Chimeric polypeptides and uses thereof

Abstract

Provided herein are chimeric polypeptides (e.g., single-chain and / or multi-chain polypeptides) that include: (i) a first target-binding domain; (ii) a soluble T cell receptor β-chain constant region (TRβC) domain; and (iii) a second target-binding domain wherein the first-target binding domain and / or the second target binding domain is an anti-mesothelin antigen-binding domain. Also provided here are methods of using these chimeric polypeptides and nucleic acids encoding these chimeric polypeptides.

Description

[0001] Attorney Docket No. 47039-0043W01

[0002] CHIMERIC POLYPEPTIDES AND USES THEREOF CROSS-REFERENCE TO RELATED APPLICATIONS

[0003] This application claims priority to U. S. Provisional Patent Application No. 63 / 744,809, filed January 13, 2025, and U. S. Provisional Patent Application No. 63 / 830,440, filed June 25, 2025, each of which is herein incorporated by reference in its entirety.

[0004] SEQUENCE LISTING

[0005] This application contains a Sequence Listing that has been submitted electronically as an XML file named “47039-0043W01_SL_ST26. XML.” The XML file, created on January 12, 2026, is 145,895 bytes in size. The material in the XML file is hereby incorporated by reference in its entirety.

[0006] TECHNICAL FIELD

[0007] The present disclosure relates to the field of biotechnology, and more specifically, to target-binding molecules.

[0008] BACKGROUND

[0009] The T cell receptor (TCR) is expressed by the majority of mature T cell lymphomas (and -30% of T cell acute lymphoblastic leukemias). The TCR comprises a heterodimeric protein complex of two chains, TCRα and TCRβ. An ancestral duplication of the human β-chain constant gene results in the expression of one of two highly homologous chains, T cell receptor β-chain constant (TRβC) domains 1 and 2, in a mutually exclusive manner, following TCR locus rearrangement.

[0010] Human TRβC1 and TRβC2 isoforms differ by only four amino acid mutations in the extracellular domain. Otherwise, the sequence is identical between the two isoforms and the folded structure remains largely unchanged. While healthy T cells randomly express either TRβC1 or TRβC2, clonally expanded mature T cell lymphomas express one of them exclusively.

[0011] Mesothelin (MSLN) is a glycoprotein which is highly expressed on the surface of many types of cancers, including mesothelioma, and cancers of the ovary, pancreas, lung, and stomach. Therapies targeting MSLN are desirable.Attorney Docket No. 47039-0043W01

[0012] SUMMARY

[0013] The present invention is based on the discovery that T-cell receptor β-chain constant region domains 1 or 2 (TRβC1 or TRβC2) can be used as a scaffold for chimeric polypeptides including a target-binding domain.

[0014] Based on this discovery provided herein are single-chain chimeric polypeptides that include: (i) a first target-binding domain; (ii) a soluble T cell receptor β-chain constant region (TRβC) domain; and (iii) a second target-binding domain, wherein the first-target binding domain and / or the second target binding domain is an anti-mesothelin antigen-binding domain. Also provided herein are compositions that include any of the single-chain chimeric polypeptides described herein, nucleic acids that encode any of the single-chain chimeric polypeptides described herein, and cells that include any of the nucleic acids that encode any of the singlechain chimeric polypeptides described herein. Also provided herein are methods of stimulating an immune cell and methods of treating a subject in need thereof that include the use of any of the single-chain chimeric polypeptides described herein. Also provided herein are methods of producing any of the single-chain chimeric polypeptides described herein. Additional uses of any of the single-chain chimeric polypeptides are described herein.

[0015] Also provided herein are multi-chain chimeric polypeptides that include: (a) a first chimeric polypeptide comprising: (i) a first target-binding domain; (ii) a soluble T cell receptor β-chain constant region (TRβC) domain; and (iii) a first domain of a pair of affinity domains; and (b) a second chimeric polypeptide comprising: (i) a second domain of a pair of affinity domains; and (ii) a second target-binding domain, wherein the first chimeric polypeptide and the second chimeric polypeptide associate through the binding of the first domain and the second domain of the pair of affinity domains, and wherein the first-target binding domain and / or the second target binding domain is an anti-mesothelin antigen-binding domain. Also provided herein are compositions that include any of the multi -chain chimeric polypeptides described herein, nucleic acids that encode any of the multi-chain chimeric polypeptides described herein, and cells that include any of the nucleic acids that encode any of the multi-chain chimeric polypeptides described herein. Also provided herein are methods of stimulating an immune cell and methods of treating a subject in need thereof that include the use of any of the multi-chain chimeric polypeptides described herein. Also provided herein are methods of producing any ofAttorney Docket No. 47039-0043W01

[0016] the multi-chain chimeric polypeptides described herein. Additional uses of any of the multi-chain chimeric polypeptides are described herein.

[0017] In some embodiments, the soluble TRβC domain is a soluble human TRβC domain. In some embodiments, the soluble TRβC domain does not bind to an MHC protein with or without a loaded antigen. In some embodiments, the soluble TRβC domain does not stimulate an immune response in a mammal. In some embodiments, the soluble TRβC domain comprises or consists of a sequence from a wildtype soluble human TRβC. In some embodiments, the soluble TRβC domain comprises a sequence that is at least 80% identical to SEQ ID NO: 1. In some embodiments, the soluble human TRβC domain comprises a sequence that is at least 80% identical to SEQ ID NO: 3.

[0018] Provided herein are multi-chain chimeric polypeptide comprising: (a) a first chimeric polypeptide comprising: (i) a first target-binding domain; (ii) a soluble T cell receptor β-chain constant region (TRβC) domain; and (iii) a first domain of a pair of affinity domains; and (b) a second chimeric polypeptide comprising: (i) a second domain of a pair of affinity domains; and (ii) a second target-binding domain, wherein the first chimeric polypeptide and the second chimeric polypeptide associate through the binding of the first domain and the second domain of the pair of affinity domains, and wherein the first-target binding domain and / or the second target binding domain is an anti-mesothelin antigen-binding domain.

[0019] In some embodiments, the first target-binding domain and the soluble TRβC domain directly abut each other in the first chimeric polypeptide. In some embodiments, the first chimeric polypeptide further comprises a linker sequence between the first target-binding domain and the soluble TRβC domain in the first chimeric polypeptide. In some embodiments, the soluble TRβC domain and the first domain of the pair of affinity domains directly abut each other in the first chimeric polypeptide. In some embodiments, the first chimeric polypeptide further comprises a linker sequence between the soluble TRβC domain and the first domain of the pair of affinity domains in the first chimeric polypeptide. In some embodiments, the second domain of the pair of affinity domains and the second target-binding domain directly abut each other in the second chimeric polypeptide. In some embodiments, the second chimeric polypeptide further comprises a linker sequence between the second domain of the pair of affinity domains and the second target-binding domain in the second chimeric polypeptide.Attorney Docket No. 47039-0043W01

[0020] In some embodiments, the first chimeric polypeptide further comprises one or more additional target-binding domains. In some embodiments, the first domain of the pair of affinity domains and one of the one or more additional target-binding domains directly abut each other in the first chimeric polypeptide. In some embodiments, the first chimeric polypeptide further comprises a linker sequence between the first domain of the pair of affinity domains and one of the one or more additional target-binding domains in the first chimeric polypeptide.

[0021] In some embodiments, the second chimeric polypeptide further comprises one or more additional target-binding domains. In some embodiments, the second domain of the pair of affinity domains and one of the one or more additional target-binding domains directly abut each other in the second chimeric polypeptide. In some embodiments, the second chimeric polypeptide further comprises a linker sequence between the second domain of the pair of affinity domains and one of the one or more additional target-binding domains in the second chimeric polypeptide.

[0022] In some embodiments, the one or more additional target-binding domains bind specifically to CD3. In some embodiments, the one or more additional target-binding domains comprises an anti-CD3 antibody.

[0023] In some embodiments, both the first target-binding domain and the second target-binding domain are an antigen-binding domain.

[0024] In some embodiments, the first target-binding domain or the second target-binding domain binds specifically to a target selected from the group consisting of tissue factor

[0025] (CD 142), CD 16a, CD28, CD3, CD33, CD20, CD 19, CD22, CD 123, IL-1R, IL-1, VEGF, IL-6R, IL-4, IL-10, PDL-1, TIGIT, PD-1, TIM3, CTLA4, MICA, MICB, IL-6, IL-8, TNFa, CD26a, CD36, ULBP2, CD30, CD200, IGF-1R, MUC4AC, MUC5AC, Trop-2, CMET, EGFR, HER1, HER2, HER3, PSMA, CEA, B7H3, EPCAM, BCMA, P-cadherin, CEACAM5, a UL16-binding protein, HLA-DR, DLL4, TYRO3, AXL, MER, CD 122, CD 155, PDGFDD, a ligand of TGF-β receptor II (TGF-βRII), a ligand of TGF-βRIII, a ligand of DNAM1, a ligand of NKp46, a ligand of NKp44, a ligand of NKG2D, a ligand of NKp30, a ligand for a scMHCI, a ligand for a scMHCII, a ligand for a scTCR, a receptor for IL-1, a receptor for IL-2, a receptor for IL-3, a receptor for IL-4, a receptor for IL-7, a receptor for IL-8, a receptor for IL- 10, a receptor for IL-12, a receptor for IL-15, a receptor for IL-17, a receptor for IL-18, a receptor for IL-21, a receptor for PDGF-DD, a receptor for stem cell factor (SCF), a receptor for stem cell-likeAttorney Docket No. 47039-0043W01

[0026] tyrosine kinase 3 ligand (FLT3L), a receptor for MICA, a receptor for MICB, a receptor for a ULP16-binding protein, a receptor for CD 155, a receptor for CD 122, and a receptor for CD28.

[0027] In some embodiments, one of the first target-binding domain and the second targetbinding domain is a soluble interleukin or cytokine protein. In some embodiments, the soluble interleukin, cytokine, or ligand protein is selected from the group consisting of: IL-1, IL-2, IL-3, IL-4, IL-7, IL-8, IL-10, IL-12, IL-15, IL-17, IL-18, IL-21, PDGF-DD, SCF, and FLT3L. In some embodiments, the soluble interleukin, soluble cytokine protein, or soluble cell surface protein is IL-7.

[0028] In some embodiments, one or both of the first target-binding domain and the second target-binding domain is a soluble interleukin or cytokine receptor. 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, a scMHCI, a scMHCII, a scTCR, a soluble CD155, or a soluble CD28. In some embodiments, the soluble interleukin receptor, the soluble cytokine receptor, or the soluble cell surface receptor is a soluble TGF-βRII.

[0029] In some embodiments, the first target-binding domain is IL-7 and the second targetbinding domain is the anti-mesothelin antigen-binding domain. In some embodiments, the second chimeric polypeptide further comprises an anti-CD3 antigen-binding domain.

[0030] In some embodiments, the first target-binding domain is a soluble TGF-βRII and the second target-binding domain comprises the anti-mesothelin antigen-binding domain. In some embodiments, the second target-binding domain further comprises an anti-tissue factor antigenbinding domain. In some embodiments, the second chimeric polypeptide further comprises an anti-CD3 antigen-binding domain.

[0031] In some embodiments, the first target-binding domain is a soluble TGF-βRII and the second target-binding domain is the anti-mesothelin antigen-binding domain. In some embodiments, the second chimeric polypeptide further comprises an anti-CD3 antigen-binding domain.

[0032] In some embodiments, one of the first target-binding domain and the second targetbinding domain is an anti-tissue factor antigen-binding domain. In some embodiments, the first target-binding domain is the anti-mesothelin antigen-binding domain and the second target-Attorney Docket No. 47039-0043W01

[0033] binding domain is an anti-tissue factor antigen-binding domain. In some embodiments, the second chimeric polypeptide further comprises an anti-CD3 antigen-binding domain.

[0034] In some embodiments, the soluble TRβC domain is a soluble human TRβC domain. In some embodiments, the soluble TRβC domain does not bind to an MHC protein with or without a loaded antigen. In some embodiments, the soluble TRβC domain does not stimulate an immune response in a mammal. In some embodiments, the soluble TRβC domain comprises or consists of a sequence from a wildtype soluble human TRβC.

[0035] In some embodiments, the pair of affinity domains is a sushi domain from an alpha chain of human IL-15 receptor (IL15Ra) and a soluble IL-15. In some embodiments, the pair of affinity domains is selected from the group consisting of: barnase and barstar, a PKA and an AKAP, adapter / docking tag modules based on mutated RNase I fragments, and SNARE modules based on interactions of the proteins syntaxin, synaptotagmin, synaptobrevin, and SNAP25.

[0036] In some embodiments, the first chimeric polypeptide and / or the second chimeric polypeptide further comprises a signal sequence at its N-terminal end.

[0037] Provided herein are compositions comprising any of the multi -chain chimeric polypeptides discussed above. In some embodiments, the composition is a pharmaceutical composition. Also provided herein are kits comprising at least one dose of any of the compositions provided here.

[0038] Provided herein are methods of stimulating an immune cell, the method comprising: contacting an immune cell with an effective amount of any of the multi -chain chimeric polypeptides discussed above or any of the compositions discussed above.

[0039] Provided herein are methods of inducing or increasing proliferation of an immune cell, the method comprising: contacting an immune cell with an effective amount of any of the multichain chimeric polypeptides discussed above or any of the compositions discussed above.

[0040] Provided herein are methods of inducing differentiation of an immune cell into a memory or memory-like immune cell, the method comprising: contacting an immune cell with an effective amount of any of the multi-chain chimeric polypeptides discussed above or any of the compositions discussed above.

[0041] In some embodiments, the immune cell is contacted in vitro. In some embodiments, the immune cell is contacted in vivo. In some embodiments, the immune cell is selected from the group consisting of: an immature thymocyte, a peripheral blood lymphocyte, a naive T cell, aAttorney Docket No. 47039-0043W01

[0042] pluripotent Th cell precursor, a lymphoid progenitor cell, a Treg cell, a Thl7 cell, a Th22 cell, a Th9 cell, a Th2 cell, a Thl cell, a Th3 cell, y8 T cell, an ap T cell, a tumor-infiltrating T cell, a CD8+ T cell, a CD4+ T cell, a natural killer T cell, a mast cell, a macrophage, a neutrophil, a dendritic cell, a basophil, an eosinophil, and a natural killer cell. In some embodiments, the CD8+T cell is a progenitor exhausted CD8+T (Tpex) cell, an effector memory CD8+T (Tern) cell, a central memory CD8+T (Tcm) cell, a memory CD8+stem (Tscm) cell, a virtual memory CD8+T (Tvm) cell, an antigen-experienced CD8+T cell, or a terminally-exhausted CD8+T (Tex) cell. In some embodiments, the immune cell has previously been genetically modified to express a chimeric antigen receptor or a recombinant T-cell receptor.

[0043] Provided herein are methods of killing a cancer cell, an infected cell, or a senescent cell in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of any of the multi-chain chimeric polypeptides discussed above or any of the compositions discussed above.

[0044] Provided herein are methods of treating a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of any of the multi-chain chimeric polypeptides discussed above or any of the compositions discussed above.

[0045] In some embodiments, the subject has been identified or diagnosed as having a cancer, an aging-related disease or condition, or an infectious disease. In some embodiments, the cancer is selected from the group consisting of: solid tumor, hematological tumor, sarcoma, osteosarcoma, 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 lymphocytic leukemia (ALL), myelodysplastic syndromes (MDS), cutaneous T-cell lymphoma, retinoblastoma, stomach cancer, urothelial carcinoma, lung cancer, renal cell carcinoma, gastic and esophageal cancer, pancreatic cancer, prostate cancer, breast cancer, colorectal cancer, ovarian cancer, non-small cell lung carcinoma, squamous cell head and neck carcinoma, endometrial cancer, cervical cancer, liver cancer, biliary cancer, and hepatocellular carcinoma. In some embodiments, the cancer is selected from lung cancer, ovarian cancer, colon cancer, pancreatic cancer, cholangiocarcinoma, glioblastoma, thymic cancer, gastric cancer, endometrial cancer, cervical cancer, breast cancer, acute myeloid leukemia, liver cancer, a head and neck cancer, biliary cancer, and mesothelioma. In someAttorney Docket No. 47039-0043W01

[0046] embodiments, the cancer is selected from triple-negative breast cancer (TNBC), malignant mesothelioma, lung adenocarcinoma, non-small cell lung cancer, pancreatic ductal adenocarcinoma, platinum-resistant ovarian cancer, colorectal cancer, uterine serous carcinoma, epithelial ovarian carcinoma.

[0047] In some embodiments, the aging-related disease or condition is selected from the group consisting of: a senescence-associated skin condition, atopic dermatitis, wrinkles, senile lentigo, age spots, photoaged skin, radiation-induced dermatitis, radiation-induced skin fibrosis / injury, cutaneous wound healing, Alzheimer’s disease, aneurysm, cystic fibrosis, fibrosis in pancreatitis, glaucoma, hypertension, idiopathic pulmonary fibrosis, inflammatory bowel disease, intervertebral disc degeneration, macular degeneration, osteoarthritis, type 2 diabetes mellitus, adipose atrophy, lipodystrophy, atherosclerosis, cataracts, COPD, idiopathic pulmonary fibrosis, kidney transplant failure, liver fibrosis, loss of bone mass, myocardial infarction, sarcopenia, wound healing, alopecia, cardiomyocyte hypertrophy, osteoarthritis, Parkinson’s disease, age-associated loss of lung tissue elasticity, macular degeneration, cachexia, glomerulosclerosis, liver cirrhosis, NAFLD, osteoporosis, amyotrophic lateral sclerosis, Huntington’s disease, spinocerebellar ataxia, multiple sclerosis, and renal dysfunction.

[0048] 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.

[0049] Provided herein are nucleic acids encoding any of the multi-chain chimeric polypeptides discussed above. Also provided herein is a vector comprising any of the nucleic acids encoding any of the multi-chain chimeric polypeptides discussed above. Also provided herein is a cell comprising any of the nucleic acids encoding any of the multi-chain chimeric polypeptides discussed above. Also provided herein are methods of producing a multi-chain chimeric polypeptide, the method comprising: culturing any of the cells discussed above in a culture medium under conditions sufficient to result in the production of the multi-chain chimeric polypeptide; and recovering the multi-chain chimeric polypeptide from the cell and / or the culture medium. In some embodiments, the method comprises producing a multi-chain chimeric polypeptide by the methods discussed above.Attorney Docket No. 47039-0043W01

[0050] Provided herein is a single-chain chimeric polypeptide comprising: (i) a first targetbinding domain; (ii) a soluble T cell receptor 0-chain constant region (TRpC) domain; and (iii) a second target-binding domain, wherein the first-target binding domain and / or the second target binding domain is an anti-mesothelin antigen-binding domain.

[0051] In some embodiments, the first target-binding domain and the soluble TRβC domain directly abut each other. In some embodiments, the single-chain chimeric polypeptide further comprises a linker sequence between the first target-binding domain and the soluble TRβC domain. In some embodiments, the soluble TRβC domain and the second target-binding domain directly abut each other. In some embodiments, the single-chain chimeric polypeptide further comprises a linker sequence between the soluble TRβC domain and the second target-binding domain.

[0052] In some embodiments, the first target-binding domain and the second target-binding domain bind specifically to the same antigen. In some embodiments, the first target-binding domain and the second target-binding domain bind specifically to the same epitope. In some embodiments, the first target-binding domain and the second target-binding domain comprise the same amino acid sequence. In some embodiments, the first target-binding domain and the second target-binding domain bind specifically to different antigens. In some embodiments, both of the first target-binding domain and the second target-binding domain are an antigen-binding domain.

[0053] In some embodiments, one of the first target-binding domain and the second targetbinding domain bind to a target selected from the group consisting of: tissue factor (CD142), 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, TNFa, CD26, CD26a, CD36, ULBP2, CD30, CD200, IGF-1R, MUC4AC, MUC5AC, Trop-2, CMET, EGFR, HER1, HER2, HER3, PSMA, CEA, B7H3, EPCAM, BCMA, P-cadherin, CEACAM5, a UL16-binding protein, HLA-DR, DLL4, TYRO3, AXL, MER, CD 122, CD 155, PDGFDD, a ligand of TGF-β receptor II (TGF-βRII), a ligand of TGF-βRIII, a ligand of DNAM1, a ligand of NKp46, a ligand of NKp44, a ligand of NKG2D, a ligand of NKp30, a ligand for a scMHCI, a ligand for a scMHCII, a ligand for a scTCR, a receptor for IL-1, a receptor for IL-2, a receptor for IL-3, a receptor for IL-4, a receptor for IL-7, a receptor for IL-8, a receptor for IL- 10, a receptor for IL-12, a receptor for IL-15, a receptor for IL-17, a receptor for IL-18, a receptor for IL-21, aAttorney Docket No. 47039-0043W01

[0054] receptor for PDGF-DD, a receptor for stem cell factor (SCF), a receptor for stem cell-like tyrosine kinase 3 ligand (FLT3L), a receptor for MICA, a receptor for MICB, a receptor for a ULP16-binding protein, a receptor for CD 155, a receptor for CD 122, and a receptor for CD28.

[0055] In some embodiments, the first target-binding domain or the second target-binding domain is a soluble interleukin, a soluble cytokine protein, or a soluble cell surface protein. 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-4, IL-7, IL-8, IL-10, IL-12, IL-15, IL-17, IL-18, IL-21, PDGF-D, SCF, FLT3L, MICA, MICB, and a ULP 16-binding protein. In some embodiments, the soluble interleukin, soluble cytokine protein, or soluble cell surface protein is IL-7.

[0056] In some embodiments, the first target-binding domain or the second target-binding domain is a soluble interleukin receptor, a soluble cytokine receptor, or a soluble cell surface receptor. In some embodiments, the soluble interleukin receptor, the soluble cytokine receptor, or the 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, a scMHCI, a scMHCII, a scTCR, a soluble CD155, or a soluble CD28. In some embodiments, the soluble interleukin receptor, the soluble cytokine receptor, or the soluble cell surface receptor is a soluble TGF-βRII.

[0057] In some embodiments, the first target-binding domain is IL-7 and the second targetbinding domain is the anti-mesothelin antigen-binding domain. In some embodiments, the first target-binding domain is a soluble TGF-βRII and the second target-binding domain is the anti-mesothelin antigen-binding domain.

[0058] In some embodiments, the single-chain chimeric polypeptide further comprises one or more additional target-binding domains at its N- and / or C-terminus.

[0059] In some embodiments, the soluble TRβC domain is a soluble human TRβC domain. In some embodiments, the soluble TRβC domain does not bind to an MHC protein with or without a loaded antigen. In some embodiments, the soluble TRβC domain does not stimulate an immune response in a mammal. In some embodiments, the soluble TRβC domain comprises or consists of a sequence from a wildtype soluble human TRβC.

[0060] Also provided herein are compositions comprising any of the single-chain chimeric polypeptides discussed above. In some embodiments, the composition is a pharmaceuticalAttorney Docket No. 47039-0043W01

[0061] composition. Also provided herein are kits comprising at least one dose of any of the compositions discussed above.

[0062] Also provided herein are methods of stimulating an immune cell, the method comprising: contacting an immune cell with an effective amount of any of the single-chain chimeric polypeptides discussed above or any of the compositions discussed above.

[0063] Also provided herein are methods of inducing or increasing proliferation of an immune cell, the method comprising: contacting an immune cell with an effective amount of any of the single-chain chimeric polypeptides discussed above or any of the compositions discussed above.

[0064] Also provided herein are methods of inducing differentiation of an immune cell into a memory or memory-like immune cell, the method comprising: contacting an immune cell with an effective amount of any of the single-chain chimeric polypeptides discussed above or any of the compositions discussed above.

[0065] In some embodiments, the immune cell is contacted in vitro. In some embodiments, the immune cell is contacted in vivo. In some embodiments, the immune cell is selected from the group consisting of: an immature thymocyte, a peripheral blood lymphocyte, a naive T cell, a pluripotent Th cell precursor, a lymphoid progenitor cell, a Treg cell, a Thl7 cell, a Th22 cell, a Th9 cell, a Th2 cell, a Thl cell, a Th3 cell, y<5 T cell, an a[3 T cell, a tumor-infiltrating T cell, a CD8+ T cell, a CD4+ T cell, a natural killer T cell, a mast cell, a macrophage, a neutrophil, a dendritic cell, a basophil, an eosinophil, and a natural killer cell. In some embodiments, the CD8+T cell is a progenitor exhausted CD8+T (Tpex) cell, an effector memory CD8+T (Tem) cell, a central memory CD8+T (Tcm) cell, a memory CD8+stem (Tscm) cell, a virtual memory CD8+T (Tvm) cell, an antigen-experienced CD8+T cell, or a terminally-exhausted CD8+T (Tex) cell. In some embodiments, the immune cell has previously been genetically modified to express a chimeric antigen receptor or a recombinant T-cell receptor. In some embodiments, the method further comprises administering the immune cell to a subject in need thereof.

[0066] Also provided herein are methods of killing a cancer cell, an infected cell, or a senescent cell in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of any of the single-chain chimeric polypeptides discussed above or any of the compositions discussed above.Attorney Docket No. 47039-0043W01

[0067] Also provided herein are methods of treating a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of any of the singlechain chimeric polypeptides discussed above or any of the compositions discussed above.

[0068] In some embodiments, the subject has been identified or diagnosed as having a cancer, an aging-related disease or condition, or an infectious disease.

[0069] In some embodiments, the cancer is selected from the group consisting of: solid tumor, hematological tumor, sarcoma, osteosarcoma, 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 lymphocytic leukemia (ALL), myelodysplastic syndromes (MDS), cutaneous T-cell lymphoma, retinoblastoma, stomach cancer, urothelial carcinoma, lung cancer, renal cell carcinoma, gastic and esophageal cancer, pancreatic cancer, prostate cancer, breast cancer, colorectal cancer, ovarian cancer, non-small cell lung carcinoma, squamous cell head and neck carcinoma, endometrial cancer, cervical cancer, liver cancer, biliary cancer, and hepatocellular carcinoma.

[0070] In some embodiments, the cancer is a mesothelin-positive cancer. For example, a mesothelin-positive cancer can include a cancer with one or more cancerous cells that express or overexpress mesothelin. Mesothelin-positive cancers can be identified using methods known in the art such as anti-mesothelin antibody staining of a biopsy sample and serum soluble mesothelin related peptide. See, e.g., Byun et al. Ann Surg. 2023 Nov 1;278(5):e 1003-el 010, which is incorporated by reference herein in its entirety.

[0071] In some embodiments, the cancer is selected from prostate cancer, lung cancer, ovarian cancer, colon cancer, pancreatic cancer, cholangiocarcinoma, glioblastoma, thymic cancer, gastric cancer, endometrial cancer, cervical cancer, breast cancer, acute myeloid leukemia, liver cancer, a head and neck cancer, biliary cancer, and mesothelioma. In some embodiments, the cancer is selected from triple-negative breast cancer (TNBC), malignant mesothelioma, lung adenocarcinoma, non-small cell lung cancer, pancreatic ductal adenocarcinoma, platinum-resistant ovarian cancer, colorectal cancer, uterine serous carcinoma, epithelial ovarian carcinoma.

[0072] In some embodiments, the aging-related disease or condition is selected from the group consisting of: a senescence-associated skin condition, atopic dermatitis, wrinkles, senile lentigo,Attorney Docket No. 47039-0043W01

[0073] age spots, photoaged skin, radiation-induced dermatitis, radiation-induced skin fibrosis / injury, cutaneous wound healing, Alzheimer’s disease, aneurysm, cystic fibrosis, fibrosis in pancreatitis, glaucoma, hypertension, idiopathic pulmonary fibrosis, inflammatory bowel disease, intervertebral disc degeneration, macular degeneration, osteoarthritis, type 2 diabetes mellitus, adipose atrophy, lipodystrophy, atherosclerosis, cataracts, COPD, idiopathic pulmonary fibrosis, kidney transplant failure, liver fibrosis, loss of bone mass, myocardial infarction, sarcopenia, wound healing, alopecia, cardiomyocyte hypertrophy, osteoarthritis, Parkinson’s disease, age-associated loss of lung tissue elasticity, macular degeneration, cachexia, glomerulosclerosis, liver cirrhosis, NAFLD, osteoporosis, amyotrophic lateral sclerosis, Huntington’s disease, spinocerebellar ataxia, multiple sclerosis, and renal dysfunction.

[0074] 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.

[0075] Provided herein is a nucleic acid encoding any of the single-chain chimeric polypeptides discussed above. Also provided herein is a vector comprising any of the nucleic acids discussed above. Provided herein is a cell comprising the nucleic acid discussed above or any of the vectors discussed above.

[0076] Also provided herein are methods of producing a single-chain chimeric polypeptide, the method comprising: culturing any of the cells discussed above in a culture medium under conditions sufficient to result in the production of the single-chain chimeric polypeptide; and recovering the single-chain chimeric polypeptide from the cell and / or the culture medium. In some embodiments, the method comprises producing a single-chain chimeric polypeptide by the methods discussed above.

[0077] A “multi-chain polypeptide” as used herein to refers to a polypeptide comprising two or more (e.g., three, four, five, six, seven, eight, nine, or ten) protein chains (e.g., at least a first chimeric polypeptide and a second polypeptide), where the two or more proteins chains associate through non-covalent bonds to form a quaternary structure.

[0078] A “single-chain polypeptide” as used herein to refers to a single protein chain.

[0079] As used herein, the term “chimeric” refers to a polypeptide that includes amino acid sequences (e.g., domains) originally derived from two different sources (e.g., two differentAttorney Docket No. 47039-0043W01

[0080] naturally-occurring proteins, e.g., from the same or different species). For example, a chimeric polypeptide can include domains from at least two different naturally occurring human proteins. In some examples, a chimeric polypeptide can include a domain that is a synthetic sequence (e.g., an scFv) and a domain that is derived from a naturally-occurring protein (e.g., a naturally-occurring human protein). In some embodiments, a chimeric polypeptide can include at least two different domains that are synthetic sequences (e.g., two different scFvs).

[0081] A “T cell receptor P-chain constant region domain” refers to a polypeptide having at least 70% identity (e.g., at least 75% identity, at least 80% identity, at least 85% identity, at least 90% identity, at least 95% identity, at least 99% identity, or 100% identical) to a segment of a constant region of a wildtype mammalian T cell receptor P-chain (e.g., a wildtype human T cell receptor P-chain). Non-limiting examples of T cell receptor P-chain constant region domains are described herein.

[0082] 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 examples, a soluble interleukin protein can include a sequence that is at least 70% identical, 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 to a wildtype mature and secreted mammalian interleukin protein (e.g., a wildtype human interleukin protein) and retains its biological activity. Nonlimiting examples of soluble interleukin proteins are described herein.

[0083] The term “soluble cytokine protein” is used herein to refer to a mature and secreted cytokine protein or a biologically active fragment thereof. In some examples, a soluble cytokine protein can include a sequence that is at least 70% identical, 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 to a wildtype mature and secreted mammalian cytokine protein (e.g., a wildtype human cytokine protein) and retains its biological activity. Non-limiting examples of soluble cytokine proteins are described herein.

[0084] The term “soluble interleukin receptor” is used herein in the broadest sense to refer to a polypeptide that lacks a transmembrane domain (and optionally an intracellular domain) that is capable of binding one or more of its natural ligands (e.g., under physiological conditions, e.g., in phosphate buffered saline at room temperature). For example, a soluble interleukin receptor can include a sequence that is at least 70% identical (e.g., at least 75% identical, at least 80%Attorney Docket No. 47039-0043W01

[0085] identical, at least 85% identical, at least 90% identical, at least 95% identical, at least 99% identical, or 100% identical) to an extracellular domain of wildtype interleukin receptor and retains its ability to specifically bind to one or more of its natural ligands, but lacks its transmembrane domain (and optionally, further lacks its intracellular domain). Non-limiting examples of soluble interleukin receptors are described herein.

[0086] The term “soluble cytokine receptor” is used herein in the broadest sense to refer to a polypeptide that lacks a transmembrane domain (and optionally an intracellular domain) that is capable of binding one or more of its natural ligands (e.g., under physiological conditions, e.g., in phosphate buffered saline at room temperature). For example, a soluble cytokine receptor can include a sequence that is at least 70% identical (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) to an extracellular domain of wildtype cytokine receptor and retains its ability to specifically bind to one or more of its natural ligands, but lacks its transmembrane domain (and optionally, further lacks its intracellular domain). Non-limiting examples of soluble cytokine receptors are described herein.

[0087] An “antigen-binding domain” is one or more protein domain(s) (e.g., formed from amino acids from a single polypeptide or formed from amino acids from two or more polypeptides (e.g., the same or different polypeptides) that is capable of specifically binding to one or more different antigen(s). In some examples, an antigen-binding domain can bind to an antigen or epitope with specificity and affinity similar to that of naturally-occurring antibodies. In some embodiments, the antigen-binding domain can be an antibody or a fragment thereof. In some embodiments, an antigen-binding domain can include an alternative scaffold. Non-limiting examples of antigen-binding domains are described herein. Additional examples of antigenbinding domains are known in the art.

[0088] 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. An antibody specifically includes, e g., intact antibodies (e.g., intact immunoglobulins), antibody fragments, and multi-specific antibodies. One example of an antigen-binding domain is an antigen binding domain formed by a VH-VL or a VL-VH. Another example of an antigen-binding domain is an antigen-binding domain formed by a VH -VL dimerAttorney Docket No. 47039-0043W01

[0089] or VL-VH dimer. Additional examples of an antibody are described herein. Additional examples of an antibody are known in the art.

[0090] The term “epitope” means a portion of an antigen that specifically binds to an antigenbinding domain. Epitopes can, e.g., consist of surface-accessible amino acid residues and / or sugar side chains and may have specific three-dimensional structural characteristics, as well as specific charge characteristics. Conformational and non-conformational epitopes are distinguished in that the binding to the former but not the latter may be lost in the presence of denaturing solvents. An epitope may comprise amino acid residues that are directly involved in the binding, and other amino acid residues, which are not directly involved in the binding.

[0091] Methods for identifying an epitope to which an antigen-binding domain binds are known in the art.

[0092] An “Fv” fragment includes a non-covalently-linked dimer of one heavy chain variable domain and one light chain variable domain.

[0093] A “Fab” fragment includes the constant domain of the light chain and the first constant domain (CHI) of the heavy chain, in addition to the heavy and light chain variable domains of the Fv fragment.

[0094] A “F(ab')2” fragment includes two Fab fragments joined, near the hinge region, by disulfide bonds.

[0095] A “dual variable domain immunoglobulin” or “DVD-Ig” refers to multivalent and multispecific binding proteins as described, e.g., in DiGiammarino et al., Methods Mol. Biol. 899:145-156, 2012; Jakob et al., MABs 5:358-363, 2013; and U. S. Patent Nos. 7,612,181;

[0096] 8,258,268; 8,586,714; 8,716,450; 8,722,855; 8,735,546; and 8,822,645, each of which is incorporated by reference in its entirety.

[0097] “Affinity” refers to the strength of the sum total of non-covalent interactions between an antigen-binding site and its binding partner (e.g., an antigen or epitope). Unless indicated otherwise, as used herein, “affinity” refers to intrinsic binding affinity, which reflects a 1: 1 interaction between members of an antigen-binding domain and an antigen or epitope. The affinity of a molecule X for its partner Y can be represented by the dissociation equilibrium constant (KD). The kinetic components that contribute to the dissociation equilibrium constant are described in more detail below. Affinity can be measured by common methods known in the art, including those described herein. Affinity can be determined, for example, using surfaceAttorney Docket No. 47039-0043W01

[0098] plasmon resonance (SPR) technology (e.g., BIACORE®) or biolayer interferometry (e.g., FORTEBIO®). Additional methods for determining the affinity for an antigen-binding domain and its corresponding antigen or epitope are known in the art.

[0099] The term “pair of affinity domains” is two different protein domain(s) that bind specifically to each other with a KD of less than of less than 1 x 10'7M (e g., less than 1 x 10'8M, less than 1 x 10‘9M, less than 1 x IO'10M, or less than 1 x 10'11M). In some examples, a pair of affinity domains can be a pair of naturally-occurring proteins. In some embodiments, a pair of affinity domains can be a pair of synthetic proteins. Non-limiting examples of pairs of affinity domains are described herein.

[0100] An “immune effector cell” refers to a cell of the immune system of a mammal that is capable, directly or indirectly, of recognizing and / or causing cytostasis or cell death of a pathogenic cell (e.g., a cancer cell) in the mammal. 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. Additional examples of immune effector cells are known in the art.

