acids, approximately 115 amino acids to approximately 120 amino acids, approximately 120 amino acids to approximately 1000 amino acids, approximately 120 amino acids to approximately 950 amino acids, approximately 120 amino acids to approximately 900 amino acids, approximately 120 amino acids to approximately 850 amino acids, approximately 120 amino acids to approximately 800 amino acids, approximately 120 amino acids to approximately 750 amino acids, approximately 120 amino acids to approximately 700 amino acids, approximately 120 amino acids to approximately 650 amino acids, approximately 120 amino acids to approximately 600 amino acids, approximately 120 amino acids to approximately 550 amino acids, approximately 120 amino acids to approximately 500 amino acids, approximately 120 amino acids to approximately 450 amino acids, approximately 120 From approximately 120 amino acids to approximately 400 amino acids, from approximately 120 amino acids to approximately 350 amino acids, from approximately 120 amino acids to approximately 300 amino acids, from approximately 120 amino acids to approximately 280 amino acids, from approximately 120 amino acids to approximately 260 amino acids, from approximately 120 amino acids to approximately 240 amino acids, from approximately 120 amino acids to approximately 220 amino acids, from approximately 120 amino acids to approximately 200 amino acids, from approximately 120 amino acids to approximately 195 amino acids, from approximately 120 amino acids to approximately 190 amino acids, from approximately 120 amino acids to approximately 185 amino acids, from approximately 120 amino acids to approximately 180 amino acids, from approximately 120 amino acids to approximately 175 amino acids, from approximately 120 amino acids to approximately 170 amino acids.Approximately 120 amino acids to approximately 165 amino acids, approximately 120 amino acids to approximately 160 amino acids, approximately 120 amino acids to approximately 155 amino acids, approximately 120 amino acids to approximately 150 amino acids, approximately 120 amino acids to approximately 145 amino acids, approximately 120 amino acids to approximately 140 amino acids, approximately 120 amino acids to approximately 135 amino acids, approximately 120 amino acids to approximately 130 amino acids, approximately 120 amino acids to approximately 125 amino acids, approximately 125 amino acids to approximately 1000 amino acids, approximately 125 amino acids to approximately 950 amino acids, approximately 125 amino acids to approximately 900 amino acids, approximately 125 amino acids to approximately 850 amino acids, approximately 125 amino acids to approximately 800 amino acids. 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 Approximately 220 amino acids, approximately 125 amino acids to approximately 200 amino acids, approximately 125 amino acids to approximately 195 amino acids, approximately 125 amino acids to approximately 190 amino acids, approximately 125 amino acids to approximately 185 amino acids, approximately 125 amino acids to approximately 180 amino acids, approximately 125 amino acids to approximately 175 amino acids, approximately 125 amino acids to approximately 170 amino acids, approximately 125 amino acids to approximately 165 amino acids, approximately 125 amino acids to approximately 160 amino acids, approximately 125 amino acids to approximately 155 amino acids, approximately 125 amino acids to approximately 150 amino acids, approximately 125 amino acids to approximately 145 amino acids, approximately 125 amino acids to approximately 140 amino acids, approximately 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 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.Approximately 130 amino acids to approximately 400 amino acids, approximately 130 amino acids to approximately 350 amino acids, approximately 130 amino acids to approximately 300 amino acids, approximately 130 amino acids to approximately 280 amino acids, approximately 130 amino acids to approximately 260 amino acids, approximately 130 amino acids to approximately 240 amino acids, approximately 130 amino acids to approximately 220 amino acids, approximately 130 amino acids to approximately 200 amino acids, approximately 130 amino acids to approximately 195 amino acids, approximately 130 amino acids to approximately 190 amino acids, approximately 130 amino acids to approximately 185 amino acids, approximately 130 amino acids to approximately 180 amino acids, approximately 130 amino acids to approximately 175 amino acids, approximately 130 amino acids to approximately 170 amino acids. Amino acids, approximately 130 amino acids to approximately 165 amino acids, approximately 130 amino acids to approximately 160 amino acids, approximately 130 amino acids to approximately 155 amino acids, approximately 130 amino acids to approximately 150 amino acids, approximately 130 amino acids to approximately 145 amino acids, approximately 130 amino acids to approximately 140 amino acids, approximately 130 amino acids to approximately 135 amino acids, approximately 135 amino acids to approximately 1000 amino acids, approximately 135 amino acids to approximately 950 amino acids, approximately 135 amino acids to approximately 900 amino acids, approximately 135 amino acids to approximately 850 amino acids, approximately 135 amino acids to approximately 800 amino acids, approximately 135 amino acids to approximately 750 amino acids, approximately 135 amino acids to Approximately 700 amino acids, approximately 135 amino acids to approximately 650 amino acids, approximately 135 amino acids to approximately 600 amino acids, approximately 135 amino acids to approximately 550 amino acids, approximately 135 amino acids to approximately 500 amino acids, approximately 135 amino acids to approximately 450 amino acids, approximately 135 amino acids to approximately 400 amino acids, approximately 135 amino acids to approximately 350 amino acids, approximately 135 amino acids to approximately 300 amino acids, approximately 135 amino acids to approximately 280 amino acids, approximately 135 amino acids to approximately 260 amino acids, approximately 135 amino acids to approximately 240 amino acids, approximately 135 amino acids to approximately 220 amino acids, approximately 135 amino acids to approximately 200 amino acids, approximately 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.Approximately 140 amino acids to approximately 900 amino acids, approximately 140 amino acids to approximately 850 amino acids, approximately 140 amino acids to approximately 800 amino acids, approximately 140 amino acids to approximately 750 amino acids, approximately 140 amino acids to approximately 700 amino acids, approximately 140 amino acids to approximately 650 amino acids, approximately 140 amino acids to approximately 600 amino acids, approximately 140 amino acids to approximately 550 amino acids, approximately 140 amino acids to approximately 500 amino acids, approximately 140 amino acids to approximately 450 amino acids, approximately 140 amino acids to approximately 400 amino acids, approximately 140 amino acids to approximately 350 amino acids, approximately 140 amino acids to approximately 300 amino acids, approximately 140 amino acids to approximately 280 amino acids. 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 Approximately 150 amino acids, approximately 140 amino acids to approximately 145 amino acids, approximately 145 amino acids to approximately 1000 amino acids, approximately 145 amino acids to approximately 950 amino acids, approximately 145 amino acids to approximately 900 amino acids, approximately 145 amino acids to approximately 850 amino acids, approximately 145 amino acids to approximately 800 amino acids, approximately 145 amino acids to approximately 750 amino acids, approximately 145 amino acids to approximately 700 amino acids, approximately 145 amino acids to approximately 650 amino acids, approximately 145 amino acids to approximately 600 amino acids, approximately 145 amino acids to approximately 550 amino acids, approximately 145 amino acids to approximately 500 amino acids, approximately 145 amino acids to approximately 450 amino acids, approximately 145 From approximately 145 amino acids to approximately 350 amino acids, from approximately 145 amino acids to approximately 300 amino acids, from approximately 145 amino acids to approximately 280 amino acids, from approximately 145 amino acids to approximately 260 amino acids, from approximately 145 amino acids to approximately 240 amino acids, from approximately 145 amino acids to approximately 220 amino acids, from approximately 145 amino acids to approximately 200 amino acids, from approximately 145 amino acids to approximately 195 amino acids, from approximately 145 amino acids to approximately 190 amino acids, from approximately 145 amino acids to approximately 185 amino acids, from approximately 145 amino acids to approximately 180 amino acids, from approximately 145 amino acids to approximately 175 amino acids, from approximately 145 amino acids to approximately 170 amino acids.Approximately 145 amino acids to approximately 165 amino acids, approximately 145 amino acids to approximately 160 amino acids, approximately 145 amino acids to approximately 155 amino acids, approximately 145 amino acids to approximately 150 amino acids, approximately 150 amino acids to approximately 1000 amino acids, approximately 150 amino acids to approximately 950 amino acids, approximately 150 amino acids to approximately 900 amino acids, approximately 150 amino acids to approximately 850 amino acids, approximately 150 amino acids to approximately 800 amino acids, approximately 150 amino acids to approximately 750 amino acids, approximately 150 amino acids to approximately 700 amino acids, approximately 150 amino acids to approximately 650 amino acids, approximately 150 amino acids to approximately 600 amino acids, approximately 150 amino acids to approximately 550 amino acids. One amino acid, approximately 150 amino acids to approximately 500 amino acids, approximately 150 amino acids to approximately 450 amino acids, approximately 150 amino acids to approximately 400 amino acids, approximately 150 amino acids to approximately 350 amino acids, approximately 150 amino acids to approximately 300 amino acids, approximately 150 amino acids to approximately 280 amino acids, approximately 150 amino acids to approximately 260 amino acids, approximately 150 amino acids to approximately 240 amino acids, approximately 150 amino acids to approximately 220 amino acids, approximately 150 amino acids to approximately 200 amino acids, approximately 150 amino acids to approximately 195 amino acids, approximately 150 amino acids to approximately 190 amino acids, approximately 150 amino acids to approximately 185 amino acids, approximately 150 amino acids to Approximately 180 amino acids, approximately 150 amino acids to approximately 175 amino acids, approximately 150 amino acids to approximately 170 amino acids, approximately 150 amino acids to approximately 165 amino acids, approximately 150 amino acids to approximately 160 amino acids, approximately 150 amino acids to approximately 155 amino