[0101] The term “treatment” means to ameliorate at least one symptom of a disorder. In some examples, the disorder being treated is cancer and to ameliorate at least one symptom of cancer includes reducing aberrant proliferation, gene expression, signaling, translation, and / or secretion of factors. Generally, the methods of treatment include administering a therapeutically effective amount of composition that reduces at least one symptom of a disorder to a subject who is in need of, or who has been determined to be in need of such treatment.

[0102] The term “subject” refers to any mammal. In some embodiments, the subject or “subject in need of treatment” may be a canine (e.g., a dog), feline (e.g., a cat), equine (e.g., a horse), ovine, bovine, porcine, caprine, primate, e.g., a simian (e.g., a monkey (e.g., marmoset, baboon), or an ape (e.g., a gorilla, chimpanzee, orangutan, or gibbon) or a human; or rodent (e.g., a mouse, a guinea pig, a hamster, or a rat). In some embodiments, the subject or “subject in need of treatment” may be a non-human mammal, especially mammals that are conventionally used as models for demonstrating therapeutic efficacy in humans (e.g., murine, lapine, porcine, canine or primate animals) may be employed.

[0103] 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 inventionAttorney Docket No. 47039-0043W01

[0104] belongs. Methods and materials are described herein for use in the present invention; other, suitable methods and materials known in the art can also be used. The materials, methods, and examples are illustrative only and not intended to be limiting. All publications, patent applications, patents, sequences, database entries, and other references mentioned herein are incorporated by reference in their entirety. In case of conflict, the present specification, including definitions, will control.

[0105] Other features and advantages of the invention will be apparent from the following detailed description and figures, and from the claims.

[0106] BRIEF DESCRIPTION OF DRAWINGS

[0107] Figures 1A and IB are schematics of an exemplary multi-chain chimeric polypeptide: (i) a first chimeric polypeptide including a first target-binding domain (A in Fig. IB), a soluble T cell receptor (TCR) P-chain constant region (TR0C) domain, a first domain of an affinity pair of domains (B in Fig. IB); and (ii) a second chimeric polypeptide including a second domain of an affinity pair of domains (D in Fig. IB) and a second target-binding domain (C in Fig. IB). The schematic in Fig. 1A depicts the association of the first and the second chimeric polypeptides through the pair of affinity domains. The schematic in Fig. IB shows the order of the domains in the first and second chimeric polypeptides.

[0108] Figures 2A and 2B are schematics of an exemplary multi-chain chimeric polypeptide: (i) a first chimeric polypeptide including a first target-binding domain (A in Fig. 2B), a soluble TRpC domain, a first domain of an affinity pair of domains (B in Fig. 2B); and (ii) a second chimeric polypeptide including a second domain of an affinity pair of domains (D in Fig. 2B), a second target-binding domain (C in Fig. 2B), and an additional target-binding domain (E in Fig.

[0109] 2B). The schematic in Fig. 2A depicts the association of the first and the second chimeric polypeptides through the pair of affinity domains. The schematic in Fig. 2B shows the order of the domains in the first and second chimeric polypeptides.

[0110] Figures 3Aand 3B are schematic diagrams of exemplary TGFpRII / hTRpCl / IL-15 and anti-mesothelin antibody / IL-15RaSu fusion proteins resulting in TGFpRII / hTRpCl / IL-15: anti-mesothelin antibody / IL-15RaSu complex. The schematic in Fig. 3 A depicts the association of the first and the second chimeric polypeptides through the pair of affinity domains. TheAttorney Docket No. 47039-0043W01

[0111] schematic in Fig. 3B shows the order of the domains in the first and second chimeric polypeptides.

[0112] Figures 4A and 4B are schematic diagrams of exemplary TGFpRII / hTRpCl / IL-15 and anti-mesothelin antibody / IL-15RaSu / anti-CD3 antibody fusion proteins resulting in TGFpRII / hTRpCl / IL-15:anti-mesothelin antibody / IL-15RaSu / anti-CD3 antibody complex. The schematic in Fig. 4A depicts the association of the first and the second chimeric polypeptides through the pair of affinity domains. The schematic in Fig. 4B shows the order of the domains in the first and second chimeric polypeptides.

[0113] Figures 5Aand 5B are schematic diagrams of exemplary IL-7 / hTRpCl / IL-15 and anti-mesothelin antibody / IL-15RaSu / anti-CD3 antibody fusion proteins resulting in IL-7 / hTRpCl / IL-15:anti-mesothelin antibody / IL-15RαSu / anti-CD3 antibody complex. The schematic in Fig. 5 A depicts the association of the first and the second chimeric polypeptides through the pair of affinity domains. The schematic in Fig. 5B shows the order of the domains in the first and second chimeric polypeptides.

[0114] Figures 6Aand 6B are schematic diagrams of exemplary IL-7 / hTRpCl / IL-15 and anti-mesothelin antibody / IL-15RaSu fusion proteins resulting in IL-7 / hTRpCl / IL-15:anti-mesothelin antibody / IL-15RaSu complex. The schematic in Fig. 6A depicts the association of the first and the second chimeric polypeptides through the pair of affinity domains. The schematic in Fig. 6B shows the order of the domains in the first and second chimeric polypeptides.

[0115] Figure 7 is a schematic of an exemplary single-chain chimeric polypeptide: (i) a first target-binding domain; (ii) a soluble T cell receptor (TCR) P-chain constant region (TRβC) domain; and (iii) a second target-binding domain.

[0116] Figure 8 is a schematic diagram of an exemplary single-chain chimeric polypeptide: (i) a first target-binding domain; (ii) a soluble T cell receptor (TCR) P-chain constant region (TRpC) domain; and (iii) a second target-binding domain.

[0117] Figure 9Ais a schematic diagram of an exemplary TGFpRII / hTRpCl / IL-15 and ot-MSLNscFv / IL-15RaSu / aCD3scFv protein complex. The schematic depicts the association of the first and the second chimeric polypeptides through the pair of affinity domains.

[0118] Figure 9B is a schematic diagram of an exemplary soluble TGFpRII / hTRpCl / IL-15 and humanized anti-MSLNscFv / IL-15RoiSu / anti-CD3scFv protein complex. The schematic depictsAttorney Docket No. 47039-0043W01

[0119] the association of the first and the second chimeric polypeptides through the pair of affinity domains.

[0120] Figure 10A is a plot showing tissue factor staining in human cancer cell lines (Jurkat, A431, AsPCl, MiaPaCa2, SW1990).

[0121] Figure 1 OB is a plot showing mesothelin staining in human cancer cell lines (Jurkat, A431, AsPCl, MiaPaCa2, SW1990).

[0122] Figure 11 A is a plot showing T-cell engager-mediated conjugation of Jurkat T cells with AsPCl tumor cells using a TGFpRII / hTRpCl / IL-15:anti-TFscFv / IL-15RaSu / anti-CD3scFV protein complex.

[0123] Figure 1 IB is a plot showing T-cell engager-mediated conjugation of Jurkat T cells with AsPCl using a TGFpRII / hTRpCl / IL-15:oc-MSLNscFV / IL-15RaSu / ocCD3scFv protein complex.

[0124] Figure 11C is a plot showing T-cell engager-mediated conjugation of Jurkat T cells with AsPCl tumor cells using a TGFpRII / hTRpCl / IL-15:anti-TFscFv / IL-15RaSu / anti-CD3scFV in combination with a TGFpRII / hTRpCl / IL-15:a-MSLNscFV / IL-15RaSu / aCD3scFv protein complex.

[0125] Figure 12A is a plot showing conjugation of Jurkat T-cells with SW1990 cancer cells using a TGFpRII / hTRpCl / IL-15:anti-TFscFv / IL-15RaSu / anti-CD3scFV fusion protein.

[0126] Figure 12B is a plot showing conjugation of Jurkat T-cells with SW1990 cancer cells using a TGFpRIVhTRpCl / IL-15:oi-MSLNscFV / IL-15RaSu / o(CD3scFv protein complex.

[0127] Figure 12C is a plot showing conjugation of Jurkat T-cells with SW1990 cancer cells using a TGFpRIVhTRpCl / IL-15:anti-TFscFv / IL-15RaSu / anti-CD3scFV protein complex in combination with a TGFpRII / hTRpCl / IL-15:a-MSLNscFV / IL-15RaSu / aCD3scFv protein complex.

[0128] Figure 13A is a plot showing the percentage of CD25+ cells within CD8+ T cell populations following incubation of human PBMCs with AsPC (TF+ / MSLN+) or A431 (TF-high / MSLN-low) tumor cells and a TGFpRII / hTRpCl / IL-15:anti-TFscFv / IL-15RaSu / anti-CD3scFV protein complex, or a TGFpRIVhTRpCl / IL-15:a-MSLNscFV / IL-15RaSu / ocCD3scFv protein complex.

[0129] Figure 13B is a plot showing the percentage of CD69+ cells within CD8+ T cell populations following incubation of human PBMCs with AsPC (TF+ / MSLN+) or A431 (TF-high / MSLN-low) tumor cells and a TGFpRII / hTRpCl / IL-15:anti-TFscFv / IL-15RaSu / anti-Attorney Docket No. 47039-0043W01

[0130] CD3scFV protein complex, or a TGFpRII / hTRpCI / IL-15:a-MSLNscFV / IL-l 5RaSu / aCD3scFv protein complex.

[0131] Figure 13C is a plot showing the percentage of CD25+ cells within CD4+ T cell populations following incubation of human PBMCs with AsPC (TF+ / MSLN+) or A431 (TF-high / MSLN-low) tumor cells and a TGFpRII / hTRpCl / IL-15:anti-TFscFv / IL-15RaSu / anti-CD3scFV protein complex, or a TGFpRIVhTRpCl / IL-15:a-MSLNscFV / IL-15RaSu / aCD3scFv protein complex.

[0132] Figure 13D is a plot showing the percentage of CD69+ cells within CD4+ T cell populations following incubation of human PBMCs with AsPC (TF+ / MSLN+) or A431 (TF-high / MSLN-low) tumor cells and a TGFpRII / hTRpCl / IL-15:anti-TFscFv / IL-15RaSu / anti-CD3scFV protein complex, or a TGFpRIVhTRpCl / IL-15:a-MSLNscFV / IL-15RaSu / aCD3scFv protein complex.

[0133] Figure 14 is a plot showing the percentage of target cancer cell killing mediated by a TGFpRIVhTRpCl / IL-15:anti-TFscFv / IL-15RaSu / anti-CD3scFV protein complex, ora TGFpRIVhTRpCl / IL-15:a-MSLNscFV / IL-15RaSu / aCD3scFv protein complex through the appearance of PI+ and CellTrace Violet+ dead cells.

[0134] Figure 15 is a plot showing anti -tumor activity of T cell Engagers (TCEs) in human T cell-infused NSG mice bearing pancreatic tumor.

[0135] Figures 16A-16B are plots showing T cell engagers enhance immune infiltration of adoptively transferred human T cells into tumor-bearing NSG mice.

[0136] Figure 17A shows a schematic of the experimental study in tumor-bearing NSG mice related to Figures 17B-17F.

[0137] Figure 17B is a plot showing probability of survival after tumor implantation of tumorbearing NSG mice treated with TCE.

[0138] Figure 17C is a plot showing tumor volume after tumor implantation of tumor-bearing NSG mice treated with TCE.

[0139] Figure 17D shows tumors from NSG mice after treatment with TCE.

[0140] Figure 17E is a plot showing tumor volume after tumor implantation of tumor-bearing NSG mice treated with TCE.

[0141] Figure 17F is a plot showing tumor weight after tumor implantation of tumor-bearing NSG mice treated with TCE.Attorney Docket No. 47039-0043W01

[0142] Figure 18A is a plot showing flow cytometry of human CD3+T cells in tumors of NSG mice treated with TCE.

[0143] Figure 18B is a plot showing flow cytometry of human CD8+T cells in tumors of NSG mice treated with TCE.

[0144] Figure 18C is a plot showing flow cytometry of human CD4+T cells in tumors of NSG mice treated with TCE.

[0145] Figure 18D is a plot showing flow cytometry of human CD8+ / TEM cells in tumors of NSG mice treated with TCE.

[0146] Figure 18E is a plot showing flow cytometry of human CD87NKG2D cells in tumors of NSG mice treated with TCE.

[0147] Figure 18F is a plot showing flow cytometry of human CD87Gzmb T cells in tumors of NSG mice treated with TCE.

[0148] Figure 18G is a plot showing flow cytometry of human CD8+ / CD25 in tumors of NSG mice treated with TCE.

[0149] Figure 18H is a plot showing flow cytometry of human CD8+ / Ki67 in tumors of NSG mice treated with TCE.

[0150] Figure 181 is a plot showing flow cytometry of human CD4+ / Ki67 in tumors of NSG mice treated with TCE.

[0151] Figure 19Ais a schematic diagram of an exemplary humanized a-MSLNAb / hTRpCl / IL-15 and anti-TF / IL-15RaSu / aCD3Ab protein complex. The schematic depicts the association of the first and the second chimeric polypeptides through the pair of affinity domains.

[0152] Figure 19B is a schematic diagram of an exemplary TGFpRII / hTRpCl / IL-15 and anti-TF-humanized-a-MesothelinAb / IL-15RaSu / aCD3Ab protein complex. The schematic depicts the association of the first and the second chimeric polypeptides through the pair of affinity domains.

[0153] DETAILED DESCRIPTION

[0154] Provided herein are single-chain chimeric polypeptides that include: (i) a first targetbinding domain (e g., any of the exemplary target-binding domains described herein or known in the art), (ii) a soluble T cell receptor P-chain constant region (TRPC) domain (e.g., any of the exemplary soluble TRpC domains described herein or known in the art), and (iii) a second target-Attorney Docket No. 47039-0043W01

[0155] binding domain (eg., any of the exemplary target-binding domains described herein or known in the art), wherein the first-target binding domain and / or the second target binding domain is an anti-mesothelin antigen-binding domain.

[0156] Also provided herein are multi-chain chimeric polypeptides that include: (a) a first chimeric polypeptide including: (i) a first target-binding domain (e.g., any of the target-binding domains described herein or known in the art); (ii) a soluble T cell receptor β-chain constant region (TRβC) domain (e.g., any of the exemplary soluble TRpC domains described herein or known in the art); and (iii) a first domain of a pair of affinity domains; and (b) a second chimeric polypeptide including: (i) a second domain of a pair of affinity domains; and (ii) a second targetbinding domain (e.g., any of the target-binding domains described herein or known in the art), where the first chimeric polypeptide and the second chimeric polypeptide associate through the binding of the first domain and the second domain of the pair of affinity domains and wherein the first-target binding domain and / or the second target binding domain is an anti-mesothelin antigen-binding domain.

[0157] Also provided herein are compositions that include any of the chimeric polypeptides described herein (e.g., any of the multi-chain chimeric polypeptides described herein and / or single-chain chimeric polypeptides described herein), nucleic acids that encode any of the chimeric polypeptides described herein, and cells that include any of the nucleic acids that encode any of the chimeric polypeptides described herein. Also provided herein are methods of stimulating an immune cell and methods of treating a subject in need thereof that include the use of any of the chimeric polypeptides described herein (e.g., any of the multi-chain chimeric polypeptides described herein and / or single-chain chimeric polypeptides described herein). Also provided herein are methods of producing any of the chimeric polypeptides described herein (e.g., any of the multi -chain chimeric polypeptides described herein and / or single-chain chimeric polypeptides described herein).

[0158] In some examples of any of the chimeric polypeptides described herein (e.g., any of the multi-chain chimeric polypeptides described herein and / or single-chain chimeric polypeptides described herein) the total length of a polypeptide (e.g., a single-chain polypeptide, a first chimeric polypeptide of a multi-chain chimeric polypeptide, or a second chimeric polypeptide of a multi-chain chimeric polypeptide) can each independently be about 50 amino acids to about 3000 amino acids, about 50 amino acids to about 2500 amino acids, about 50 amino acids toAttorney Docket No. 47039-0043W01

[0159] 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, 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 480 amino acids, about 50 amino acids to about 460 amino acids, about 50 amino acids to about 440 amino acids, about 50 amino acids to about 420 amino acids, about 50 amino acids to about 400 amino acids, about 50 amino acids to about 380 amino acids, about 50 amino acids to about 360 amino acids, about 50 amino acids to about 340 amino acids, about 50 amino acids to about 320 amino acids, about 50 amino acids to about 300 amino acids, about 50 amino acids to about 280 amino acids, about 50 amino acids to about 260 amino acids, about 50 amino acids to about 240 amino acids, about 50 amino acids to about 220 amino acids, about 50 amino acids to about 200 amino acids, about 50 amino acids to about 150 amino acids, about 50 amino acids to about 100 amino acids, about 100 amino acids to about 3000 amino acids, about 100 amino acids to about 2500 amino acids, about 100 amino acids to about 2000 amino acids, about 100 amino acids to about 1500 amino acids, about 100 amino acids to about 1000 amino acids, about 100 amino acids to about 950 amino acids, about 100 amino acids to about 900 amino acids, about 100 amino acids to about 850 amino acids, about 100 amino acids to about 800 amino acids, about 100 amino acids to about 750 amino acids, about 100 amino acids to about 700 amino acids, about 100 amino acids to about 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, about 100 amino acids to about 340 amino acids, about 100 amino acids to about 320 amino acids, about 100 amino acids to about 300 amino acids, about 100 amino acids to about 280 amino acids, about 100 amino acids to about 260 amino acids, about 100 amino acids to about 240 amino acids, about 100 amino acids to about 220 amino acids, about 100 amino acids to about 200 amino acids, about 100 amino acids to aboutAttorney Docket No. 47039-0043W01

[0160] 150 amino acids, about 150 amino acids to about 3000 amino acids, about 150 amino acids to about 2500 amino acids, about 150 amino acids to about 2000 amino acids, about 150 amino acids to about 1500 amino acids, about 150 amino acids to about 1000 amino acids, about 150 amino acids to about 950 amino acids, about 150 amino acids to about 900 amino acids, about 150 amino acids to about 850 amino acids, about 150 amino acids to about 800 amino acids, about 150 amino acids to about 750 amino acids, about 150 amino acids to about 700 amino acids, about 150 amino acids to about 650 amino acids, about 150 amino acids to about 600 amino acids, about 150 amino acids to about 550 amino acids, about 150 amino acids to about 500 amino acids, about 150 amino acids to about 480 amino acids, about 150 amino acids to about 460 amino acids, about 150 amino acids to about 440 amino acids, about 150 amino acids to about 420 amino acids, about 150 amino acids to about 400 amino acids, about 150 amino acids to about 380 amino acids, about 150 amino acids to about 360 amino acids, about 150 amino acids to about 340 amino acids, about 150 amino acids to about 320 amino acids, about 150 amino acids to about 300 amino acids, about 150 amino acids to about 280 amino acids, about 150 amino acids to about 260 amino acids, about 150 amino acids to about 240 amino acids, about 150 amino acids to about 220 amino acids, about 150 amino acids to about 200 amino acids, about 200 amino acids to about 3000 amino acids, about 200 amino acids to about 2500 amino acids, about 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, about 200 amino acids to about 460 amino acids, about 200 amino acids to about 440 amino acids, about 200 amino acids to about 420 amino acids, about 200 amino acids to about 400 amino acids, about 200 amino acids to about 380 amino acids, about 200 amino acids to about 360 amino acids, about 200 amino acids to about 340 amino acids, about 200 amino acids to about 320 amino acids, about 200 amino acids to about 300 amino acids, about 200 amino acids to about 280 amino acids, about 200 amino acids to about 260 amino acids, about 200 amino acids to about 240 amino acids,Attorney Docket No. 47039-0043W01

[0161] about 200 amino acids to about 220 amino acids, about 220 amino acids to about 3000 amino acids, about 220 amino acids to about 2500 amino acids, about 220 amino acids to about 2000 amino acids, about 220 amino acids to about 1500 amino acids, about 220 amino acids to about 1000 amino acids, about 220 amino acids to about 950 amino acids, about 220 amino acids to about 900 amino acids, about 220 amino acids to about 850 amino acids, about 220 amino acids to about 800 amino acids, about 220 amino acids to about 750 amino acids, about 220 amino acids to about 700 amino acids, about 220 amino acids to about 650 amino acids, about 220 amino acids to about 600 amino acids, about 220 amino acids to about 550 amino acids, about 220 amino acids to about 500 amino acids, about 220 amino acids to about 480 amino acids, about 220 amino acids to about 460 amino acids, about 220 amino acids to about 440 amino acids, about 220 amino acids to about 420 amino acids, about 220 amino acids to about 400 amino acids, about 220 amino acids to about 380 amino acids, about 220 amino acids to about 360 amino acids, about 220 amino acids to about 340 amino acids, about 220 amino acids to about 320 amino acids, about 220 amino acids to about 300 amino acids, about 220 amino acids to about 280 amino acids, about 220 amino acids to about 260 amino acids, about 220 amino acids to about 240 amino acids, about 240 amino acids to about 3000 amino acids, about 240 amino acids to about 2500 amino acids, about 240 amino acids to about 2000 amino acids, about 240 amino acids to about 1500 amino acids, about 240 amino acids to about 1000 amino acids, about 240 amino acids to about 950 amino acids, about 240 amino acids to about 900 amino acids, about 240 amino acids to about 850 amino acids, about 240 amino acids to about 800 amino acids, about 240 amino acids to about 750 amino acids, about 240 amino acids to about 700 amino acids, about 240 amino acids to about 650 amino acids, about 240 amino acids to about 600 amino acids, about 240 amino acids to about 550 amino acids, about 240 amino acids to about 500 amino acids, about 240 amino acids to about 480 amino acids, about 240 amino acids to about 460 amino acids, about 240 amino acids to about 440 amino acids, about 240 amino acids to about 420 amino acids, about 240 amino acids to about 400 amino acids, about 240 amino acids to about 380 amino acids, about 240 amino acids to about 360 amino acids, about 240 amino acids to about 340 amino acids, about 240 amino acids to about 320 amino acids, about 240 amino acids to about 300 amino acids, about 240 amino acids to about 280 amino acids, about 240 amino acids to about 260 amino acids, about 260 amino acids to about 3000 amino acids, about 260 amino acids to about 2500 amino acids, about 260 amino acids toAttorney Docket No. 47039-0043W01

[0162] about 2000 amino acids, about 260 amino acids to about 1500 amino acids, about 260 amino acids to about 1000 amino acids, about 260 amino acids to about 950 amino acids, about 260 amino acids to about 900 amino acids, about 260 amino acids to about 850 amino acids, about 260 amino acids to about 800 amino acids, about 260 amino acids to about 750 amino acids, about 260 amino acids to about 700 amino acids, about 260 amino acids to about 650 amino acids, about 260 amino acids to about 600 amino acids, about 260 amino acids to about 550 amino acids, about 260 amino acids to about 500 amino acids, about 260 amino acids to about 480 amino acids, about 260 amino acids to about 460 amino acids, about 260 amino acids to about 440 amino acids, about 260 amino acids to about 420 amino acids, about 260 amino acids to about 400 amino acids, about 260 amino acids to about 380 amino acids, about 260 amino acids to about 360 amino acids, about 260 amino acids to about 340 amino acids, about 260 amino acids to about 320 amino acids, about 260 amino acids to about 300 amino acids, about 260 amino acids to about 280 amino acids, about 280 amino acids to about 3000 amino acids, about 280 amino acids to about 2500 amino acids, about 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, about 280 amino acids to about 460 amino acids, about 280 amino acids to about 440 amino acids, about 280 amino acids to about 420 amino acids, about 280 amino acids to about 400 amino acids, about 280 amino acids to about 380 amino acids, about 280 amino acids to about 360 amino acids, about 280 amino acids to about 340 amino acids, about 280 amino acids to about 320 amino acids, about 280 amino acids to about 300 amino acids, about 300 amino acids to about 3000 amino acids, about 300 amino acids to about 2500 amino acids, about 300 amino acids to about 2000 amino acids, about 300 amino acids to about 1500 amino acids, about 300 amino acids to about 1000 amino acids, about 300 amino acids to about 950 amino acids, about 300 amino acids to about 900 amino acids, about 300 amino acids to about 850 amino acids, about 300 amino acids to about 800 amino acids, about 300 amino acids to about 750 aminoAttorney Docket No. 47039-0043W01

[0163] acids, about 300 amino acids to about 700 amino acids, about 300 amino acids to about 650 amino acids, about 300 amino acids to about 600 amino acids, about 300 amino acids to about 550 amino acids, about 300 amino acids to about 500 amino acids, about 300 amino acids to about 480 amino acids, about 300 amino acids to about 460 amino acids, about 300 amino acids to about 440 amino acids, about 300 amino acids to about 420 amino acids, about 300 amino acids to about 400 amino acids, about 300 amino acids to about 380 amino acids, about 300 amino acids to about 360 amino acids, about 300 amino acids to about 340 amino acids, about 300 amino acids to about 320 amino acids, about 320 amino acids to about 3000 amino acids, about 320 amino acids to about 2500 amino acids, about 320 amino acids to about 2000 amino acids, about 320 amino acids to about 1500 amino acids, about 320 amino acids to about 1000 amino acids, about 320 amino acids to about 950 amino acids, about 320 amino acids to about 900 amino acids, about 320 amino acids to about 850 amino acids, about 320 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, about 320 amino acids to about 340 amino acids, about 340 amino acids to about 3000 amino acids, about 340 amino acids to about 2500 amino acids, about 340 amino acids to about 2000 amino acids, about 340 amino acids to about 1500 amino acids, about 340 amino acids to about 1000 amino acids, about 340 amino acids to about 950 amino acids, about 340 amino acids to about 900 amino acids, about 340 amino acids to about 850 amino acids, about 340 amino acids to about 800 amino acids, about 340 amino acids to about 750 amino acids, about 340 amino acids to about 700 amino acids, about 340 amino acids to about 650 amino acids, about 340 amino 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 aminoAttorney Docket No. 47039-0043W01

[0164] 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, about 360 amino acids to about 1000 amino acids, about 360 amino acids to about 950 amino acids, about 360 amino acids to about 900 amino acids, about 360 amino acids to about 850 amino acids, about 360 amino acids to about 800 amino acids, about 360 amino acids to about 750 amino acids, about 360 amino acids to about 700 amino acids, about 360 amino acids to about 650 amino acids, about 360 amino acids to about 600 amino acids, about 360 amino acids to about 550 amino acids, about 360 amino acids to about 500 amino acids, about 360 amino acids to about 480 amino acids, about 360 amino acids to about 460 amino acids, about 360 amino acids to about 440 amino acids, about 360 amino acids to about 420 amino acids, about 360 amino acids to about 400 amino acids, about 360 amino acids to about 380 amino acids, about 380 amino acids to about 3000 amino acids, about 380 amino acids to about 2500 amino acids, about 380 amino acids to about 2000 amino acids, about 380 amino acids to about 1500 amino acids, about 380 amino acids to about 1000 amino acids, about 380 amino acids to about 950 amino acids, about 380 amino acids to about 900 amino acids, about 380 amino acids to about 850 amino acids, about 380 amino acids to about 800 amino acids, about 380 amino acids to about 750 amino acids, about 380 amino acids to about 700 amino acids, about 380 amino acids to about 650 amino acids, about 380 amino acids to about 600 amino acids, about 380 amino acids to about 550 amino acids, about 380 amino acids to about 500 amino acids, about 380 amino acids to about 480 amino acids, about 380 amino acids to about 460 amino acids, about 380 amino acids to about 440 amino acids, about 380 amino acids to about 420 amino acids, about 380 amino acids to about 400 amino acids, about 400 amino acids to about 3000 amino acids, about 400 amino acids to about 2500 amino acids, about 400 amino acids to about 2000 amino acids, about 400 amino acids to about 1500 amino acids, about 400 amino acids to about 1000 amino acids, about 400 amino acids to about 950 amino acids, about 400 amino acids to about 900 amino acids, about 400 amino acids to about 850 amino acids, about 400 amino acids to about 800 amino acids, about 400 amino acids to about 750 amino acids, about 400 amino acids to about 700 amino acids, about 400 amino acids to about 650 amino acids, about 400 amino acids to about 600 amino acids, about 400 amino acids to about 550 amino acids, about 400 amino acids to about 500 amino acids, about 400 amino acids to about 480 amino acids, about 400 amino acids to about 460 aminoAttorney Docket No. 47039-0043W01

[0165] acids, about 400 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, about 420 amino acids to about 600 amino acids, about 420 amino acids to about 550 amino acids, about 420 amino acids to about 500 amino acids, about 420 amino acids to about 480 amino acids, about 420 amino acids to about 460 amino acids, about 420 amino acids to about 440 amino acids, about 440 amino acids to about 3000 amino acids, about 440 amino acids to about 2500 amino acids, about 440 amino acids to about 2000 amino acids, about 440 amino acids to about 1500 amino acids, about 440 amino acids to about 1000 amino acids, about 440 amino acids to about 950 amino acids, about 440 amino acids to about 900 amino acids, about 440 amino acids to about 850 amino acids, about 440 amino acids to about 800 amino acids, about 440 amino acids to about 750 amino acids, about 440 amino acids to about 700 amino acids, about 440 amino acids to about 650 amino acids, about 440 amino acids to about 600 amino acids, about 440 amino acids to about 550 amino acids, about 440 amino acids to about 500 amino acids, about 440 amino acids to about 480 amino acids, about 440 amino acids to about 460 amino acids, about 460 amino acids to about 3000 amino acids, about 460 amino acids to about 2500 amino acids, about 460 amino acids to about 2000 amino acids, about 460 amino acids to about 1500 amino acids, about 460 amino acids to about 1000 amino acids, about 460 amino acids to about 950 amino acids, about 460 amino acids to about 900 amino acids, about 460 amino acids to about 850 amino acids, about 460 amino acids to about 800 amino acids, about 460 amino acids to about 750 amino acids, about 460 amino acids to about 700 amino acids, about 460 amino acids to about 650 amino acids, about 460 amino acids to about 600 amino acids, about 460 amino acids to about 550 amino acids, about 460 amino acids to about 500 amino acids, about 460 amino acids to about 480 amino acids, about 480 amino acids to about 3000 amino acids, about 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 480Attorney Docket No. 47039-0043W01

[0166] 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, about 500 amino acids to about 3000 amino acids, about 500 amino acids to about 2500 amino acids, about 500 amino acids to about 2000 amino acids, about 500 amino acids to about 1500 amino acids, about 500 amino acids to about 1000 amino acids, about 500 amino acids to about 950 amino acids, about 500 amino acids to about 900 amino acids, about 500 amino acids to about 850 amino acids, about 500 amino acids to about 800 amino acids, about 500 amino acids to about 750 amino acids, about 500 amino acids to about 700 amino acids, about 500 amino acids to about 650 amino acids, about 500 amino acids to about 600 amino acids, about 500 amino acids to about 550 amino acids, about 550 amino acids to about 3000 amino acids, about 550 amino acids to about 2500 amino acids, about 550 amino acids to about 2000 amino acids, about 550 amino acids to about 1500 amino acids, about 550 amino acids to about 1000 amino acids, about 550 amino acids to about 950 amino acids, about 550 amino acids to about 900 amino acids, about 550 amino acids to about 850 amino acids, about 550 amino acids to about 800 amino acids, about 550 amino acids to about 750 amino acids, about 550 amino acids to about 700 amino acids, about 550 amino acids to about 650 amino acids, about 550 amino acids to about 600 amino acids, about 600 amino acids to about 3000 amino acids, about 600 amino acids to about 2500 amino acids, about 600 amino acids to about 2000 amino acids, about 600 amino acids to about 1500 amino acids, about 600 amino acids to about 1000 amino acids, about 600 amino acids to about 950 amino acids, about 600 amino acids to about 900 amino acids, about 600 amino acids to about 850 amino acids, about 600 amino acids to about 800 amino acids, about 600 amino acids to about 750 amino acids, about 600 amino acids to about 700 amino acids, about 600 amino acids to about 650 amino acids, about 650 amino acids to about 3000 amino acids, about 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 toAttorney Docket No. 47039-0043W01

[0167] 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 about 1500 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 3000 amino acids, about 750 amino acids to about 2500 amino acids, about 750 amino acids to about 2000 amino acids, about 750 amino acids to about 1500 amino acids, about 750 amino acids to about 1000 amino acids, about 750 amino acids to about 950 amino acids, about 750 amino acids to about 900 amino acids, about 750 amino acids to about 850 amino acids, about 750 amino acids to about 800 amino acids, about 800 amino acids to about 3000 amino acids, about 800 amino acids to about 2500 amino acids, about 800 amino acids to about 2000 amino acids, about 800 amino acids to about 1500 amino acids, about 800 amino acids to about 1000 amino acids, about 800 amino acids to about 950 amino acids, about 800 amino acids to about 900 amino acids, about 800 amino acids to about 850 amino acids, about 850 amino acids to about 3000 amino acids, about 850 amino acids to about 2500 amino acids, about 850 amino acids to about 2000 amino acids, about 850 amino acids to about 1500 amino acids, about 850 amino acids to about 1000 amino acids, about 850 amino acids to about 950 amino acids, about 850 amino acids to about 900 amino acids, about 900 amino acids to about 3000 amino acids, about 900 amino acids to about 2500 amino acids, about 900 amino acids to about 2000 amino acids, about 900 amino acids to about 1500 amino acids, about 900 amino acids to about 1000 amino acids, about 900 amino acids to about 950 amino acids, about 950 amino acids to about 3000 amino acids, about 950 amino acids to about 2500 amino acids, about 950 amino acids to about 2000 amino acids, about 950 amino acids to about 1500 amino acids, about 950 amino acids to about 1000 amino acids, about 1000 amino acids to about 3000 amino acids, about 1000 amino acids to about 2500 amino acids, about 1000 amino acids to about 2000 amino acids, about 1000 amino acids to about 1500 amino acids, about 1500 amino acids to about 3000 amino acids, about 1500 amino acids to about 2500 amino acids, about 1500 amino acids to about 2000 amino acids, about 2000 amino acids to about 3000 amino acids, about 2000 amino acids to about 2500 amino acids, or about 2500 amino acids to about 3000 amino acids.Attorney Docket No. 47039-0043W01

[0168] In some embodiments of any of the single-chain chimeric polypeptides described herein, the first target-binding domain (e.g., any of the exemplary target-binding domains described herein or known in the art) and the soluble TR0C domain (e.g., any of the exemplary soluble TRpC domains described herein) directly abut each other. In some embodiments of any of the single-chain chimeric polypeptides described herein, the single-chain chimeric polypeptide further comprises a linker sequence (e.g., any of the exemplary linker sequences described herein or known in the art) between the first target-binding domain (e.g., any of the exemplary targetbinding domains described herein or known in the art) and the soluble TR0C domain (e.g., any of the exemplary soluble TR C domains described herein). In some embodiments of any of the single-chain chimeric polypeptides described herein, the soluble TRpC domain (e.g., any of the exemplary soluble TRpC domains described herein) and the second target-binding domain (e.g., any of the exemplary target-binding domains described herein or known in the art) directly abut each other. In some embodiments of any of the single-chain chimeric polypeptides described herein, the single-chain chimeric polypeptide further comprises a linker sequence (e.g., any of the exemplary linker sequences described herein or known in the art) between the soluble TRpC domain (e.g., any of the exemplary soluble TRpC domains described herein) and the second target-binding domain (e.g., any of the exemplary target-binding domains described herein or known in the art).

[0169] In some embodiments of any of the single-chain chimeric polypeptides described herein, the first target-binding domain (e.g., any of the exemplary target-binding domains described herein or known in the art) and the second target-binding domain (e g., any of the exemplary target-binding domains described herein or known in the art) directly abut each other. In some embodiments of any of the single-chain chimeric polypeptides described herein, the single-chain chimeric polypeptide further comprises a linker sequence (e.g., any of the exemplary linker sequences described herein or known in the art) between the first target-binding domain (e.g., any of the exemplary target-binding domains described herein or known in the art) and the second target-binding domain (e.g., any of the exemplary target-binding domains described herein or known in the art). In some embodiments of any of the single-chain chimeric polypeptides described herein, the second target-binding domain (e.g., any of the exemplary target-binding domains described herein or known in the art) and the soluble TRpC domain (e.g., any of the exemplary soluble TR0C domains described herein) directly abut each other. In someAttorney Docket No. 47039-0043W01

[0170] embodiments of any of the single-chain chimeric polypeptides described herein, the single-chain chimeric polypeptide further comprises a linker sequence (e.g., any of the exemplary linker sequences described herein or known in the art) between the second target-binding domain (e.g., any of the exemplary target-binding domains described herein or known in the art) and the soluble TR C domain (e.g., any of the exemplary soluble TR0C domains described herein or known in the art).

[0171] Diagrams of exemplary single-chain chimeric polypeptides provided herein are depicted in Figures 1A-1B and 2A-2B.