acids, approximately 155 amino acids to approximately 1000 amino acids, approximately 155 amino acids to approximately 950 amino acids, approximately 155 amino acids to approximately 900 amino acids, approximately 155 amino acids to approximately 850 amino acids, approximately 155 amino acids to approximately 800 amino acids, approximately 155 amino acids to approximately 750 amino acids, approximately 155 amino acids to approximately 700 amino acids, approximately 155 amino acids to approximately 650 amino acids, approximately 155 From approximately 155 amino acids to approximately 600 amino acids, from approximately 155 amino acids to approximately 550 amino acids, from approximately 155 amino acids to approximately 500 amino acids, from approximately 155 amino acids to approximately 450 amino acids, from approximately 155 amino acids to approximately 400 amino acids, from approximately 155 amino acids to approximately 350 amino acids, from approximately 155 amino acids to approximately 300 amino acids, from approximately 155 amino acids to approximately 280 amino acids, from approximately 155 amino acids to approximately 260 amino acids, from approximately 155 amino acids to approximately 240 amino acids, from approximately 155 amino acids to approximately 220 amino acids, from approximately 155 amino acids to approximately 200 amino acids, from approximately 155 amino acids to approximately 195 amino acids, from approximately 155 amino acids to approximately 190 amino acids.Approximately 155 amino acids to approximately 185 amino acids, approximately 155 amino acids to approximately 180 amino acids, approximately 155 amino acids to approximately 175 amino acids, approximately 155 amino acids to approximately 170 amino acids, approximately 155 amino acids to approximately 165 amino acids, approximately 155 amino acids to approximately 160 amino acids, approximately 160 amino acids to approximately 1000 amino acids, approximately 160 amino acids to approximately 950 amino acids, approximately 160 amino acids to approximately 900 amino acids, approximately 160 amino acids to approximately 850 amino acids, approximately 160 amino acids to approximately 800 amino acids, approximately 160 amino acids to approximately 750 amino acids, approximately 160 amino acids to approximately 700 amino acids, approximately 160 amino acids to approximately 650 amino acids. 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 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 Approximately 190 amino acids, approximately 160 amino acids to approximately 185 amino acids, approximately 160 amino acids to approximately 180 amino acids, approximately 160 amino acids to approximately 175 amino acids, approximately 160 amino acids to approximately 170 amino acids, approximately 160 amino acids to approximately 165 amino acids, approximately 165 amino acids to approximately 1000 amino acids, approximately 165 amino acids to approximately 950 amino acids, approximately 165 amino acids to approximately 900 amino acids, approximately 165 amino acids to approximately 850 amino acids, approximately 165 amino acids to approximately 800 amino acids, approximately 165 amino acids to approximately 750 amino acids, approximately 165 amino acids to approximately 700 amino acids, approximately 165 amino acids to approximately 650 amino acids, approximately 165 From approximately 165 amino acids to approximately 550 amino acids, from approximately 165 amino acids to approximately 500 amino acids, from approximately 165 amino acids to approximately 450 amino acids, from approximately 165 amino acids to approximately 400 amino acids, from approximately 165 amino acids to approximately 350 amino acids, from approximately 165 amino acids to approximately 300 amino acids, from approximately 165 amino acids to approximately 280 amino acids, from approximately 165 amino acids to approximately 260 amino acids, from approximately 165 amino acids to approximately 240 amino acids, from approximately 165 amino acids to approximately 220 amino acids, from approximately 165 amino acids to approximately 200 amino acids, from approximately 165 amino acids to approximately 195 amino acids, from approximately 165 amino acids to approximately 190 amino acids.Approximately 165 amino acids to approximately 185 amino acids, approximately 165 amino acids to approximately 180 amino acids, approximately 165 amino acids to approximately 175 amino acids, approximately 165 amino acids to approximately 170 amino acids, approximately 170 amino acids to approximately 1000 amino acids, approximately 170 amino acids to approximately 950 amino acids, approximately 170 amino acids to approximately 900 amino acids, approximately 170 amino acids to approximately 850 amino acids, approximately 170 amino acids to approximately 800 amino acids, approximately 170 amino acids to approximately 750 amino acids, approximately 170 amino acids to approximately 700 amino acids, approximately 170 amino acids to approximately 650 amino acids, approximately 170 amino acids to approximately 600 amino acids, approximately 170 amino acids to approximately 550 amino acids. 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 Approximately 180 amino acids, approximately 170 amino acids to approximately 175 amino acids, approximately 175 amino acids to approximately 1000 amino acids, approximately 175 amino acids to approximately 950 amino acids, approximately 175 amino acids to approximately 900 amino acids, approximately 175 amino acids to approximately 850 amino acids, approximately 175 amino acids to approximately 800 amino acids, approximately 175 amino acids to approximately 750 amino acids, approximately 175 amino acids to approximately 700 amino acids, approximately 175 amino acids to approximately 650 amino acids, approximately 175 amino acids to approximately 600 amino acids, approximately 175 amino acids to approximately 550 amino acids, approximately 175 amino acids to approximately 500 amino acids, approximately 175 amino acids to approximately 450 amino acids, approximately 175 From approximately 175 amino acids to approximately 400 amino acids, from approximately 175 amino acids to approximately 350 amino acids, from approximately 175 amino acids to approximately 300 amino acids, from approximately 175 amino acids to approximately 280 amino acids, from approximately 175 amino acids to approximately 260 amino acids, from approximately 175 amino acids to approximately 240 amino acids, from approximately 175 amino acids to approximately 220 amino acids, from approximately 175 amino acids to approximately 200 amino acids, from approximately 175 amino acids to approximately 195 amino acids, from approximately 175 amino acids to approximately 190 amino acids, from approximately 175 amino acids to approximately 185 amino acids, from approximately 175 amino acids to approximately 180 amino acids, from approximately 180 amino acids to approximately 1000 amino acids, from approximately 180 amino acids to approximately 950 amino acids.Approximately 180 amino acids to approximately 900 amino acids, approximately 180 amino acids to approximately 850 amino acids, approximately 180 amino acids to approximately 800 amino acids, approximately 180 amino acids to approximately 750 amino acids, approximately 180 amino acids to approximately 700 amino acids, approximately 180 amino acids to approximately 650 amino acids, approximately 180 amino acids to approximately 600 amino acids, approximately 180 amino acids to approximately 550 amino acids, approximately 180 amino acids to approximately 500 amino acids, approximately 180 amino acids to approximately 450 amino acids, approximately 180 amino acids to approximately 400 amino acids, approximately 180 amino acids to approximately 350 amino acids, approximately 180 amino acids to approximately 300 amino acids, approximately 180 amino acids to approximately 280 amino acids. Amino acids, approximately 180 amino acids to approximately 260 amino acids, approximately 180 amino acids to approximately 240 amino acids, approximately 180 amino acids to approximately 220 amino acids, approximately 180 amino acids to approximately 200 amino acids, approximately 180 amino acids to approximately 195 amino acids, approximately 180 amino acids to approximately 190 amino acids, approximately 180 amino acids to approximately 185 amino acids, approximately 185 amino acids to approximately 1000 amino acids, approximately 185 amino acids to approximately 950 amino acids, approximately 185 amino acids to approximately 900 amino acids, approximately 185 amino acids to approximately 850 amino acids, approximately 185 amino acids to approximately 800 amino acids, approximately 185 amino acids to approximately 750 amino acids, approximately 185 amino acids to Approximately 700 amino acids, approximately 185 amino acids to approximately 650 amino acids, approximately 185 amino acids to approximately 600 amino acids, approximately 185 amino acids to approximately 550 amino acids, approximately 185 amino acids to approximately 500 amino acids, approximately 185 amino acids to approximately 450 amino acids, approximately 185 amino acids to approximately 400 amino acids, approximately 185 amino acids to approximately 350 amino acids, approximately 185 amino acids to approximately 300 amino acids, approximately 185 amino acids to approximately 280 amino acids, approximately 185 amino acids to approximately 260 amino acids, approximately 185 amino acids to approximately 240 amino acids, approximately 185 amino acids to approximately 220 amino acids, approximately 185 amino acids to approximately 200 amino acids, approximately 185 Amino acids to about 195 amino acids, about 185 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.Approximately 190 amino acids to approximately 400 amino acids, approximately 190 amino acids to approximately 350 amino acids, approximately 190 amino acids to approximately 300 amino acids, approximately 190 amino acids to approximately 280 amino acids, approximately 190 amino acids to approximately 260 amino acids, approximately 190 amino acids to approximately 240 amino acids, approximately 190 amino acids to approximately 220 amino acids, approximately 190 amino acids to approximately 200 amino acids, approximately 190 amino acids to approximately 195 amino acids, approximately 195 amino acids to approximately 1000 amino acids, approximately 195 amino acids to approximately 950 amino acids, approximately 195 amino acids to approximately 900 amino acids, approximately 195 amino acids to approximately 850 amino acids, approximately 195 amino acids to approximately 800 amino acids. 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 Approximately 220 amino acids, approximately 195 amino acids to approximately 200 amino acids, approximately 200 amino acids to approximately 1000 amino acids, approximately 200 amino acids to approximately 950 amino acids, approximately 200 amino acids to approximately 900 amino acids, approximately 200 amino acids to approximately 850 amino acids, approximately 200 amino acids to approximately 800 amino acids, approximately 200 amino acids to approximately 750 amino acids, approximately 200 amino acids to approximately 700 amino acids, approximately 200 amino acids to approximately 650 amino acids, approximately 200 amino acids to approximately 600 amino acids, approximately 200 amino acids to approximately 550 amino acids, approximately 200 amino acids to approximately 500 amino acids, approximately 200 amino acids to approximately 450 amino acids, approximately 200 The numbers appear to be a series of sequences related to amino acids: approximately 1 to approximately 400 amino acids, approximately 200 to approximately 350 amino acids, approximately 200 to approximately 300 amino acids, approximately 200 to approximately 280 amino acids, approximately 200 to approximately 260 amino acids, approximately 200 to approximately 240 amino acids, approximately 200 to approximately 220 amino acids, approximately 220 to approximately 1000 amino acids, approximately 220 to approximately 950 amino acids, approximately 220 to approximately 900 amino acids, approximately 220 to approximately 850 amino acids, approximately 220 to approximately 800 amino acids, approximately 220 to approximately 750 amino acids, and approximately 220 to approximately 700 amino acids.Approximately 220 amino acids to approximately 650 amino acids, approximately 220 amino acids to approximately 600 amino acids, approximately 220 amino acids to approximately 550 amino acids, approximately 220 amino acids to approximately 500 amino acids, approximately 220 amino acids to approximately 450 amino acids, approximately 220 amino acids to approximately 400 amino acids, approximately 220 amino acids to approximately 350 amino acids, approximately 220 amino acids to approximately 300 amino acids, approximately 220 amino acids to approximately 280 amino acids, approximately 220 amino acids to approximately 260 amino acids, approximately 220 amino acids to approximately 240 amino acids, approximately 240 amino acids to approximately 1000 amino acids, approximately 240 amino acids to approximately 950 amino acids, approximately 240 amino acids to approximately 900 amino acids. 1 amino acid, approximately 240 amino acids to approximately 850 amino acids, approximately 240 amino acids to approximately 800 amino acids, approximately 240 amino acids to approximately 750 amino acids, approximately 240 amino acids to approximately 700 amino acids, approximately 240 amino acids to approximately 650 amino acids, approximately 240 amino acids to approximately 600 amino acids, approximately 240 amino acids to approximately 550 amino acids, approximately 240 amino acids to approximately 500 amino acids, approximately 240 amino acids to approximately 450 amino acids, approximately 240 amino acids to approximately 400 amino acids, approximately 240 amino acids to approximately 350 amino acids, approximately 240 amino acids to approximately 300 amino acids, approximately 240 amino acids to approximately 280 amino acids, approximately 240 amino acids to Approximately 260 amino acids, approximately 260 amino acids to approximately 1000 amino acids, approximately 260 amino acids to approximately 950 amino acids, approximately 260 amino acids to approximately 900 amino acids, approximately 260 amino acids to approximately 850 amino acids, approximately 260 amino acids to approximately 800 amino acids, approximately 260 amino acids to approximately 750 amino acids, approximately 260 amino acids to approximately 700 amino acids, approximately 260 amino acids to approximately 650 amino acids, approximately 260 amino acids to approximately 600 amino acids, approximately 260 amino acids to approximately 550 amino acids, approximately 260 amino acids to approximately 500 amino acids, approximately 260 amino acids to approximately 450 amino acids, approximately 260 amino acids to approximately 400 amino acids, approximately 260 From approximately 350 amino acids, from approximately 260 amino acids to approximately 300 amino acids, from approximately 260 amino acids to approximately 280 amino acids, from approximately 280 amino acids to approximately 1000 amino acids, from approximately 280 amino acids to approximately 950 amino acids, from approximately 280 amino acids to approximately 900 amino acids, from approximately 280 amino acids to approximately 850 amino acids, from approximately 280 amino acids to approximately 800 amino acids, from approximately 280 amino acids to approximately 750 amino acids, from approximately 280 amino acids to approximately 700 amino acids, from approximately 280 amino acids to approximately 650 amino acids, from approximately 280 amino acids to approximately 600 amino acids, from approximately 280 amino acids to approximately 550 amino acids, from approximately 280 amino acids to approximately 500 amino acids.Approximately 280 amino acids to approximately 450 amino acids, approximately 280 amino acids to approximately 400 amino acids, approximately 280 amino acids to approximately 350 amino acids, approximately 280 amino acids to approximately 300 amino acids, approximately 300 amino acids to approximately 1000 amino acids, approximately 300 amino acids to approximately 950 amino acids, approximately 300 amino acids to approximately 900 amino acids, approximately 300 amino acids to approximately 850 amino acids, approximately 300 amino acids to approximately 800 amino acids, approximately 300 amino acids to approximately 750 amino acids, approximately 300 amino acids to approximately 700 amino acids, approximately 300 amino acids to approximately 650 amino acids, approximately 300 amino acids to approximately 600 amino acids, approximately 300 amino acids to approximately 550 amino acids. One amino acid, approximately 300 amino acids to approximately 500 amino acids, approximately 300 amino acids to approximately 450 amino acids, approximately 300 amino acids to approximately 400 amino acids, approximately 300 amino acids to approximately 350 amino acids, approximately 350 amino acids to approximately 1000 amino acids, approximately 350 amino acids to approximately 950 amino acids, approximately 350 amino acids to approximately 900 amino acids, approximately 350 amino acids to approximately 850 amino acids, approximately 350 amino acids to approximately 800 amino acids, approximately 350 amino acids to approximately 750 amino acids, approximately 350 amino acids to approximately 700 amino acids, approximately 350 amino acids to approximately 650 amino acids, approximately 350 amino acids to approximately 600 amino acids, approximately 350 amino acids to Approximately 550 amino acids, approximately 350 amino acids to approximately 500 amino acids, approximately 350 amino acids to approximately 450 amino acids, approximately 350 amino acids to approximately 400 amino acids, approximately 400 amino acids to approximately 1000 amino acids, approximately 400 amino acids to approximately 950 amino acids, approximately 400 amino acids to approximately 900 amino acids, approximately 400 amino acids to approximately 850 amino acids, approximately 400 amino acids to approximately 800 amino acids, approximately 400 amino acids to approximately 750 amino acids, approximately 400 amino acids to approximately 700 amino acids, approximately 400 amino acids to approximately 650 amino acids, approximately 400 amino acids to approximately 600 amino acids, approximately 400 amino acids to approximately 550 amino acids, approximately 400 Amino acids to about 500 amino acids, about 400 amino acids to about 450 amino acids, about 450 amino acids to about 1000 amino acids, about 450 amino acids to about 950 amino acids, about 450 amino acids to about 900 amino acids, about 450 amino acids to about 850 amino acids, about 450 amino acids to about 800 amino acids, about 450 amino acids to about 750 amino acids, about 450 amino acids to about 700 amino acids, about 450 amino acids to about 650 amino acids, about 450 amino acids to about 600 amino acids, about 450 amino acids to about 550 amino acids, about 450 amino acids to about 500 amino acids, about 500 amino acids to about 1000 amino acids.Approximately 500 amino acids to approximately 950 amino acids, approximately 500 amino acids to approximately 900 amino acids, approximately 500 amino acids to approximately 850 amino acids, approximately 500 amino acids to approximately 800 amino acids, approximately 500 amino acids to approximately 750 amino acids, approximately 500 amino acids to approximately 700 amino acids, approximately 500 amino acids to approximately 650 amino acids, approximately 500 amino acids to approximately 600 amino acids, approximately 500 amino acids to approximately 550 amino acids, approximately 550 amino acids to approximately 1000 amino acids, approximately 550 amino acids to approximately 950 amino acids, approximately 550 amino acids to approximately 900 amino acids, approximately 550 amino acids to approximately 850 amino acids, approximately 550 amino acids to approximately 800 amino acids, approximately 550 amino acids to approximately 750 amino acids, approximately 550 amino acids to approximately 700 amino acids, approximately 550 amino acids to approximately 650 amino acids, approximately 550 amino acids to approximately 600 amino acids, approximately 600 amino acids to approximately 1000 amino acids, approximately 600 amino acids to approximately 950 amino acids, approximately 600 amino acids to approximately 900 amino acids, approximately 600 amino acids to approximately 850 amino acids, approximately 600 amino acids to approximately 800 amino acids, approximately 600 amino acids to approximately 750 amino acids, approximately 600 amino acids to approximately 700 amino acids, approximately 600 amino acids to approximately 650 amino acids, approximately 650 amino acids to approximately 1000 amino acids, approximately 6 50 amino acids to about 950 amino acids, about 650 amino acids to about 900 amino acids, about 650 amino acids to about 850 amino acids, about 650 amino acids to about 800 amino acids, about 650 amino acids to about 750 amino acids, about 650 amino acids to about 700 amino acids, about 700 amino acids to about 1000 amino acids, about 700 amino acids to about 950 amino acids, about 700 amino acids to about 900 amino acids, about 700 amino acids to about 850 amino acids, about 700 amino acids to about 800 amino acids, about 700 amino acids to about 750 amino acids, about 750 amino acids to about 1000 amino acids, about 750 amino acids to about 9 50 amino acids, approximately 750 amino acids to approximately 900 amino acids, approximately 750 amino acids to approximately 850 amino acids, approximately 750 amino acids to approximately 800 amino acids, approximately 800 amino acids to approximately 1000 amino acids, approximately 800 amino acids to approximately 950 amino acids, approximately 800 amino acids to approximately 900 amino acids, approximately 800 amino acids to approximately 850 amino acids, approximately 850 amino acids to approximately 1000 amino acids, approximately 850 amino acids to approximately 950 amino acids, approximately 850 amino acids to approximately 900 amino acids, approximately 900 amino acids to approximately 1000 amino acids, approximately 900 amino acids to approximately 950 amino acids, or approximately 950 amino acids to approximately 1000 amino acids.