[0172] In some embodiments of any of the multi-chain chimeric polypeptides described herein, the first target-binding domain (e.g., any of the first target-binding domains described herein) and the soluble TRpC domain (e.g., any of the exemplary soluble TRpC domains described herein) directly abut each other in the first chimeric polypeptide. In some embodiments of any of the multi-chain chimeric polypeptides described herein, the first chimeric polypeptide further comprises a linker sequence (e.g., any of the exemplary linker sequences described herein or known in the art) between the first target-binding domain (e.g., any of the exemplary first targetbinding domains described herein) and the soluble TRpC domain (e.g., any of the exemplary soluble TRPC domains described herein) in the first chimeric polypeptide.

[0173] In some embodiments of any of the multi-chain chimeric polypeptides described herein, the soluble TRpC domain (e.g., any of the exemplary soluble TRpC domains described herein) and the first domain of the pair of affinity domains (e.g., any of the exemplary first domains of any of the exemplary pairs of affinity domains described herein) directly abut each other in the first chimeric polypeptide.

[0174] In some embodiments of any of the multi-chain chimeric polypeptides described herein, the first chimeric polypeptide further comprises a linker sequence (e.g., any of the exemplary linker sequences described herein or known in the art) between the soluble TR C domain (e.g., any of the exemplary soluble TRpC domains described herein) and the first domain of the pair of affinity domains (e.g., any of the exemplary first domains of any of the exemplary pairs of affinity domains described herein) in the first chimeric polypeptide.

[0175] In some embodiments of any of the multi-chain chimeric polypeptides described herein, the second domain of the pair of affinity domains (e.g., any of the exemplary second domains of any of the exemplary pairs of affinity domains described herein) and the second target-bindingAttorney Docket No. 47039-0043W01

[0176] domain (e g., any of the exemplary second target-binding domains described herein) directly abut each other in the second chimeric polypeptide.

[0177] In some embodiments of any of the multi-chain chimeric polypeptides described herein, the second chimeric polypeptide further comprises a linker sequence (e.g., any of the exemplary linker sequences described herein or known in the art) between the second domain of the pair of affinity domains (e.g., any of the exemplary second domains of any of the exemplary pairs of affinity domains described herein) and the second target-binding domain (e.g., any of the exemplary second target-binding domains described herein) in the second chimeric polypeptide.

[0178] Diagrams of exemplary multi-chain chimeric polypeptides provided herein are depicted in Figures 8A-8B.

[0179] Non-limiting aspects of these chimeric polypeptides, nucleic acids, vectors, cells, and methods are described below, and can be used in any combination without limitation. Additional aspects of these chimeric polypeptides, nucleic acids, vectors, cells, and methods are known in the art.

[0180] T Cell Receptor p-Chain Constant Region (TR C)

[0181] The T-cell receptor (TCR) is a heterodimer of two chains, an a-chain and a P-chain or a y-chain and a 8-chain. An individual T cell can express either an aP heterodimer or a y8 heterodimer as its receptor. The TCR is expressed with cluster of differentiation 3 (CD3) polypeptides and forms a complex comprising the TCR and six CD3 polypeptides (the TCR-CD3 complex). Three dimers of CD3 proteins, 8e and ys heterodimers and homodimers, are present in the TCR-CD3 complex. TCR-CD3 complexes can be found on the surface of T cells where they recognize self or foreign antigens presented by major histocompatibility complex (MHC) molecules and can be involved in immune synapse formation, inducing intracellular signaling, and initiating target cell killing. The TCR is involved in antigen recognition, and the CD3 polypeptides contain intracellular signaling domains that couple the TCR-CD3 complex to the downstream signaling machinery.

[0182] TCR chains include an extracellular region, a transmembrane region, and a shorter cytoplasmic tail. The extracellular regions of TCR chains contain a variable (V) region and a constant (C) region. TCR diversity can be generated by somatic V(D)J recombination in which variable (V) regions and joining (J) regions or variable (V) regions, joining (J) regions, andAttorney Docket No. 47039-0043W01

[0183] diversity (D) regions can be combined during the production of a TCR chain (e.g., V, D and J for P and 8 chains; V and J for a and y chains). Cells in a clonal T cell population express the same unique TCR.

[0184] Different species have 1 to 5 different genes encoding the T cell receptor P-chain constant region domain (TRBC or BC). Human and mice have genes encoding the TRpCl (TRBC1 gene) or TRPC2 (TRBC2 gene) isoforms. Human TRpCl and TRPC2 differ by only four amino acid mutations in the extracellular domain. The surrounding sequence is identical between the two isoforms and the folded structure remains largely unchanged. Each TCR (and therefore each T cell) selects a TCR P-chain constant region encoded by either TRpCl or TRPC2 for expression in a mutually exclusive manner. Hence, a population of normal T cells will comprise a mixture of individual cells, some expressing TRpCl and others expressing TRPC2, whereas the entire population of a T cell cancer will express either TRpCl or TRPC2.

[0185] Soluble T cell Receptor P-chain Constant Region (TR C) Domains

[0186] In some embodiments of any of the single-chain chimeric polypeptides, multi-chain chimeric polypeptides, compositions, or methods described herein, the soluble TRpC domain can be a soluble human TRpC domain. In some embodiments, the soluble TRpC domain can be a soluble human TRpC l domain. In some embodiments, the soluble TRPC domain can be a soluble human TRPC2 domain. Non-limiting examples of soluble TRpC domains are shown below.

[0187] In some embodiments, a soluble T cell receptor P-chain constant region (TRpC) domain can comprise or consist of a sequence that is at least 80%, 85%, 90%, 92%, 95%, 96%, 98%, or 100% identical to SEQ ID NO: 1 or 3. In some embodiments, a soluble T cell receptor P-chain constant region (TRpC) domain is encoded by a nucleic acid that is at least 80%, 85%, 90%, 92%, 95%, 96%, 98%, or 100% identical to SEQ ID NO: 2.

[0188] Exemplary Soluble Human TR Cl Domain (SEQ ID NO: 1 )

[0189] EDLNKVFPPEVAVFEPSEAEISHTQKATLVCLATGFFPDHVELSWWVNGKEVHSGV STDPQPLKEQPALNDSRYCLSSRLRVSATFWQNPRNHFRCQVQFYGLSENDEWTQD RAKPVTQIVSAEAWGRADAttorney Docket No. 47039-0043W01

[0190] Exemplary Nucleic Acid Encoding Soluble Human TRftCl Domain (SEQ ID NO: 2)

[0191] GAGGACCTGAACAAGGTGTTCCCACCCGAGGTCGCTGTGTTTGAGCCATCAGAAG CAGAGATCTCCCACACCCAAAAGGCCACACTGGTGTGCCTGGCCACAGGCTTCTT CCCCGACCACGTGGAGCTGAGCTGGTGGGTGAATGGGAAGGAGGTGCACAGTGGG GTCAGCACGGACCCGCAGCCCCTCAAGGAGCAGCCCGCCCTCAATGACTCCAGAT ACTGCCTGAGCAGCCGCCTGAGGGTCTCGGCCACCTTCTGGCAGAACCCCCGCAA CCACTTCCGCTGTCAAGTCCAGTTCTACGGGCTCTCGGAGAATGACGAGTGGACC CAGGATAGGGCCAAACCCGTCACCCAGATCGTCAGCGCCGAGGCCTGGGGTAGAG CAGAC

[0192] Exemplary Soluble Human TRpC2 Domain (SEQ ID NO: 3)

[0193] EDLKNVFPPEVAVFEPSEAEISHTQKATLVCLATGFYPDHVELSWWVNGKEVHS GVSTDPQPLKEQPALNDSRYCLSSRLRVSATFWQNPRNHFRCQVQFYGLSENDE WTQDRAKPVTQIVSAEAWGRAD

[0194] As can be appreciated in the art, one skilled in the art would understand that mutation of amino acids that are conserved between different mammalian species is more likely to decrease the activity and / or structural stability of the protein, while mutation of amino acids that are not conserved between different mammalian species is less likely to decrease the activity and / or structural stability of the protein.

[0195] In some embodiments, the soluble tissue factor domain can comprise or consist of a soluble wildtype human TRPC2 domain (or any sequence therefrom).

[0196] In some embodiments, the soluble TRpC domain does not bind to an MHC protein with or without a loaded antigen. In some embodiments, the soluble TR0C domain does not stimulate an immune response in a mammal.

[0197] In some embodiments, the soluble TRpC domain can have a total length of 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, acids, about 20 amino acids to about 155 amino acids, about 20 amino acids to about 150 amino acids, about 20 amino acids to about 145 amino acids, about 20 amino acids to about 140 amino acids, about 20 amino acids to about 135 amino acids, about 20 amino acids to about 130 amino acids, about 20 amino acids to about 125 amino acids, about 20 amino acids to about 120 amino acids, about 20 amino acids to about 115 amino acids,Attorney Docket No. 47039-0043W01

[0198] 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 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 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, acids, about 30 amino acids to about 155 amino acids, about 30 amino acids to about 150 amino acids, about 30 amino acids to about 145 amino acids, about 30 amino acids to about 140 amino acids, about 30 amino acids to about 135 amino acids, about 30 amino acids to about 130 amino acids, about 30 amino acids to about 125 amino acids, about 30 amino acids to about 120 amino acids, about 30 amino acids to about 115 amino acids, about 30 amino acids to about 110 amino acids, about 30 amino acids to about 105 amino acids, about 30 amino acids to about 100 amino acids, about 30 amino acids to about 95 amino acids, about 30 amino acids to about 90 amino acids, about 30 amino acids to about 85 amino acids, about 30 amino acids to about 80 amino acids, about 30 amino acids to about 75 amino acids, about 30 amino acids to about 70 amino acids, about 30 amino acids to about 60 amino acids, about 30 amino acids to about 50 amino acids, about 30 amino acids to about 40 amino acids, about 40 amino acids to about 200 amino acids, about 40 amino acids to about 195 amino acids, about 40 amino acids to about 190 amino acids, about 40 amino acids to about 185 amino acids, about 40 amino acids to about 180 amino acids, about 40 amino acids to about 175 amino acids, about 40 amino acids to about 170 amino acids, about 40 amino acids to about 165 amino acids, about 40 amino acids to about 160 amino acids, acids, about 40 amino acids to about 155 amino acids, about 40 amino acids to about 150 amino acids, about 40 amino acids to about 145 amino acids, about 40 amino acids to about 140 amino acids, about 40 amino acids to about 135 amino acids, about 40 amino acids to about 130 amino acids, about 40 amino acids to about 125 amino acids, about 40 amino acids to about 120 amino acids, about 40 amino acids to about 115 amino acids, about 40 amino acids to about 110Attorney Docket No. 47039-0043W01

[0199] amino acids, about 40 amino acids to about 105 amino acids, about 40 amino acids to about 100 amino acids, about 40 amino acids to about 95 amino acids, about 40 amino acids to about 90 amino acids, about 40 amino acids to about 85 amino acids, about 40 amino acids to about 80 amino acids, about 40 amino acids to about 75 amino acids, about 40 amino acids to about 70 amino acids, about 40 amino acids to about 60 amino acids, about 40 amino acids to about 50 amino acids, about 50 amino acids to about 200 amino acids, about 50 amino acids to about 195 amino acids, about 50 amino acids to about 190 amino acids, about 50 amino acids to about 185 amino acids, about 50 amino acids to about 180 amino acids, about 50 amino acids to about 175 amino acids, about 50 amino acids to about 170 amino acids, about 50 amino acids to about 165 amino acids, about 50 amino acids to about 160 amino acids, acids, about 50 amino acids to about 155 amino acids, about 50 amino acids to about 150 amino acids, about 50 amino acids to about 145 amino acids, about 50 amino acids to about 140 amino acids, about 50 amino acids to about 135 amino acids, about 50 amino acids to about 130 amino acids, about 50 amino acids to about 125 amino acids, about 50 amino acids to about 120 amino acids, about 50 amino acids to about 115 amino acids, about 50 amino acids to about 110 amino acids, about 50 amino acids to about 105 amino acids, about 50 amino acids to about 100 amino acids, about 50 amino acids to about 95 amino acids, about 50 amino acids to about 90 amino acids, about 50 amino acids to about 85 amino acids, about 50 amino acids to about 80 amino acids, about 50 amino acids to about 75 amino acids, about 50 amino acids to about 70 amino acids, about 50 amino acids to about 60 amino acids, about 60 amino acids to about 200 amino acids, about 60 amino acids to about 195 amino acids, about 60 amino acids to about 190 amino acids, about 60 amino acids to about 185 amino acids, about 60 amino acids to about 180 amino acids, about 60 amino acids to about 175 amino acids, about 60 amino acids to about 170 amino acids, about 60 amino acids to about 165 amino acids, about 60 amino acids to about 160 amino acids, acids, about 60 amino acids to about 155 amino acids, about 60 amino acids to about 150 amino acids, about 60 amino acids to about 145 amino acids, about 60 amino acids to about 140 amino acids, about 60 amino acids to about 135 amino acids, about 60 amino acids to about 130 amino acids, about 60 amino acids to about 125 amino acids, about 60 amino acids to about 120 amino acids, about 60 amino acids to about 115 amino acids, about 60 amino acids to about 110 amino acids, about 60 amino acids to about 105 amino acids, about 60 amino acids to about 100 amino acids, about 60 amino acids to about 95 amino acids, about 60 amino acids to about 90 amino acids, about 60 aminoAttorney Docket No. 47039-0043W01

[0200] acids to about 85 amino acids, about 60 amino acids to about 80 amino acids, about 60 amino acids to about 75 amino acids, about 60 amino acids to about 70 amino acids, about 70 amino acids to about 200 amino acids, about 70 amino acids to about 195 amino acids, about 70 amino acids to about 190 amino acids, about 70 amino acids to about 185 amino acids, about 70 amino acids to about 180 amino acids, about 70 amino acids to about 175 amino acids, about 70 amino acids to about 170 amino acids, about 70 amino acids to about 165 amino acids, about 70 amino acids to about 160 amino acids, acids, about 70 amino acids to about 155 amino acids, about 70 amino acids to about 150 amino acids, about 70 amino acids to about 145 amino acids, about 70 amino acids to about 140 amino acids, about 70 amino acids to about 135 amino acids, about 70 amino acids to about 130 amino acids, about 70 amino acids to about 125 amino acids, about 70 amino acids to about 120 amino acids, about 70 amino acids to about 115 amino acids, about 70 amino acids to about 110 amino acids, about 70 amino acids to about 105 amino acids, about 70 amino acids to about 100 amino acids, about 70 amino acids to about 95 amino acids, about 70 amino acids to about 90 amino acids, about 70 amino acids to about 85 amino acids, about 70 amino acids to about 80 amino acids, about 80 amino acids to about 200 amino acids, about 80 amino acids to about 195 amino acids, about 80 amino acids to about 190 amino acids, about 80 amino acids to about 185 amino acids, about 80 amino acids to about 180 amino acids, about 80 amino acids to about 175 amino acids, about 80 amino acids to about 170 amino acids, about 80 amino acids to about 165 amino acids, about 80 amino acids to about 160 amino acids, acids, about 80 amino acids to about 155 amino acids, about 80 amino acids to about 150 amino acids, about 80 amino acids to about 145 amino acids, about 80 amino acids to about 140 amino acids, about 80 amino acids to about 135 amino acids, about 80 amino acids to about 130 amino acids, about 80 amino acids to about 125 amino acids, about 80 amino acids to about 120 amino acids, about 80 amino acids to about 115 amino acids, about 80 amino acids to about 110 amino acids, about 80 amino acids to about 105 amino acids, about 80 amino acids to about 100 amino acids, about 80 amino acids to about 95 amino acids, about 80 amino acids to about 90 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, acids, about 90 amino acids to about 155 aminoAttorney Docket No. 47039-0043W01

[0201] acids, about 90 amino acids to about 150 amino acids, about 90 amino acids to about 145 amino acids, about 90 amino acids to about 140 amino acids, about 90 amino acids to about 135 amino acids, about 90 amino acids to about 130 amino acids, about 90 amino acids to about 125 amino acids, about 90 amino acids to about 120 amino acids, about 90 amino acids to about 115 amino acids, about 90 amino acids to about 110 amino acids, about 90 amino acids to about 105 amino acids, about 90 amino acids to about 100 amino acids, about 100 amino acids to about 200 amino acids, about 100 amino acids to about 195 amino acids, about 100 amino acids to about 190 amino acids, about 100 amino acids to about 185 amino acids, about 100 amino acids to about 180 amino acids, about 100 amino acids to about 175 amino acids, about 100 amino acids to about 170 amino acids, about 100 amino acids to about 165 amino acids, about 100 amino acids to about 160 amino acids, 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 110 amino acids to about 200 amino acids, about 110 amino acids to about 195 amino acids, about 110 amino acids to about 190 amino acids, about 110 amino acids to about 185 amino acids, about 110 amino acids to about 180 amino acids, about 110 amino acids to about 175 amino acids, about 110 amino acids to about 170 amino acids, about 110 amino acids to about 165 amino acids, about 110 amino acids to about 160 amino acids, acids, about 110 amino acids to about 155 amino acids, about 110 amino acids to about 150 amino acids, about 110 amino acids to about 145 amino acids, about 110 amino acids to about 140 amino acids, about 110 amino acids to about 135 amino acids, about 110 amino acids to about 130 amino acids, about 110 amino acids to about 125 amino acids, about 110 amino acids to about 120 amino acids, about 110 amino acids to about 115 amino acids, about 115 amino acids to about 200 amino acids, about 115 amino acids to about 195 amino acids, about 115 amino acids to about 190 amino acids, about 115 amino acids to about 185 amino acids, about 115 amino acids to about 180 amino acids, about 115 amino acids to about 175 amino acids, about 115 amino acids to about 170 amino acids, about 115 amino acids to about 165 amino acids, about 115 amino acids to about 160 amino acids, acids, about 115 amino acids to about 155 amino acids, about 115 amino acids to about 150Attorney Docket No. 47039-0043W01

[0202] 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 200 amino acids, about 120 amino acids to about 195 amino acids, about 120 amino acids to about 190 amino acids, about 120 amino acids to about 185 amino acids, about 120 amino acids to about 180 amino acids, about 120 amino acids to about 175 amino acids, about 120 amino acids to about 170 amino acids, about 120 amino acids to about 165 amino acids, about 120 amino acids to about 160 amino acids, acids, about 120 amino acids to about 155 amino acids, about 120 amino acids to about 150 amino acids, about 120 amino acids to about 145 amino acids, about 120 amino acids to about 140 amino acids, about 120 amino acids to about 135 amino acids, about 120 amino acids to about 130 amino acids, about 120 amino acids to about 125 amino acids, about 125 amino acids to about 200 amino acids, about 125 amino acids to about 195 amino acids, about 125 amino acids to about 190 amino acids, about 125 amino acids to about 185 amino acids, about 125 amino acids to about 180 amino acids, about 125 amino acids to about 175 amino acids, about 125 amino acids to about 170 amino acids, about 125 amino acids to about 165 amino acids, about 125 amino acids to about 160 amino acids, acids, about 125 amino acids to about 155 amino acids, about 125 amino acids to about 150 amino acids, about 125 amino acids to about 145 amino acids, about 125 amino acids to about 140 amino acids, about 125 amino acids to about 135 amino acids, about 125 amino acids to about 130 amino acids, about 130 amino acids to about 200 amino acids, about 130 amino acids to about 195 amino acids, about 130 amino acids to about 190 amino acids, about 130 amino acids to about 185 amino acids, about 130 amino acids to about 180 amino acids, about 130 amino acids to about 175 amino acids, about 130 amino acids to about 170 amino acids, about 130 amino acids to about 165 amino acids, about 130 amino acids to about 160 amino acids, 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 200 amino acids, about 135 amino acids to about 195 amino acids, about 135 amino acids to about 190 amino acids, about 135 amino acids to about 185 amino acids, about 135 amino acids to about 180 amino acids, about 135 amino acids to about 175 amino acids, about 135 amino acids to about 170 amino acids,Attorney Docket No. 47039-0043W01

[0203] about 135 amino acids to about 165 amino acids, about 135 amino acids to about 160 amino acids, 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 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, acids, about 140 amino acids to about 155 amino acids, about 140 amino acids to about 150 amino acids, about 140 amino acids to about 145 amino acids, about 145 amino acids to about 200 amino acids, about 145 amino acids to about 195 amino acids, about 145 amino acids to about 190 amino acids, about 145 amino acids to about 185 amino acids, about 145 amino acids to about 180 amino acids, about 145 amino acids to about 175 amino acids, about 145 amino acids to about 170 amino acids, about 145 amino acids to about 165 amino acids, about 145 amino acids to about 160 amino acids, acids, about 145 amino acids to about 155 amino acids, about 145 amino acids to about 150 amino acids, about 150 amino acids to about 200 amino acids, about 150 amino acids to about 195 amino acids, about 150 amino acids to about 190 amino acids, about 150 amino acids to about 185 amino acids, about 150 amino acids to about 180 amino acids, about 150 amino acids to about 175 amino acids, about 150 amino acids to about 170 amino acids, about 150 amino acids to about 165 amino acids, about 150 amino acids to about 160 amino acids, acids, about 150 amino acids to about 155 amino acids, about 155 amino acids to about 200 amino acids, about 155 amino acids to about 195 amino acids, about 155 amino acids to about 190 amino acids, about 155 amino acids to about 185 amino acids, about 155 amino acids to about 180 amino acids, about 155 amino acids to about 175 amino acids, about 155 amino acids to about 170 amino acids, about 155 amino acids to about 165 amino acids, about 155 amino acids to about 160 amino acids, acids, about 160 amino acids to about 200 amino acids, about 160 amino acids to about 195 amino acids, about 160 amino acids to about 190 amino acids, about 160 amino acids to about 185 amino acids, about 160 amino acids to about 180 amino acids, about 160 amino acids to about 175 amino acids, about 160 amino acids to about 170 amino acids, about 160 amino acids to about 165 amino acids, about 165 amino acids to about 200 amino acids, about 165 amino acids to about 195 amino acids,Attorney Docket No. 47039-0043W01

[0204] about 165 amino acids to about 190 amino acids, about 165 amino acids to about 185 amino acids, about 165 amino acids to about 180 amino acids, about 165 amino acids to about 175 amino acids, about 165 amino acids to about 170 amino acids, about 170 amino acids to about 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 200 amino acids, about 175 amino acids to about 195 amino acids, about 175 amino acids to about 190 amino acids, about 175 amino acids to about 185 amino acids, about 175 amino acids to about 180 amino acids, about 180 amino acids to about 200 amino acids, about 180 amino acids to about 195 amino acids, about 180 amino acids to about 190 amino acids, about 180 amino acids to about 185 amino acids, about 185 amino acids to about 200 amino acids, about 185 amino acids to about 195 amino acids, about 185 amino acids to about 190 amino acids, about 190 amino acids to about 200 amino acids, about 190 amino acids to about 195 amino acids, or about 195 amino acids to about 200 amino acids.

[0205] Target-Binding Domains

[0206] In some embodiments, the first target-binding domain, the second target-binding domain, and / or the one or more additional target-binding domains can be an antigen-binding domain (e.g., any of the exemplary antigen-binding domains described herein or known in the art), a soluble interleukin or cytokine protein (e.g., any of the exemplary soluble interleukin proteins or soluble cytokine proteins described herein), and a soluble interleukin or cytokine receptor (e.g., any of the exemplary soluble interleukin receptors or soluble cytokine receptors described herein).

[0207] In some embodiments, a polypeptide described herein (e.g., one or more of a single-chain polypeptide, a first chimeric polypeptide of a multi -chain chimeric polypeptide, or a second chimeric polypeptide of a multi-chain chimeric polypeptide) further includes one or more (e.g., two, three, four, five, six, seven, eight, nine, or ten) additional target-binding domain(s) (e.g., any of the exemplary target-binding domains described herein or known in the art) at the N- and / or C-terminus of the polypeptide.

[0208] In some embodiments, at least one of the one or more additional target-binding domain(s) is positioned between the soluble TRpC domain (e.g., any of the exemplary soluble TRpCAttorney Docket No. 47039-0043W01

[0209] domains described herein or known in the art) and the first domain of the pair of affinity domains of a multi-chain chimeric polypeptide (e.g., any of the exemplary first domains of any of the exemplary pairs of affinity domains described herein). In some embodiments, the first chimeric polypeptide can further include a linker sequence (e.g., any of the exemplary linker sequences described herein or known in the art) between the soluble TR0C domain (e g., any of the exemplary soluble TRPC domains described herein) and the at least one of the one or more additional target-binding domain(s) (e.g., any of the exemplary target-binding domains described herein or known in the art), and / or a linker sequence (e.g., any of the exemplary linker sequences described herein or known in the art) between the at least one of the one or more additional target-binding domain(s) (e.g., any of the exemplary target-binding domains described herein or known in the art) and the first domain of the pair of affinity domains (e.g., any of the exemplary first domains described herein of any of the exemplary pairs of affinity domains described herein).

[0210] In some embodiments of any of the chimeric polypeptides described herein (e.g., any of the multi-chain chimeric polypeptides described herein and / or single-chain chimeric polypeptides described herein), one or more of the first target-binding domain (e.g., any of the exemplary first target-binding domains described herein or known in the art), the second targetbinding domain (e.g., any of the exemplary second target-binding domains described herein or known in the art), and the one or more additional target-binding domains can each, independently, bind specifically to a target selected from the group of bind specifically to a target selected from the group consisting of: tissue factor (CD142), CD16a, CD28, CD3 (e.g., one or more of CD3a, CD3£, CD35, CD38, and CD3y), 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, TNFa, CD26a, CD36, ULBP2, CD30, CD200, IGF-1R, MUC4AC, MUC5AC, Trop-2, CMET, EGFR, HER1, HER2, HER3, PSMA, CEA, B7H3, EPCAM, BCMA, P-cadherin, CEACAM5, a UL16-binding protein (e.g, ULBP1, ULBP2, ULBP3, ULBP4, ULBP5, and ULBP6), HLA-DR, DLL4, TYRO3, AXL, MER, CD 122, CD 155, PDGF-DD, a ligand of TGF-p receptor II (TGF-|3RII), a ligand of TGF-0RIII, a ligand of DNAM1, a ligand of NKp46, a ligand of NKp44, a ligand of NKG2D, a ligand of NKp30, a ligand for a scMHCI, a ligand for a scMHCII, a ligand for a scTCR, a receptor for IL-1, a receptor for IL-2, a receptor for IL-3, a receptor for IL-4, a receptor for IL-7, a receptor for IL-8, a receptor for IL- 10, a receptor for IL-Attorney Docket No. 47039-0043W01

[0211] 12, a receptor for IL-15, a receptor for IL-17, a receptor for IL-18, a receptor for IL-21, a receptor for PDGF-DD, a receptor for stem cell factor (SCF), a receptor for stem cell-like tyrosine kinase 3 ligand (FLT3L), a receptor for MICA, a receptor for MICB, a receptor for a ULP16-binding protein, a receptor for CD155, a receptor for CD122, and a receptor for CD28. In some embodiments, at least one of the first target-binding domain and the second target-binding domain bind specifically to mesothelin.

[0212] In some embodiments of any of the chimeric polypeptides described herein (e.g., any of the multi-chain chimeric polypeptides described herein and / or single-chain chimeric polypeptides described herein), the first target-binding domain, the second target-binding domain, and / or the one or more additional target-binding domains can each independently have a total number of amino acids of 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 amino acids to about 450 amino acids, about 5 amino acids to about 400 amino acids, about 5 amino acids to about 350 amino acids, about 5 amino acids to about 300 amino acids, about 5 amino acids to about 280 amino acids, about 5 amino acids to about 260 amino acids, about 5 amino acids to about 240 amino acids, about 5 amino acids to about 220 amino acids, about 5 amino acids to about 200 amino acids, about 5 amino acids to about 195 amino acids, about 5 amino acids to about 190 amino acids, about 5 amino acids to about 185 amino acids, about 5 amino acids to about 180 amino acids, about 5 amino acids to about 175 amino acids, about 5 amino acids to about 170 amino acids, about 5 amino acids to about 165 amino acids, about 5 amino acids to about 160 amino acids, about 5 amino acids to about 155 amino acids, about 5 amino acids to about 150 amino acids, about 5 amino acids to about 145 amino acids, about 5 amino acids to about 140 amino acids, about 5 amino acids to about 135 amino acids, about 5 amino acids to about 130 amino acids, about 5 amino acids to about 125 amino acids, about 5 amino acids to about 120 amino acids, about 5 amino acids to about 115 amino acids, about 5 amino acids to about 110 amino acids, about 5 amino acids to about 105 amino acids, about 5 amino acids to about 100 amino acids, about 5 amino acids to about 95 amino acids, about 5 aminoAttorney Docket No. 47039-0043W01

[0213] acids to about 90 amino acids, about 5 amino acids to about 85 amino acids, about 5 amino acids to about 80 amino acids, about 5 amino acids to about 75 amino acids, about 5 amino acids to about 70 amino acids, about 5 amino acids to about 65 amino acids, about 5 amino acids to about 60 amino acids, about 5 amino acids to about 55 amino acids, about 5 amino acids to about 50 amino acids, about 5 amino acids to about 45 amino acids, about 5 amino acids to about 40 amino acids, about 5 amino acids to about 35 amino acids, about 5 amino acids to about 30 amino acids, about 5 amino acids to about 25 amino acids, about 5 amino acids to about 20 amino acids, about 5 amino acids to about 15 amino acids, about 5 amino acids to about 10 amino acids, about 10 amino acids to about 1000 amino acids, about 10 amino acids to about 950 amino acids, about 10 amino acids to about 900 amino acids, about 10 amino acids to about 850 amino acids, about 10 amino acids to about 800 amino acids, about 10 amino acids to about 750 amino acids, about 10 amino acids to about 700 amino acids, about 10 amino acids to about 650 amino acids, about 10 amino acids to about 600 amino acids, about 10 amino acids to about 550 amino acids, about 10 amino acids to about 500 amino acids, about 10 amino acids to about 450 amino acids, about 10 amino acids to about 400 amino acids, about 10 amino acids to about 350 amino acids, about 10 amino acids to about 300 amino acids, about 10 amino acids to about 280 amino acids, about 10 amino acids to about 260 amino acids, about 10 amino acids to about 240 amino acids, about 10 amino acids to about 220 amino acids, about 10 amino acids to about 200 amino acids, about 10 amino acids to about 195 amino acids, about 10 amino acids to about 190 amino acids, about 10 amino acids to about 185 amino acids, about 10 amino acids to about 180 amino acids, about 10 amino acids to about 175 amino acids, about 10 amino acids to about 170 amino acids, about 10 amino acids to about 165 amino acids, about 10 amino acids to about 160 amino acids, about 10 amino acids to about 155 amino acids, about 10 amino acids to about 150 amino acids, about 10 amino acids to about 145 amino acids, about 10 amino acids to about 140 amino acids, about 10 amino acids to about 135 amino acids, about 10 amino acids to about 130 amino acids, about 10 amino acids to about 125 amino acids, about 10 amino acids to about 120 amino acids, about 10 amino acids to about 115 amino acids, about 10 amino acids to about 110 amino acids, about 10 amino acids to about 105 amino acids, about 10 amino acids to about 100 amino acids, about 10 amino acids to about 95 amino acids, about 10 amino acids to about 90 amino acids, about 10 amino acids to about 85 amino acids, about 10 amino acids to about 80 amino acids, about 10 amino acids to about 75 amino acids, about 10 amino acids to about 70Attorney Docket No. 47039-0043W01

[0214] amino acids, about 10 amino acids to about 65 amino acids, about 10 amino acids to about 60 amino acids, about 10 amino acids to about 55 amino acids, about 10 amino acids to about 50 amino acids, about 10 amino acids to about 45 amino acids, about 10 amino acids to about 40 amino acids, about 10 amino acids to about 35 amino acids, about 10 amino acids to about 30 amino acids, about 10 amino acids to about 25 amino acids, about 10 amino acids to about 20 amino acids, about 10 amino acids to about 15 amino acids, about 15 amino acids to about 1000 amino acids, about 15 amino acids to about 950 amino acids, about 15 amino acids to about 900 amino acids, about 15 amino acids to about 850 amino acids, about 15 amino acids to about 800 amino acids, about 15 amino acids to about 750 amino acids, about 15 amino acids to about 700 amino acids, about 15 amino acids to about 650 amino acids, about 15 amino acids to about 600 amino acids, about 15 amino acids to about 550 amino acids, about 15 amino acids to about 500 amino acids, about 15 amino acids to about 450 amino acids, about 15 amino acids to about 400 amino acids, about 15 amino acids to about 350 amino acids, about 15 amino acids to about 300 amino acids, about 15 amino acids to about 280 amino acids, about 15 amino acids to about 260 amino acids, about 15 amino acids to about 240 amino acids, about 15 amino acids to about 220 amino acids, about 15 amino acids to about 200 amino acids, about 15 amino acids to about 195 amino acids, about 15 amino acids to about 190 amino acids, about 15 amino acids to about 185 amino acids, about 15 amino acids to about 180 amino acids, about 15 amino acids to about 175 amino acids, about 15 amino acids to about 170 amino acids, about 15 amino acids to about 165 amino acids, about 15 amino acids to about 160 amino acids, about 15 amino acids to about 155 amino acids, about 15 amino acids to about 150 amino acids, about 15 amino acids to about 145 amino acids, about 15 amino acids to about 140 amino acids, about 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, about 15 amino acids to about 60 amino acids, about 15 amino acids to about 55 amino acids, about 15 amino acids to about 50 amino acids, about 15 amino acids to about 45Attorney Docket No. 47039-0043W01

[0215] amino acids, about 15 amino acids to about 40 amino acids, about 15 amino acids to about 35 amino acids, about 15 amino acids to about 30 amino acids, about 15 amino acids to about 25 amino acids, about 15 amino acids to about 20 amino acids, about 20 amino acids to about 1000 amino acids, about 20 amino acids to about 950 amino acids, about 20 amino acids to about 900 amino acids, about 20 amino acids to about 850 amino acids, about 20 amino acids to about 800 amino acids, about 20 amino acids to about 750 amino acids, about 20 amino acids to about 700 amino acids, about 20 amino acids to about 650 amino acids, about 20 amino acids to about 600 amino acids, about 20 amino acids to about 550 amino acids, about 20 amino acids to about 500 amino acids, about 20 amino acids to about 450 amino acids, about 20 amino acids to about 400 amino acids, about 20 amino acids to about 350 amino acids, about 20 amino acids to about 300 amino acids, about 20 amino acids to about 280 amino acids, about 20 amino acids to about 260 amino acids, about 20 amino acids to about 240 amino acids, about 20 amino acids to about 220 amino acids, about 20 amino acids to about 200 amino acids, about 20 amino acids to about 195 amino acids, about 20 amino acids to about 190 amino acids, about 20 amino acids to about 185 amino acids, about 20 amino acids to about 180 amino acids, about 20 amino acids to about 175 amino acids, about 20 amino acids to about 170 amino acids, about 20 amino acids to about 165 amino acids, about 20 amino acids to about 160 amino acids, about 20 amino acids to about 155 amino acids, about 20 amino acids to about 150 amino acids, about 20 amino acids to about 145 amino acids, about 20 amino acids to about 140 amino acids, about 20 amino acids to about 135 amino acids, about 20 amino acids to about 130 amino acids, about 20 amino acids to about 125 amino acids, about 20 amino acids to about 120 amino acids, about 20 amino acids to about 115 amino acids, about 20 amino acids to about 110 amino acids, about 20 amino acids to about 105 amino acids, about 20 amino acids to about 100 amino acids, about 20 amino acids to about 95 amino acids, about 20 amino acids to about 90 amino acids, about 20 amino acids to about 85 amino acids, about 20 amino acids to about 80 amino acids, about 20 amino acids to about 75 amino acids, about 20 amino acids to about 70 amino acids, about 20 amino acids to about 65 amino acids, about 20 amino acids to about 60 amino acids, about 20 amino acids to about 55 amino acids, about 20 amino acids to about 50 amino acids, about 20 amino acids to about 45 amino acids, about 20 amino acids to about 40 amino acids, about 20 amino acids to about 35 amino acids, about 20 amino acids to about 30 amino acids, about 20 amino acids to about 25 amino acids, about 25 amino acids to about 1000 amino acids, about 25 amino acids to about 950Attorney Docket No. 47039-0043W01