[0133] The target binding domain described in this article can achieve a binding rate of less than 1 × 10⁻⁶. -7 M, less than 1×10 -8 M, less than 1×10 -9 M, less than 1×10 -10 M, less than 1×10 -11 M, less than 1×10 -12 M or less than 1×10 -13 The dissociation equilibrium constant of M (K) D ) binds to its target. In some embodiments, the antigen-binding protein constructs provided herein are capable of binding to their targets at approximately 1 × 10⁻⁶. -3 M to approximately 1×10 - 5 M, approximately 1×10 -4 M to approximately 1×10 -6 M, approximately 1×10 -5 M to approximately 1×10 -7 M, approximately 1×10 -6 M to approximately 1×10 -8 M, approximately 1×10 -7 M to approximately 1×10 -9 M, approximately 1×10 -8 M to approximately 1×10 -10 M, or approximately 1×10 -9 M to approximately 1×10 -11 M (including endpoints) of K D It binds to the identified antigen.

[0134] Any of the target-binding domains described in this article can bind to its target, and its K... DBetween approximately 1 pM and approximately 30 nM (e.g., approximately 1 pM to approximately 25 nM, approximately 1 pM to approximately 20 nM, approximately 1 pM to approximately 15 nM, approximately 1 pM to approximately 10 nM, approximately 1 pM to approximately 5 nM, approximately 1 pM to approximately 2 nM, approximately 1 pM to approximately 1 nM, approximately 1 pM to approximately 950 pM, approximately 1 pM to approximately 900 pM, approximately 1 pM to approximately 850 pM, approximately 1 pM to approximately 800 pM, approximately 1 pM to approximately 750 pM, approximately 1 pM to approximately 700 pM, approximately 1 pM to approximately 650 pM, approximately 1 pM to approximately 600 pM, approximately 1 pM to approximately 550 pM, approximately 1 pM to approximately 500 pM, approximately 1 pM to approximately 450 pM, approximately 1 pM to approximately 400 pM, approximately 1 pM to approximately 350 pM, approximately 1 p M to approximately 300 pM, approximately 1 pM to approximately 250 pM, approximately 1 pM to approximately 200 pM, approximately 1 pM to approximately 150 pM, approximately 1 pM to approximately 100 pM, approximately 1 pM to approximately 90 pM, approximately 1 pM to approximately 80 pM, approximately 1 pM to approximately 70 pM, approximately 1 pM to approximately 60 pM, approximately 1 pM to approximately 50 pM, approximately 1 pM to approximately 40 pM, approximately 1 pM to approximately 30 pM, approximately 1 pM to approximately 20 pM, approximately 1 pM to approximately 10 pM, approximately 1 pM to approximately 5 pM, approximately 1 pM to approximately 4 pM, approximately 1 pM to approximately 3 pM, approximately 1 pM to approximately 2 pM, approximately 2 pM to approximately 30 nM, approximately 2 pM to approximately 25 nM, approximately 2 pM to approximately 20 nM, approximately 2 pM to approximately 15 nM, approximately 2 pM to approximately 10 nM M, approximately 2 pM to approximately 5 nM, approximately 2 pM to approximately 2 nM, approximately 2 pM to approximately 1 nM, approximately 2 pM to approximately 950 pM, approximately 2 pM to approximately 900 pM, approximately 2 pM to approximately 850 pM, approximately 2 pM to approximately 800 pM, approximately 2 pM to approximately 750 pM, approximately 2 pM to approximately 700 pM, approximately 2 pM to approximately 650 pM, approximately 2 pM to approximately 600 pM, approximately 2 pM to approximately 550 pM, approximately 2 pM to approximately 500 pM, approximately 2 pM to approximately 450 pM, approximately 2 pM to approximately 400 pM, approximately 2 pM to approximately 350 pM, approximately 2 pM to approximately 300 pM, approximately 2 pM to approximately 250 pM, approximately 2 pM to approximately 200 pM, approximately 2 pM to approximately 150 pM, approximately 2 pM to approximately 100 pM, approximately 2pM to about 90pM, about 2pM to about 80pM, about 2pM to about 70pM, about 2pM to about 60pM, about 2pM to about 50pM, about 2pM to about 40pM, about 2pM to about 30pM, about 2pM to about 20pM, about 2pM to about 10pM, about 2pM to about 5pM, about 2pM to about 4pM, about 2pM to about 3pM, about 5pM to about 30nm, about 5pM to about 25nm, about 5pM to about 20nm, about 5pM to about 15nm, about 5pM to about 10nm, about 5pM to about 5nm, about 5pM to about 2nm, about 5pM to about 1nm, about 5pM to about 950pM, about 5pM to about 900pM, about 5pM to about 850pM.Approximately 5 pM to approximately 800 pM, approximately 5 pM to approximately 750 pM, approximately 5 pM to approximately 700 pM, approximately 5 pM to approximately 650 pM, approximately 5 pM to approximately 600 pM, approximately 5 pM to approximately 550 pM, approximately 5 pM to approximately 500 pM, approximately 5 pM to approximately 450 pM, approximately 5 pM to approximately 400 pM, approximately 5 pM to approximately 350 pM, approximately 5 pM to approximately 300 pM, approximately 5 pM to approximately 250 pM, approximately 5 pM to approximately 200 pM, approximately 5 pM to approximately 150 pM, approximately 5 pM to approximately 100 pM, approximately 5 pM to approximately 90 pM, approximately 5 pM to approximately 80 pM, approximately 5 pM to approximately 70 pM, approximately 5 pM to approximately 60 pM, approximately 5 pM to approximately 50 pM, approximately 5 pM to approximately 40 pM, approximately 5pM to about 30pM, about 5pM to about 20pM, about 5pM to about 10pM, about 10pM to about 30nM, about 10pM to about 25nM, about 10pM to about 20nM, about 10pM to about 15nM, about 10pM to about 10nM, about 10pM to about 5nM, about 10pM to about 2nM, about 10pM to about 1nM, about 10pM to about 950pM, about 10pM to about 900pM, about 10pM to about 850pM, about 10pM to about 800pM, about 10pM to about 750pM, about 10pM to about 700pM, about 10pM to about 650pM, about 10pM to about 600pM, about 10pM to about 550pM, about 10pM to Approximately 500 pM, approximately 10 pM to approximately 450 pM, approximately 10 pM to approximately 400 pM, approximately 10 pM to approximately 350 pM, approximately 10 pM to approximately 300 pM, approximately 10 pM to approximately 250 pM, approximately 10 pM to approximately 200 pM, approximately 10 pM to approximately 150 pM, approximately 10 pM to approximately 100 pM, approximately 10 pM to approximately 90 pM, approximately 10 pM to approximately 80 pM, approximately 10 pM to approximately 70 pM, approximately 10 pM to approximately 60 pM, approximately 10 pM to approximately 50 pM, approximately 10 pM to approximately 40 pM, approximately 10 pM to approximately 30 pM, approximately 10 pM to approximately 20 pM, approximately 15 pM to approximately 30 nM, approximately 15 pM to approximately 25 nM, approximately 15 pM to approximately 20 nM, approximately 15 p M to approximately 15 nm, approximately 15 pM to approximately 10 nm, approximately 15 pM to approximately 5 nm, approximately 15 pM to approximately 2 nm, approximately 15 pM to approximately 1 nm, approximately 15 pM to approximately 950 pM, approximately 15 pM to approximately 900 pM, approximately 15 pM to approximately 850 pM, approximately 15 pM to approximately 800 pM, approximately 15 pM to approximately 750 pM, approximately 15 pM to approximately 700 pM, approximately 15 pM to approximately 650 pM, approximately 15 pM to approximately 600 pM, approximately 15 pM to approximately 550 pM, approximately 15 pM to approximately 500 pM, approximately 15 pM to approximately 450 pM, approximately 15 pM to approximately 400 pM, approximately 15 pM to approximately 350 pM, approximately 15 pM to approximately 300 pM, approximately 15 pM to approximately 250 pM.Approximately 15 pM to approximately 200 pM, approximately 15 pM to approximately 150 pM, approximately 15 pM to approximately 100 pM, approximately 15 pM to approximately 90 pM, approximately 15 pM to approximately 80 pM, approximately 15 pM to approximately 70 pM, approximately 15 pM to approximately 60 pM, approximately 15 pM to approximately 50 pM, approximately 15 pM to approximately 40 pM, approximately 15 pM to approximately 30 pM, approximately 15 pM to approximately 20 pM, approximately 20 pM to approximately 30 nM, approximately 20 pM to approximately 25 nM, approximately 20 pM to approximately 20 nM, approximately 20 pM to approximately 15 nM, approximately 20 pM to approximately 10 nM, approximately 20 pM to approximately 5 nM, approximately 20 pM to approximately 2 nM, approximately 20 pM to approximately 1 nM, approximately 20 pM to approximately 950 pM, approximately 20 pM to approximately 900pM, approximately 20pM to approximately 850pM, approximately 20pM to approximately 800pM, approximately 20pM to approximately 750pM, approximately 20pM to approximately 700pM, approximately 20pM to approximately 650pM, approximately 20pM to approximately 600pM, approximately 20pM to approximately 550pM, approximately 20pM to approximately 500pM, approximately 20pM to approximately 450pM, approximately 20pM to approximately 400pM, approximately 20pM to approximately 350pM, approximately 20pM to approximately 300pM, approximately 20pM to approximately 250pM, approximately 20pM to approximately 20pM, approximately 200pM to approximately 150pM, approximately 20pM to approximately 100pM, approximately 20pM to approximately 90pM, approximately 20pM to approximately 80pM, approximately 20pM to approximately 70pM M, approximately 20 pM to approximately 60 pM, approximately 20 pM to approximately 50 pM, approximately 20 pM to approximately 40 pM, approximately 20 pM to approximately 30 pM, approximately 30 pM to approximately 30 nM, approximately 30 pM to approximately 25 nM, approximately 30 pM to approximately 30 nM, approximately 30 pM to approximately 15 nM, approximately 30 pM to approximately 10 nM, approximately 30 pM to approximately 5 nM, approximately 30 pM to approximately 2 nM, approximately 30 pM to approximately 1 nM, approximately 30 pM to approximately 950 pM, approximately 30 pM to approximately 900 pM, approximately 30 pM to approximately 850 pM, approximately 30 pM to approximately 800 pM, approximately 30 pM to approximately 750 pM, approximately 30 pM to approximately 700 pM, approximately 30 pM to approximately 650 pM, approximately 30 pM to approximately 600 pM, approximately 30pM to approximately 550pM, approximately 30pM to approximately 500pM, approximately 30pM to approximately 450pM, approximately 30pM to approximately 400pM, approximately 30pM to approximately 350pM, approximately 30pM to approximately 300pM, approximately 30pM to approximately 250pM, approximately 30pM to approximately 200pM, approximately 30pM to approximately 150pM, approximately 30pM to approximately 100pM, approximately 30pM to approximately 90pM, approximately 30pM to approximately 80pM, approximately 30pM to approximately 70pM, approximately 30pM to approximately 60pM, approximately 30pM to approximately 50pM, approximately 30pM to approximately 40pM, approximately 40pM to approximately 30nM, approximately 40pM to approximately 25nM, approximately 40pM to approximately 30nM, approximately 40pM to approximately 15nM.Approximately 40 pM to approximately 10 nm, approximately 40 pM to approximately 5 nm, approximately 40 pM to approximately 2 nm, approximately 40 pM to approximately 1 nm, approximately 40 pM to approximately 950 pM, approximately 40 pM to approximately 900 pM, approximately 40 pM to approximately 850 pM, approximately 40 pM to approximately 800 pM, approximately 40 pM to approximately 750 pM, approximately 40 pM to approximately 700 pM, approximately 40 pM to approximately 650 pM, approximately 40 pM to approximately 600 pM, approximately 40 pM to approximately 550 pM, approximately 40 pM to approximately 500 pM, approximately 40 pM to approximately 450 pM, approximately 40 pM to approximately 400 pM, approximately 40 pM to approximately 350 pM, approximately 40 pM to approximately 300 pM, approximately 40 pM to approximately 250 pM, approximately 40 pM to approximately 200pM, approximately 40pM to approximately 150pM, approximately 40pM to approximately 100pM, approximately 40pM to approximately 90pM, approximately 40pM to approximately 80pM, approximately 40pM to approximately 70pM, approximately 40pM to approximately 60pM, approximately 40pM to approximately 50pM, approximately 50pM to approximately 30nM, approximately 50pM to approximately 25nM, approximately 50pM to approximately 30nM, approximately 50pM to approximately 15nM, approximately 50pM to approximately 10nM, approximately 50pM to approximately 5nM, approximately 50pM to approximately 2nM, approximately 50pM to approximately 1nM, approximately 50pM to approximately 950pM, approximately 50pM to approximately 900pM, approximately 50pM to approximately 850pM, approximately 50pM to approximately 800pM, approximately 50pM to approximately 750pM pM, approximately 50 pM to approximately 700 pM, approximately 50 pM to approximately 650 pM, approximately 50 pM to approximately 600 pM, approximately 50 pM to approximately 550 pM, approximately 50 pM to approximately 500 pM, approximately 50 pM to approximately 450 pM, approximately 50 pM to approximately 400 pM, approximately 50 pM to approximately 350 pM, approximately 50 pM to approximately 300 pM, approximately 50 pM to approximately 250 pM, approximately 50 pM to