[0216] amino acids, about 25 amino acids to about 900 amino acids, about 25 amino acids to about 850 amino acids, about 25 amino acids to about 800 amino acids, about 25 amino acids to about 750 amino acids, about 25 amino acids to about 700 amino acids, about 25 amino acids to about 650 amino acids, about 25 amino acids to about 600 amino acids, about 25 amino acids to about 550 amino acids, about 25 amino acids to about 500 amino acids, about 25 amino acids to about 450 amino acids, about 25 amino acids to about 400 amino acids, about 25 amino acids to about 350 amino acids, about 25 amino acids to about 300 amino acids, about 25 amino acids to about 280 amino acids, about 25 amino acids to about 260 amino acids, about 25 amino acids to about 240 amino acids, about 25 amino acids to about 220 amino acids, about 25 amino acids to about 200 amino acids, about 25 amino acids to about 195 amino acids, about 25 amino acids to about 190 amino acids, about 25 amino acids to about 185 amino acids, about 25 amino acids to about 180 amino acids, about 25 amino acids to about 175 amino acids, about 25 amino acids to about 170 amino acids, about 25 amino acids to about 165 amino acids, about 25 amino acids to about 160 amino acids, about 25 amino acids to about 155 amino acids, about 25 amino acids to about 150 amino acids, about 25 amino acids to about 145 amino acids, about 25 amino acids to about 140 amino acids, about 25 amino acids to about 135 amino acids, about 25 amino acids to about 130 amino acids, about 25 amino acids to about 125 amino acids, about 25 amino acids to about 120 amino acids, about 25 amino acids to about 115 amino acids, about 25 amino acids to about 110 amino acids, about 25 amino acids to about 105 amino acids, about 25 amino acids to about 100 amino acids, about 25 amino acids to about 95 amino acids, about 25 amino acids to about 90 amino acids, about 25 amino acids to about 85 amino acids, about 25 amino acids to about 80 amino acids, about 25 amino acids to about 75 amino acids, about 25 amino acids to about 70 amino acids, about 25 amino acids to about 65 amino acids, about 25 amino acids to about 60 amino acids, about 25 amino acids to about 55 amino acids, about 25 amino acids to about 50 amino acids, about 25 amino acids to about 45 amino acids, about 25 amino acids to about 40 amino acids, about 25 amino acids to about 35 amino acids, about 25 amino acids to about 30 amino acids, about 30 amino acids to about 1000 amino acids, about 30 amino acids to about 950 amino acids, about 30 amino acids to about 900 amino acids, about 30 amino acids to about 850 amino acids, about 30 amino acids to about 800 amino acids, about 30 amino acids to about 750 amino acids, about 30 amino acids to about 700 amino acids, about 30 amino acids to about 650 amino acids, about 30 amino acids to about 600 amino acids, about 30 amino acids to about 550Attorney Docket No. 47039-0043W01

[0217] amino acids, about 30 amino acids to about 500 amino acids, about 30 amino acids to about 450 amino acids, about 30 amino acids to about 400 amino acids, about 30 amino acids to about 350 amino acids, about 30 amino acids to about 300 amino acids, about 30 amino acids to about 280 amino acids, about 30 amino acids to about 260 amino acids, about 30 amino acids to about 240 amino acids, about 30 amino acids to about 220 amino acids, about 30 amino acids to about 200 amino acids, about 30 amino acids to about 195 amino acids, about 30 amino acids to about 190 amino acids, about 30 amino acids to about 185 amino acids, about 30 amino acids to about 180 amino acids, about 30 amino acids to about 175 amino acids, about 30 amino acids to about 170 amino acids, about 30 amino acids to about 165 amino acids, about 30 amino acids to about 160 amino acids, about 30 amino acids to about 155 amino acids, about 30 amino acids to about 150 amino acids, about 30 amino acids to about 145 amino acids, about 30 amino acids to about 140 amino acids, about 30 amino acids to about 135 amino acids, about 30 amino acids to about 130 amino acids, about 30 amino acids to about 125 amino acids, about 30 amino acids to about 120 amino acids, about 30 amino acids to about 115 amino acids, about 30 amino acids to about 110 amino acids, about 30 amino acids to about 105 amino acids, about 30 amino acids to about 100 amino acids, about 30 amino acids to about 95 amino acids, about 30 amino acids to about 90 amino acids, about 30 amino acids to about 85 amino acids, about 30 amino acids to about 80 amino acids, about 30 amino acids 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 to about 700 amino acids, about 35 amino acids to about 650 amino acids, about 35 amino acids to about 600 amino acids, about 35 amino acids to about 550 amino acids, about 35 amino acids to about 500 amino acids, about 35 amino acids to about 450 amino acids, about 35 amino acids to about 400 amino acids, about 35 amino acids to about 350 amino acids, about 35 amino acids to about 300 amino acids, about 35 amino acids to about 280 amino acids, about 35 amino acids to about 260 amino acids, about 35 amino acids to about 240 amino acids, about 35 amino acids to about 220Attorney Docket No. 47039-0043W01

[0218] amino acids, about 35 amino acids to about 200 amino acids, about 35 amino acids to about 195 amino acids, about 35 amino acids to about 190 amino acids, about 35 amino acids to about 185 amino acids, about 35 amino acids to about 180 amino acids, about 35 amino acids to about 175 amino acids, about 35 amino acids to about 170 amino acids, about 35 amino acids to about 165 amino acids, about 35 amino acids to about 160 amino acids, about 35 amino acids to about 155 amino acids, about 35 amino acids to about 150 amino acids, about 35 amino acids to about 145 amino acids, about 35 amino acids to about 140 amino acids, about 35 amino acids to about 135 amino acids, about 35 amino acids to about 130 amino acids, about 35 amino acids to about 125 amino acids, about 35 amino acids to about 120 amino acids, about 35 amino acids to about 115 amino acids, about 35 amino acids to about 110 amino acids, about 35 amino acids to about 105 amino acids, about 35 amino acids to about 100 amino acids, about 35 amino acids to about 95 amino acids, about 35 amino acids to about 90 amino acids, about 35 amino acids to about 85 amino acids, about 35 amino acids to about 80 amino acids, about 35 amino acids to about 75 amino acids, about 35 amino acids to about 70 amino acids, about 35 amino acids to about 65 amino acids, about 35 amino acids to about 60 amino acids, about 35 amino acids to about 55 amino acids, about 35 amino acids to about 50 amino acids, about 35 amino acids to about 45 amino acids, about 35 amino acids to about 40 amino acids, about 40 amino acids to about 1000 amino acids, about 40 amino acids to about 950 amino acids, about 40 amino acids to about 900 amino acids, about 40 amino acids to about 850 amino acids, about 40 amino acids to about 800 amino acids, about 40 amino acids to about 750 amino acids, about 40 amino acids to about 700 amino acids, about 40 amino acids to about 650 amino acids, about 40 amino acids to about 600 amino acids, about 40 amino acids to about 550 amino acids, about 40 amino acids to about 500 amino acids, about 40 amino acids to about 450 amino acids, about 40 amino acids to about 400 amino acids, about 40 amino acids to about 350 amino acids, about 40 amino acids to about 300 amino acids, about 40 amino acids to about 280 amino acids, about 40 amino acids to about 260 amino acids, about 40 amino acids to about 240 amino acids, about 40 amino acids to about 220 amino acids, about 40 amino acids to about 200 amino acids, about 40 amino acids to about 195 amino acids, about 40 amino acids to about 190 amino acids, about 40 amino acids to about 185 amino acids, about 40 amino acids to about 180 amino acids, about 40 amino acids to about 175 amino acids, about 40 amino acids to about 170 amino acids, about 40 amino acids to about 165 amino acids, about 40 amino acids to about 160 amino acids, about 40 amino acids to about 155Attorney Docket No. 47039-0043W01

[0219] amino acids, about 40 amino acids to about 150 amino acids, about 40 amino acids to about 145 amino acids, about 40 amino acids to about 140 amino acids, about 40 amino acids to about 135 amino acids, about 40 amino acids to about 130 amino acids, about 40 amino acids to about 125 amino acids, about 40 amino acids to about 120 amino acids, about 40 amino acids to about 115 amino acids, about 40 amino acids to about 110 amino acids, about 40 amino acids to about 105 amino acids, about 40 amino acids to about 100 amino acids, about 40 amino acids to about 95 amino acids, about 40 amino acids to about 90 amino acids, about 40 amino acids to about 85 amino acids, about 40 amino acids to about 80 amino acids, about 40 amino acids to about 75 amino acids, about 40 amino acids to about 70 amino acids, about 40 amino acids to about 65 amino acids, about 40 amino acids to about 60 amino acids, about 40 amino acids to about 55 amino acids, about 40 amino acids to about 50 amino acids, about 40 amino acids to about 45 amino acids, about 45 amino acids to about 1000 amino acids, about 45 amino acids to about 950 amino acids, about 45 amino acids to about 900 amino acids, about 45 amino acids to about 850 amino acids, about 45 amino acids to about 800 amino acids, about 45 amino acids to about 750 amino acids, about 45 amino acids to about 700 amino acids, about 45 amino acids to about 650 amino acids, about 45 amino acids to about 600 amino acids, about 45 amino acids to about 550 amino acids, about 45 amino acids to about 500 amino acids, about 45 amino acids to about 450 amino acids, about 45 amino acids to about 400 amino acids, about 45 amino acids to about 350 amino acids, about 45 amino acids to about 300 amino acids, about 45 amino acids to about 280 amino acids, about 45 amino acids to about 260 amino acids, about 45 amino acids to about 240 amino acids, about 45 amino acids to about 220 amino acids, about 45 amino acids to about 200 amino acids, about 45 amino acids to about 195 amino acids, about 45 amino acids to about 190 amino acids, about 45 amino acids to about 185 amino acids, about 45 amino acids to about 180 amino acids, about 45 amino acids to about 175 amino acids, about 45 amino acids to about 170 amino acids, about 45 amino acids to about 165 amino acids, about 45 amino acids to about 160 amino acids, about 45 amino acids to about 155 amino acids, about 45 amino acids to about 150 amino acids, about 45 amino acids to about 145 amino acids, about 45 amino acids to about 140 amino acids, about 45 amino acids to about 135 amino acids, about 45 amino acids to about 130 amino acids, about 45 amino acids to about 125 amino acids, about 45 amino acids to about 120 amino acids, about 45 amino acids to about 115 amino acids, about 45 amino acids to about 110 amino acids, about 45 amino acids to about 105 amino acids, about 45 amino acids to about 100Attorney Docket No. 47039-0043W01

[0220] amino acids, about 45 amino acids to about 95 amino acids, about 45 amino acids to about 90 amino acids, about 45 amino acids to about 85 amino acids, about 45 amino acids to about 80 amino acids, about 45 amino acids to about 75 amino acids, about 45 amino acids to about 70 amino acids, about 45 amino acids to about 65 amino acids, about 45 amino acids to about 60 amino acids, about 45 amino acids to about 55 amino acids, about 45 amino acids to about 50 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, 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 280 amino acids, about 50 amino acids to about 260 amino acids, about 50 amino acids to about 240 amino acids, about 50 amino acids to about 220 amino acids, about 50 amino acids to about 200 amino acids, about 50 amino acids to about 195 amino acids, about 50 amino acids to about 190 amino acids, about 50 amino acids to about 185 amino acids, about 50 amino acids to about 180 amino acids, about 50 amino acids to about 175 amino acids, about 50 amino acids to about 170 amino acids, about 50 amino acids to about 165 amino acids, about 50 amino acids to about 160 amino acids, about 50 amino acids to about 155 amino acids, about 50 amino acids to about 150 amino acids, about 50 amino acids to about 145 amino acids, about 50 amino acids to about 140 amino acids, about 50 amino acids to about 135 amino acids, about 50 amino acids to about 130 amino acids, about 50 amino acids to about 125 amino acids, about 50 amino acids to about 120 amino acids, about 50 amino acids to about 115 amino acids, about 50 amino acids to about 110 amino acids, about 50 amino acids to about 105 amino acids, about 50 amino acids to about 100 amino acids, about 50 amino acids to about 95 amino acids, about 50 amino acids to about 90 amino acids, about 50 amino acids to about 85 amino acids, about 50 amino acids to about 80 amino acids, about 50 amino acids to about 75 amino acids, about 50 amino acids to about 70 amino acids, about 50 amino acids to about 65 amino acids, about 50 amino acids to about 60 amino acids, about 50 amino acids to about 55 amino acids, about 55 amino acids to about 1000 amino acids, about 55 amino acids to about 950 amino acids, about 55 amino acids to about 900Attorney Docket No. 47039-0043W01

[0221] amino acids, about 55 amino acids to about 850 amino acids, about 55 amino acids to about 800 amino acids, about 55 amino acids to about 750 amino acids, about 55 amino acids to about 700 amino acids, about 55 amino acids to about 650 amino acids, about 55 amino acids to about 600 amino acids, about 55 amino acids to about 550 amino acids, about 55 amino acids to about 500 amino acids, about 55 amino acids to about 450 amino acids, about 55 amino acids to about 400 amino acids, about 55 amino acids to about 350 amino acids, about 55 amino acids to about 300 amino acids, about 55 amino acids to about 280 amino acids, about 55 amino acids to about 260 amino acids, about 55 amino acids to about 240 amino acids, about 55 amino acids to about 220 amino acids, about 55 amino acids to about 200 amino acids, about 55 amino acids to about 195 amino acids, about 55 amino acids to about 190 amino acids, about 55 amino acids to about 185 amino acids, about 55 amino acids to about 180 amino acids, about 55 amino acids to about 175 amino acids, about 55 amino acids to about 170 amino acids, about 55 amino acids to about 165 amino acids, about 55 amino acids to about 160 amino acids, about 55 amino acids to about 155 amino acids, about 55 amino acids to about 150 amino acids, about 55 amino acids to about 145 amino acids, about 55 amino acids to about 140 amino acids, about 55 amino acids to about 135 amino acids, about 55 amino acids to about 130 amino acids, about 55 amino acids to about 125 amino acids, about 55 amino acids to about 120 amino acids, about 55 amino acids to about 115 amino acids, about 55 amino acids to about 110 amino acids, about 55 amino acids to about 105 amino acids, about 55 amino acids to about 100 amino acids, about 55 amino acids to about 95 amino acids, about 55 amino acids to about 90 amino acids, about 55 amino acids to about 85 amino acids, about 55 amino acids to about 80 amino acids, about 55 amino acids to about 75 amino acids, about 55 amino acids to about 70 amino acids, about 55 amino acids to about 65 amino acids, about 55 amino acids to about 60 amino acids, about 60 amino acids to about 1000 amino acids, about 60 amino acids to about 950 amino acids, about 60 amino acids to about 900 amino acids, about 60 amino acids to about 850 amino acids, about 60 amino acids to about 800 amino acids, about 60 amino acids to about 750 amino acids, about 60 amino acids to about 700 amino acids, about 60 amino acids to about 650 amino acids, about 60 amino acids to about 600 amino acids, about 60 amino acids to about 550 amino acids, about 60 amino acids to about 500 amino acids, about 60 amino acids to about 450 amino acids, about 60 amino acids to about 400 amino acids, about 60 amino acids to about 350 amino acids, about 60 amino acids to about 300 amino acids, about 60 amino acids to about 280 amino acids, about 60 amino acids to about 260Attorney Docket No. 47039-0043W01

[0222] amino acids, about 60 amino acids to about 240 amino acids, about 60 amino acids to about 220 amino acids, about 60 amino acids to about 200 amino acids, about 60 amino acids to about 195 amino acids, about 60 amino acids to about 190 amino acids, about 60 amino acids to about 185 amino acids, about 60 amino acids to about 180 amino acids, about 60 amino acids to about 175 amino acids, about 60 amino acids to about 170 amino acids, about 60 amino acids to about 165 amino acids, about 60 amino acids to about 160 amino acids, about 60 amino acids to about 155 amino acids, about 60 amino acids to about 150 amino acids, about 60 amino acids to about 145 amino acids, about 60 amino acids to about 140 amino acids, about 60 amino acids to about 135 amino acids, about 60 amino acids to about 130 amino acids, about 60 amino acids to about 125 amino acids, about 60 amino acids to about 120 amino acids, about 60 amino acids to about 115 amino acids, about 60 amino acids to about 110 amino acids, about 60 amino acids to about 105 amino acids, about 60 amino acids to about 100 amino acids, about 60 amino acids to about 95 amino acids, about 60 amino acids to about 90 amino acids, about 60 amino acids to about 85 amino acids, about 60 amino acids to about 80 amino acids, about 60 amino acids to about 75 amino acids, about 60 amino acids to about 70 amino acids, about 60 amino acids to about 65 amino acids, about 65 amino acids to about 1000 amino acids, about 65 amino acids to about 950 amino acids, about 65 amino acids to about 900 amino acids, about 65 amino acids to about 850 amino acids, about 65 amino acids to about 800 amino acids, about 65 amino acids to about 750 amino acids, about 65 amino acids to about 700 amino acids, about 65 amino acids to about 650 amino acids, about 65 amino acids to about 600 amino acids, about 65 amino acids to about 550 amino acids, about 65 amino acids to about 500 amino acids, about 65 amino acids to about 450 amino acids, about 65 amino acids to about 400 amino acids, about 65 amino acids to about 350 amino acids, about 65 amino acids to about 300 amino acids, about 65 amino acids to about 280 amino acids, about 65 amino acids to about 260 amino acids, about 65 amino acids to about 240 amino acids, about 65 amino acids to about 220 amino acids, about 65 amino acids to about 200 amino acids, about 65 amino acids to about 195 amino acids, about 65 amino acids to about 190 amino acids, about 65 amino acids to about 185 amino acids, about 65 amino acids to about 180 amino acids, about 65 amino acids to about 175 amino acids, about 65 amino acids to about 170 amino acids, about 65 amino acids to about 165 amino acids, about 65 amino acids to about 160 amino acids, about 65 amino acids to about 155 amino acids, about 65 amino acids to about 150 amino acids, about 65 amino acids to about 145 amino acids, about 65 amino acids to about 140Attorney Docket No. 47039-0043W01

[0223] amino acids, about 65 amino acids to about 135 amino acids, about 65 amino acids to about 130 amino acids, about 65 amino acids to about 125 amino acids, about 65 amino acids to about 120 amino acids, about 65 amino acids to about 115 amino acids, about 65 amino acids to about 110 amino acids, about 65 amino acids to about 105 amino acids, about 65 amino acids to about 100 amino acids, about 65 amino acids to about 95 amino acids, about 65 amino acids to about 90 amino acids, about 65 amino acids to about 85 amino acids, about 65 amino acids to about 80 amino acids, about 65 amino acids to about 75 amino acids, about 65 amino acids to about 70 amino acids, about 70 amino acids to about 1000 amino acids, about 70 amino acids to about 950 amino acids, about 70 amino acids to about 900 amino acids, about 70 amino acids to about 850 amino acids, about 70 amino acids to about 800 amino acids, about 70 amino acids to about 750 amino acids, about 70 amino acids to about 700 amino acids, about 70 amino acids to about 650 amino acids, about 70 amino acids to about 600 amino acids, about 70 amino acids to about 550 amino acids, about 70 amino acids to about 500 amino acids, about 70 amino acids to about 450 amino acids, about 70 amino acids to about 400 amino acids, about 70 amino acids to about 350 amino acids, about 70 amino acids to about 300 amino acids, about 70 amino acids to about 280 amino acids, about 70 amino acids to about 260 amino acids, about 70 amino acids to about 240 amino acids, about 70 amino acids to about 220 amino acids, about 70 amino acids to about 200 amino acids, about 70 amino acids to about 195 amino acids, about 70 amino acids to about 190 amino acids, about 70 amino acids to about 185 amino acids, about 70 amino acids to about 180 amino acids, about 70 amino acids to about 175 amino acids, about 70 amino acids to about 170 amino acids, about 70 amino acids to about 165 amino acids, about 70 amino acids to about 160 amino acids, about 70 amino acids to about 155 amino acids, about 70 amino acids to about 150 amino acids, about 70 amino acids to about 145 amino acids, about 70 amino acids to about 140 amino acids, about 70 amino acids to about 135 amino acids, about 70 amino acids to about 130 amino acids, about 70 amino acids to about 125 amino acids, about 70 amino acids to about 120 amino acids, about 70 amino acids to about 115 amino acids, about 70 amino acids to about 110 amino acids, about 70 amino acids to about 105 amino acids, about 70 amino acids to about 100 amino acids, about 70 amino acids to about 95 amino acids, about 70 amino acids to about 90 amino acids, about 70 amino acids to about 85 amino acids, about 70 amino acids to about 80 amino acids, about 70 amino acids to about 75 amino acids, about 75 amino acids to about 1000 amino acids, about 75 amino acids to about 950 amino acids, about 75 amino acids to about 900Attorney Docket No. 47039-0043W01

[0224] amino acids, about 75 amino acids to about 850 amino acids, about 75 amino acids to about 800 amino acids, about 75 amino acids to about 750 amino acids, about 75 amino acids to about 700 amino acids, about 75 amino acids to about 650 amino acids, about 75 amino acids to about 600 amino acids, about 75 amino acids to about 550 amino acids, about 75 amino acids to about 500 amino acids, about 75 amino acids to about 450 amino acids, about 75 amino acids to about 400 amino acids, about 75 amino acids to about 350 amino acids, about 75 amino acids to about 300 amino acids, about 75 amino acids to about 280 amino acids, about 75 amino acids to about 260 amino acids, about 75 amino acids to about 240 amino acids, about 75 amino acids to about 220 amino acids, about 75 amino acids to about 200 amino acids, about 75 amino acids to about 195 amino acids, about 75 amino acids to about 190 amino acids, about 75 amino acids to about 185 amino acids, about 75 amino acids to about 180 amino acids, about 75 amino acids to about 175 amino acids, about 75 amino acids to about 170 amino acids, about 75 amino acids to about 165 amino acids, about 75 amino acids to about 160 amino acids, about 75 amino acids to about 155 amino acids, about 75 amino acids to about 150 amino acids, about 75 amino acids to about 145 amino acids, about 75 amino acids to about 140 amino acids, about 75 amino acids to about 135 amino acids, about 75 amino acids to about 130 amino acids, about 75 amino acids to about 125 amino acids, about 75 amino acids to about 120 amino acids, about 75 amino acids to about 115 amino acids, about 75 amino acids to about 110 amino acids, about 75 amino acids to about 105 amino acids, about 75 amino acids to about 100 amino acids, about 75 amino acids to about 95 amino acids, about 75 amino acids to about 90 amino acids, about 75 amino acids to about 85 amino acids, about 75 amino acids to about 80 amino acids, about 80 amino acids to about 1000 amino acids, about 80 amino acids to about 950 amino acids, about 80 amino acids to about 900 amino acids, about 80 amino acids to about 850 amino acids, about 80 amino acids to about 800 amino acids, about 80 amino acids to about 750 amino acids, about 80 amino acids to about 700 amino acids, about 80 amino acids to about 650 amino acids, about 80 amino acids to about 600 amino acids, about 80 amino acids to about 550 amino acids, about 80 amino acids to about 500 amino acids, about 80 amino acids to about 450 amino acids, about 80 amino acids to about 400 amino acids, about 80 amino acids to about 350 amino acids, about 80 amino acids to about 300 amino acids, about 80 amino acids to about 280 amino acids, about 80 amino acids to about 260 amino acids, about 80 amino acids to about 240 amino acids, about 80 amino acids to about 220 amino acids, about 80 amino acids to about 200 amino acids, about 80 amino acids to about 195Attorney Docket No. 47039-0043W01

[0225] amino acids, about 80 amino acids to about 190 amino acids, about 80 amino acids to about 185 amino acids, about 80 amino acids to about 180 amino acids, about 80 amino acids to about 175 amino acids, about 80 amino acids to about 170 amino acids, about 80 amino acids to about 165 amino acids, about 80 amino acids to about 160 amino acids, about 80 amino acids to about 155 amino acids, about 80 amino acids to about 150 amino acids, about 80 amino acids to about 145 amino acids, about 80 amino acids to about 140 amino acids, about 80 amino acids to about 135 amino acids, about 80 amino acids to about 130 amino acids, about 80 amino acids to about 125 amino acids, about 80 amino acids to about 120 amino acids, about 80 amino acids to about 115 amino acids, about 80 amino acids to about 110 amino acids, about 80 amino acids to about 105 amino acids, about 80 amino acids to about 100 amino acids, about 80 amino acids to about 95 amino acids, about 80 amino acids to about 90 amino acids, about 80 amino acids to about 85 amino acids, about 85 amino acids to about 1000 amino acids, about 85 amino acids to about 950 amino acids, about 85 amino acids to about 900 amino acids, about 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, about 85 amino acids to about 220 amino acids, about 85 amino acids to about 200 amino acids, about 85 amino acids to about 195 amino acids, about 85 amino acids to about 190 amino acids, about 85 amino acids to about 185 amino acids, about 85 amino acids to about 180 amino acids, about 85 amino acids to about 175 amino acids, about 85 amino acids to about 170 amino acids, about 85 amino acids to about 165 amino acids, about 85 amino acids to about 160 amino acids, about 85 amino acids to about 155 amino acids, about 85 amino acids to about 150 amino acids, about 85 amino acids to about 145 amino acids, about 85 amino acids to about 140 amino acids, about 85 amino acids to about 135 amino acids, about 85 amino acids to about 130 amino acids, about 85 amino acids to about 125 amino acids, about 85 amino acids to about 120 amino acids, about 85 amino acids to about 115 amino acids, about 85 amino acids to about 110 amino acids, about 85 amino acids to about 105 amino acids, about 85 amino acids to about 100Attorney Docket No. 47039-0043W01

[0226] amino acids, about 85 amino acids to about 95 amino acids, about 85 amino acids to about 90 amino acids, about 90 amino acids to about 1000 amino acids, about 90 amino acids to about 950 amino acids, about 90 amino acids to about 900 amino acids, about 90 amino acids to about 850 amino acids, about 90 amino acids to about 800 amino acids, about 90 amino acids to about 750 amino acids, about 90 amino acids to about 700 amino acids, about 90 amino acids to about 650 amino acids, about 90 amino acids to about 600 amino acids, about 90 amino acids to about 550 amino acids, about 90 amino acids to about 500 amino acids, about 90 amino acids to about 450 amino acids, about 90 amino acids to about 400 amino acids, about 90 amino acids to about 350 amino acids, about 90 amino acids to about 300 amino acids, about 90 amino acids to about 280 amino acids, about 90 amino acids to about 260 amino acids, about 90 amino acids to about 240 amino acids, about 90 amino acids to about 220 amino acids, about 90 amino acids to about 200 amino acids, about 90 amino acids to about 195 amino acids, about 90 amino acids to about 190 amino acids, about 90 amino acids to about 185 amino acids, about 90 amino acids to about 180 amino acids, about 90 amino acids to about 175 amino acids, about 90 amino acids to about 170 amino acids, about 90 amino acids to about 165 amino acids, about 90 amino acids to about 160 amino acids, about 90 amino acids to about 155 amino acids, about 90 amino acids to about 150 amino acids, about 90 amino acids to about 145 amino acids, about 90 amino acids to about 140 amino acids, about 90 amino acids to about 135 amino acids, about 90 amino acids to about 130 amino acids, about 90 amino acids to about 125 amino acids, about 90 amino acids to about 120 amino acids, about 90 amino acids to about 115 amino acids, about 90 amino acids to about 110 amino acids, about 90 amino acids to about 105 amino acids, about 90 amino acids to about 100 amino acids, about 90 amino acids to about 95 amino acids, about 95 amino acids to about 1000 amino acids, about 95 amino acids to about 950 amino acids, about 95 amino acids to about 900 amino acids, about 95 amino acids to about 850 amino acids, about 95 amino acids to about 800 amino acids, about 95 amino acids to about 750 amino acids, about 95 amino acids to about 700 amino acids, about 95 amino acids to about 650 amino acids, about 95 amino acids to about 600 amino acids, about 95 amino acids to about 550 amino acids, about 95 amino acids to about 500 amino acids, about 95 amino acids to about 450 amino acids, about 95 amino acids to about 400 amino acids, about 95 amino acids to about 350 amino acids, about 95 amino acids to about 300 amino acids, about 95 amino acids to about 280 amino acids, about 95 amino acids to about 260 amino acids, about 95 amino acids to about 240 amino acids, about 95 amino acids to about 220Attorney Docket No. 47039-0043W01

[0227] amino acids, about 95 amino acids to about 200 amino acids, about 95 amino acids to about 195 amino acids, about 95 amino acids to about 190 amino acids, about 95 amino acids to about 185 amino acids, about 95 amino acids to about 180 amino acids, about 95 amino acids to about 175 amino acids, about 95 amino acids to about 170 amino acids, about 95 amino acids to about 165 amino acids, about 95 amino acids to about 160 amino acids, about 95 amino acids to about 155 amino acids, about 95 amino acids to about 150 amino acids, about 95 amino acids to about 145 amino acids, about 95 amino acids to about 140 amino acids, about 95 amino acids to about 135 amino acids, about 95 amino acids to about 130 amino acids, about 95 amino acids to about 125 amino acids, about 95 amino acids to about 120 amino acids, about 95 amino acids to about 115 amino acids, about 95 amino acids to about 110 amino acids, about 95 amino acids to about 105 amino acids, about 95 amino acids to about 100 amino acids, about 100 amino acids to about 1000 amino acids, about 100 amino acids to about 950 amino acids, about 100 amino acids to about 900 amino acids, about 100 amino acids to about 850 amino acids, about 100 amino acids to about 800 amino acids, about 100 amino acids to about 750 amino acids, about 100 amino acids to about 700 amino acids, about 100 amino acids to about 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 450 amino acids, about 100 amino acids to about 400 amino acids, about 100 amino acids to about 350 amino acids, about 100 amino acids to about 300 amino acids, about 100 amino acids to about 280 amino acids, about 100 amino acids to about 260 amino acids, about 100 amino acids to about 240 amino acids, about 100 amino acids to about 220 amino acids, about 100 amino acids to about 200 amino acids, about 100 amino acids to about 195 amino acids, about 100 amino acids to about 190 amino acids, about 100 amino acids to about 185 amino acids, about 100 amino acids to about 180 amino acids, about 100 amino acids to about 175 amino acids, about 100 amino acids to about 170 amino acids, about 100 amino acids to about 165 amino acids, about 100 amino acids to about 160 amino acids, about 100 amino acids to about 155 amino acids, about 100 amino acids to about 150 amino acids, about 100 amino acids to about 145 amino acids, about 100 amino acids to about 140 amino acids, about 100 amino acids to about 135 amino acids, about 100 amino acids to about 130 amino acids, about 100 amino acids to about 125 amino acids, about 100 amino acids to about 120 amino acids, about 100 amino acids to about 115 amino acids, about 100 amino acids to about 110 amino acids, about 100 amino acidsAttorney Docket No. 47039-0043W01

[0228] to about 105 amino acids, about 105 amino acids to about 1000 amino acids, about 105 amino acids to about 950 amino acids, about 105 amino acids to about 900 amino acids, about 105 amino acids to about 850 amino acids, about 105 amino acids to about 800 amino acids, about 105 amino acids to about 750 amino acids, about 105 amino acids to about 700 amino acids, about 105 amino acids to about 650 amino acids, about 105 amino acids to about 600 amino acids, about 105 amino acids to about 550 amino acids, about 105 amino acids to about 500 amino acids, about 105 amino acids to about 450 amino acids, about 105 amino acids to about 400 amino acids, about 105 amino acids to about 350 amino acids, about 105 amino acids to about 300 amino acids, about 105 amino acids to about 280 amino acids, about 105 amino acids to about 260 amino acids, about 105 amino acids to about 240 amino acids, about 105 amino acids to about 220 amino acids, about 105 amino acids to about 200 amino acids, about 105 amino acids to about 195 amino acids, about 105 amino acids to about 190 amino acids, about 105 amino acids to about 185 amino acids, about 105 amino acids to about 180 amino acids, about 105 amino acids to about 175 amino acids, about 105 amino acids to about 170 amino acids, about 105 amino acids to about 165 amino acids, about 105 amino acids to about 160 amino acids, about 105 amino acids to about 155 amino acids, about 105 amino acids to about 150 amino acids, about 105 amino acids to about 145 amino acids, about 105 amino acids to about 140 amino acids, about 105 amino acids to about 135 amino acids, about 105 amino acids to about 130 amino acids, about 105 amino acids to about 125 amino acids, about 105 amino acids to about 120 amino acids, about 105 amino acids to about 115 amino acids, about 105 amino acids to about 110 amino acids, about 110 amino acids to about 1000 amino acids, about 110 amino acids to about 950 amino acids, about 110 amino acids to about 900 amino acids, about 110 amino acids to about 850 amino acids, about 110 amino acids to about 800 amino acids, about 110 amino acids to about 750 amino acids, about 110 amino acids to about 700 amino acids, about 110 amino acids to about 650 amino acids, about 110 amino acids to about 600 amino acids, about 110 amino acids to about 550 amino acids, about 110 amino acids to about 500 amino acids, about 110 amino acids to about 450 amino acids, about 110 amino acids to about 400 amino acids, about 110 amino acids to about 350 amino acids, about 110 amino acids to about 300 amino acids, about 110 amino acids to about 280 amino acids, about 110 amino acids to about 260 amino acids, about 110 amino acids to about 240 amino acids, about 110 amino acids to about 220 amino acids, about 110 amino acids to about 200 amino acids,Attorney Docket No. 47039-0043W01

[0229] about 110 amino acids to about 195 amino acids, about 110 amino acids to about 190 amino acids, about 110 amino acids to about 185 amino acids, about 110 amino acids to about 180 amino acids, about 110 amino acids to about 175 amino acids, about 110 amino acids to about 170 amino acids, about 110 amino acids to about 165 amino acids, about 110 amino acids to about 160 amino acids, about 110 amino acids to about 155 amino acids, about 110 amino acids to about 150 amino acids, about 110 amino acids to about 145 amino acids, about 110 amino acids to about 140 amino acids, about 110 amino acids to about 135 amino acids, about 110 amino acids to about 130 amino acids, about 110 amino acids to about 125 amino acids, about 110 amino acids to about 120 amino acids, about 110 amino acids to about 115 amino acids, about 115 amino acids to about 1000 amino acids, about 115 amino acids to about 950 amino acids, about 115 amino acids to about 900 amino acids, about 115 amino acids to about 850 amino acids, about 115 amino acids to about 800 amino acids, about 115 amino acids to about 750 amino acids, about 115 amino acids to about 700 amino acids, about 115 amino acids to about 650 amino acids, about 115 amino acids to about 600 amino acids, about 115 amino acids to about 550 amino acids, about 115 amino acids to about 500 amino acids, about 115 amino acids to about 450 amino acids, about 115 amino acids to about 400 amino acids, about 115 amino acids to about 350 amino acids, about 115 amino acids to about 300 amino acids, about 115 amino acids to about 280 amino acids, about 115 amino acids to about 260 amino acids, about 115 amino acids to about 240 amino acids, about 115 amino acids to about 220 amino acids, about 115 amino acids to about 200 amino acids, about 115 amino acids to about 195 amino acids, about 115 amino acids to about 190 amino acids, about 115 amino acids to about 185 amino acids, about 115 amino acids to about 180 amino acids, about 115 amino acids to about 175 amino acids, about 115 amino acids to about 170 amino acids, about 115 amino acids to about 165 amino acids, about 115 amino acids to about 160 amino acids, about 115 amino acids to about 155 amino acids, about 115 amino acids to about 150 amino acids, about 115 amino acids to about 145 amino acids, about 115 amino acids to about 140 amino acids, about 115 amino acids to about 135 amino acids, about 115 amino acids to about 130 amino acids, about 115 amino acids to about 125 amino acids, about 115 amino acids to about 120 amino acids, about 120 amino acids to about 1000 amino acids, about 120 amino acids to about 950 amino acids, about 120 amino acids to about 900 amino acids, about 120 amino acids to about 850 amino acids, about 120 amino acids to about 800 amino acids, about 120 amino acids toAttorney Docket No. 47039-0043W01

[0230] about 750 amino acids, about 120 amino acids to about 700 amino acids, about 120 amino acids to about 650 amino acids, about 120 amino acids to about 600 amino acids, about 120 amino acids to about 550 amino acids, about 120 amino acids to about 500 amino acids, about 120 amino acids to about 450 amino acids, about 120 amino acids to about 400 amino acids, about 120 amino acids to about 350 amino acids, about 120 amino acids to about 300 amino acids, about 120 amino acids to about 280 amino acids, about 120 amino acids to about 260 amino acids, about 120 amino acids to about 240 amino acids, about 120 amino acids to about 220 amino acids, about 120 amino acids to about 200 amino acids, about 120 amino acids to about 195 amino acids, about 120 amino acids to about 190 amino acids, about 120 amino acids to about 185 amino acids, about 120 amino acids to about 180 amino acids, about 120 amino acids to about 175 amino acids, about 120 amino acids to about 170 amino acids, about 120 amino acids to about 165 amino acids, about 120 amino acids to about 160 amino acids, about 120 amino acids to about 155 amino acids, about 120 amino acids to about 150 amino acids, about 120 amino acids to about 145 amino acids, about 120 amino acids to about 140 amino acids, about 120 amino acids to about 135 amino acids, about 120 amino acids to about 130 amino acids, about 120 amino acids to about 125 amino acids, about 125 amino acids to about 1000 amino acids, about 125 amino acids to about 950 amino acids, about 125 amino acids to about 900 amino acids, about 125 amino acids to about 850 amino acids, about 125 amino acids to about 800 amino acids, about 125 amino acids to about 750 amino acids, about 125 amino acids to about 700 amino acids, about 125 amino acids to about 650 amino acids, about 125 amino acids to about 600 amino acids, about 125 amino acids to about 550 amino acids, about 125 amino acids to about 500 amino acids, about 125 amino acids to about 450 amino acids, about 125 amino acids to about 400 amino acids, about 125 amino acids to about 350 amino acids, about 125 amino acids to about 300 amino acids, about 125 amino acids to about 280 amino acids, about 125 amino acids to about 260 amino acids, about 125 amino acids to about 240 amino acids, about 125 amino acids to about 220 amino acids, about 125 amino acids to about 200 amino acids, about 125 amino acids to about 195 amino acids, about 125 amino acids to about 190 amino acids, about 125 amino acids to about 185 amino acids, about 125 amino acids to about 180 amino acids, about 125 amino acids to about 175 amino acids, about 125 amino acids to about 170 amino acids, about 125 amino acids to about 165 amino acids, about 125 amino acids to about 160 amino acids, about 125 amino acids to about 155 amino acids, aboutAttorney Docket No. 47039-0043W01

[0231] 125 amino acids to about 150 amino acids, about 125 amino acids to about 145 amino acids, about 125 amino acids to about 140 amino acids, about 125 amino acids to about 135 amino acids, about 125 amino acids to about 130 amino acids, about 130 amino acids to about 1000 amino acids, about 130 amino acids to about 950 amino acids, about 130 amino acids to about 900 amino acids, about 130 amino acids to about 850 amino acids, about 130 amino acids to about 800 amino acids, about 130 amino acids to about 750 amino acids, about 130 amino acids to about 700 amino acids, about 130 amino acids to about 650 amino acids, about 130 amino acids to about 600 amino acids, about 130 amino acids to about 550 amino acids, about 130 amino acids to about 500 amino acids, about 130 amino acids to about 450 amino acids, about 130 amino acids to about 400 amino acids, about 130 amino acids to about 350 amino acids, about 130 amino acids to about 300 amino acids, about 130 amino acids to about 280 amino acids, about 130 amino acids to about 260 amino acids, about 130 amino acids to about 240 amino acids, about 130 amino acids to about 220 amino acids, about 130 amino acids to about 200 amino acids, about 130 amino acids to about 195 amino acids, about 130 amino acids to about 190 amino acids, about 130 amino acids to about 185 amino acids, about 130 amino acids to about 180 amino acids, about 130 amino acids to about 175 amino acids, about 130 amino acids to about 170 amino acids, about 130 amino acids to about 165 amino acids, about 130 amino acids to about 160 amino acids, about 130 amino acids to about 155 amino acids, about 130 amino acids to about 150 amino acids, about 130 amino acids to about 145 amino acids, about 130 amino acids to about 140 amino acids, about 130 amino acids to about 135 amino acids, about 135 amino acids to about 1000 amino acids, about 135 amino acids to about 950 amino acids, about 135 amino acids to about 900 amino acids, about 135 amino acids to about 850 amino acids, about 135 amino acids to about 800 amino acids, about 135 amino acids to about 750 amino acids, about 135 amino acids to about 700 amino acids, about 135 amino acids to about 650 amino acids, about 135 amino acids to about 600 amino acids, about 135 amino acids to about 550 amino acids, about 135 amino acids to about 500 amino acids, about 135 amino acids to about 450 amino acids, about 135 amino acids to about 400 amino acids, about 135 amino acids to about 350 amino acids, about 135 amino acids to about 300 amino acids, about 135 amino acids to about 280 amino acids, about 135 amino acids to about 260 amino acids, about 135 amino acids to about 240 amino acids, about 135 amino acids to about 220 amino acids, about 135 amino acids to about 200 amino acids, about 135 amino acids to aboutAttorney Docket No. 47039-0043W01

[0232] 195 amino acids, about 135 amino acids to about 190 amino acids, about 135 amino acids to about 185 amino acids, about 135 amino acids to about 180 amino acids, about 135 amino acids to about 175 amino acids, about 135 amino acids to about 170 amino acids, about 135 amino acids to about 165 amino acids, about 135 amino acids to about 160 amino acids, about 135 amino acids to about 155 amino acids, about 135 amino acids to about 150 amino acids, about 135 amino acids to about 145 amino acids, about 135 amino acids to about 140 amino acids, about 140 amino acids to about 1000 amino acids, about 140 amino acids to about 950 amino acids, about 140 amino acids to about 900 amino acids, about 140 amino acids to about 850 amino acids, about 140 amino acids to about 800 amino acids, about 140 amino acids to about 750 amino acids, about 140 amino acids to about 700 amino acids, about 140 amino acids to about 650 amino acids, about 140 amino acids to about 600 amino acids, about 140 amino acids to about 550 amino acids, about 140 amino acids to about 500 amino acids, about 140 amino acids to about 450 amino acids, about 140 amino acids to about 400 amino acids, about 140 amino acids to about 350 amino acids, about 140 amino acids to about 300 amino acids, about 140 amino acids to about 280 amino acids, about 140 amino acids to about 260 amino acids, about 140 amino acids to about 240 amino acids, about 140 amino acids to about 220 amino acids, about 140 amino acids to about 200 amino acids, about 140 amino acids to about 195 amino acids, about 140 amino acids to about 190 amino acids, about 140 amino acids to about 185 amino acids, about 140 amino acids to about 180 amino acids, about 140 amino acids to about 175 amino acids, about 140 amino acids to about 170 amino acids, about 140 amino acids to about 165 amino acids, about 140 amino acids to about 160 amino acids, about 140 amino acids to about 155 amino acids, about 140 amino acids to about 150 amino acids, about 140 amino acids to about 145 amino acids, about 145 amino acids to about 1000 amino acids, about 145 amino acids to about 950 amino acids, about 145 amino acids to about 900 amino acids, about 145 amino acids to about 850 amino acids, about 145 amino acids to about 800 amino acids, about 145 amino acids to about 750 amino acids, about 145 amino acids to about 700 amino acids, about 145 amino acids to about 650 amino acids, about 145 amino acids to about 600 amino acids, about 145 amino acids to about 550 amino acids, about 145 amino acids to about 500 amino acids, about 145 amino acids to about 450 amino acids, about 145 amino acids to about 400 amino acids, about 145 amino acids to about 350 amino acids, about 145 amino acids to about 300 amino acids, about 145 amino acids to about 280 amino acids, about 145Attorney Docket No. 47039-0043W01

[0233] amino acids to about 260 amino acids, about 145 amino acids to about 240 amino acids, about 145 amino acids to about 220 amino acids, about 145 amino acids to about 200 amino acids, about 145 amino acids to about 195 amino acids, about 145 amino acids to about 190 amino acids, about 145 amino acids to about 185 amino acids, about 145 amino acids to about 180 amino acids, about 145 amino acids to about 175 amino acids, about 145 amino acids to about 170 amino acids, about 145 amino acids to about 165 amino acids, about 145 amino acids to about 160 amino acids, about 145 amino acids to about 155 amino acids, about 145 amino acids to about 150 amino acids, about 150 amino acids to about 1000 amino acids, about 150 amino acids to about 950 amino acids, about 150 amino acids to about 900 amino acids, about 150 amino acids to about 850 amino acids, about 150 amino acids to about 800 amino acids, about 150 amino acids to about 750 amino acids, about 150 amino acids to about 700 amino acids, about 150 amino acids to about 650 amino acids, about 150 amino acids to about 600 amino acids, about 150 amino acids to about 550 amino acids, about 150 amino acids to about 500 amino acids, about 150 amino acids to about 450 amino acids, about 150 amino acids to about 400 amino acids, about 150 amino acids to about 350 amino acids, about 150 amino acids to about 300 amino acids, about 150 amino acids to about 280 amino acids, about 150 amino acids to about 260 amino acids, about 150 amino acids to about 240 amino acids, about 150 amino acids to about 220 amino acids, about 150 amino acids to about 200 amino acids, about 150 amino acids to about 195 amino acids, about 150 amino acids to about 190 amino acids, about 150 amino acids to about 185 amino acids, about 150 amino acids to about 180 amino acids, about 150 amino acids to about 175 amino acids, about 150 amino acids to about 170 amino acids, about 150 amino acids to about 165 amino acids, about 150 amino acids to about 160 amino acids, about 150 amino acids to about 155 amino acids, about 155 amino acids to about 1000 amino acids, about 155 amino acids to about 950 amino acids, about 155 amino acids to about 900 amino acids, about 155 amino acids to about 850 amino acids, about 155 amino acids to about 800 amino acids, about 155 amino acids to about 750 amino acids, about 155 amino acids to about 700 amino acids, about 155 amino acids to about 650 amino acids, about 155 amino acids to about 600 amino acids, about 155 amino acids to about 550 amino acids, about 155 amino acids to about 500 amino acids, about 155 amino acids to about 450 amino acids, about 155 amino acids to about 400 amino acids, about 155 amino acids to about 350 amino acids, about 155 amino acids to about 300 amino acids, about 155 amino acids to about 280Attorney Docket No. 47039-0043W01

[0234] amino acids, about 155 amino acids to about 260 amino acids, about 155 amino acids to about 240 amino acids, about 155 amino acids to about 220 amino acids, about 155 amino acids to about 200 amino acids, about 155 amino acids to about 195 amino acids, about 155 amino acids to about 190 amino acids, about 155 amino acids to about 185 amino acids, about 155 amino acids to about 180 amino acids, about 155 amino acids to about 175 amino acids, about 155 amino acids to about 170 amino acids, about 155 amino acids to about 165 amino acids, about 155 amino acids to about 160 amino acids, about 160 amino acids to about 1000 amino acids, about 160 amino acids to about 950 amino acids, about 160 amino acids to about 900 amino acids, about 160 amino acids to about 850 amino acids, about 160 amino acids to about 800 amino acids, about 160 amino acids to about 750 amino acids, about 160 amino acids to about 700 amino acids, about 160 amino acids to about 650 amino acids, about 160 amino acids to about 600 amino acids, about 160 amino acids to about 550 amino acids, about 160 amino acids to about 500 amino acids, about 160 amino acids to about 450 amino acids, about 160 amino acids to about 400 amino acids, about 160 amino acids to about 350 amino acids, about 160 amino acids to about 300 amino acids, about 160 amino acids to about 280 amino acids, about 160 amino acids to about 260 amino acids, about 160 amino acids to about 240 amino acids, about 160 amino acids to about 220 amino acids, about 160 amino acids to about 200 amino acids, about 160 amino acids to about 195 amino acids, about 160 amino acids to about 190 amino acids, about 160 amino acids to about 185 amino acids, about 160 amino acids to about 180 amino acids, about 160 amino acids to about 175 amino acids, about 160 amino acids to about 170 amino acids, about 160 amino acids to about 165 amino acids, about 165 amino acids to about 1000 amino acids, about 165 amino acids to about 950 amino acids, about 165 amino acids to about 900 amino acids, about 165 amino acids to about 850 amino acids, about 165 amino acids to about 800 amino acids, about 165 amino acids to about 750 amino acids, about 165 amino acids to about 700 amino acids, about 165 amino acids to about 650 amino acids, about 165 amino acids to about 600 amino acids, about 165 amino acids to about 550 amino acids, about 165 amino acids to about 500 amino acids, about 165 amino acids to about 450 amino acids, about 165 amino acids to about 400 amino acids, about 165 amino acids to about 350 amino acids, about 165 amino acids to about 300 amino acids, about 165 amino acids to about 280 amino acids, about 165 amino acids to about 260 amino acids, about 165 amino acids to about 240 amino acids, about 165 amino acids to about 220 amino acids, about 165 aminoAttorney Docket No. 47039-0043W01

[0235] acids to about 200 amino acids, about 165 amino acids to about 195 amino acids, about 165 amino acids to about 190 amino acids, about 165 amino acids to about 185 amino acids, about 165 amino acids to about 180 amino acids, about 165 amino acids to about 175 amino acids, about 165 amino acids to about 170 amino acids, about 170 amino acids to about 1000 amino acids, about 170 amino acids to about 950 amino acids, about 170 amino acids to about 900 amino acids, about 170 amino acids to about 850 amino acids, about 170 amino acids to about 800 amino acids, about 170 amino acids to about 750 amino acids, about 170 amino acids to about 700 amino acids, about 170 amino acids to about 650 amino acids, about 170 amino acids to about 600 amino acids, about 170 amino acids to about 550 amino acids, about 170 amino acids to about 500 amino acids, about 170 amino acids to about 450 amino acids, about 170 amino acids to about 400 amino acids, about 170 amino acids to about 350 amino acids, about 170 amino acids to about 300 amino acids, about 170 amino acids to about 280 amino acids, about 170 amino acids to about 260 amino acids, about 170 amino acids to about 240 amino acids, about 170 amino acids to about 220 amino acids, about 170 amino acids to about 200 amino acids, about 170 amino acids to about 195 amino acids, about 170 amino acids to about 190 amino acids, about 170 amino acids to about 185 amino acids, about 170 amino acids to about 180 amino acids, about 170 amino acids to about 175 amino acids, about 175 amino acids to about 1000 amino acids, about 175 amino acids to about 950 amino acids, about 175 amino acids to about 900 amino acids, about 175 amino acids to about 850 amino acids, about 175 amino acids to about 800 amino acids, about 175 amino acids to about 750 amino acids, about 175 amino acids to about 700 amino acids, about 175 amino acids to about 650 amino acids, about 175 amino acids to about 600 amino acids, about 175 amino acids to about 550 amino acids, about 175 amino acids to about 500 amino acids, about 175 amino acids to about 450 amino acids, about 175 amino acids to about 400 amino acids, about 175 amino acids to about 350 amino acids, about 175 amino acids to about 300 amino acids, about 175 amino acids to about 280 amino acids, about 175 amino acids to about 260 amino acids, about 175 amino acids to about 240 amino acids, about 175 amino acids to about 220 amino acids, about 175 amino acids to about 200 amino acids, about 175 amino acids to about 195 amino acids, about 175 amino acids to about 190 amino acids, about 175 amino acids to about 185 amino acids, about 175 amino acids to about 180 amino acids, about 180 amino acids to about 1000 amino acids, about 180 amino acids to about 950 amino acids, about 180 amino acids to about 900 aminoAttorney Docket No. 47039-0043W01

[0236] acids, about 180 amino acids to about 850 amino acids, about 180 amino acids to about 800 amino acids, about 180 amino acids to about 750 amino acids, about 180 amino acids to about 700 amino acids, about 180 amino acids to about 650 amino acids, about 180 amino acids to about 600 amino acids, about 180 amino acids to about 550 amino acids, about 180 amino acids to about 500 amino acids, about 180 amino acids to about 450 amino acids, about 180 amino acids to about 400 amino acids, about 180 amino acids to about 350 amino acids, about 180 amino acids to about 300 amino acids, about 180 amino acids to about 280 amino acids, about 180 amino acids to about 260 amino acids, about 180 amino acids to about 240 amino acids, about 180 amino acids to about 220 amino acids, about 180 amino acids to about 200 amino acids, about 180 amino acids to about 195 amino acids, about 180 amino acids to about 190 amino acids, about 180 amino acids to about 185 amino acids, about 185 amino acids to about 1000 amino acids, about 185 amino acids to about 950 amino acids, about 185 amino acids to about 900 amino acids, about 185 amino acids to about 850 amino acids, about 185 amino acids to about 800 amino acids, about 185 amino acids to about 750 amino acids, about 185 amino acids to about 700 amino acids, about 185 amino acids to about 650 amino acids, about 185 amino acids to about 600 amino acids, about 185 amino acids to about 550 amino acids, about 185 amino acids to about 500 amino acids, about 185 amino acids to about 450 amino acids, about 185 amino acids to about 400 amino acids, about 185 amino acids to about 350 amino acids, about 185 amino acids to about 300 amino acids, about 185 amino acids to about 280 amino acids, about 185 amino acids to about 260 amino acids, about 185 amino acids to about 240 amino acids, about 185 amino acids to about 220 amino acids, about 185 amino acids to about 200 amino acids, about 185 amino acids to about 195 amino acids, about 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, about 190 amino acids to about 400 amino acids, about 190 amino acids to about 350 amino acids, about 190 amino acids to about 300 amino acids, about 190 amino acids to about 280 amino acids, about 190 amino acidsAttorney Docket No. 47039-0043W01

[0237] to about 260 amino acids, about 190 amino acids to about 240 amino acids, about 190 amino acids to about 220 amino acids, about 190 amino acids to about 200 amino acids, about 190 amino acids to about 195 amino acids, about 195 amino acids to about 1000 amino acids, about 195 amino acids to about 950 amino acids, about 195 amino acids to about 900 amino acids, about 195 amino acids to about 850 amino acids, about 195 amino acids to about 800 amino acids, about 195 amino acids to about 750 amino acids, about 195 amino acids to about 700 amino acids, about 195 amino acids to about 650 amino acids, about 195 amino acids to about 600 amino acids, about 195 amino acids to about 550 amino acids, about 195 amino acids to about 500 amino acids, about 195 amino acids to about 450 amino acids, about 195 amino acids to about 400 amino acids, about 195 amino acids to about 350 amino acids, about 195 amino acids to about 300 amino acids, about 195 amino acids to about 280 amino acids, about 195 amino acids to about 260 amino acids, about 195 amino acids to about 240 amino acids, about 195 amino acids to about 220 amino acids, about 195 amino acids to about 200 amino acids, about 200 amino acids to about 1000 amino acids, about 200 amino acids to about 950 amino acids, about 200 amino acids to about 900 amino acids, about 200 amino acids to about 850 amino acids, about 200 amino acids to about 800 amino acids, about 200 amino acids to about 750 amino acids, about 200 amino acids to about 700 amino acids, about 200 amino acids to about 650 amino acids, about 200 amino acids to about 600 amino acids, about 200 amino acids to about 550 amino acids, about 200 amino acids to about 500 amino acids, about 200 amino acids to about 450 amino acids, about 200 amino acids to about 400 amino acids, about 200 amino acids to about 350 amino acids, about 200 amino acids to about 300 amino acids, about 200 amino acids to about 280 amino acids, about 200 amino acids to about 260 amino acids, about 200 amino acids to about 240 amino acids, about 200 amino acids to about 220 amino acids, about 220 amino acids to about 1000 amino acids, about 220 amino acids to about 950 amino acids, about 220 amino acids to about 900 amino acids, about 220 amino acids to about 850 amino acids, about 220 amino acids to about 800 amino acids, about 220 amino acids to about 750 amino acids, about 220 amino acids to about 700 amino acids, about 220 amino acids to about 650 amino acids, about 220 amino acids to about 600 amino acids, about 220 amino acids to about 550 amino acids, about 220 amino acids to about 500 amino acids, about 220 amino acids to about 450 amino acids, about 220 amino acids to about 400 amino acids, about 220 amino acids to about 350 amino acids, about 220 amino acids to about 300 amino acids,Attorney Docket No. 47039-0043W01

[0238] about 220 amino acids to about 280 amino acids, about 220 amino acids to about 260 amino acids, about 220 amino acids to about 240 amino acids, about 240 amino acids to about 1000 amino acids, about 240 amino acids to about 950 amino acids, about 240 amino acids to about 900 amino acids, about 240 amino acids to about 850 amino acids, about 240 amino acids to about 800 amino acids, about 240 amino acids to about 750 amino acids, about 240 amino acids to about 700 amino acids, about 240 amino acids to about 650 amino acids, about 240 amino acids to about 600 amino acids, about 240 amino acids to about 550 amino acids, about 240 amino acids to about 500 amino acids, about 240 amino acids to about 450 amino acids, about 240 amino acids to about 400 amino acids, about 240 amino acids to about 350 amino acids, about 240 amino acids to about 300 amino acids, about 240 amino acids to about 280 amino acids, about 240 amino acids to about 260 amino acids, about 260 amino acids to about 1000 amino acids, about 260 amino acids to about 950 amino acids, about 260 amino acids to about 900 amino acids, about 260 amino acids to about 850 amino acids, about 260 amino acids to about 800 amino acids, about 260 amino acids to about 750 amino acids, about 260 amino acids to about 700 amino acids, about 260 amino acids to about 650 amino acids, about 260 amino acids to about 600 amino acids, about 260 amino acids to about 550 amino acids, about 260 amino acids to about 500 amino acids, about 260 amino acids to about 450 amino acids, about 260 amino acids to about 400 amino acids, about 260 amino acids to about 350 amino acids, about 260 amino acids to about 300 amino acids, about 260 amino acids to about 280 amino acids, about 280 amino acids to about 1000 amino acids, about 280 amino acids to about 950 amino acids, about 280 amino acids to about 900 amino acids, about 280 amino acids to about 850 amino acids, about 280 amino acids to about 800 amino acids, about 280 amino acids to about 750 amino acids, about 280 amino acids to about 700 amino acids, about 280 amino acids to about 650 amino acids, about 280 amino acids to about 600 amino acids, about 280 amino acids to about 550 amino acids, about 280 amino acids to about 500 amino acids, about 280 amino acids to about 450 amino acids, about 280 amino acids to about 400 amino acids, about 280 amino acids to about 350 amino acids, about 280 amino acids to about 300 amino acids, about 300 amino acids to about 1000 amino acids, about 300 amino acids to about 950 amino acids, about 300 amino acids to about 900 amino acids, about 300 amino acids to about 850 amino acids, about 300 amino acids to about 800 amino acids, about 300 amino acids to about 750 amino acids, about 300 amino acids to about 700 amino acids, about 300 amino acids toAttorney Docket No. 47039-0043W01

[0239] about 650 amino acids, about 300 amino acids to about 600 amino acids, about 300 amino acids to about 550 amino acids, about 300 amino acids to about 500 amino acids, about 300 amino acids to about 450 amino acids, about 300 amino acids to about 400 amino acids, about 300 amino acids to about 350 amino acids, about 350 amino acids to about 1000 amino acids, about 350 amino acids to about 950 amino acids, about 350 amino acids to about 900 amino acids, about 350 amino acids to about 850 amino acids, about 350 amino acids to about 800 amino acids, about 350 amino acids to about 750 amino acids, about 350 amino acids to about 700 amino acids, about 350 amino acids to about 650 amino acids, about 350 amino acids to about 600 amino acids, about 350 amino acids to about 550 amino acids, about 350 amino acids to about 500 amino acids, about 350 amino acids to about 450 amino acids, about 350 amino acids to about 400 amino acids, about 400 amino acids to about 1000 amino acids, about 400 amino acids to about 950 amino acids, about 400 amino acids to about 900 amino acids, about 400 amino acids to about 850 amino acids, about 400 amino acids to about 800 amino acids, about 400 amino acids to about 750 amino acids, about 400 amino acids to about 700 amino acids, about 400 amino acids to about 650 amino acids, about 400 amino acids to about 600 amino acids, about 400 amino acids to about 550 amino acids, about 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, about 500 amino acids to about 950 amino acids, about 500 amino acids to about 900 amino acids, about 500 amino acids to about 850 amino acids, about 500 amino acids to about 800 amino acids, about 500 amino acids to about 750 amino acids, about 500 amino acids to about 700 amino acids, about 500 amino acids to about 650 amino acids, about 500 amino acids to about 600 amino acids, about 500 amino acids to about 550 amino acids, about 550 amino acids to about 1000 amino acids, about 550 amino acids to about 950 amino acids, about 550 amino acids to about 900 amino acids, about 550 amino acids to about 850 amino acids, about 550 amino acids to about 800 amino acids, about 550 amino acids to about 750 amino acids, aboutAttorney Docket No. 47039-0043W01

[0240] 550 amino acids to about 700 amino acids, about 550 amino acids to about 650 amino acids, about 550 amino acids to about 600 amino acids, about 600 amino acids to about 1000 amino acids, about 600 amino acids to about 950 amino acids, about 600 amino acids to about 900 amino acids, about 600 amino acids to about 850 amino acids, about 600 amino acids to about 800 amino acids, about 600 amino acids to about 750 amino acids, about 600 amino acids to about 700 amino acids, about 600 amino acids to about 650 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 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 950 amino acids, about 750 amino acids to about 900 amino acids, about 750 amino acids to about 850 amino acids, about 750 amino acids to about 800 amino acids, about 800 amino acids to about 1000 amino acids, about 800 amino acids to about 950 amino acids, about 800 amino acids to about 900 amino acids, about 800 amino acids to about 850 amino acids, about 850 amino acids to about 1000 amino acids, about 850 amino acids to about 950 amino acids, about 850 amino acids to about 900 amino acids, about 900 amino acids to about 1000 amino acids, about 900 amino acids to about 950 amino acids, or about 950 amino acids to about 1000 amino acids.

[0241] Any of the target-binding domains described herein can bind to its target with a dissociation equilibrium constant (KD) of less than 1 x 10'7M, less than 1 x 10'8M, less than 1 x 10'9M, less than 1 x 10'10M, less than 1 x 10'11M, less than 1 x 10'12M, or less than 1 x 10'13M. In some embodiments, the antigen-binding protein construct provided herein can bind to an identifying antigen with a KD of about 1 x 10'3M to about 1 x 10'5M, about 1 x 10'4M to about 1 x 10'6M, about 1 x 10'5M to about 1 x 10’7M, about 1 x 10’6M to about 1 x 10'8M, about 1 x IO’7M to about 1 x 10‘9M, about 1 x 10’8M to about 1 x 10'10M, or about 1 x 10'9M to about 1 x 10'11M (inclusive).

[0242] Any of the target-binding domains described herein can bind to its target with a KD of between about 1 pM to about 30 nM (e.g., about 1 pM to about 25 nM, about 1 pM to about 20Attorney Docket No. 47039-0043W01

[0243] nM, about 1 pM to about 15 nM, about 1 pM to about 10 nM, about 1 pM to about 5 nM, about 1 pM to about 2 nM, about 1 pM to about 1 nM, about 1 pM to about 950 pM, about 1 pM to about 900 pM, about 1 pM to about 850 pM, about 1 pM to about 800 pM, about 1 pM to about 750 pM, about 1 pM to about 700 pM, about 1 pM to about 650 pM, about 1 pM to about 600 pM, about 1 pM to about 550 pM, about 1 pM to about 500 pM, about 1 pM to about 450 pM, about 1 pM to about 400 pM, about 1 pM to about 350 pM, about 1 pM to about 300 pM, about 1 pM to about 250 pM, about 1 pM to about 200 pM, about 1 pM to about 150 pM, about 1 pM to about 100 pM, about 1 pM to about 90 pM, about 1 pM to about 80 pM, about 1 pM to about 70 pM, about 1 pM to about 60 pM, about 1 pM to about 50 pM, about 1 pM to about 40 pM, about 1 pM to about 30 pM, about 1 pM to about 20 pM, about 1 pM to about 10 pM, about 1 pM to about 5 pM, about 1 pM to about 4 pM, about 1 pM to about 3 pM, about 1 pM to about 2 pM, about 2 pM to about 30 nM, about 2 pM to about 25 nM, about 2 pM to about 20 nM, about 2 pM to about 15 nM, about 2 pM to about 10 nM, about 2 pM to about 5 nM, about 2 pM to about 2 nM, about 2 pM to about 1 nM, about 2 pM to about 950 pM, about 2 pM to about 900 pM, about 2 pM to about 850 pM, about 2 pM to about 800 pM, about 2 pM to about 750 pM, about 2 pM to about 700 pM, about 2 pM to about 650 pM, about 2 pM to about 600 pM, about 2 pM to about 550 pM, about 2 pM to about 500 pM, about 2 pM to about 450 pM, about 2 pM to about 400 pM, about 2 pM to about 350 pM, about 2 pM to about 300 pM, about 2 pM to about 250 pM, about 2 pM to about 200 pM, about 2 pM to about 150 pM, about 2 pM to about 100 pM, about 2 pM to about 90 pM, about 2 pM to about 80 pM, about 2 pM to about 70 pM, about 2 pM to about 60 pM, about 2 pM to about 50 pM, about 2 pM to about 40 pM, about 2 pM to about 30 pM, about 2 pM to about 20 pM, about 2 pM to about 10 pM, about 2 pM to about 5 pM, about 2 pM to about 4 pM, about 2 pM to about 3 pM, about 5 pM to about 30 nM, about 5 pM to about 25 nM, about 5 pM to about 20 nM, about 5 pM to about 15 nM, about 5 pM to about 10 nM, about 5 pM to about 5 nM, about 5 pM to about 2 nM, about 5 pM to about 1 nM, about 5 pM to about 950 pM, about 5 pM to about 900 pM, about 5 pM to about 850 pM, about 5 pM to about 800 pM, about 5 pM to about 750 pM, about 5 pM to about 700 pM, about 5 pM to about 650 pM, about 5 pM to about 600 pM, about 5 pM to about 550 pM, about 5 pM to about 500 pM, about 5 pM to about 450 pM, about 5 pM to about 400 pM, about 5 pM to about 350 pM, about 5 pM to about 300 pM, about 5 pM to about 250 pM, about 5 pM to about 200 pM, about 5 pM to about 150 pM, about 5 pM to about 100 pM, about 5 pM to about 90 pM, about 5Attorney Docket No. 47039-0043W01

[0244] pM to about 80 pM, about 5 pM to about 70 pM, about 5 pM to about 60 pM, about 5 pM to about 50 pM, about 5 pM to about 40 pM, about 5 pM to about 30 pM, about 5 pM to about 20 pM, about 5 pM to about 10 pM, about 10 pM to about 30 nM, about 10 pM to about 25 nM, about 10 pM to about 20 nM, about 10 pM to about 15 nM, about 10 pM to about 10 nM, about 10 pM to about 5 nM, about 10 pM to about 2 nM, about 10 pM to about 1 nM, about 10 pM to about 950 pM, about 10 pM to about 900 pM, about 10 pM to about 850 pM, about 10 pM to about 800 pM, about 10 pM to about 750 pM, about 10 pM to about 700 pM, about 10 pM to about 650 pM, about 10 pM to about 600 pM, about 10 pM to about 550 pM, about 10 pM to about 500 pM, about 10 pM to about 450 pM, about 10 pM to about 400 pM, about 10 pM to about 350 pM, about 10 pM to about 300 pM, about 10 pM to about 250 pM, about 10 pM to about 200 pM, about 10 pM to about 150 pM, about 10 pM to about 100 pM, about 10 pM to about 90 pM, about 10 pM to about 80 pM, about 10 pM to about 70 pM, about 10 pM to about 60 pM, about 10 pM to about 50 pM, about 10 pM to about 40 pM, about 10 pM to about 30 pM, about 10 pM to about 20 pM, about 15 pM to about 30 nM, about 15 pM to about 25 nM, about 15 pM to about 20 nM, about 15 pM to about 15 nM, about 15 pM to about 10 nM, about 15 pM to about 5 nM, about 15 pM to about 2 nM, about 15 pM to about 1 nM, about 15 pM to about 950 pM, about 15 pM to about 900 pM, about 15 pM to about 850 pM, about 15 pM to about 800 pM, about 15 pM to about 750 pM, about 15 pM to about 700 pM, about 15 pM to about 650 pM, about 15 pM to about 600 pM, about 15 pM to about 550 pM, about 15 pM to about 500 pM, about 15 pM to about 450 pM, about 15 pM to about 400 pM, about 15 pM to about 350 pM, about 15 pM to about 300 pM, about 15 pM to about 250 pM, about 15 pM to about 200 pM, about 15 pM to about 150 pM, about 15 pM to about 100 pM, about 15 pM to about 90 pM, about 15 pM to about 80 pM, about 15 pM to about 70 pM, about 15 pM to about 60 pM, about 15 pM to about 50 pM, about 15 pM to about 40 pM, about 15 pM to about 30 pM, about 15 pM to about 20 pM, about 20 pM to about 30 nM, about 20 pM to about 25 nM, about 20 pM to about 20 nM, about 20 pM to about 15 nM, about 20 pM to about 10 nM, about 20 pM to about 5 nM, about 20 pM to about 2 nM, about 20 pM to about 1 nM, about 20 pM to about 950 pM, about 20 pM to about 900 pM, about 20 pM to about 850 pM, about 20 pM to about 800 pM, about 20 pM to about 750 pM, about 20 pM to about 700 pM, about 20 pM to about 650 pM, about 20 pM to about 600 pM, about 20 pM to about 550 pM, about 20 pM to about 500 pM, about 20 pM to about 450 pM, about 20 pM to about 400 pM, about 20 pM to about 350 pM,Attorney Docket No. 47039-0043W01

[0245] about 20 pM to about 300 pM, about 20 pM to about 250 pM, about 20 pM to about 20 pM, about 200 pM to about 150 pM, about 20 pM to about 100 pM, about 20 pM to about 90 pM, about 20 pM to about 80 pM, about 20 pM to about 70 pM, about 20 pM to about 60 pM, about 20 pM to about 50 pM, about 20 pM to about 40 pM, about 20 pM to about 30 pM, about 30 pM to about 30 nM, about 30 pM to about 25 nM, about 30 pM to about 30 nM, about 30 pM to about 15 nM, about 30 pM to about 10 nM, about 30 pM to about 5 nM, about 30 pM to about 2 nM, about 30 pM to about 1 nM, about 30 pM to about 950 pM, about 30 pM to about 900 pM, about 30 pM to about 850 pM, about 30 pM to about 800 pM, about 30 pM to about 750 pM, about 30 pM to about 700 pM, about 30 pM to about 650 pM, about 30 pM to about 600 pM, about 30 pM to about 550 pM, about 30 pM to about 500 pM, about 30 pM to about 450 pM, about 30 pM to about 400 pM, about 30 pM to about 350 pM, about 30 pM to about 300 pM, about 30 pM to about 250 pM, about 30 pM to about 200 pM, about 30 pM to about 150 pM, about 30 pM to about 100 pM, about 30 pM to about 90 pM, about 30 pM to about 80 pM, about 30 pM to about 70 pM, about 30 pM to about 60 pM, about 30 pM to about 50 pM, about 30 pM to about 40 pM, about 40 pM to about 30 nM, about 40 pM to about 25 nM, about 40 pM to about 30 nM, about 40 pM to about 15 nM, about 40 pM to about 10 nM, about 40 pM to about 5 nM, about 40 pM to about 2 nM, about 40 pM to about 1 nM, about 40 pM to about 950 pM, about 40 pM to about 900 pM, about 40 pM to about 850 pM, about 40 pM to about 800 pM, about 40 pM to about 750 pM, about 40 pM to about 700 pM, about 40 pM to about 650 pM, about 40 pM to about 600 pM, about 40 pM to about 550 pM, about 40 pM to about 500 pM, about 40 pM to about 450 pM, about 40 pM to about 400 pM, about 40 pM to about 350 pM, about 40 pM to about 300 pM, about 40 pM to about 250 pM, about 40 pM to about 200 pM, about 40 pM to about 150 pM, about 40 pM to about 100 pM, about 40 pM to about 90 pM, about 40 pM to about 80 pM, about 40 pM to about 70 pM, about 40 pM to about 60 pM, about 40 pM to about 50 pM, about 50 pM to about 30 nM, about 50 pM to about 25 nM, about 50 pM to about 30 nM, about 50 pM to about 15 nM, about 50 pM to about 10 nM, about 50 pM to about 5 nM, about 50 pM to about 2 nM, about 50 pM to about 1 nM, about 50 pM to about 950 pM, about 50 pM to about 900 pM, about 50 pM to about 850 pM, about 50 pM to about 800 pM, about 50 pM to about 750 pM, about 50 pM to about 700 pM, about 50 pM to about 650 pM, about 50 pM to about 600 pM, about 50 pM to about 550 pM, about 50 pM to about 500 pM, about 50 pM to about 450 pM, about 50 pM to about 400 pM, about 50 pM to about 350Attorney Docket No. 47039-0043W01