approximately 200 pM, approximately 50 pM to approximately 150 pM, approximately 50 pM to approximately 100 pM, approximately 50 pM to approximately 90 pM, approximately 50 pM to approximately 80 pM, approximately 50 pM to approximately 70 pM, approximately 50 pM to approximately 60 pM, approximately 60 pM to approximately 30 nM, approximately 60 pM to approximately 25 nM, approximately 60 pM to about 30 nmM, about 60 pM to about 15 nmM, about 60 pM to about 10 nmM, about 60 pM to about 5 nmM, about 60 pM to about 2 nmM, about 60 pM to about 1 nmM, about 60 pM to about 950 pM, about 60 pM to about 900 pM, about 60 pM to about 850 pM, about 60 pM to about 800 pM, about 60 pM to about 750 pM, about 60 pM to about 700 pM, about 60 pM to about 650 pM, about 60 pM to about 600 pM, about 60 pM to about 550 pM, about 60 pM to about 500 pM, about 60 pM to about 450 pM, about 60 pM to about 400 pM, about 60 pM to about 350 pM, about 60 pM to about 300 pMApproximately 60 pM to approximately 250 pM, approximately 60 pM to approximately 200 pM, approximately 60 pM to approximately 150 pM, approximately 60 pM to approximately 100 pM, approximately 60 pM to approximately 90 pM, approximately 60 pM to approximately 80 pM, approximately 60 pM to approximately 70 pM, approximately 70 pM to approximately 30 nM, approximately 70 pM to approximately 25 nM, approximately 70 pM to approximately 30 nM, approximately 70 pM to approximately 15 nM, approximately 70 pM to approximately 10 nM, approximately 70 pM to approximately 5 nM, approximately 70 pM to approximately 2 nM, approximately 70 pM to approximately 1 nM, approximately 70 pM to approximately 950 pM, approximately 70 pM to approximately 900 pM, approximately 70 pM to approximately 850 pM, approximately 70 pM to approximately 800 pM, approximately 70 pM to approximately 750 pM, approximately 70 p M to approximately 700 pM, approximately 70 pM to approximately 650 pM, approximately 70 pM to approximately 600 pM, approximately 70 pM to approximately 550 pM, approximately 70 pM to approximately 500 pM, approximately 70 pM to approximately 450 pM, approximately 70 pM to approximately 400 pM, approximately 70 pM to approximately 350 pM, approximately 70 pM to approximately 300 pM, approximately 70 pM to approximately 250 pM, approximately 70 pM to approximately 200 pM, approximately 70 pM to approximately 150 pM, approximately 70 pM to approximately 100 pM, approximately 70 pM to approximately 90 pM, approximately 70 pM to approximately 80 pM, approximately 80 pM to approximately 30 nM, approximately 80 pM to approximately 25 nM, approximately 80 pM to approximately 30 nM, approximately 80 pM to approximately 15 nM, approximately 80 pM to approximately 10 nM, approximately 80pM to about 5nM, about 80pM to about 2nM, about 80pM to about 1nM, about 80pM to about 950pM, about 80pM to about 900pM, about 80pM to about 850pM, about 80pM to about 800pM, about 80pM to about 750pM, about 80pM to about 700pM, about 80pM to about 650pM, about 80pM to about 600pM, about 80pM to about 550pM, about 80pM to about 500pM, about 80pM to about 450pM, about 80pM to about 400pM, about 80pM to about 350pM, about 80pM to about 300pM, about 80pM to about 250pM, about 80pM to about 200pM, about 80pM to about 150pM pM, approximately 80 pM to approximately 100 pM, approximately 80 pM to approximately 90 pM, approximately 90 pM to approximately 30 nM, approximately 90 pM to approximately 25 nM, approximately 90 pM to approximately 30 nM, approximately 90 pM to approximately 15 nM, approximately 90 pM to approximately 10 nM, approximately 90 pM to approximately 5 nM, approximately 90 pM to approximately 2 nM, approximately 90 pM to approximately 1 nM, approximately 90 pM to approximately 950 pM, approximately 90 pM to approximately 900 pM, approximately 90 pM to approximately 850 pM, approximately 90 pM to approximately 800 pM, approximately 90 pM to approximately 750 pM, approximately 90 pM to approximately 700 pM, approximately 90 pM to approximately 650 pM, approximately 90 pM to approximately 600 pM, approximately 90 pM to approximately 550 pM, approximately 90 pM to approximately 500 pM.Approximately 90 pM to approximately 450 pM, approximately 90 pM to approximately 400 pM, approximately 90 pM to approximately 350 pM, approximately 90 pM to approximately 300 pM, approximately 90 pM to approximately 250 pM, approximately 90 pM to approximately 200 pM, approximately 90 pM to approximately 150 pM, approximately 90 pM to approximately 100 pM, approximately 100 pM to approximately 30 nM, approximately 100 pM to approximately 25 nM, approximately 100 pM to approximately 30 nM, approximately 100 pM to approximately 15 nM, approximately 100 pM to approximately 10 nM, approximately 100 pM to approximately 5 nM, approximately 100 pM to approximately 2 nM, approximately 100 pM to approximately 1 nM, approximately 100 pM to approximately 950 pM, approximately 100 pM to approximately 900 pM, approximately 100 pM to approximately 850 pM, approximately 100pM to about 800pM, about 100pM to about 750pM, about 100pM to about 700pM, about 100pM to about 650pM, about 100pM to about 600pM, about 100pM to about 550pM, about 100pM to about 500pM, about 100pM to about 450pM, about 100pM to about 400pM, about 100pM to about 350pM, about 100pM to about 300pM, about 100pM to about 250pM, about 100pM to about 200pM, about 100pM to about 150pM, about 150pM to about 30nM, about 150pM to about 25nM, about 150pM to about 30nM, about 150pM to about 15nM, about 1 50pM to about 10nM, about 150pM to about 5nM, about 150pM to about 2nM, about 150pM to about 1nM, about 150pM to about 950pM, about 150pM to about 900pM, about 150pM to about 850pM, about 150pM to about 800pM, about 150pM to about 750pM, about 150pM to about 700pM, about 150pM to about 650pM, about 150pM to about 600pM, about 150pM to about 550pM, about 150pM to about 500pM, about 150pM to about 450pM, about 150pM to about 400pM, about 150pM to about 350pM, about 150pM to about 300pM, about 150pM From approximately 250 pM, from approximately 150 pM to approximately 200 pM, from approximately 200 pM to approximately 30 nM, from approximately 200 pM to approximately 25 nM, from approximately 200 pM to approximately 30 nM, from approximately 200 pM to approximately 15 nM, from approximately 200 pM to approximately 10 nM, from approximately 200 pM to approximately 5 nM, from approximately 200 pM to approximately 2 nM, from approximately 200 pM to approximately 1 nM, from approximately 200 pM to approximately 950 pM, from approximately 200 pM to approximately 900 pM, from approximately 200 pM to approximately 850 pM, from approximately 200 pM to approximately 800 pM, from approximately 200 pM to approximately 750 pM, from approximately 200 pM to approximately 700 pM, from approximately 200 pM to approximately 650 pM, from approximately 200 pM to approximately 600 pM, from approximately 200 pM to approximately 550 pM.Approximately 200 pM to approximately 500 pM, approximately 200 pM to approximately 450 pM, approximately 200 pM to approximately 400 pM, approximately 200 pM to approximately 350 pM, approximately 200 pM to approximately 300 pM, approximately 200 pM to approximately 250 pM, approximately 300 pM to approximately 30 nM, approximately 300 pM to approximately 25 nM, approximately 300 pM to approximately 30 nM, approximately 300 pM to approximately 15 nM, approximately 300 pM to approximately 10 nM, approximately 300 pM to approximately 5 nM, approximately 300 pM to approximately 2 nM, approximately 300 pM to approximately 1 nM, approximately 300 pM to approximately 950 pM, approximately 300 pM to approximately 900 pM, approximately 300 pM to approximately 850 pM, approximately 300 pM to approximately 800 pM, approximately 300 pM to Approximately 750 pM, approximately 300 pM to approximately 700 pM, approximately 300 pM to approximately 650 pM, approximately 300 pM to approximately 600 pM, approximately 300 pM to approximately 550 pM, approximately 300 pM to approximately 500 pM, approximately 300 pM to approximately 450 pM, approximately 300 pM to approximately 400 pM, approximately 300 pM to approximately 350 pM, approximately 400 pM to approximately 30 nm, approximately 400 pM to approximately 25 nm, approximately 400 pM to approximately 30 nm, approximately 400 pM to approximately 15 nm, approximately 400 pM to approximately 10 nm, approximately 400 pM to approximately 5 nm, approximately 400 pM to approximately 2 nm, approximately 400 pM to approximately 1 nm, approximately 400 pM to approximately 950 pM, approximately 400 pM to approximately 900 pM. Approximately 400 pM to approximately 850 pM, approximately 400 pM to approximately 800 pM, approximately 400 pM to approximately 750 pM, approximately 400 pM to approximately 700 pM, approximately 400 pM to approximately 650 pM, approximately 400 pM to approximately 600 pM, approximately 400 pM to approximately 550 pM, approximately 400 pM to approximately 500 pM, approximately 500 pM to approximately 30 nM, approximately 500 pM to approximately 25 nM, approximately 500 pM to approximately 30 nM, approximately 500 pM to approximately 15 nM, approximately 500 pM to approximately 10 nM, approximately 500 pM to approximately 5 nM, approximately 500 pM to approximately 2 nM, approximately 500 pM to approximately 1 nM, approximately 500 pM to approximately 950 pM, approximately 500 pM to approximately 900 pM, approximately 500 pM to Approximately 850 pM, approximately 500 pM to approximately 800 pM, approximately 500 pM to approximately 750 pM, approximately 500 pM to approximately 700 pM, approximately 500 pM to approximately 650 pM, approximately 500 pM to approximately 600 pM, approximately 500 pM to approximately 550 pM, approximately 600 pM to approximately 30 nM, approximately 600 pM to approximately 25 nM, approximately 600 pM to approximately 30 nM, approximately 600 pM to approximately 15 nM, approximately 600 pM to approximately 10 nM, approximately 600 pM to approximately 5 nM, approximately 600 pM to approximately 2 nM, approximately 600 pM to approximately 1 nM, approximately 600 pM to approximately 950 pM, approximately 600 pM to approximately 900 pM, approximately 600 pM to approximately 850 pM, approximately 600 pM to approximately 800 pMApproximately 600 pM to approximately 750 pM, approximately 600 pM to approximately 700 pM, approximately 600 pM to approximately 650 pM, approximately 700 pM to approximately 30 nM, approximately 700 pM to approximately 25 nM, approximately 700 pM to approximately 30 nM, approximately 700 pM to approximately 15 nM, approximately 700 pM to approximately 10 nM, approximately 700 pM to approximately 5 nM, approximately 700 pM to approximately 2 nM, approximately 700 pM to approximately 1 nM, approximately 700 pM to approximately 950 pM, approximately 700 pM to approximately 900 pM, approximately 700 pM to approximately 850 pM, approximately 700 pM to approximately 800 pM, approximately 700 pM to approximately 750pM, approximately 800pM to approximately 30nM, approximately 800pM to approximately 25nM, approximately 800pM to approximately 30nM, approximately 800pM to approximately 15nM, approximately 800pM to approximately 10nM, approximately 800pM to approximately 5nM, approximately 800pM to approximately 2nM, approximately 800pM to approximately 1nM, approximately 800pM to approximately 950pM, approximately 800pM to approximately 900pM, approximately 800pM to approximately 850pM, approximately 900pM to approximately 30nM, approximately 900pM to approximately 25nM, approximately 900pM to approximately 30nM, approximately 900pM to approximately 15nM, approximately 900pM to Approximately 10 nm, approximately 900 pM to approximately 5 nm, approximately 900 pM to approximately 2 nm, approximately 900 pM to approximately 1 nm, approximately 900 pM to approximately 950 pM, approximately 1 nm to approximately 30 nm, approximately 1 nm to approximately 25 nm, approximately 1 nm to approximately 20 nm, approximately 1 nm to approximately 15 nm, approximately 1 nm to approximately 10 nm, approximately 1 nm to approximately 5 nm, approximately 2 nm to approximately 30 nm, approximately 2 nm to approximately 25 nm, approximately 2 nm to approximately 20 nm, approximately 2 nm to approximately 15 nm, approximately 2 nm to approximately 10 nm, approximately 2 nm to approximately 5 nm, approximately 4 nm to approximately 30 nm, approximately 4 nm to approximately 25 nm. Approximately 4 nm to approximately 20 nm, approximately 4 nm to approximately 15 nm, approximately 4 nm to approximately 10 nm, approximately 4 nm to approximately 5 nm, approximately 5 nm to approximately 30 nm, approximately 5 nm to approximately 25 nm, approximately 5 nm to approximately 20 nm, approximately 5 nm to approximately 15 nm, approximately 5 nm to approximately 10 nm, approximately 10 nm to approximately 30 nm, approximately 10 nm to approximately 25 nm, approximately 10 nm to approximately 20 nm, approximately 10 nm to approximately 15 nm, approximately 15 nm to approximately 30 nm, approximately 15 nm to approximately 25 nm, approximately 15 nm to approximately 20 nm, approximately 20 nm to approximately 30 nm, and approximately 20 nm to approximately 25 nm).

[0135] Any of the target-binding domains described in this article can bind to its target, and its K... DBetween approximately 1 nm and approximately 10 nm (e.g., approximately 1 nm to approximately 9 nm, approximately 1 nm to approximately 8 nm, approximately 1 nm to approximately 7 nm, approximately 1 nm to approximately 6 nm, approximately 1 nm to approximately 5 nm, approximately 1 nm to approximately 4 nm, approximately 1 nm to approximately 3 nm, approximately 1 nm to approximately 2 nm, approximately 2 nm to approximately 10 nm, approximately 2 nm to approximately 9 nm, approximately 2 nm to approximately 8 nm, approximately 2 nm to approximately 7 nm, approximately 2 nm to approximately 6 nm, approximately 2 nm to approximately 5 nm, approximately 2 nm to approximately 4 nm, approximately 2 nm to approximately 3 nm, approximately 3 nm to approximately 10 nm, approximately 3 nm to approximately 9 nm, approximately 3 nm to approximately 8 nm, approximately 3 nm to approximately 7 nm, approximately 3 nm to approximately 6 nm, approximately 3 nm to... About 5 nM, about 3 nM to about 4 nM, about 4 nM to about 10 nM, about 4 nM to about 9 nM, about 4 nM to about 8 nM, about 4 nM to about 7 nM, about 4 nM to about 6 nM, about 4 nM to about 5 nM, about 5 nM to about 10 nM, about 5 nM to about 9 nM, about 5 nM to about 8 nM, about 5 nM to about 7 nM, about 5 nM to about 6 nM, about 6 nM to about 10 nM, about 6 nM to about 9 nM, about 6 nM to about 8 nM, about 6 nM to about 7 nM, about 7 nM to about 10 nM, about 7 nM to about 9 nM, about 7 nM to about 8 nM, about 8 nM to about 10 nM, about 8 nM to about 9 nM, and about 9 nM to about 10 nM).