[0246] pM, about 50 pM to about 300 pM, about 50 pM to about 250 pM, about 50 pM to about 200 pM, about 50 pM to about 150 pM, about 50 pM to about 100 pM, about 50 pM to about 90 pM, about 50 pM to about 80 pM, about 50 pM to about 70 pM, about 50 pM to about 60 pM, about 60 pM to about 30 nM, about 60 pM to about 25 nM, about 60 pM to about 30 nM, about 60 pM to about 15 nM, about 60 pM to about 10 nM, about 60 pM to about 5 nM, about 60 pM to about 2 nM, about 60 pM to about 1 nM, 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 pM, about 60 pM to about 250 pM, about 60 pM to about 200 pM, about 60 pM to about 150 pM, about 60 pM to about 100 pM, about 60 pM to about 90 pM, about 60 pM to about 80 pM, about 60 pM to about 70 pM, about 70 pM to about 30 nM, about 70 pM to about 25 nM, about 70 pM to about 30 nM, about 70 pM to about 15 nM, about 70 pM to about 10 nM, about 70 pM to about 5 nM, about 70 pM to about 2 nM, about 70 pM to about 1 nM, about 70 pM to about 950 pM, about 70 pM to about 900 pM, about 70 pM to about 850 pM, about 70 pM to about 800 pM, about 70 pM to about 750 pM, about 70 pM to about 700 pM, about 70 pM to about 650 pM, about 70 pM to about 600 pM, about 70 pM to about 550 pM, about 70 pM to about 500 pM, about 70 pM to about 450 pM, about 70 pM to about 400 pM, about 70 pM to about 350 pM, about 70 pM to about 300 pM, about 70 pM to about 250 pM, about 70 pM to about 200 pM, about 70 pM to about 150 pM, about 70 pM to about 100 pM, about 70 pM to about 90 pM, about 70 pM to about 80 pM, about 80 pM to about 30 nM, about 80 pM to about 25 nM, about 80 pM to about 30 nM, about 80 pM to about 15 nM, about 80 pM to about 10 nM, about 80 pM to about 5 nM, about 80 pM to about 2 nM, about 80 pM to about 1 nM, about 80 pM to about 950 pM, about 80 pM to about 900 pM, about 80 pM to about 850 pM, about 80 pM to about 800 pM, about 80 pM to about 750 pM, about 80 pM to about 700 pM, about 80 pM to about 650 pM, about 80 pM to about 600 pM, about 80 pM to about 550 pM, about 80 pM to about 500 pM, about 80 pM to about 450 pM, about 80 pM to about 400 pM, about 80 pM to about 350 pM, about 80 pM to about 300 pM, about 80 pM to about 250 pM, about 80 pM to about 200 pM, about 80 pM to about 150 pM, about 80 pM to about 100 pM, about 80 pM to about 90 pM, about 90 pM to about 30 nM, about 90 pM to about 25 nM, about 90 pM to about 30 nM, aboutAttorney Docket No. 47039-0043W01

[0247] 90 pM to about 15 nM, about 90 pM to about 10 nM, about 90 pM to about 5 nM, about 90 pM to about 2 nM, about 90 pM to about 1 nM, about 90 pM to about 950 pM, about 90 pM to about 900 pM, about 90 pM to about 850 pM, about 90 pM to about 800 pM, about 90 pM to about 750 pM, about 90 pM to about 700 pM, about 90 pM to about 650 pM, about 90 pM to about 600 pM, about 90 pM to about 550 pM, about 90 pM to about 500 pM, about 90 pM to about 450 pM, about 90 pM to about 400 pM, about 90 pM to about 350 pM, about 90 pM to about 300 pM, about 90 pM to about 250 pM, about 90 pM to about 200 pM, about 90 pM to about 150 pM, about 90 pM to about 100 pM, about 100 pM to about 30 nM, about 100 pM to about 25 nM, about 100 pM to about 30 nM, about 100 pM to about 15 nM, about 100 pM to about 10 nM, about 100 pM to about 5 nM, about 100 pM to about 2 nM, about 100 pM to about 1 nM, about 100 pM to about 950 pM, about 100 pM to about 900 pM, about 100 pM to about 850 pM, about 100 pM to about 800 pM, about 100 pM to about 750 pM, about 100 pM to about 700 pM, about 100 pM to about 650 pM, about 100 pM to about 600 pM, about 100 pM to about 550 pM, about 100 pM to about 500 pM, about 100 pM to about 450 pM, about 100 pM to about 400 pM, about 100 pM to about 350 pM, about 100 pM to about 300 pM, about 100 pM to about 250 pM, about 100 pM to about 200 pM, about 100 pM to about 150 pM, about 150 pM to about 30 nM, about 150 pM to about 25 nM, about 150 pM to about 30 nM, about 150 pM to about 15 nM, about 150 pM to about 10 nM, about 150 pM to about 5 nM, about 150 pM to about 2 nM, about 150 pM to about 1 nM, about 150 pM to about 950 pM, about 150 pM to about 900 pM, about 150 pM to about 850 pM, about 150 pM to about 800 pM, about 150 pM to about 750 pM, about 150 pM to about 700 pM, about 150 pM to about 650 pM, about 150 pM to about 600 pM, about 150 pM to about 550 pM, about 150 pM to about 500 pM, about 150 pM to about 450 pM, about 150 pM to about 400 pM, about 150 pM to about 350 pM, about 150 pM to about 300 pM, about 150 pM to about 250 pM, about 150 pM to about 200 pM, about 200 pM to about 30 nM, about 200 pM to about 25 nM, about 200 pM to about 30 nM, about 200 pM to about 15 nM, about 200 pM to about 10 nM, about 200 pM to about 5 nM, about 200 pM to about 2 nM, about 200 pM to about 1 nM, about 200 pM to about 950 pM, about 200 pM to about 900 pM, about 200 pM to about 850 pM, about 200 pM to about 800 pM, about 200 pM to about 750 pM, about 200 pM to about 700 pM, about 200 pM to about 650 pM, about 200 pM to about 600 pM, about 200 pM to about 550 pM, about 200 pM to about 500 pM, about 200 pM to about 450 pM, about 200 pM to about 400 pM, about 200 pM to about 350 pM, about 200 pM to about 300 pM, about 200 pM toAttorney Docket No. 47039-0043W01

[0248] about 250 pM, about 300 pM to about 30 nM, about 300 pM to about 25 nM, about 300 pM to about 30 nM, about 300 pM to about 15 nM, about 300 pM to about 10 nM, about 300 pM to about 5 nM, about 300 pM to about 2 nM, about 300 pM to about 1 nM, about 300 pM to about 950 pM, about 300 pM to about 900 pM, about 300 pM to about 850 pM, about 300 pM to about 800 pM, about 300 pM to about 750 pM, about 300 pM to about 700 pM, about 300 pM to about 650 pM, about 300 pM to about 600 pM, about 300 pM to about 550 pM, about 300 pM to about 500 pM, about 300 pM to about 450 pM, about 300 pM to about 400 pM, about 300 pM to about 350 pM, about 400 pM to about 30 nM, about 400 pM to about 25 nM, about 400 pM to about 30 nM, about 400 pM to about 15 nM, about 400 pM to about 10 nM, about 400 pM to about 5 nM, about 400 pM to about 2 nM, about 400 pM to about 1 nM, about 400 pM to about 950 pM, about 400 pM to about 900 pM, about 400 pM to about 850 pM, about 400 pM to about 800 pM, about 400 pM to about 750 pM, about 400 pM to about 700 pM, about 400 pM to about 650 pM, about 400 pM to about 600 pM, about 400 pM to about 550 pM, about 400 pM to about 500 pM, about 500 pM to about 30 nM, about 500 pM to about 25 nM, about 500 pM to about 30 nM, about 500 pM to about 15 nM, about 500 pM to about 10 nM, about 500 pM to about 5 nM, about 500 pM to about 2 nM, about 500 pM to about 1 nM, about 500 pM to about 950 pM, about 500 pM to about 900 pM, about 500 pM to about 850 pM, about 500 pM to about 800 pM, about 500 pM to about 750 pM, about 500 pM to about 700 pM, about 500 pM to about 650 pM, about 500 pM to about 600 pM, about 500 pM to about 550 pM, about 600 pM to about 30 nM, about 600 pM to about 25 nM, about 600 pM to about 30 nM, about 600 pM to about 15 nM, about 600 pM to about 10 nM, about 600 pM to about 5 nM, about 600 pM to about 2 nM, about 600 pM to about 1 nM, about 600 pM to about 950 pM, about 600 pM to about 900 pM, about 600 pM to about 850 pM, about 600 pM to about 800 pM, about 600 pM to about 750 pM, about 600 pM to about 700 pM, about 600 pM to about 650 pM, about 700 pM to about 30 nM, about 700 pM to about 25 nM, about 700 pM to about 30 nM, about 700 pM to about 15 nM, about 700 pM to about 10 nM, about 700 pM to about 5 nM, about 700 pM to about 2 nM, about 700 pM to about 1 nM, about 700 pM to about 950 pM, about 700 pM to about 900 pM, about 700 pM to about 850 pM, about 700 pM to about 800 pM, about 700 pM to about 750 pM, about 800 pM to about 30 nM, about 800 pM to about 25 nM, about 800 pM to about 30 nM, about 800 pM to about 15 nM, about 800 pM to about 10 nM, about 800 pM to about 5 nM, about 800 pM to about 2 nM, about 800 pM to about 1 nM, about 800 pM to about 950 pM, about 800 pM toAttorney Docket No. 47039-0043W01

[0249] about 900 pM, about 800 pM to about 850 pM, about 900 pM to about 30 nM, about 900 pM to about 25 nM, about 900 pM to about 30 nM, about 900 pM to about 15 nM, about 900 pM to about 10 nM, about 900 pM to about 5 nM, about 900 pM to about 2 nM, about 900 pM to about 1 nM, about 900 pM to about 950 pM, about 1 nM to about 30 nM, about 1 nM to about 25 nM, about 1 nM to about 20 nM, about 1 nM to about 15 nM, about 1 nM to about 10 nM, about 1 nM to about 5 nM, about 2 nM to about 30 nM, about 2 nM to about 25 nM, about 2 nM to about 20 nM, about 2 nM to about 15 nM, about 2 nM to about 10 nM, about 2 nM to about 5 nM, about 4 nM to about 30 nM, about 4 nM to about 25 nM, about 4 nM to about 20 nM, about 4 nM to about 15 nM, about 4 nM to about 10 nM, about 4 nM to about 5 nM, about 5 nM to about 30 nM, about 5 nM to about 25 nM, about 5 nM to about 20 nM, about 5 nM to about 15 nM, about 5 nM to about 10 nM, about 10 nM to about 30 nM, about 10 nM to about 25 nM, about 10 nM to about 20 nM, about 10 nM to about 15 nM, about 15 nM to about 30 nM, about 15 nM to about 25 nM, about 15 nM to about 20 nM, about 20 nM to about 30 nM, and about 20 nM to about 25 nM).

[0250] Any of the target-binding domains described herein can bind to its target with a KD of between about 1 nM to about 10 nM (e.g., about 1 nM to about 9 nM, about 1 nM to about 8 nM, about 1 nM to about 7 nM, about 1 nM to about 6 nM, about 1 nM to about 5 nM, about 1 nM to about 4 nM, about 1 nM to about 3 nM, about 1 nM to about 2 nM, about 2 nM to about 10 nM, about 2 nM to about 9 nM, about 2 nM to about 8 nM, about 2 nM to about 7 nM, about 2 nM to about 6 nM, about 2 nM to about 5 nM, about 2 nM to about 4 nM, about 2 nM to about 3 nM, about 3 nM to about 10 nM, about 3 nM to about 9 nM, about 3 nM to about 8 nM, about 3 nM to about 7 nM, about 3 nM to about 6 nM, about 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).

[0251] A variety of different methods known in the art can be used to determine the KD values of any of the antigen-binding protein constructs described herein (e.g., an electrophoretic mobilityAttorney Docket No. 47039-0043W01

[0252] shift assay, a filter binding assay, surface plasmon resonance, and a biomolecular binding kinetics assay, etc.).

[0253] Soluble Interleukin or Cytokine Protein

[0254] In some embodiments, the first target-binding domain or the second target-binding domain can be a soluble interleukin protein, soluble cytokine protein, or soluble cell surface protein. In some embodiments, the soluble interleukin protein or soluble cytokine protein, is selected from the group of: IL-2, IL-3, IL-4, IL-7, IL-8, IL-10, IL-12, IL-15, IL-17, IL-18, IL-21, PDGF-DD, SCF, FLT3L. Non-limiting examples of soluble IL-7 and IL-21 are provided below.

[0255] In some embodiments, a target-binding domain can comprise a sequence that is at least 80%, at least 85%, at least 90%, at least 92%, at least 94%, at least 95%, at least 96%, at least 98%, at least 99%, or 100% identical to:

[0256] Human Soluble IL-7 (SEQ ID NO: 4) DCDIEGKDGKQYESVLMVSIDQLLDSMKEIGSNCLNNEFNFFKRHICDANKEGMF LFRAARKLRQFLKMNSTGDFDLHLLKVSEGTTILLNCTGQVKGRKPAALGEAQPT KSLEENKSLKEQKKLNDLCFLKRLLQEIKTCWNKILMGTKEH

[0257] In some embodiments, a soluble IL-7 protein can include a sequence that is at least 70% identical (e.g., at least 75% identical, at least 80% identical, at least 85% identical, at least 90% identical, at least 92%, at least 94%, at least 95% identical, at least 96% identical, at least 98% identical, at least 99% identical, or 100% identical) to SEQ ID NO: 4.

[0258] In some embodiments, a soluble human IL-7 is encoded by a sequence that is at least 80% identical (e.g., 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: 90.

[0259] Nucleic Acid Encoding Soluble IL-7 (SEQ ID NO: 90)

[0260] GAT T G T GAT AT T GAAGG T AAGAT G G CAAAC AAT AT GAGAG TGTTCTAATGGTC AGCATCGATCAATTATTGGACAGCATGAAAGAAATTGGTAGCAATTGCCTGAAT AAT GAAT T TAAC T T T T T TAAAAGACATAT C T GTGAT GC TAATAAG GAAGG TAT G T T T T TAT T CCGT GC T GC T CGCAAGT T GAGGCAAT T T C T TAAAATGAATAGCAC T GG T GAT T T T GAT C T C CAC T TAT T AAAAGT T T CAGAAGGCACAACAAT AC T G T T G AACTGCACTGGCCAGGTTAAAGGAAGAAAACCAGCTGCCCTGGGTGAAGCCCAA CCAACAAAGAGT T T GGAAGAAAATAAAT C T T TAAAGGAACAGAAAAAAC TGAATAttorney Docket No. 47039-0043W01

[0261] GAC TTGTGTTTCC T AAGAG C TAT T AC AAGAGAT AAAAAC T T G T T G GAAT AAA AT T T TGAT GGGCAC TAAAGAACAC

[0262] Additional examples of soluble interleukin proteins and soluble cytokine proteins are known in the art.

[0263] Soluble Interleukin or Cytokine Receptor

[0264] In some embodiments, the first target-binding domain or the second target-binding domain is a soluble interleukin receptor, a soluble cytokine receptor, or a soluble cell surface receptor. In some embodiments, the soluble receptor is a soluble TGF-P receptor II (TGF-PRII) (see, e.g., those described in Yung et al., Am. J. Resp. Crit. Care Med. 194(9): 1140-1151, 2016).

[0265] Exemplary Soluble TGFpRII Receptor

[0266] In some embodiments, a target-binding domain includes or is a soluble TGFpRII receptor. In some embodiments, a soluble TGFpRII receptor can include a first sequence that is at least 70% identical (e.g., at least 75% identical, at least 80% identical, at least 85% identical, at least 90% identical, at least 92% identical, at least 94% identical, at least 95% identical, at least 96% identical, at least 98% identical, at least 99%, or 100% identical) to:

[0267] IPPHVQKSVNNDMIVTDNNGAVKFPQLCKFCDVRFSTCDNQKSCMSNCSITSICE KPQEVCVAVWRKNDENITLETVCHDPKLPYHDFILEDAASPKCIMKEKKKPGETF FMCSCSSDECNDNI I FSEEYNTSNPD (SEQ ID NO: 5),

[0268] and a second sequence that is at least 70% identical (e.g., at least 75% identical, at least 80% identical, at least 85% identical, at least 90% identical, at least 92% identical, at least 95% identical, at least 96% identical, at least 98% identical, at least 99%, or 100% identical) to:

[0269] IPPHVQKSVNNDMIVTDNNGAVKFPQLCKFCDVRFSTCDNQKSCMSNCSITSIC EKPQEVCVAVWRKNDENITLETVCHDPKLPYHDFILEDAASPKCIMKEKKKPGE TFFMCSCSSDECNDNI I FSEEYNTSNPD (SEQ ID NO: 5).

[0270] In some embodiments, the soluble TGFpRII receptor can further include a linker sequence (e.g., any of the exemplary linker sequences described herein or known in the art) between the first sequence and the second sequence.

[0271] In some embodiments, a soluble TGFpRII receptor is encoded by a first nucleic acid encoding a first sequence at least 70% identical (e.g., at least 75% identical, at least 80%Attorney Docket No. 47039-0043W01

[0272] identical, at least 85% identical, at least 90% identical, at least 92% identical, at least 94% identical, at least 95% identical, at least 96% identical, at least 98% identical, at least 99%, or 100% identical) to:

[0273] ATCCCGCCACACGTGCAAAAGAGCGTGAACAACGACATGATCGTTACGGACAAC AATGGGGCCGTGAAGTTCCCTCAGTTATGCAAATTCTGTGACGTGAGATTCAGC ACGTGCGACAATCAGAAGAGCTGCATGTCCAATTGCTCAATCACAAGCATCTGC GAGAAACCCCAAGAAGTTTGCGTGGCCGTGTGGAGAAAGAATGACGAGAACATC ACCCTGGAAACCGTGTGCCATGATCCTAAACTCCCTTACCATGACTTTATCTTG GAGGATGCTGCTAGCCCAAAGTGCATCATGAAAGAGAAGAAGAAACCGGGGGAA ACTTTTTTCATGTGTTCATGCAGCTCTGACGAGTGTAATGATAACATTATCTTC AGCGAAGAGTATAACACTAGCAATCCTGAT (SEQ ID NO: 6), or

[0274] AT C C C C C C C CAT G T G C AAAAGAG C G T GAAC AAC GAT AT GAT C G T GAG C GACAAC AACGGCGCCGTGAAGTTTCCCCAGCTCTGCAAGTTCTGCGATGTCAGGTTCAGC ACCTGCGATAATCAGAAGTCCTGCATGTCCAACTGCACGATCACCTCCATCTGC GAGAAGCCCCAAGAAGTGTGCGTGGCCGTGTGGCGGAAAAATGACGAGAACATC ACCCTGGAGACCGTGTGTCACGACCCCAAGCTCCCTTATCACGACTTCATTCTG GAGGACGCTGCCTCCCCCAAATGCATCATGAAGGAGAAGAAGAAGCCCGGAGAG ACCTTCTTTATGTGTTCCTGTAGCAGCGACGAGTGTAACGACAACATCATCTTC AGCGAAGAGTACAACACCAGCAACCCTGAT (SEQ ID NO: 7),

[0275] and a second nucleic acid sequence encoding a second sequence that is at least 70% identical (e g., at least 75% identical, at least 80% identical, at least 85% identical, at least 90% identical, at least 92% identical, at least 94% identical, at least 95% identical, at least 96% identical, at least 98% identical, at least 99%, or 100% identical) to:

[0276] ATCCCCCCC CAT G T G C AAAAAT C AG T GAAC AAT GAC T GAT C G T GAC C GAT AAT AACGGGGCGGTAAAGTTTCCTCAACTGTGTAAATTTTGCGACGTCAGATTCAGC ACATGCGACAATCAAAAGTCCTGCATGAGCAACTGCAGCATCACCTCTATCTGT GAGAAACCACAAGAAGTATGTGTGGCGGTATGGCGGAAGAACGATGAAAATATC ACGCTGGAGACCGTTTGCCATGATCCTAAGCTGCCATATCACGATTTCATCCTG GAGGACGCCGCTAGCCCTAAGTGTATCATGAAGGAGAAGAAGAAGCCCGGTGAG ACCTTTTTCATGTGCTCATGCAGCTCCGACGAGTGCAACGACAACATCATCTTC AGCGAGGAGTACAATACAAGCAATCCTGAC (SEQ ID NO: 8), or

[0277] AT C C C C C C C CAT G T G C AAAAGAG C G T GAAC AAC GAT AT GAT C G T GAC C GACAAC AACGGCGCCGTGAAGTTTCCCCAGCTCTGCAAGTTCTGCGATGTCAGGTTCAGC ACCTGCGATAATCAGAAGTCCTGCATGTCCAACTGCACGATCACCTCCATCTGC GAGAAGCCCCAAGAAGTGTGCGTGGCCGTGTGGCGGAAAAATGACGAGAACATC ACCCTGGAGACCGTGTGTCACGACCCCAAGCTCCCTTATCACGACTTCATTCTG GAGGACGCTGCCTCCCCCAAATGCATCATGAAGGAGAAGAAGAAGCCCGGAGAG ACCTTCTTTATGTGTTCCTGTAGCAGCGACGAGTGTAACGACAACATCATCTTC AGCGAAGAGTACAACACCAGCAACCCTGAT (SEQ ID NO: 7).Attorney Docket No. 47039-0043W01

[0278] In some embodiments, the nucleic acid encoding a soluble TGFpRII receptor further includes a nucleic acid sequence encoding a linker sequence (e.g., any of the exemplary linker sequences described herein or known in the art) between the first nucleic acid and the second nucleic acid.

[0279] In some embodiments, a soluble TGFpRII receptor includes a sequence that is at least 70% identical (e.g., at least 75% identical, at least 80% identical, at least 85% identical, at least 90% identical, at least 92% identical, at least 94% identical, at least 95% identical, at least 96% identical, at least 98% identical, at least 99%, or 100% identical) to:

[0280] IPPHVQKSVNNDMIVTDNNGAVKFPQLCKFCDVRFSTCDNQKSCMSNCSITSIC EKPQEVCVAVWRKNDENITLETVCHDPKLPYHDFILEDAASPKCIMKEKKKPGE TFFMCSCSSDECNDNIIFSEEYNTSNPDGGGGSIPPHVQKSVNNDMIVTDNNGA VKFPQLCKFCDVRFSTCDNQKSCMSNCSITSICEKPQEVCVAVWRKNDENITLE TVCHDPKLPYHDFILEDAASPKCIMKEKKKPGETFFMCSCSSDECNDNI IFSEE YNTSNPD (SEQ ID NO: 9), or IPPHVQKSVNNDMIVTDNNGAVKFPQLCKFCDVRFSTCDNQKSCMSNCSITSIC EKPQEVCVAVWRKNDENITLETVCHDPKLPYHDFILEDAASPKCIMKEKKKPGE TFFMCSCSSDECNDNI I FSEEYNTSNPDGGGGSGGGGSGGGGS IPPHVQKSVNN DMIVTDNNGAVKFPQLCKFCDVRFSTCDNQKSCMSNCSITSICEKPQEVCVAVW RKNDENITLETVCHDPKLPYHDFILEDAASPKCIMKEKKKPGETFFMCSCSSDE CNDNI I FSEEYNTSNPD (SEQ ID NO: 10).

[0281] In some embodiments, a soluble TGFpRII receptor is encoded by a nucleic acid including a sequence at least 70% identical (e.g., at least 75% identical, at least 80% identical, at least 85% identical, at least 90% identical, at least 92% identical, at least 94% identical, at least 95% identical, at least 96% identical, at least 98% identical, at least 99%, or 100% identical) to:

[0282] ATCCCGCCACACGTGCAAAAGAGCGTGAACAACGACATGATCGTTACGGACAAC AATGGGGCCGTGAAGTTCCCTCAGTTATGCAAATTCTGTGACGTGAGATTCAGC ACGTGCGACAATCAGAAGAGCTGCATGTCCAATTGCTCAATCACAAGCATCTGC GAGAAACCCCAAGAAGTTTGCGTGGCCGTGTGGAGAAAGAATGACGAGAACATC ACCCTGGAAACCGTGTGCCATGATCCTAAACTCCCTTACCATGACTTTATCTTG GAGGATGCTGCTAGCCCAAAGTGCATCATGAAAGAGAAGAAGAAACCGGGGGAA ACTTTTTTCATGTGTTCATGCAGCTCTGACGAGTGTAATGATAACATTATCTTC AGCGAAGAGTATAACACTAGCAATCCTGATGGAGGTGGCGGATCTATCCCCCCC CATGTGCAAAAATCAGTGAACAATGACATGATCGTGACCGATAATAACGGGGCG GTAAAGTTTCCTCAACTGTGTAAATTTTGCGACGTCAGATTCAGCACATGCGAC AAT C AAAAG T C C T G C AT GAG C AAC T G GAG C AT C AC C T C T AT C T G T GAGAAAC C A CAAGAAGTATGTGTGGCGGTATGGCGGAAGAACGATGAAAATATCACGCTGGAG ACCGTTTGCCATGATCCTAAGCTGCCATATCACGATTTCATCCTGGAGGACGCC GCTAGCCCTAAGTGTATCATGAAGGAGAAGAAGAAGCCCGGTGAGACCTTTTTC ATGTGCTCATGCAGCTCCGACGAGTGCAACGACAACATCATCTTCAGCGAGGAG TACAATACAAGCAATCCTGAC (SEQ ID NO: 11), orAttorney Docket No. 47039-0043W01

[0283] AT C C C C C C C CAT G T G C AAAAGAG C G T GAAC AC GAT AT GAT C G T GAC C GACAAC AACGGCGCCGTGAAGTTTCCCCAGCTCTGCAAGTTCTGCGATGTCAGGTTCAGC ACCTGCGATAATCAGAAGTCCTGCATGTCCAACTGCACGATCACCTCCATCTGC GAGAAGCCCCAAGAAGTGTGCGTGGCCGTGTGGCGGAAAAATGACGAGAACATC ACCCTGGAGACCGTGTGTCACGACCCCAAGCTCCCTTATCACGACTTCATTCTG GAGGACGCTGCCTCCCCCAAATGCATCATGAAGGAGAAGAAGAAGCCCGGAGAG ACCTTCTTTATGTGTTCCTGTAGCAGCGACGAGTGTAACGACAACATCATCTTC AGCGAAGAGTACAACACCAGCAACCCTGATGGAGGTGGCGGATCCGGAGGTGGA GGTTCTGGTGGAGGTGGGAGTATTCCTCCCCACGTGCAGAAGAGCGTGAATAAT GACATGATCGTGACCGATAACAATGGCGCCGTGAAATTTCCCCAGCTGTGCAAA TTCTGCGATGTGAGGTTTTCCACCTGCGACAACCAGAAGTCCTGTATGAGCAAC TGCACAATCACCTCCATCTGTGAGAAGCCTCAGGAGGTGTGCGTGGCTGTCTGG CGGAAGAATGACGAGAATATCACCCTGGAAACCGTCTGCCACGATCCCAAGCTG CCCTACCACGATTTCATCCTGGAAGACGCCGCCAGCCCTAAGTGCATCATGAAA GAGAAAAAGAAGCCTGGCGAGACCTTTTTCATGTGCTCCTGCAGCAGCGACGAA T G GAAC GAC AAT AT C AT C T T T AG C GAG GAAT AC AAT C C AG C AAC C C C GAC

[0284] (SEQ ID NO: 12), or

[0285] ATTCCTCCT CAC G T G C AGAAAAG C G T GAAC AAC GAC AT GAT AG T GAC C GAC AAT AACGGCGCGGTCAAGTTCCCACAGCTCTGCAAATTTTGCGACGTGAGATTTAGT ACCT GC GAC AAT C AGAAAT C C T G C AT GAG C AAT T G TAG CAT CAC T AG C AT C T G T GAAAAGCCCCAAGAGGTCTGCGTAGCAGTGTGGAGAAAAAATGACGAGAACATC ACACTAGAGACCGTGTGCCATGACCCTAAGCTGCCTTATCATGATTTCATCCTT GAAGATGCTGCTAGCCCTAAGTGCATTATGAAAGAGAAAAAGAAGCCTGGCGAA ACCTTTTTCATGTGTAGCTGTAGTAGCGACGAGTGCAACGATAACATCATCTTC TCGGAGGAGTATAATACATCGAATCCTGATGGCGGCGGCGGCAGCATCCCTCCT C AT G T T C AGAAGAG C G T GAAT AAC GACAT GAT T G T GAC GGAT AAC AAT G GC G C C GTAAAGTTCCCTCAGCTGTGTAAGTTCTGCGATGTGCGATTTAGCACCTGCGAC AAT C AGAAGAG CTGTATGTC C AAT T G T T C AAT C AC AAG C AT C T G T GAGAAG C C T CAAGAGGTGTGCGTGGCCGTTTGGAGAAAGAACGACGAGAACATTACCCTTGAG ACCGTATGCCACGATCCTAAGCTGCCGTATCACGACTTCATACTGGAGGATGCC G C T AGC C C T AAAT G CAT CAT GAAAGAAAAAAAAAAG C C T G G T GAGAC T T T T T T ATGTGTAGCTGCTCGTCTGACGAGTGCAATGACAATATCATTTTCAGCGAAGAA TATAACACCTCAAACCCTGAC (SEQ ID NO: 13).

[0286] Additional examples of soluble interleukin receptors and soluble cytokine receptors are known in the art.Attorney Docket No. 47039-0043W01

[0287] Antigen-Binding Domains

[0288] In some embodiments, one or both of the first target-binding domain and the second target-binding domain is an antigen-binding domain. In some embodiments, the first targetbinding domain and the second target-binding domain are each antigen-binding domains.

[0289] In some embodiments, the first target-binding domain and the second target-binding domain bind specifically to the same antigen. In some embodiments, the first target-binding domain and the second target-binding domain bind specifically to the same epitope. In some embodiments, the first target-binding domain and the second target-binding domain include the same amino acid sequence.

[0290] In some embodiments, the first target-binding domain and the second target-binding domain bind specifically to different antigens.

[0291] In some embodiments of any of the antigen-binding domains described herein can bind to an antigen selected from the group consisting of a protein, a carbohydrate, a lipid, and a combination thereof.

[0292] In some examples, an antigen-binding domain (e.g., any of the antigen-binding domains described herein) can bind specifically to any one of tissue factor (CD 142) (see e.g., those described in U. S. Patent Nos. 9,714,297; 8,951,525; 11,447,566; and 9,708,410), CD16a (see, e.g., those described in U. S. Patent No. 9,035,026), CD28 (see, e.g., those described in U. S. Patent No. 7,723,482), CD3 (see, e.g., those described in U. S. Patent No. 9,226,962), CD33 (see, e.g., those described in U. S. Patent No. 8,759,494), CD20 (see, e.g., those described in WO 2014 / 026054), CD19 (see, e.g., those described in U. S. Patent No. 9,701,758), CD22 (see, e.g., those described in WO 2003 / 104425), CD123 (see, e.g., those described in WO 2014 / 130635), IL-1R (see, e.g., those described in U. S. Patent No. 8,741,604), IL-1 (see, e.g., those described in WO 2014 / 095808), VEGF (see, e.g., those described in U. S. Patent No. 9,090,684), IL-6R (see, e.g., those described in U. S. Patent No. 7,482,436), IL-4 (see, e.g., those described in U. S. Patent Application Publication No. 2012 / 0171197), IL-10 (see, e.g., those described in U. S. Patent Application Publication No. 2016 / 0340413), PDL-1 (see, e.g., those described in Drees et al., Protein Express. Purif. 94:60-66, 2014), TIGIT (see, e.g., those described in U. S. Patent Application Publication No. 2017 / 0198042), PD-1 (see, e.g., those described in U. S. Patent No.

[0293] 7,488,802), TIM3 (see, e.g., those described in U. S. Patent No. 8,552,156), CTLA4 (see, e.g., those described in WO 2012 / 120125), MICA (see, e.g., those described in WO 2016 / 154585),Attorney Docket No. 47039-0043W01

[0294] MICB (see, e.g., those described in U. S. Patent No. 8,753,640), IL-6 (see, e.g., those described in Gejima et al., Human Antibodies 11(4):121-129, 2002), IL-8 (see, e.g., those described in U. S. Patent No. 6,117,980), TNFa (see, e.g., those described in Geng et al., Immunol. Res. 62(3):371-385, 2015, CD26a (see, e.g., those described in WO 2017 / 189526), CD36 (see, e.g., those described in U. S. Patent Application Publication No. 2015 / 0259429), ULBP2 (see, e.g., those described in U. S. Patent No. 9,273,136), CD30 (see, e.g., those described in Homach et al., Scand. J. Immunol. 48(5):497-501, 1998), CD200 (see, e.g., those described in U. S. Patent No.

[0295] 9,085,623), IGF-1R (see, e.g., those described in U. S. Patent Application Publication No.

[0296] 2017 / 0051063), MUC4AC (see, e.g., those described in WO 2012 / 170470), MUC5AC (see, e.g., those described in U. S. Patent No. 9,238,084), Trop-2 (see, e.g., those described in WO 2013 / 068946), CMET (see, e g., those described in Edwardraja et al., Biotechnol. Bioeng.

[0297] 106(3):367-375, 2010), EGFR (see, e.g., those described in Akbari et al., Protein Expr. Purif. 127:8-15, 2016), HER1 (see, e.g., those described in U. S. Patent Application Publication No. 2013 / 0274446), HER2 (see, e.g., those described in Cao et al., Biotechnol. Lett. 37(7):1347-1354, 2015), HER3 (see, e.g., those described in U. S. Patent No. 9,505,843), PSMA (see, e.g., those described in Parker et al., Protein Expr. Purif. 89(2): 136-145, 2013), CEA (see, e.g., those described in WO 1995 / 015341), B7H3 (see, e.g., those described in U. S. Patent No. 9,371,395), EPCAM (see, e.g., those described in WO 2014 / 159531), BCMA (see, e.g., those described in Smith et al., Mol. Ther. 26(6): 1447-1456, 2018), P-cadherin (see, e.g., those described in U. S. Patent No. 7,452,537), CEACAM5 (see, e.g., those described in U. S. Patent No. 9,617,345), a UL16-binding protein (see, e.g., those described in WO 2017 / 083612), HLA-DR (see, e.g., Pistillo et al., Exp. Clin. Immunogenet. 14(2): 123-130, 1997), DLL4 (see, e.g., those described in WO 2014 / 007513), TYRO3 (see, e.g., those described in WO 2016 / 166348), AXL (see, e.g., those described in WO 2012 / 175692), MER (see, e g., those described in WO 2016 / 106221), CD122 (see, e.g., those described in U. S. Patent Application Publication No. 2016 / 0367664), CD155 (see, e.g., those described in WO 2017 / 149538), orPDGF-DD (see, e.g., those described in U. S. Patent No. 9,441,034).

[0298] In some embodiments of any of the multi-chain chimeric polypeptides described herein, the antigen-binding domain includes or is a scFv or a single domain antibody (e.g., a VHH or a VNAR domain). In some embodiments, the antigen-binding domains present in any of the multi-Attorney Docket No. 47039-0043W01

[0299] chain chimeric polypeptides described herein are each independently selected from the group consisting of: a VHH domain, a VNAR domain, and a scFv.

[0300] A VHH domain is a single monomeric variable antibody domain that can be found in camelids. A VNAR domain is a single monomeric variable antibody domain that can be found in cartilaginous fish. Non-limiting aspects of VHH domains and VNAR domains are described in, e.g., Cromie et al., Curr. Top. Med. (Them. 15:2543-2557, 2016; De Genst et al., Dev. Comp. Immunol. 30:187-198, 2006; De Meyer et al., Trends Biotechnol. 32:263-270, 2014; Kijanka et al., Nanomedicine 10:161-174, 2015; Kovaleva et al., Expert. Opin. Biol. Then 14:1527-1539, 2014; Krah et al., Immunopharmacol. Immunotoxicol. 38:21-28, 2016; Mujic-Delic et al., Trends Pharmacol. Sci. 35:247-255, 2014; Muyldermans, J. Biotechnol. 74:277-302, 2001;

[0301] Muyldermans et al., Trends Biochem. Sci. 26:230-235, 2001; Muyldermans, Ann. Rev. Biochem.

[0302] 82:775-797, 2013; Rahbarizadeh et al., Immunol. Invest. 40:299-338, 2011; Van Audenhove et al., EBioMedicine 8:40-48, 2016; Van Bockstaele et al., Curr. Opin. Investig. Drugs 10:1212- 1224, 2009; Vincke et a\. Methods Mol. Biol. 911:15-26, 2012; and Wesolowski et al., Med. Microbiol. Immunol. 198:157-174, 2009.

[0303] In some embodiments, each of the antigen-binding domains in the multi-chain chimeric polypeptides described herein are both VHH domains, or at least one antigen-binding domain is a VHH domain. In some embodiments, each of the antigen-binding domains in the multi-chain chimeric polypeptides described herein are both VNAR domains, or at least one antigen-binding domain is a VNAR domain. In some embodiments, each of the antigen-binding domains in the multi-chain chimeric polypeptides described herein are both scFv domains, or at least one antigen-binding domain is a scFv domain.