[0136] K can be determined using a variety of different methods known in the art for any of the antigen-binding protein constructs described herein. D Values ​​(e.g., electrophoretic mobility migration analysis, membrane binding analysis, surface plasmon resonance and biomolecular binding kinetics analysis, etc.).

[0137] antigen-binding domain

[0138] In some embodiments of any of the single-chain chimeric peptides described herein, the first target-binding domain and the second target-binding domain specifically bind to the same antigen. In some embodiments of these single-chain chimeric peptides, the first target-binding domain and the second target-binding domain specifically bind to the same epitope. In some embodiments of these single-chain chimeric peptides, the first target-binding domain and the second target-binding domain comprise the same amino acid sequence.

[0139] In some embodiments of any of the single-chain chimeric peptides described herein, the first target-binding domain and the second target-binding domain specifically bind to different antigens.

[0140] In some embodiments of any single-chain chimeric polypeptide described herein, one or both of the first target-binding domain and the second target-binding domain are antigen-binding domains.

[0141] In some embodiments of any single-chain chimeric polypeptide described herein, the antigen-binding domain includes either scFv or a single-domain antibody (e.g., V). H H or V NAR domain).

[0142] In some instances, the antigen-binding domain (e.g., any of the antigen-binding domains described herein) may specifically bind to any of the following: CD16a (see, for example, those described in U.S. Patent No. 9,035,026), CD28 (see, for example, those described in U.S. Patent No. 7,723,482), CD3 (see, for example, those described in U.S. Patent No. 9,226,962), CD33 (see, for example, those described in U.S. Patent No. 8,759,494), CD20 (see, for example, those described in WO 2014 / 026054), CD19 (see, for example, those described in U.S. Patent No. 9,701,758), CD22 (see, for example, those described in WO 2003 / 104425), CD123 (see, for example, those described in WO 2014 / 130635), IL-1R (see, for example, those described in U.S. Patent No. 8,741,604), IL-1 (see, for example, ...026054), CD19 (see, for example, those described in U.S. Patent No. 9,701,758), CD22 (see, for example, those described in WO 2003 / 130635), IL-1R (see, for example, those described in U.S. Patent No. 8,741,604), IL-1 (see, for example, WO 2014 / 026054), CD19 (see, for example, those described in U. The following are examples of IL-6R (see, for example, those described in U.S. Patent No. 7,482,436), IL-4 (see, for example, those described in U.S. Patent Application Publication No. 2012 / 0171197), IL-10 (see, for example, those described in U.S. Patent Application Publication No. 2016 / 0340413), and PDL-1 (see, for example, Dreees et al., "Protein Expression and Purification"). Those described in Express. Purif. 94:60-66, 2014), TIGIT (see, for example, those described in U.S. Patent Application Publication No. 2017 / 0198042), PD-1 (see, for example, those described in U.S. Patent No. 7,488,802), TIM3 (see, for example, those described in U.S. Patent No. 8,552,156), CTLA4 (see, for example, those described in WO 2012 / 120125), MICA (see, for example, those described in WO 2016 / 154585), MICB (see, for example, those described in U.S. Patent No. 8,753,640), IL-6 (see, for example, Gejima et al., Human Antibodies) Those described in Antibodies, 11(4):121-129, 2002), IL-8 (see, for example, those described in U.S. Patent No. 6,117,980), TNFα (see, for example, Geng et al., Immunological Research (Immunol.Res.)).The following are listed in WO 2017 / 189526: 377-385, 2015: CD26a (see, for example, those listed in WO 2017 / 189526), ​​CD36 (see, for example, those listed in U.S. Patent Application Publication No. 2015 / 0259429), ULBP2 (see, for example, those listed in U.S. Patent No. 9,273,136), CD30 (see, for example, those listed in Homach et al., Scandinavia Journal of Immunology 48(5):497-501, 1998), CD200 (see, for example, those listed in U.S. Patent No. 9,085,623), IGF-1R (see, for example, those listed in U.S. Patent Application Publication No. 2017 / 0051063), MUC4AC (see, for example, WO 2017 / 189526), ​​CD26a (see, for example, those listed in WO 2017 / 189526), ​​CD36a (see, for example, those listed in U.S. Patent Application Publication No. 2015 / 0259429), ULBP2 (see, for example, those listed in U.S. Patent No. 9,273,136), CD30 (see, for example, those listed in Homach et al., Scandinavia Journal of Immunology 48(5):497-501, 1998), CD200 (see, for example, those listed in U.S. Patent No. 9,085,623), IGF-1R (see, for example, those listed in U.S. Patent Application Publication No. 2017 / 0051063), MUC4AC (see, for example, WO 2017 / 189526), ​​CD Those described in 2012 / 170470), MUC5AC (see, for example, those described in U.S. Patent No. 9,238,084), Trop-2 (see, for example, WO The following are examples of proteins described in 2013 / 068946: CMET (see, for example, Edwardraja et al., Biotechnol. Bioeng. 106(3):367-375, 2010), EGFR (see, for example, Akbari et al., Protein Expression and Purification 127:8-15, 2016), HER1 (see, for example, U.S. Patent Application Publication No. 2013 / 0274446), HER2 (see, for example, Cao et al., Biotechnol. Lett. 37(7):1347-1354, 2015), HER3 (see, for example, U.S. Patent No. 9,505,843), PSMA (see, for example, Parker et al., Protein Expression and Purification 89(2):136-145, 2013), CEA (see, for example, WO The following are examples of proteins described in 1995 / 015341, B7H3 (see, for example, those described in U.S. Patent No. 9,371,395), EPCAM (see, for example, those described in WO 2014 / 159531), BCMA (see, for example, those described in Smith et al., Molecular Therapy 26(6):1447-1456, 2018), P-cadherin (see, for example, those described in U.S. Patent No. 7,452,537), CEACAM5 (see, for example, those described in U.S. Patent No. 9,617,345), UL16 binding protein (see, for example, those described in WO 2017 / 083612), and HLA-DR (see, for example, Pistillo et al., Experimental and Clinical Immunogenetics).Those described in Clin. Immunogenet. 14(2):123-130, 1997), DLL4 (see, for example, those described in WO 2014 / 007513), TYRO3 (see, for example, those described in WO 2016 / 166348), AXL (see, for example, those described in WO 2012 / 175692), MER (see, for example, those described in WO 2016 / 106221), CD122 (see, for example, those described in U.S. Patent Application Publication No. 2016 / 0367664), CD155 (see, for example, those described in WO 2017 / 149538), or PDGF-DD (see, for example, those described in U.S. Patent No. 9,441,034).