[0304] In some embodiments, any of the antigen-binding domains described herein is a BiTE, a (SCFV)2, a nanobody, a nanobody-HSA, a DART, a TandAb, a scDiabody, a scDiabody-CH3, scFv-CH-CL-scFv, a HSAbody, scDiabody -HAS, or a tandem-scFv. DARTs are described in, e.g., Garber, Nature Reviews Drug Discovery 13:799-801, 2014. Additional examples of antigenbinding domains that can be used in any of the multi-chain chimeric polypeptide are known in the art.

[0305] In some embodiments, any of the antigen-binding domains described herein is an antigen-binding fragment of an antibody (e.g., any of the antigen-binding fragments of an antibody described herein), a VHH-scAb, a VHH-Fab, a Dual scFab, a F(ab’)2, a diabody, aAttorney Docket No. 47039-0043W01

[0306] crossMab, a DAF (two-in-one), a DAF (four-in-one), a DutaMab, a DT-IgG, a knobs-in-holes common light chain, a knobs-in-holes assembly, a charge pair, a Fab-arm exchange, a SEEDbody, a LUZ-Y, a Fcab, a K / .-body, an orthogonal Fab, a DVD-IgG, a IgG(H)-scFv, a scFv-(H)IgG, IgG(L)-scFv, scFv-(L)IgG, IgG(L, H)-Fv, IgG(H)-V, V(H)-IgG, IgG(L)-V, V(L)-IgG, KIH IgG-scFab, 2scFv-IgG, IgG-2scFv, scFv4-Ig, Zybody, DVI-IgG, Diabody-CH3, a triple body, a miniantibody, a minibody, a TriBi minibody, scFv-CH3 KIH, Fab-scFv, aF(ab’)2-scFv2, a scFv-KIH, a Fab-scFv-Fc, a tetravalent HCAb, a scDiabody-Fc, a Diabody-Fc, a tandem scFv-Fc, an Intrabody, a dock and lock, a ImmTAC, an IgG-IgG conjugate, a Cov-X-Body, and a scFvl-PEG-scFv2. See, e.g., Spiess et al., Mol. Immunol. 67:95-106, 2015, incorporated in its entirety herewith, for a description of these elements. In some embodiments, two or more polypeptides present in the multi-chain chimeric polypeptide assemble (e.g., non-covalently assemble) to form any of the antigen-binding domains described herein. In some embodiments, the two or more polypeptides present in the multi-chain chimeric polypeptide are connected with a peptide linker sequence. For example, two or more polypeptide chains connected by a linker can assemble (e.g., non-covalently assemble) to form any of the antigenbinding domains described herein.

[0307] Non-limiting examples of an antigen-binding fragment of an antibody include an Fv fragment, a Fab fragment, aF(ab')2 fragment, and a Fab1fragment. Additional examples of an antigen-binding fragment of an antibody is an antigen-binding fragment of an IgG (e.g., an antigen-binding fragment of IgGl, IgG2, IgG3, or IgG4) (e.g., an antigen-binding fragment of a human or humanized IgG, e.g., human or humanized IgGl, IgG2, IgG3, or IgG4); an antigenbinding fragment of an IgA (e.g., an antigen-binding fragment of IgAl or IgA2) (e.g., an antigenbinding fragment of a human or humanized IgA, e.g., a human or humanized IgAl or IgA2); an antigen-binding fragment of an IgD (e.g., an antigen-binding fragment of a human or humanized IgD); an antigen-binding fragment of an IgE (e.g., an antigen-binding fragment of a human or humanized IgE); or an antigen-binding fragment of an IgM (e.g., an antigen-binding fragment of a human or humanized IgM).

[0308] Anti-Mesothelin Antigen-Binding Domains

[0309] In some embodiments, at least one of the first target-binding domain and the second target-binding domain includes or is an anti-mesothelin antigen binding domain. In someAttorney Docket No. 47039-0043W01

[0310] embodiments, the first target-binding domain, the second target-binding domain, and / or one or more of the one or more additional target-binding domains includes or is an anti-mesothelin antigen binding domain. In some embodiments, the anti-mesothelin antigen binding domain includes or is an anti-mesothelin antibody. Non-limiting examples of an anti-mesothelin antigen binding domain include those described in, e.g., U. S. Publication Nos. US2018 / 0258181;

[0311] US2024 / 0277865; US2024 / 100157; US2020 / 181280; US2018 / 194862; US2020 / 095332;

[0312] US2025 / 092154; US2020 / 095332; US 2017 / 267755; US2006 / 204506; US2012 / 237509;

[0313] US2013 / 177571; US2014 / 127237; US2021 / 309755; US2023 / 192843; US2023 / 192843;

[0314] US2021 / 024648; US2019 / 388553; US2021 / 113706; US2024 / 165249; US2018 / 338940;

[0315] US2019 / 060362; US2019 / 167617; US2020 / 069622; US2021 / 113506; US2020 / 270362;

[0316] US2017 / 267755; US2018 / 002439; U. S. Patent Nos. 11,866,508; 11,981,745; International Publication Nos. WO2018 / 064215; WO2023 / 218027; W02021 / 160708; WO2025 / 114357; W02020 / 234114; W02020 / 092631; WO2024 / 165567; WO2024 / 165567; WO2023 / 107825; WO2022 / 067224; W0202 / 4042104; W02024 / 042105; WO2019 / 087425; WO2018 / 215835; WO2018 / 060480; WO2017 / 156461; WO2018 / 049118; WO2014 / 052064; WO2014 / 043523; WO2012 / 154530; WO2013 / 063419; WO2010 / 111282; W02009 / 120769; WO2025 / 234736; WO2025 / 230359; WO2025 / 161620; WO2025 / 143287; W02025 / 002318; WO2024 / 131751; WO2024 / 119065; WO2024 / 076927; WO2024 / 233583; WO2024 / 076927; WO2024 / 089551; WO2024 / 094775; W02024 / 020703; WO2024 / 026358; WO2024 / 012395; WO2023 / 218027; WO2023 / 179740; WO2023 / 177974; WO2023 / 116637; WO2023 / 116637; WO2023 / 125813; WO2023 / 115528; W02023 / 091420; WO2023 / 088295; WO2023 / 088295; WO2023 / 164627; WO2023 / 066133; WO2023 / 064876; WO2023 / 064876; W02023 / 060157; W02023 / 006391; W02023 / 006390; W02023 / 006390; W02023 / 006391; WO2022 / 257984; WO2022 / 262859; WO2022 / 242703; WO2022 / 143550; WO2022 / 121941; W02022 / 120033; WO2022 / 096716; WO2021 / 239987; W02022 / 084440; WO2022 / 034562; WO2021 / 249549; WO2021 / 239987; WO2021 / 247812; WO2021 / 130535; WO2021 / 127528; WO2020 / 247854; W02020 / 204305; W02020 / 204305; WO2021 / 231969; WO2020 / 123425; W02020 / 043152; W02020 / 011970; W02020 / 011972; W02020 / 011976; WO2019 / 231879; WO2019 / 078698; WO2019 / 062518; WO2019 / 043026; WO2019 / 005208; W02019 / 005208; WO2019 / 005208; WO2019 / 005208; WO2018 / 209304; WO2022 / 256498; WO2018 / 186706; WO2017 / 177337; W02009 / 045957;Attorney Docket No. 47039-0043W01

[0317] Liu et al. Proc Natl Acad Sci USA. 2024; 121 (4):e2317283121; Zhang et al. SciRep. 2015;

[0318] 5:9928; Chakraborty et al. Mol Cancer Ther. 2024; 23(7): 1021-1030; and Matsuura et al.

[0319] Cancer Sci. 2019; 110(9):2722-2733, the anti-mesothelin antigen binding domains in each of which are herein incorporated by reference. In some embodiments, the anti-mesothelin antigen binding domain includes or is a single chain anti-mesothelin antibody. Non-limiting examples of a single chain anti-mesothelin antibody include those described in, e.g., U. S. Publication No. 2024 / 0277865 and Matsuura et al. Cancer Sci. 2019; 110(9):2722-2733, each of which is incorporated by reference in its entirety.

[0320] In some embodiments, the anti-mesothelin antigen binding domain is humanized.

[0321] In some embodiments, the anti-mesothelin antigen binding domain is an scFv. In some embodiments, a scFv (e.g., any of the scFvs described herein) can include a linker sequence (e.g., any of the exemplary linker sequences described herein or known in the art) between the heavy chain variable domain and the light chain variable domain.

[0322] In some embodiments, an anti-mesothelin scFv includes a heavy chain variable domain including a sequence that is at least 70% identical (e.g., at least 75% identical, at least 80% identical, at least 85% identical, at least 90% identical, at least 92% identical, at least 94% identical, at least 95% identical, at least 96% identical, at least 98% identical, or 100% identical) to:

[0323] EVQLQQSGPVLVKPGASVKISCKASGYSFTGYYMHWVRQSLVKRLEWIGRINPY TGVPSYKHNFKDKASLTVDKSSSTAYMELHSLTSEDSAVYYCARELGGYWGQGT TLTVSS ( SEQ ID NO: 96)

[0324] and / or a light chain variable domain including a sequence that is at least 70% identical (e.g., at least 75% identical, at least 80% identical, at least 85% identical, at least 90% identical, at least 92% identical, at least 94% identical, at least 95% identical, at least 96% identical, at least 98% identical, or 100% identical) to:

[0325] QAWTQESALTTSPGETVTLTCRSSTGAVTTGNYPNWVQEKPDHLFTGLIAGTN NRAPGVPARFSGSLIGDKAALTITGAQTEDEAIYFCALWFSSHWVFGGGTKLTV

[0326] L ( SEQ ID NO: 97).

[0327] In some embodiments, an anti-mesothelin antigen binding domain includes a sequence that is at least 70% identical (e.g., at least 75% identical, at least 80% identical, at least 85%Attorney Docket No. 47039-0043W01

[0328] identical, at least 90% identical, at least 92% identical, at least 94% identical, at least 95% identical, at least 96% identical, at least 98% identical, or 100% identical) to:

[0329] QAWTQESALTTSPGETVTLTCRSSTGAVTTGNYPNWVQEKPDHLFTGLIAGTN NRAPGVPARFSGSLIGDKAALTITGAQTEDEAIYFCALWFSSHWVFGGGTKLTV LGGGGSGGGGSGGGGSEVQLQQSGPVLVKPGASVKISCKASGYSFTGYYMHWVR QSLVKRLEWIGRINPYTGVPSYKHNFKDKASLTVDKSSSTAYMELHSLTSEDSA VYYCARELGGYWGQGTTLTVSS (SEQ ID NO: 14).

[0330] In some embodiments, an anti-mesothelin antigen binding domain is encoded by a nucleic acid encoding a first sequence at least 70% identical (e.g., at least 75% identical, at least 80% identical, at least 85% identical, at least 90% identical, at least 92% identical, at least 94% identical, at least 95% identical, at least 96% identical, at least 98% identical, or 100% identical) to:

[0331] CAAGCCGTGGTGACCCAAGAGAGCGCCCTGACCACAAGCCCTGGCGAGACCGTG ACCCTGACCTGCAGAAGCAGCACCGGCGCCGTGACCACCGGCAACTACCCTAAC TGGGTGCAAGAGAAGCCTGACCACCTGTTCACCGGCCTGATCGCCGGCACCAAC AACAGAGCCCCTGGCGTGCCTGCTAGATTCAGCGGCAGCCTGATCGGCGACAAG GCCGCCCTGACCATCACCGGCGCTCAGACCGAGGACGAGGCCATCTACTTCTGC GCCCTGTGGTTCAGCAGCCATTGGGTATTTGGCGGCGGCACCAAGCTCACAGTC CTGGGTGGGGGTGGTTCCGGTGGCGGCGGGAGTGGTGGCGGTGGCTCTGAAGTG CAGCTGCAGCAGAGCGGCCCTGTGCTGGTGAAGCCTGGCGCTAGCGTGAAGATC AGCTGCAAGGCTAGCGGCTACAGCTTCACCGGCTACTACATGCACTGGGTGAGA CAGAGCCTGGTGAAGAGACTGGAGTGGATCGGCAGAATCAACCCTTACACCGGC GTGCCTAGCTACAAGCACAACTTCAAGGACAAGGCTAGCCTGACCGTGGACAAG AGCAGCAGCACCGCCTACATGGAGCTGCACAGCCTGACAAGCGAGGACAGCGCC GTGTACTACTGCGCTAGAGAGCTGGGCGGCTACTGGGGCCAAGGCACCACCCTG ACCGTGAGCAGC (SEQ ID NO: 15).

[0332] In some embodiments, an anti-mesothelin antigen binding domain includes a heavy chain variable domain including a sequence that is at least 70% identical (e.g., at least 75% identical, at least 80% identical, at least 85% identical, at least 90% identical, at least 92% identical, at least 94% identical, at least 95% identical, at least 96% identical, at least 98% identical, or 100% identical) to:

[0333] QVQLVQSGAEVKKPGASVKVSCKASGYSFTGYYMHWVRQAPGQGLEWMGRINPY TGVPSYKHKFQGRVTMTVDKSTSTAYMELSSLTSEDTAVYYCARELGGYWGQGT TLTVSS (SEQ ID NO: 104)Attorney Docket No. 47039-0043W01

[0334] and / or a light chain variable domain including a sequence that is at least 70% identical (e.g., at least 75% identical, at least 80% identical, at least 85% identical, at least 90% identical, at least 92% identical, at least 94% identical, at least 95% identical, at least 96% identical, at least 98% identical, or 100% identical) to:

[0335] QAWTQEPSLTVSPGGTVTLTCASSTGAVTTGNYPNWFQQKPGQAFRGLIAGTN NKASWTPARFSGSLIGDKAALTLSGVQPEDEAEYYCALWFSSHWVFGGGTKLTV

[0336] L (SEQ IDNO: 105).

[0337] In some embodiments, an anti-mesothelin antigen binding domain includes a heavy chain variable domain including a sequence that is at least 70% identical (e.g., at least 75% identical, at least 80% identical, at least 85% identical, at least 90% identical, at least 92% identical, at least 94% identical, at least 95% identical, at least 96% identical, at least 98% identical, or 100% identical) to:

[0338] QQQLEESGGGLVKPEGSLTLTCKASGDLGFYFYACWVRQAPGKGLEWIACI YTA GSGSTYYASWAKGRFTISKASSTTVTLQMTSLAAADTATYFCARSTANTRSTYY LNLWGPGTLVTVSS ( SEQ ID NO: 110)

[0339] and / or a light chain variable domain including a sequence that is at least 70% identical (e.g., at least 75% identical, at least 80% identical, at least 85% identical, at least 90% identical, at least 92% identical, at least 94% identical, at least 95% identical, at least 96% identical, at least 98% identical, or 100% identical) to:

[0340] DWMTQTPASVSEPVGGTVTIKCQASQRISSYLSWYQQKPGQRPKLLIFGASTL ASGVPSRFKGSGSGTEYTLTISDLECADAATYYCQSYAYFDSNNWHAFGGGTEV

[0341] W ( SEQ ID NO: 111).

[0342] In some embodiments, an anti-mesothelin antigen binding domain includes a heavy chain variable domain including a sequence that is at least 70% identical (e.g., at least 75% identical, at least 80% identical, at least 85% identical, at least 90% identical, at least 92% identical, at least 94% identical, at least 95% identical, at least 96% identical, at least 98% identical, or 100% identical) to:

[0343] QVQLVQSGAEVKKPGASVKVSCKASGYSFTGYTMNWVRQAPGQGLEWMGLITPY NGASSYNQKFRGKATMTVDTSTSTVYMELSSLRSEDTAVYYCARGGYDGRGFDY WGQGTLVTVSS ( SEQ ID NO: 114)

[0344] and / or a light chain variable domain including a sequence that is at least 70% identical (e.g., at least 75% identical, at least 80% identical, at least 85% identical, at least 90% identical, at leastAttorney Docket No. 47039-0043W01

[0345] 92% identical, at least 94% identical, at least 95% identical, at least 96% identical, at least 98% identical, or 100% identical) to:

[0346] DIQMTQSPSSLSASVGDRVTITCSASSSVSYMHWYQQKSGKAPKLLIYDTSKLA SGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQWSKHPLTFGQGTKLEIK

[0347] (SEQ IDNO: 115).

[0348] In some embodiments, an anti-mesothelin antigen binding domain includes a sequence that is at least 70% identical (e.g., at least 75% identical, at least 80% identical, at least 85% identical, at least 90% identical, at least 92% identical, at least 94% identical, at least 95% identical, at least 96% identical, at least 98% identical, or 100% identical) to:

[0349] QAWTQEPSLTVSPGGTVTLTCASSTGAVTTGNYPNWFQQKPGQAFRGLIAGTN NKASWTPARFSGSLIGDKAALTLSGVQPEDEAEYYCALWFSSHWVFGGGTKLTV LGGGGSGGGGSGGGGSQVQLVQSGAEVKKPGASVKVSCKASGYSFTGYYMHWVR QAPGQGLEWMGRINPYTGVPSYKHKFQGRVTMTVDKSTSTAYMELSSLRSEDTA VYYCARELGGYWGQGTTLTVSS (SEQ ID NO: 16).

[0350] In some embodiments, an anti-mesothelin antigen binding domain is encoded by a nucleic acid encoding a sequence at least 70% identical (e.g., at least 75% identical, at least 80% identical, at least 85% identical, at least 90% identical, at least 92% identical, at least 94% identical, at least 95% identical, at least 96% identical, at least 98% identical, or 100% identical) to:

[0351] CAAGCCGTGGTGACCCAAGAGCCTAGCCTGACAGTCAGCCCTGGCGGCACAGTC ACCCTGACCTGCGCTAGCAGCACCGGCGCCGTGACCACCGGCAACTACCCTAAC TGGTTTCAGCAGAAGCCTGGCCAAGCCTTCAGAGGCCTGATCGCCGGCACCAAC AACAAGGCTAGCTGGACCCCTGCTAGATTCAGCGGCAGCCTGATCGGCGACAAG GCCGCCCTGACCCTGAGCGGCGTGCAGCCTGAGGACGAGGCCGAGTACTACTGC GCCCTCTGGTTCAGCAGCCACTGGGTGTTTGGCGGCGGAACCAAGCTGACAGTC CTGGGCGGCGGAGGCAGCGGCGGAGGCGGCTCCGGAGGAGGAGGCAGCCAAGTG CAGCTGGTGCAGAGCGGCGCCGAGGTGAAGAAGCCTGGCGCTAGCGTGAAGGTG AGCTGCAAGGCTAGCGGCTACAGCTTCACCGGCTACTACATGCACTGGGTGAGA CAAGCCCCTGGCCAAGGCCTGGAGTGGATGGGCAGAATCAACCCTTACACCGGC GTGCCTAGCTACAAGCACAAGTTCCAAGGCAGAGTGACCATGACCGTGGACAAG AGCACAAGCACCGCCTACATGGAGCTGAGCAGCCTGAGAAGCGAGGACACCGCC GTGTACTATTGCGCTAGAGAGCTGGGCGGCTACTGGGGCCAAGGCACCACCCTG ACCGTGAGCAGC (SEQ ID NO: 17).

[0352] In some embodiments, an anti-mesothelin antigen binding domain includes a heavy chain variable domain including a sequence that is at least 70% identical (e.g., at least 75% identical, atAttorney Docket No. 47039-0043W01

[0353] least 80% identical, at least 85% identical, at least 90% identical, at least 92% identical, at least 94% identical, at least 95% identical, at least 96% identical, at least 98% identical, or 100% identical) to:

[0354] QVQLVQSGAEVKKPGASVKVSCKASGYSFTGYTMNWVRQAPGQGLEWMGLITPY NGASSYNQKFRGKATMTVDTSTSTVYMELSSLRSEDTAVYYCARGGYDGRGFDY WGQGTLVTVSS (SEQ ID NO: 116),

[0355] and / or a light chain variable domain including a sequence that is at least 70% identical (e.g., at least 75% identical, at least 80% identical, at least 85% identical, at least 90% identical, at least 92% identical, at least 94% identical, at least 95% identical, at least 96% identical, at least 98% identical, or 100% identical) to:

[0356] DIQMTQSPSSLSASVGDRVTITCSASSSVSYMHWYQQKSGKAPKLLIYDTSKLA SGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQWSKHPLTFGQGTKLEIK

[0357] (SEQ ID NO: 117).

[0358] In some embodiments, an anti-mesothelin antigen binding domain includes a heavy chain variable domain including a sequence that is at least 70% identical (e.g., at least 75% identical, at least 80% identical, at least 85% identical, at least 90% identical, at least 92% identical, at least 94% identical, at least 95% identical, at least 96% identical, at least 98% identical, or 100% identical) to:

[0359] EVQLLESGGGLVQPGGSLRLSCAASGLTFRSYAMTWVRQAPGKGLEWVSGISVS GGI T YYAS SVKGRFT I SRDNSKNTLYLQMNSLRAEDTAVYYCAKRGAAVGS FDY WGQGTLVTVSS (SEQ ID NO: 118),

[0360] and / or a light chain variable domain including a sequence that is at least 70% identical (e.g., at least 75% identical, at least 80% identical, at least 85% identical, at least 90% identical, at least 92% identical, at least 94% identical, at least 95% identical, at least 96% identical, at least 98% identical, or 100% identical) to:

[0361] DIQMTQSPSSLSASVGDRVTITCRSSQGIGSWLAWYQQKPEKAPQSLIYAASS LQSGVPSRFSGSGSGTDFTLTISNLQPEDFATYYCQQYKSYPLTFGGGTKVEI

[0362] K (SEQ ID NO: 119).

[0363] In some embodiments, an anti-mesothelin antigen binding domain includes a sequence that is at least 70% identical (e.g., at least 75% identical, at least 80% identical, at least 85% identical, at least 90% identical, at least 92% identical, at least 94% identical, at least 95% identical, at least 96% identical, at least 98% identical, or 100% identical) to:Attorney Docket No. 47039-0043W01

[0364] QVQLQESGPGLVKPSETLSLTCTVSGGSISSSSYFWGWIRQPPGKCLEWIGNIY YSGSSNYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCARLPRGDRDAF DIWGQGTMVTVSSGGGGQGGGGQGGGGQDIVMTQSPSSLSASVGDRVTITCRAS QGISNYLAWYQQKPGKVPKLLIYAASTLQSGVPSRFSGSGSGTDFTLTISSLQP EDFATYYCQQSYSTPFTFGCGTKVEIK (SEQ ID NO: 120).

[0365] In some embodiments, an anti-mesothelin antigen binding domain includes a sequence that is at least 70% identical (e.g., at least 75% identical, at least 80% identical, at least 85% identical, at least 90% identical, at least 92% identical, at least 94% identical, at least 95% identical, at least 96% identical, at least 98% identical, or 100% identical) to:

[0366] QPVLTQPASLSASPGASASLTCTLRSGINVRDYRI FWYQQKPGSPPQYLLRYKS ASDKQQGSGVPSRFSGSKDASANAGILLISGLQSEDEADYYCMIWHHDSEGHAF VFGGGTKLTVL (SEQ ID NO: 121).

[0367] In some embodiments, an anti-mesothelin antigen binding domain includes a heavy chain variable domain including a sequence that is at least 70% identical (e.g., at least 75% identical, at least 80% identical, at least 85% identical, at least 90% identical, at least 92% identical, at least 94% identical, at least 95% identical, at least 96% identical, at least 98% identical, or 100% identical) to:

[0368] QVQLVQSGAEVKKPGASVKVSCKASGYTFSLYWMHWVRQAPGQGLEWMGMIHPN SGSTNYNEKFKSRVTMTRDTSTSTAYMELSSLRSEDTAVYYCARRLGYGGRYEY FDVWGKGT TVTVS S (SEQ ID NO: 122),

[0369] and / or a light chain variable domain including a sequence that is at least 70% identical (e.g., at least 75% identical, at least 80% identical, at least 85% identical, at least 90% identical, at least 92% identical, at least 94% identical, at least 95% identical, at least 96% identical, at least 98% identical, or 100% identical) to:

[0370] DIVMTQSPATLSVSPGERATLSCSASSSVSYMHWYQQKPGKAPKLLI YDTSRL ASGVPSRFSGSGSGTDFTLTISSLQAEDFATYYCQQWSSYPLTFGQGTKLEIK

[0371] R (SEQ ID NO: 123).

[0372] In some embodiments, where the first target-binding domain includes an anti-mesothelin antigen binding domain, the first target-binding domain further includes an anti-tissue factor binding domain (e.g., any of the anti -tissue factor antigen-binding domains provided herein). In some embodiments, where the second target-binding domain includes an anti-mesothelin antigenAttorney Docket No. 47039-0043W01

[0373] binding domain, the second target-binding domain further includes an anti -tissue factor binding domain (e.g., any of the anti-tissue factor antigen-binding domains provided herein).

[0374] Anti-Tissue Factor Antigen-Binding Domains

[0375] In some embodiments, the first target-binding domain, the second target-binding domain, and / or one or more of the one or more additional target-binding domains includes or is an antitissue factor antigen-binding domain. In some embodiments of any of the multi-chain chimeric polypeptides described herein, the additional target-binding domain includes or is an anti-tissue factor antigen-binding domain.

[0376] In some embodiments, an anti-tissue factor antigen-binding domain can include a sequence that is at least 70% identical (e.g., at least 75% identical, at least 80% identical, at least 85% identical, at least 90% identical, at least 92% identical, at least 94% identical, at least 95% identical, at least 96% identical, at least 98% identical, or 100% identical) to:

[0377] DIQMTQSPASLSASVGDRVTITCLASQTIDTWLAWYLQKPGKSPQLLIYAATNL ADGVPSRFSGSGSGTDFSFTISSLQPEDFATYYCQQVYSSPFTFGQGTKLEIKG GGGSGGGGSGGGGSQIQLVQSGGEVKKPGASVRVSCKASGYSFTDYNVYWVRQS PGKGLEWIGYIDPYNGITIYDQNFKGKATLTVDKSTSTAYMELSSLRSEDTAVY FCARDVTTALDFWGQGT TVTVS S (SEQ ID NO: 91).

[0378] In some embodiments, an anti-tissue factor antibody can include a heavy chain variable sequence that is at least 70% identical (e.g., at least 75% identical, at least 80% identical, at least 85% identical, at least 90% identical, at least 92% identical, at least 94% identical, at least 95% identical, at least 96% identical, at least 98% identical, or 100% identical) to:

[0379] Q I QLVQS GGEVKKPGAS VRVSCKAS GYS FTDYNVYWVRQS PGKGLEW I GYI DP Y NGITIYDQNFKGKATLTVDKSTSTAYMELSSLRSEDTAVYFCARDVTTALDFWG QGTTVTVSS (SEQ ID NO: 92),

[0380] and a light chain variable sequence that is at least 70% identical (e.g., at least 75% identical, at least 80% identical, at least 85% identical, at least 90% identical, at least 92% identical, at least 95% identical, at least 96% identical, at least 98% identical, or 100% identical) to:

[0381] DIQMTQSPASLSASVGDRVTITCLASQTIDTWLAWYLQKPGKSPQLLIYAATNL ADGVPSRFSGSGSGTDFSFTISSLQPEDFATYYCQQVYSSPFTFGQGTKLEIK

[0382] (SEQ ID NO: 93).Attorney Docket No. 47039-0043W01

[0383] In some embodiments, the first target-binding domain, the second target-binding domain, and / or one or more of the one or more additional target-binding domains includes or is an antitissue factor single chain antibody. In some embodiments of any of the multi-chain chimeric polypeptides described herein, the additional target-binding domain includes or is an anti-tissue factor single chain antibody. In some embodiments, an anti-tissue factor single chain antibody includes a sequence that is at least 70% identical (e.g., at least 75% identical, at least 80% identical, at least 85% identical, at least 90% identical, at least 92% identical, at least 94% identical, at least 95% identical, at least 96% identical, at least 98% identical, or 100% identical) to:

[0384] DIQMTQSPASLSASVGDRVTITCLASQTIDTWLAWYLQKPGKSPQLLIYAATNL ADGVPSRFSGSGSGTDFSFTISSLQPEDFATYYCQQVYSSPFTFGQGTKLEIKG GGGSGGGGSGGGGSQIQLVQSGGEVKKPGASVRVSCKASGYSFTDYNVYWVRQS PGKGLEWIGYIDPYNGITIYDQNFKGKATLTVDKSTSTAYMELSSLRSEDTAVY FCARDVTTALDFWGQGT TVTVS S (SEQ ID NO: 91).

[0385] In some embodiments, an anti-tissue factor single chain antibody is encoded by a nucleic acid including a sequence at least 70% identical (e.g., at least 75% identical, at least 80% identical, at least 85% identical, at least 90% identical, at least 92% identical, at least 94% identical, at least 95% identical, at least 96% identical, at least 98% identical, or 100% identical) to:

[0386] GACATACAGATGACCCAAAGCCCTGCTAGCCTGAGCGCGAGCGTGGGCGACAGA GTGACCATCACCTGCCTCGCCTCTCAGACGATTGACACCTGGCTGGCTTGGTAT TTGCAGAAACCTGGGAAAAGCCCTCAGCTGCTGATCTACGCAGCCACCAATTTA GCCGATGGCGTCCCTTCTAGATTTAGCGGCTCGGGATCCGGCACCGATTTTAGC TTTACAATTTCATCTCTACAGCCTGAGGACTTCGCCACCTATTATTGTCAACAA GTATATAGCAGCCCTTTCACCTTCGGCCAAGGCACCAAGTTAGAGATCAAGGGC GGTGGCGGCTCCGGGGGCGGGGGAAGTGGAGGTGGGGGGTCACAGATTCAGCTG GTTCAAAGCGGCGGCGAGGTGAAGAAACCTGGCGCCTCGGTGAGAGTTAGCTGC AAAGCTAGCGGATATTCGTTTACTGACTACAACGTGTATTGGGTGAGGCAGAGC CCGGGCAAGGGACTGGAGTGGATTGGGTACATTGACCCCTACAACGGCATAACC ATATAT GAT CAGAAC T T CAAGGGCAAAGCCACGC T GACCGT GGATAAAAGCACA AGCACCGCTTATATGGAGTTGAGCTCCCTCCGTAGCGAAGACACGGCAGTATAC TTTTGCGCACGGGATGTGACCACGGCCCTAGATTTCTGGGGACAAGGGACAACC GTTACTGTTAGTTCC (SEQ IDNO: 124).

[0387] In some embodiments, an anti-tissue factor antigen-binding domain can include a sequence that is at least 70% identical (e.g., at least 75% identical, at least 80% identical, at leastAttorney Docket No. 47039-0043W01

[0388] 85% identical, at least 90% identical, at least 92% identical, at least 94% identical, at least 95% identical, at least 96% identical, at least 98% identical, or 100% identical) to:

[0389] DIQMTQSPASLSASVGDRVTITCLASQTIDTWLAWYLQKPGKSPQLLIYAATNL ADGVPSRFSGSGSGTDFSFTISSLQPEDFATYYCQQVYSSPFTFGQGTKLEIKG GGGSQIQLVQSGGEVKKPGASVRVSCKASGYSFTDYNVYWVRQSPGKGLEWIGY IDPYNGITIYDQNFKGKATLTVDKSTSTAYMELSSLRSEDTAVYFCARDVTTAL DFWGQGTTVTVSS (SEQ ID NO: 18).

[0390] In some embodiments, an anti-tissue factor antibody can include a first sequence that is at least 70% identical (e.g., at least 75% identical, at least 80% identical, at least 85% identical, at least 90% identical, at least 92% identical, at least 94% identical, at least 95% identical, at least 96% identical, at least 98% identical, or 100% identical) to:

[0391] DIQMTQSPASLSASVGDRVTITCLASQTIDTWLAWYLQKPGKSPQLLIYAATNL ADGVPSRFSGSGSGTDFSFTISSLQPEDFATYYCQQVYSSPFTFGQGTKLEIKG GGGSQIQLVQSGGEVKKPGASVRVSCKASGYSFTDYNVYWVRQSPGKGLEWIGY IDPYNGITIYDQNFKGKATLTVDKSTSTAYMELSSLRSEDTAVYFCARDVTTAL DFWGQGTTVTVSS (SEQ ID NO: 18),

[0392] and a second sequence that is at least 70% identical (e g., at least 75% identical, at least 80% identical, at least 85% identical, at least 90% identical, at least 92% identical, at least 95% identical, at least 96% identical, at least 98% identical, or 100% identical) to:

[0393] DIQMTQSPASLSASVGDRVTITCLASQTIDTWLAWYLQKPGKSPQLLIYAATNL ADGVPSRFSGSGSGTDFSFTISSLQPEDFATYYCQQVYSSPFTFGQGTKLEIKG GGGSQIQLVQSGGEVKKPGASVRVSCKASGYSFTDYNVYWVRQSPGKGLEWIGY IDPYNGITIYDQNFKGKATLTVDKSTSTAYMELSSLRSEDTAVYFCARDVTTAL DFWGQGTTVTVSS (SEQ ID NO: 18).

[0394] In some embodiments, the anti-tissue factor antigen-binding domain can further include a linker sequence (e.g., any of the exemplary linker sequences described herein or known in the art) between the first sequence and the second sequence.

[0395] In some embodiments, the first target-binding domain, the second target-binding domain, and / or one or more of the one or more additional target-binding domains includes or is an antitissue factor diabody. In some embodiments of any of the multi -chain chimeric polypeptides described herein, the additional target-binding domain includes or is an anti-tissue factor diabody. In some embodiments, an anti-tissue factor diabody includes a sequence that is at least 70% identical (e.g., at least 75% identical, at least 80% identical, at least 85% identical, at leastAttorney Docket No. 47039-0043W01

[0396] 90% identical, at least 92% identical, at least 94% identical, at least 95% identical, at least 96% identical, at least 98% identical, or 100% identical) to:

[0397] DIQMTQSPASLSASVGDRVTITCLASQTIDTWLAWYLQKPGKSPQLLIYAATNL ADGVPSRFSGSGSGTDFSFTISSLQPEDFATYYCQQVYSSPFTFGQGTKLEIKG GGGSQIQLVQSGGEVKKPGASVRVSCKASGYSFTDYNVYWVRQSPGKGLEWIGY IDPYNGITIYDQNFKGKATLTVDKSTSTAYMELSSLRSEDTAVYFCARDVTTAL DFWGQGTTVTVSSGGGGSGGGGSGGGGSDIQMTQSPASLSASVGDRVTITCLAS QTIDTWLAWYLQKPGKSPQLLIYAATNLADGVPSRFSGSGSGTDFSFTISSLQP EDFATYYCQQVYSSPFTFGQGTKLEIKGGGGSQIQLVQSGGEVKKPGASVRVSC KASGYSFTDYNVYWVRQSPGKGLEWIGYIDPYNGITIYDQNFKGKATLTVDKST STAYMELSSLRSEDTAVYFCARDVTTALDFWGQGTTVTVSS (SEQ IDNO: 19).