[0143] The antigen-binding domains present in any of the single-chain chimeric polypeptides described herein are each independently selected from the following group: VHH domain, VNAR domain, and scFv.

[0144] In some embodiments, one or more of the first target binding domain, the second target binding domain, and / or one or more other antigen binding domains may be anti-CD3 scFv. In some embodiments, anti-CD3 scFv may include a heavy chain variable domain and / or a light chain variable domain, wherein the heavy chain variable domain comprises a sequence that is at least 70% identical (e.g., at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100% identical) to QVQLQQSGAELARPGASVKMSCKASGYTFTRYTMHWVKQRPGQGLEWIGYINPSRGYTNYNQKFKDKATLTTDKSSSTAYMQLSSLTSEDSAVYYCARYYDDHYCLDYWGQGTTLTVSS (SEQ ID NO:16), and the light chain variable domain comprises a sequence that is at least 70% identical (e.g., at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100% identical) to QIVLTQSPAIMSASPGEKVTMTCSASSSVSYMNWYQQKSGTSPKRWIYDTSKLASGVPAHFRGSGSGTSYSLTISGMEAEDAATYYCQQWSSNPFTFGSGTKLEINR (SEQ ID NO:16). NO:17) at least 70% congruent (e.g., at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100% congruent). In some embodiments, the scFv (e.g., any of the scFvs described herein) may include a linker sequence (e.g., any exemplary linker sequence described herein or known in the art) between a heavy chain variable domain and a light chain variable domain. In some embodiments, the anti-CD3 scFv may include: a heavy chain variable domain encoded by a nucleic acid comprising a sequence that is at least 70% congruent (e.g., at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 9...

Claims

1. A single-chain chimeric polypeptide comprising, from the N-terminus to the C-terminus: (i) First target binding domain; (ii) a soluble tissue factor domain, wherein the soluble tissue factor domain comprises the sequence of SEQ ID NO:9; and (iii) The second target binding domain, wherein: (a) The first target binding domain is composed of the sequence of SEQ ID NO:28, and the second target binding domain is composed of the sequence of SEQ ID NO:28; (b) The first target binding domain is composed of the first sequence of SEQ ID NO: 16 and the second sequence of SEQ ID NO: 17, and the second target binding domain is composed of the first sequence of SEQ ID NO: 22 and the second sequence of SEQ ID NO: 23, or (c) The first target binding domain is composed of the first sequence of SEQ ID NO: 22 and the second sequence of SEQ ID NO: 23, and the second target binding domain is composed of the first sequence of SEQ ID NO: 16 and the second sequence of SEQ ID NO:

17.

2. The single-chain chimeric polypeptide according to claim 1, wherein the first target-binding domain and the soluble tissue factor domain are directly adjacent to each other.

3. The single-chain chimeric polypeptide of claim 1, wherein the single-chain chimeric polypeptide further comprises a linker sequence between the first target-binding domain and the soluble tissue factor domain.

4. The single-chain chimeric polypeptide according to claim 1, wherein the soluble tissue factor domain and the second target-binding domain are directly adjacent to each other.

5. The single-chain chimeric polypeptide of claim 1, wherein the single-chain chimeric polypeptide further comprises a linker sequence between the soluble tissue factor domain and the second target-binding domain.

6. The single-chain chimeric polypeptide according to claim 1, wherein the single-chain chimeric polypeptide comprises the sequence of SEQ ID NO:108 or SEQ ID NO:

110.

7. The single-chain chimeric polypeptide according to claim 1, wherein: The first target binding domain consists of the first sequence of SEQ ID NO:16 and the second sequence of SEQ ID NO:17; The soluble tissue factor domain is composed of the sequence SEQ ID NO:9; and The second target binding domain consists of the first sequence of SEQ ID NO:22 and the second sequence of SEQ ID NO:

23.

8. The single-chain chimeric polypeptide according to claim 1, wherein: The first target binding domain further includes a connector between the first sequence and the second sequence of the first target binding domain; and The second target binding domain also includes a connector between the first and second sequences of the second target binding domain.

9. The single-chain chimeric polypeptide according to claim 8, wherein: The linker between the first and second sequences of the first target-binding domain is composed of the sequence of SEQ ID NO:13; and The linker between the first and second sequences of the second target binding domain is composed of the sequence of SEQ ID NO:

13.

10. The single-chain chimeric polypeptide of claim 1, wherein the single-chain chimeric polypeptide comprises the sequence of SEQ ID NO:1 or SEQ ID NO:

3.

11. The single-chain chimeric polypeptide of claim 1, wherein the soluble tissue factor domain does not induce blood clotting.

12. The single-chain chimeric polypeptide according to claim 1, wherein: The first target binding domain consists of the first sequence of SEQ ID NO:22 and the second sequence of SEQ ID NO:23; The soluble tissue factor domain is composed of the sequence SEQ ID NO:9; and The second target binding domain consists of the first sequence of SEQ ID NO:16 and the second sequence of SEQ ID NO:

17.

13. The single-chain chimeric polypeptide according to claim 12, wherein: The first target binding domain further includes a connector between the first sequence and the second sequence of the first target binding domain; and The second target binding domain also includes a connector between the first sequence and the second sequence of the second target binding domain.

14. The single-chain chimeric polypeptide according to claim 13, wherein: The linker between the first and second sequences of the first target binding domain has the sequence of SEQ ID NO:13; and The linker between the first and second sequences of the second target binding domain has the sequence of SEQ ID NO:

13.

15. The single-chain chimeric polypeptide according to claim 1, wherein the single-chain chimeric polypeptide comprises the sequence of SEQ ID NO:5 or SEQ ID NO:

7.

16. A composition comprising a single-chain chimeric polypeptide according to any one of claims 1 to 15.

17. The composition according to claim 16, wherein the composition is a pharmaceutical composition.

18. A kit comprising at least one dose of the composition according to claim 16.

19. Use of the composition according to claim 16 for in vitro stimulation of immune cells.

20. Use of the composition according to claim 16 for in vitro induction or enhancement of immune cell proliferation.

21. Use of the composition according to claim 16 for in vitro induction of immune cell differentiation into memory or memory-like immune cells.

22. The use according to any one of claims 19 to 21, wherein the immune cells are selected from the group consisting of: CD8 + T cells, CD4 + T cells and natural killer cells.

23. The use according to any one of claims 19 to 21, wherein the immune cells have been genetically modified in advance to express chimeric antigen receptors or recombinant T-cell receptors.

24. Use of a single-chain chimeric polypeptide in the preparation of a medicament for treating type 2 diabetes or atherosclerosis, wherein the single-chain chimeric polypeptide comprises, from the N-terminus to the C-terminus: (i) First target binding domain; (ii) a soluble tissue factor domain, wherein the soluble tissue factor domain comprises the sequence of SEQ ID NO:9; and (iii) The second target binding domain, wherein: The first target binding domain is composed of the sequence of SEQ ID NO:28, and the second target binding domain is composed of the sequence of SEQ ID NO:

28.

25. A nucleic acid encoding any single-chain chimeric polypeptide according to any one of claims 1 to 15.

26. A vector comprising the nucleic acid according to claim 25.

27. A cell comprising the nucleic acid according to claim 25.

28. A method for generating a single-chain chimeric polypeptide, the method comprising: The cells according to claim 27 are cultured in a culture medium under conditions sufficient to induce the production of the single-chain chimeric polypeptide. and The single-chain chimeric polypeptide is recovered from the cells and / or culture medium.

29. A single-chain chimeric polypeptide generated by the method according to claim 28.

Images