[0398] In some embodiments, an anti-tissue factor diabody is encoded by a nucleic acid including a sequence at least 70% identical (e.g., at least 75% identical, at least 80% identical, at least 85% identical, at least 90% identical, at least 92% identical, at least 94% identical, at least 95% identical, at least 96% identical, at least 98% identical, or 100% identical) to:

[0399] GATATTCAGATGACACAGTCACCTGCCTCACTGTCAGCGTCTGTGGGGGACCGC GTGACAATTACGTGCCTGGCTAGCCAAACCATAGATACCTGGCTGGCGTGGTAC TTGCAGAAACCTGGAAAGTCGCCACAGCTGCTGATCTATGCGGCAACTAACCTG GCGGACGGCGTACCCTCGCGTTTTTCCGGGTCTGGCAGCGGTACCGATTTCTCC TTCACTATTAGCAGCCTTCAGCCGGAGGACTTTGCTACATATTATTGTCAGCAA GTGTACTCATCCCCTTTCACCTTCGGCCAAGGCACCAAACTAGAAATAAAAGGC GGGGGCGGTTCACAAATTCAGCTGGTGCAAAGCGGCGGCGAAGTGAAGAAGCCT GGAGCTAGCGTGCGTGTGAGCTGTAAGGCATCTGGCTACTCCTTCACTGATTAC AACGTGTATTGGGTTAGACAGTCCCCTGGTAAAGGGCTTGAGTGGATTGGCTAT AT AGAC OCT T AT AAC GG C AT TAG CAT AT AC GAT C AAAAC T T C AAAG G T AAG G C C AC C C T GAC C G T T GAC AAG T C AAC AAG CAC C G C C T AC AT GGAG C T GAG C AGC C TA AGAAGCGAGGACACCGCTGTCTATTTCTGTGCTAGAGACGTAACCACAGCCCTG GATTTCTGGGGCCAAGGCACAACGGTGACAGTCAGCTCGGGTGGCGGCGGTTCG GGTGGGGGAGGAAGCGGGGGGGGGGGATCCGACATTCAAATGACGCAAAGCCCT GCTAGCCTTTCCGCTAGCGTGGGAGACCGCGTTACGATCACATGTCTGGCCTCT CAGACCATCGACACTTGGCTGGCCTGGTACCTACAAAAACCTGGCAAAAGCCCA CAACTACTGATCTATGCTGCTACGAATCTGGCCGACGGCGTACCGTCCCGCTTC AGCGGCTCCGGTTCTGGGACAGACTTCTCTTTCACCATTTCATCTCTGCAGCCT GAAGATTTCGCCACATATTATTGTCAACAAGTGTATAGCAGCCCGTTTACCTTT GGGCAAGGCACGAAGTTAGAAATCAAGGGCGGAGGCGGATCCCAAATTCAGCTC GTGCAGAGTGGCGGGGAAGTCAAGAAACCTGGCGCTAGCGTGAGAGTGTCTTGT AAGGCAAGTGGCTACAGCTTTACCGACTATAATGTGTATTGGGTGAGACAATCA CCCGGCAAGGGGCTGGAGTGGATCGGCTACATCGACCCGTATAACGGCATTACG ATCTACGACCAAAACTTTAAAGGCAAGGCCACCCTAACGGTCGACAAGTCGACC TCCACCGCTTATATGGAGCTTAGCTCATTGAGAAGCGAGGATACCGCGGTGTAC TTCTGCGCTAGAGACGTGACAACCGCCCTGGACTTCTGGGGGCAAGGCACAACC GTCACCGTGTCGTCC (SEQ ID NO: 20).Attorney Docket No. 47039-0043W01

[0400] Anti-CD3 Antigen-Binding Domains

[0401] In some embodiments, the first target-binding domain, the second target-binding domain, and / or one or more of the one or more additional target-binding domains includes or is an anti-CD3 antibody. In some embodiments, the anti-CD3 antibody can include a heavy chain that includes a sequence that is at least 70% identical (e.g., at least 75% identical, at least 80% identical, at least 85% identical, at least 90% identical, at least 92% identical, at least 94% identical, at least 95% identical, at least 96% identical, at least 98% identical, or 100% identical) to:

[0402] EVQLVESGGGLVQPGGSLRLSCAASGYSFTGYTMNWVRQAPGKGLEWVALINPY KGVSTYNQKFKDRFTISVDKSKNTAYLQMNSLRAEDTAVYYCARSGYYGDSDWY FDVWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVS WNSGALTSGVHTFPAVLQSSGLYSLSSWTVPSSSLGTQTYICNVNHKPSNTKV DKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVWD VSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRWSVLTVLHQDWLNGKEY KCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFY PSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSV MHEALHNHYTQKSLSLSPGK (SEQ IDNO: 94),

[0403] and / or a light chain that includes a sequence that is at least 70% identical (e.g., at least 75% identical, at least 80% identical, at least 85% identical, at least 90% identical, at least 92% identical, at least 94% identical, at least 95% identical, at least 96% identical, at least 98% identical, or 100% identical) to:

[0404] DIQMTQSPSSLSASVGDRVTITCRASQDIRNYLNWYQQKPGKAPKLLIYYTSRL ESGVPSRFSGSGSGTDYTLTISSLQPEDFATYYCQQGNTLPWTFGQGTKVEIKR TVAAPSVFI FPPSDEQLKSGTASWCLLNNFYPREAKVQWKVDNALQSGNSQES VTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC

[0405] (SEQ ID NO: 95).

[0406] In some embodiments, the anti-CD3 antibody can include a heavy chain that includes a sequence that is at least 70% identical (e.g., at least 75% identical, at least 80% identical, at least 85% identical, at least 90% identical, at least 92% identical, at least 94% identical, at least 95% identical, at least 96% identical, at least 98% identical, or 100% identical) to:

[0407] QVQLVQSGGGWQPGRSLRLSCKASGYTFTRYTMHWVRQAPGKGLEWIGYINPS RGYTNYNQKVKDRFTISRDNSKNTAFLQMDSLRPEDTGVYFCARYYDDHYCLDY WGQGTPVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNS GALTSGVHTFPAVLQSSGLYSLSSWTVPSSSLGTQTYICNVNHKPSNTKVDKK VEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCWVDVSH EDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRWSVLTVLHQDWLNGKEYKCKAttorney Docket No. 47039-0043W01

[0408] VSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSD IAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHE ALHNHYTQKSLSLSPGK (SEQ ID NO: 21),

[0409] and / or a light chain that includes a sequence that is at least 70% identical (e.g., at least 75% identical, at least 80% identical, at least 85% identical, at least 90% identical, at least 92% identical, at least 94% identical, at least 95% identical, at least 96% identical, at least 98% identical, or 100% identical) to:

[0410] DIQMTQSPSSLSASVGDRVTITCSASSSVSYMNWYQQTPGKAPKRWIYDTSKLA SGVPSRFSGSGSGTDYTFTISSLQPEDIATYYCQQWSSNPFTFGQGTKLQITRT VAAPSVFIFPPSDEQLKSGTASWCLLNNFYPREAKVQWKVDNALQSGNSQESV TEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC

[0411] (SEQ ID NO: 22).

[0412] In some embodiments of any of the multi-chain chimeric polypeptides described herein, the additional target-binding domain includes or is an anti-CD3 antibody. In some embodiments, the anti-CD3 antibody can include a heavy chain that includes a sequence that is at least 70% identical (e.g., at least 75% identical, at least 80% identical, at least 85% identical, at least 90% identical, at least 92% identical, at least 94% identical, at least 95% identical, at least 96% identical, at least 98% identical, or 100% identical) to:

[0413] QVQLQQSGAELARPGASVKMSCKASGYTFTRYTMHWVKQRPGQGLEWIGYINPSRGYTN YNQKFKDKATLTTDKSSSTAYMQLSSLTSEDSAVYYCARYYDDHYCLDYWGQGTTLTVS SAKTTAPSVYPLAPVCGGTTGSSVTLGCLVKGYFPEPVTLTWNSGSLSSGVHTFPAVLQ SDLYTLSSSVTVTSSTWPSQSITCNVAHPASSTKVDKKIEPRPKSCDKTHTCPPCPAPE LLGGPSVFLFPPKPKDTLMISRTPEVTCVWDVSHEDPEVKFNWYVDGVEVHNAKTKPR EEQYNSTYRWSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTL PPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKL TVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK (SEQ ID NO: 23),

[0414] and / or a light chain that includes a sequence that is at least 70% identical (e.g., at least 75% identical, at least 80% identical, at least 85% identical, at least 90% identical, at least 92% identical, at least 94% identical, at least 95% identical, at least 96% identical, at least 98% identical, or 100% identical) to:

[0415] QIVLTQSPAIMSASPGEKVTMTCSASSSVSYMNWYQQKSGTSPKRWI YDTSKLA SGVPAHFRGSGSGTSYSLTISGMEAEDAATYYCQQWSSNPFTFGSGTKLEINRA DTAPTVS I FPPSSEQLTSGGASWCFLNNFYPKDINVKWKIDGSERQNGVLNSW TDQDSKDSTYSMSSTLTLTKDEYERHNSYTCEATHKTSTSPIVKSFNRNEC

[0416] (SEQ ID NO: 24).Attorney Docket No. 47039-0043W01

[0417] scCD3 Antibody

[0418] In some embodiments, the first target-binding domain, the second target-binding domain, and / or one or more of the one or more additional target-binding domains includes or is a singlechain anti-CD3 antibody. In some embodiments, a single chain anti-CD3 (scCD3) antibody includes a sequence that is at least 70% identical (e.g., at least 75% identical, at least 80% identical, at least 85% identical, at least 90% identical, at least 92% identical, at least 94% identical, at least 95% identical, at least 96% identical, at least 98% identical, or 100% identical) to:

[0419] DIQMTQSPSSLSASVGDRVTITCRASQDIRNYLNWYQQKPGKAPKLLIYYTSRL ESGVPSRFSGSGSGTDYTLTISSLQPEDFATYYCQQGNTLPWTFGQGTKVEIKG GGGSGGGGSGGGGSEVQLVESGGGLVQPGGSLRLSCAASGYSFTGYTMNWVRQA PGKGLEWVALINPYKGVSTYNQKFKDRFTISVDKSKNTAYLQMNSLRAEDTAVY YCARSGYYGDSDWYFDVWGQGTLVTVSS (SEQ ID NO: 25).

[0420] In some embodiments, a scCD3 antibody is encoded by a nucleic acid including a sequence at least 70% identical (e.g., at least 75% identical, at least 80% identical, at least 85% identical, at least 90% identical, at least 92% identical, at least 94% identical, at least 95% identical, at least 96% identical, at least 98% identical, or 100% identical) to:

[0421] GATATTCAGATGACGCAGTCGCCAAGCTCACTCTCGGCCTCCGTAGGAGACAGA G T TAG CAT C AC C T G C AGAG C T AG C C AGAC AT C AGAAAC T AC C T GAAC T GG T AT CAGCAGAAGCCTGGCAAGGCCCCTAAGCTCCTGATCTACTACACGAGCAGACTT GAGAGCGGGGTACCATCCCGCTTCAGCGGATCTGGAAGCGGCACAGACTACACC CTGACCATCTCTAGTCTGCAGCCTGAGGACTTCGCCACTTACTATTGTCAGCAA GGCAACACCCTGCCTTGGACCTTTGGCCAAGGCACCAAAGTTGAGATAAAGGGA GGGGGAGGGAGCGGTGGCGGAGGCTCGGGTGGGGGTGGCTCAGAAGTACAGCTG GTTGAGTCCGGCGGTGGTCTGGTGCAGCCTGGCGGAAGCCTGAGACTGAGCTGC GCCGCAAGCGGCTATAGCTTCACTGGCTATACCATGAACTGGGTGCGCCAAGCC CCTGGCAAGGGCCTGGAATGGGTGGCCCTGATCAACCCTTACAAGGGCGTGAGC AC C T AC AAT CAGAAG T T C AAG GAC AGAT T C AC CAT C AG C G T G GAC AAG GC AAG AACACCGCCTACCTGCAGATGAACAGCCTGAGAGCCGAGGACACTGCGGTCTAT TACTGTGCTAGAAGCGGCTACTACGGCGACAGCGACTGGTACTTCGACGTGTGG GGGCAAGGCACCTTGGTAACGGTAAGCTCC (SEQ ID NO: 26).

[0422] In some embodiments, a scCD3 antibody includes a sequence that is at least 70% identical (e.g., at least 75% identical, at least 80% identical, at least 85% identical, at least 90% identical, at least 92% identical, at least 94% identical, at least 95% identical, at least 96% identical, at least 98% identical, or 100% identical) to:Attorney Docket No. 47039-0043W01

[0423] DIKLQQSGAELARPGASVKMSCKTSGYTFTRYTMHWVKQRPGQGLEWIGYINPS RGYTNYNQKFKDKATLTTDKSSSTAYMQLSSLTSEDSAVYYCARYYDDHYCLDY WGQGTTLTVSSVEGGSGGSGGSGGSGGVDDIQLTQSPAIMSASPGEKVTMTCRA SSSVSYMNWYQQKSGTSPKRWIYDTSKVASGVPYRFSGSGSGTSYSLTISSMEA EDAATYYCQQWSSNPLTFGAGTKLELK (SEQ ID NO: 27).

[0424] In some embodiments, a scCD3 antibody is encoded by a nucleic acid including a sequence at least 70% identical (e.g., at least 75% identical, at least 80% identical, at least 85% identical, at least 90% identical, at least 92% identical, at least 94% identical, at least 95%> identical, at least 96% identical, at least 98% identical, or 100% identical) to:

[0425] GACATCAAGCTGCAGCAGAGCGGCGCCGAGCTGGCTAGACCTGGGGCTAGCGTC AAGATGAGC TGCAAGACAAGCGGC TACAC T T T CACAAGATACACCAT GCAC T GG GTGAAGCAGAGACCTGGCCAAGGCCTGGAATGGATCGGTTATATCAACCCTAGC AGAG GC TACAC C AAC T AC AAT C AGAAG T T C AAGGAC AAAG C C AC C C T GAC AAC T GATAAGAGCAGCAGCACCGCCTATATGCAACTGTCAAGCCTCACCTCAGAGGAC AGCGCCGTGTACTACTGCGCTAGATACTACGACGACCACTACTGCCTGGACTAT TGGGGCCAAGGCACTACTTTGACTGTTAGCTCGGTCGAGGGAGGCTCCGGCGGG AGTGGTGGTTCTGGCGGCAGCGGCGGAGTTGACGACATTCAGCTGACACAAAGT CCTGCCATAATGAGCGCTAGCCCTGGCGAGAAGGTGACCATGACCTGCAGAGCT AGCAGCAGCGTGAGCTACATGAACTGGTATCAGCAGAAGAGCGGCACAAGCCCT AAAAGATGGATCTACGACACAAGCAAGGTGGCTAGCGGCGTGCCTTACCGGTTT AGCGGCTCTGGTAGCGGCACAAGCTATAGCCTCACGATTAGCTCAATGGAGGCC GAGGACGCCGCCACGTATTATTGCCAACAGTGGAGCAGCAACCCTCTGACCTTC GGCGCCGGGACCAAGCTGGAACTTAAG (SEQ ID NO: 28).

[0426] Anti-CD3 scFv

[0427] In some embodiments, the first target-binding domain, the second target-binding domain, and / or one or more of the one or more additional target-binding domains includes or is an anti-CD3 scFv. In some embodiments, the anti-CD3 scFv can include a heavy chain variable domain including a sequence that is at least 70% identical (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) to

[0428] EVQLVESGGGLVQPGGSLRLSCAASGYSFTGYTMNWVRQAPGKGLEWVALINPYK GVSTYNQKFKDRFTISVDKSKNTAYLQMNSLRAEDTAVYYCARSGYYGDSDWYFD VWGQGTLVTVSS (SEQ ID NO: 98),Attorney Docket No. 47039-0043W01

[0429] and / or a light chain variable domain including a sequence that is at least 70% identical (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) to:

[0430] DIQMTQSPSSLSASVGDRVTITCRASQDIRNYLNWYQQKPGKAPKLLIYYTSRLE SGVPSRFSGSGSGTDYTLTISSLQPEDFATYYCQQGNTLPWTFGQGTKVEIK

[0431] (SEQ ID NO: 99).

[0432] In some embodiments, a scFv (e.g., any of the scFvs described herein) can include a linker sequence (e.g., any of the exemplary linker sequences described herein or known in the art) between the heavy chain variable domain and the light chain variable domain.

[0433] In some embodiments, the anti-CD3 scFv can include a heavy chain variable domain encoded by a nucleic acid including a sequence that is at least 70% identical (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) to:

[0434] GAAGTACAGCTGGTTGAGTCCGGCGGTGGTCTGGTGCAGCCTGGCGGAAGCCTGA GACTGAGCTGCGCCGCAAGCGGCTATAGCTTCACTGGCTATACCATGAACTGGGT GCGCCAAGCCCCTGGCAAGGGCCTGGAATGGGTGGCCCTGATCAACCCTTACAAG GGC G T GAGCAC C TACAAT CAGAAG T T CAAGGACAGAT T GAG CAT CAGC G T GGACA AGAGCAAGAACACCGCCTACCTGCAGATGAACAGCCTGAGAGCCGAGGACACTGC GGTCTATTACTGTGCTAGAAGCGGCTACTACGGCGACAGCGACTGGTACTTCGAC GTGTGGGGGCAAGGCACCTTGGTAACGGTAAGCTCC (SEQ ID NO: 100)

[0435] and / or a light chain variable domain encoded by a nucleic acid including a sequence that is at least 70% identical (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) to:

[0436] GATATTCAGATGACGCAGTCGCCAAGCTCACTCTCGGCCTCCGTAGGAGACAGA G T TAG CAT C AC C T G C AGAG C T AG C C AAGAC AT C AGAAAC T AC C T GAAC T GG T AT CAGCAGAAGCCTGGCAAGGCCCCTAAGCTCCTGATCTACTACACGAGCAGACTT GAGAGCGGGGTACCATCCCGCTTCAGCGGATCTGGAAGCGGCACAGACTACACC CTGACCATCTCTAGTCTGCAGCCTGAGGACTTCGCCACTTACTATTGTCAGCAA GGCAACACCCTGCCTTGGACCTTTGGCCAAGGCACCAAAGTTGAGATAAAG

[0437] (SEQ ID NO: 101).

[0438] In some embodiments, the anti-CD3 scFv can include a heavy chain variable domain including a sequence that is at least 70% identical (e.g., at least 75% identical, at least 80%Attorney Docket No. 47039-0043W01

[0439] identical, at least 85% identical, at least 90% identical, at least 95% identical, at least 99% identical, or 100% identical) to QVQLQQSGAELARPGASVKMSCKASGYTFTRYTMHWVKQRPGQGLEWIGYINPSR GYTNYNQKFKDKATLTTDKSSSTAYMQLSSLTSEDSAVYYCARYYDDHYCLDYWG QGTTLTVSS (SEQ ID NO: 29),

[0440] and / or a light chain variable domain including a sequence that is at least 70% identical (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) to QIVLTQSPAIMSASPGEKVTMTCSASSSVSYMNWYQQKSGTSPKRWIYDTSKLAS GVPAHFRGSGSGTSYSLTISGMEAEDAATYYCQQWSSNPFTFGSGTKLEINR

[0441] (SEQ ID NO: 30).

[0442] In some embodiments, a scFv (e.g., any of the scFvs described herein) can include a linker sequence (e.g., any of the exemplary linker sequences described herein or known in the art) between the heavy chain variable domain and the light chain variable domain. In some embodiments, the anti-CD3 scFv can include a heavy chain variable domain encoded by a nucleic acid including a sequence that is at least 70% identical (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) to CAAGTTCAGCTCCAGCAAAGCGGCGCCGAACTCGCTCGGCCCGGCGCTTCCGTGA AGATGTCTTGTAAGGCCTCCGGCTATACCTTCACCCGGTACACAATGCACTGGGT CAAGCAACGGCCCGGTCAAGGTTTAGAGTGGATTGGCTATATCAACCCCTCCCGG GGCTATACCAACTACAACCAGAAGTTCAAGGACAAAGCCACCCTCACCACCGACA AGTCCAGCAGCACCGCTTACATGCAGCTGAGCTCTTTAACATCCGAGGATTCCGC CGTGTACTACTGCGCTCGGTACTACGACGATCATTACTGCCTCGATTACTGGGGC CAAGGTACCACCTTAACAGTCTCCTCC (SEQ ID NO: 31),

[0443] and / or a light chain variable domain encoded by a nucleic acid including a sequence that is at least 70% identical (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) to CAGATCGTGCTGACCCAGTCCCCCGCTATTATGAGCGCTAGCCCCGGTGAAAAGG TGACTATGACATGCAGCGCCAGCTCTTCCGTGAGCTACATGAACTGGTATCAGCA GAAGTCCGGCACCAGCCCTAAAAGGTGGATCTACGACACCAGCAAGCTGGCCAGC GGCGTCCCCGCTCACTTTCGGGGCTCCGGCTCCGGAACAAGCTACTCTCTGACCA TCAGCGGCATGGAAGCCGAGGATGCCGCTACCTATTACTGTCAGCAGTGGAGCTC CAACCCCTTCACCTTTGGATCCGGCACCAAGCTCGAGATTAATCGT (SEQ ID

[0444] NO: 32).Attorney Docket No. 47039-0043W01

[0445] In some embodiments, an anti-CD3 scFv can include a sequence that is at least 70% identical (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) to QIVLTQSPAIMSASPGEKVTMTCSASSSVSYMNWYQQKSGTSPKRWIYDTSKLAS GVPAHFRGSGSGTSYSLTISGMEAEDAATYYCQQWSSNPFTFGSGTKLEINRGGG GSGGGGSGGGGSQVQLQQSGAELARPGASVKMSCKASGYTFTRYTMHWVKQRPGQ GLEWIGYINPSRGYTNYNQKFKDKATLTTDKSSSTAYMQLSSLTSEDSAVYYCAR YYDDHYCLDYWGQGTTLTVSS (SEQ ID NO: 33).

[0446] In some embodiments, an anti-CD3 scFv can include a sequence encoded by a nucleic acid sequence that is at least 70% identical (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) to CAGATCGTGCTGACCCAGTCCCCCGCTATTATGAGCGCTAGCCCCGGTGAAAAG GTGACTATGACATGCAGCGCCAGCTCTTCCGTGAGCTACATGAACTGGTATCAG CAGAAGTCCGGCACCAGCCCTAAAAGGTGGATCTACGACACCAGCAAGCTGGCC AGCGGCGTCCCCGCTCACTTTCGGGGCTCCGGCTCCGGAACAAGCTACTCTCTG ACCATCAGCGGCATGGAAGCCGAGGATGCCGCTACCTATTACTGTCAGCAGTGG AGCTCCAACCCCTTCACCTTTGGATCCGGCACCAAGCTCGAGATTAATCGTGGA GGCGGAGGTAGCGGAGGAGGCGGATCCGGCGGTGGAGGTAGCCAAGTTCAGCTC CAGCAAAGCGGCGCCGAACTCGCTCGGCCCGGCGCTTCCGTGAAGATGTCTTGT AAGGCCTCCGGCTATACCTTCACCCGGTACACAATGCACTGGGTCAAGCAACGG CCCGGTCAAGGTTTAGAGTGGATTGGCTATATCAACCCCTCCCGGGGCTATACC AACTACAACCAGAAGTTCAAGGACAAAGCCACCCTCACCACCGACAAGTCCAGC AGCACCGCTTACATGCAGCTGAGCTCTTTAACATCCGAGGATTCCGCCGTGTAC TACTGCGCTCGGTACTACGACGATCATTACTGCCTCGATTACTGGGGCCAAGGT ACCACCTTAACAGTCTCCTCC (SEQ ID NO: 34).

[0447] Pairs of Affinity Domains

[0448] In some embodiments, a multi-chain chimeric polypeptide includes: 1) a first chimeric polypeptide that includes a first domain of a pair of affinity domains, and 2) a second chimeric polypeptide that includes a second domain of a pair of affinity domains such that the first chimeric polypeptide and the second chimeric polypeptide associate through the binding of the first domain and the second domain of the pair of affinity domains. In some embodiments, the pair of affinity domains is a sushi domain from an alpha chain of human IL- 15 receptor (IL15Ra) and a soluble IL- 15. A sushi domain, also known as a short consensus repeat or type 1 glycoprotein motif, is a common motif in protein-protein interaction. Sushi domains have beenAttorney Docket No. 47039-0043W01

[0449] identified on a number of protein-binding molecules, including complement components Clr, Cis, factor H, and C2m, as well as the nonimmunologic molecules factor XIII and 02-gly coprotein. A typical Sushi domain has approximately 60 amino acid residues and contains four cysteines (Ranganathan, Pac. Symp Biocomput. 2000:155-67). The first cysteine can form a disulfide bond with the third cysteine, and the second cysteine can form a disulfide bridge with the fourth cysteine. In some embodiments in which one member of the pair of affinity domains is a soluble IL- 15, the soluble IL- 15 has a D8N or D8A amino acid substitution. In some embodiments in which one member of the pair of affinity domains is an alpha chain of human IL-15 receptor (IL15Ra), the human IL15Ra is a mature full-length IL15Ra. In some embodiments, the pair of affinity domains is barnase and barstar. In some embodiments, the pair of affinity domains is a PKA and an AKAP. In some embodiments, the pair of affinity domains is an adapter / docking tag module based on mutated RNase I fragments (Rossi, Proc Natl Acad Sci USA. 103:6841-6846, 2006; Sharkey et al., Cancer Res. 68:5282-5290, 2008; Rossi et al., Trends Pharmacol Sci. 33:474-481, 2012) or SNARE modules based on interactions of the proteins syntaxin, synaptotagmin, synaptobrevin, and SNAP25 (Deyev et al., Nat Biotechnol. 1486-1492, 2003).

[0450] In some embodiments, a first chimeric polypeptide of a multi-chain chimeric polypeptide includes a first domain of a pair of affinity domains and a second chimeric polypeptide of the multi-chain chimeric polypeptide includes a second domain of a pair of affinity domains, wherein the first domain of the pair of affinity domains and the second domain of the pair of affinity domains bind to each other with a dissociation equilibrium constant (KD) of less than 1 x 10’7M, less than 1 x 10’8M, less than 1 x 10’9M, less than 1 x 1010M, less than 1 x 1011M, less than 1 x 10'12M, or less than 1 x 10'13M. In some embodiments, the first domain of the pair of affinity domains and the second domain of the pair of affinity domains bind to each other with a KD of about 1 x 10‘4M to about 1 x IO'6M, about 1 x IO'5M to about 1 x 10’7M, about l x 10’6M to about 1 x 10'8M, about 1 x 10'7M to about 1 x 10'9M, about 1 x 10'8M to about 1 x 10‘10M, about 1 x 10’9M to about 1 x 10'11M, about 1 x 10’10M to about 1 x 10'12M, about 1 x 10'11M to about 1 x 10'13M, about 1 x 10'4M to about 1 x 10’5M, about 1 x 10'3M to about l x 10'6M, about 1 x 10'6M to about 1 x 10'7M, about 1 x 10'7M to about 1 x 10'8M, about 1 x 10'8M to about 1 x 10'9M, about 1 x 10'9M to about 1 x 10’10M, about 1 x 10'10M to about 1 x 10'11M, about 1 x 10'11M to about 1 x 10'12M, or about 1 x ICT12M to about 1 x 10'13M (inclusive).Attorney Docket No. 47039-0043W01

[0451] Any of a variety of different methods known in the art can be used to determine the KD value of the binding of the first domain of the pair of affinity domains and the second domain of the pair of affinity domains (e.g., an electrophoretic mobility shift assay, a filter binding assay, surface plasmon resonance, and a biomolecular binding kinetics assay, etc.).

[0452] In some embodiments, a first chimeric polypeptide of a multi-chain chimeric polypeptide includes a first domain of a pair of affinity domains and a second chimeric polypeptide of the multi-chain chimeric polypeptide includes a second domain of a pair of affinity domains, wherein the first domain of the pair of affinity domains, the second domain of the pair of affinity domains, or both is about 10 to 100 amino acids in length. For example, a first domain of a pair of affinity domains, a second domain of a pair of affinity domains, or both can be about 10 to 100 amino acids in length, about 15 to 100 amino acids in length, about 20 to 100 amino acids in length, about 25 to 100 amino acids in length, about 30 to 100 amino acids in length, about 35 to 100 amino acids in length, about 40 to 100 amino acids in length, about 45 to 100 amino acids in length, about 50 to 100 amino acids in length, about 55 to 100 amino acids in length, about 60 to 100 amino acids in length, about 65 to 100 amino acids in length, about 70 to 100 amino acids in length, about 75 to 100 amino acids in length, about 80 to 100 amino acids in length, about 85 to 100 amino acids in length, about 90 to 100 amino acids in length, about 95 to 100 amino acids in length, about 10 to 95 amino acids in length, about 10 to 90 amino acids in length, about 10 to 85 amino acids in length, about 10 to 80 amino acids in length, about 10 to 75 amino acids in length, about 10 to 70 amino acids in length, about 10 to 65 amino acids in length, about 10 to 60 amino acids in length, about 10 to 55 amino acids in length, about 10 to 50 amino acids in length, about 10 to 45 amino acids in length, about 10 to 40 amino acids in length, about 10 to 35 amino acids in length, about 10 to 30 amino acids in length, about 10 to 25 amino acids in length, about 10 to 20 amino acids in length, about 10 to 15 amino acids in length, about 20 to 30 amino acids in length, about 30 to 40 amino acids in length, about 40 to 50 amino acids in length, about 50 to 60 amino acids in length, about 60 to 70 amino acids in length, about 70 to 80 amino acids in length, about 80 to 90 amino acids in length, about 90 to 100 amino acids in length, about 20 to 90 amino acids in length, about 30 to 80 amino acids in length, about 40 to 70 amino acids in length, about 50 to 60 amino acids in length, or any range in between. In some embodiments, a first domain of a pair of affinity domains, a second domain of a pair of affinity domains, or both isAttorney Docket No. 47039-0043W01

[0453] about 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, or 100 amino acids in length.

[0454] In some embodiments, any of the first and / or second domains of a pair of affinity domains disclosed herein can include one or more additional amino acids (e.g., 1, 2, 3, 5, 6, 7, 8, 9, 10, or more amino acids) at its N-terminus and / or C-terminus, so long as the function of the first and / or second domains of a pair of affinity domains remains intact. For example, a sushi domain from an alpha chain of human IL- 15 receptor (IL15Ra) can include one or more additional amino acids at the N-terminus and / or the C-terminus, while still retaining the ability to bind to a soluble IL-15. Additionally or alternatively, a soluble IL- 15 can include one or more additional amino acids at the N-terminus and / or the C-terminus, while still retaining the ability to bind to a sushi domain from an alpha chain of human IL- 15 receptor (IL15Ra).

[0455] Exemplary Pairs of Affinity Domains

[0456] In some embodiments, a first affinity domain and a second affinity domain of a pair of affinity domains can be a sushi domain from an alpha chain IL- 15 receptor alpha and a soluble IL-15.

[0457] Anon-limiting example of a sushi domain from an alpha chain of IL- 15 receptor (IL15Roi) can include a sequence that is at least 70% identical, 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 to:

[0458] ITCPPPMSVEHADIWVKSYSLYSRERYICNSGFKRKAGTSSLTECVLNKATNVAH WTTPSLKCIR (SEQ ID NO: 35).

[0459] In some embodiments, a sushi domain from an alpha chain of IL15Ra can be encoded by a nucleic acid including:

[0460] ATCACGTGCCCTCCCCCCATGTCCGTGGAACACGCAGACATCTGGGTCAAGAGC TACAGCTTGTACTCCAGGGAGCGGTACATTTGTAACTCTGGTTTCAAGCGTAAA GCCGGCACGTCCAGCCTGACGGAGTGCGTGTTGAACAAGGCCACGAATGTCGCC CACTGGACAACCCCCAGTCTCAAATGCATTAGAG (SEQ ID NO: 36), or ATTACATGCCCCCCTCCCATGAGCGTGGAGCACGCCGACATCTGGGTGAAGAGC TATAGCCTCTACAGCCGGGAGAGGTATATCTGTAACAGCGGCTTCAAGAGGAAG GCCGGCACCAGCAGCCTCACCGAGTGCGTGCTGAATAAGGCTACCAACGTGGCT CACTGGACAACACCCTCTTTAAAGTGCATCCGG (SEQ ID NO: 37).Attorney Docket No. 47039-0043W01

[0461] In some embodiments, a sushi domain from an alpha chain of IL- 15 receptor alpha (IL15Ra) can include a sequence that is at least 70% identical, 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 to:

[0462] ITCPPPMSVEHADIWVKSYSLYSRERYICNSGFKRKAGTSSLTECVLNKATNVA HWTTPSLKCIRDPALVHQR (SEQ ID NO: 38).

[0463] In some embodiments, a sushi domain from an alpha chain of IL15Ra can be encoded by a nucleic acid including:

[0464] ATCACGTGCCCTCCCCCCATGTCCGTGGAACACGCAGACATCTGGGTCAAGAGC TACAGCTTGTACTCCAGGGAGCGGTACATTTGTAACTCTGGTTTCAAGCGTAAA GCCGGCACGTCCAGCCTGACGGAGTGCGTGTTGAACAAGGCCACGAATGTCGCC CACTGGACAACCCCCAGTCTCAAATGCATTAGAGACCCTGCCCTGGTTCACCAA AGG (SEQ IDNO: 39).

[0465] In some embodiments, a soluble IL- 15 can include a sequence that is at least 70% identical, 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 to:

[0466] NWVNVISDLKKIEDLIQSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLES GDASIHDTVENLI ILANNSLSSNGNVTESGCKECEELEEKNIKEFLQSFVHIVQ MFINTS (SEQ ID NO: 40).

[0467] In some embodiments, a soluble IL- 15 can be encoded by a nucleic acid including the sequence of:

[0468] AAC T GG G T GAAT G T AAT AAG T GAT T T GAAAAAAAT T GAAGAT C T T AT T C AAT C T AT G CAT AT T GAT GCTACTTTATATACG GAAAG T GAT G T T C AC C C C AG T T GC AAA GTAACAGCAATGAAGTGCTTTCTCTTGGAGTTACAAGTTATTTCACTTGAGTCC GGAGAT GCAAGTAT T CAT GATACAGTAGAAAATC T GAT CAT CC TAGCAAACAAC AGTTTGTCTTCTAATGGGAATGTAACAGAATCTGGATGCAAAGAATGTGAGGAA C T G GAG GAAAAAAAT AT T AAGAAT T T T T G C AGAG T T T T G TAG AT AT T G T C C A ATGTTCATCAACACTTCT (SEQ ID NO: 41), or

[0469] AAC T GG G T GAAC G T GAT C AG C GAC C T GAAGAAGAT C GAGGAC C T GAT T C AGAG C ATGCACATCGACGCCACCCTGTACACCGAGAGCGACGTGCACCCTAGCTGCAAG GTGACCGCCATGAAGTGCTTCCTGCTGGAGCTGCAAGTGATCAGCCTGGAGAGC GGCGACGCTAGCATCCACGACACCGTGGAGAACCTGATCATCCTGGCCAACAAC AGCCTGAGCAGCAACGGCAACGTGACCGAGAGCGGCTGCAAGGAGTGCGAGGAG CTGGAGGAGAAGAACATCAAGGAGTTCCTGCAGAGCTTCGTGCACATCGTGCAG ATGTTCATCAACACAAGC (SEQ ID NO: 42).Attorney Docket No. 47039-0043W01

[0470] Linker Sequences

[0471] In some embodiments, a chimeric polypeptide described herein (e.g., one or more of a single-chain polypeptide, a first chimeric polypeptide of a multi-chain chimeric polypeptide, or a second chimeric polypeptide of a multi-chain chimeric polypeptide) can include one, two, three, four, five, six, seven, eight, nine, or ten linker sequence(s) (e.g., the same or different linker sequences, e.g., any of the exemplary linker sequences described herein or known in the art). In some embodiments, a chimeric polypeptide described herein (e.g., one or more of a single-chain polypeptide, a first chimeric polypeptide of a multi-chain chimeric polypeptide, or a second chimeric polypeptide of a multi-chain chimeric polypeptide) can include one, two, three, four, five, six, seven, eight, nine, or ten linker sequence(s) (e.g., the same or different linker sequences, e.g., any of the exemplary linker sequences described herein or known in the art).

[0472] In some embodiments, the linker sequence can be a flexible linker sequence. Nonlimiting examples of linker 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 Sci., 2013 Feb; 22(2):153-167. In some examples, the linker sequence is a synthetic linker sequence.

[0473] In some embodiments, a linker sequence can have a total length of 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 to about 14 amino acids, 1 amino acid to about 12 amino acids, 1 amino acid to about 10 amino acids, 1 amino acid to about 8 amino acids, 1 amino acid to about 6 amino acids, 1 amino acid to about 4 amino acids, about 2 amino acids to about 100 amino acids, about 2 amino acids to about 90 amino acids, about 2 amino acids to about 80 amino acids, about 2 amino acids to about 70 amino acids, about 2 amino acids to about 60 amino acids, about 2 amino acids to about 50 amino acids, about 2 amino acids to about 45 amino acids, about 2 amino acids to about 40 amino acids, about 2 amino acids to about 35 amino acids, about 2 amino acids to about 30 amino acids, about 2 amino acids to about 25 amino acids, about 2Attorney Docket No. 47039-0043W01

[0474] amino acids to about 24 amino acids, about ...

Claims

Attorney Docket No. 47039-0043W01WHAT TS CLAIMED IS:

1. A multi-chain chimeric polypeptide comprising:(a) a first chimeric polypeptide comprising:(i) a first target-binding domain;(ii) a soluble T cell receptor -chain constant region (TRpC) domain; and (iii) a first domain of a pair of affinity domains; and(b) a second chimeric polypeptide comprising:(i) a second domain of a pair of affinity domains; and(ii) a second target-binding domain,wherein the first chimeric polypeptide and the second chimeric polypeptide associate through the binding of the first domain and the second domain of the pair of affinity domains, and wherein the first-target binding domain and / or the second target binding domain is an anti-mesothelin antigen-binding domain.

2. The multi-chain chimeric polypeptide of claim 1, wherein the first target-binding domain and the soluble TRpC domain directly abut each other in the first chimeric polypeptide.

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

4. The multi-chain chimeric polypeptide of any one of claims 1-3, wherein the soluble TRpC domain and the first domain of the pair of affinity domains directly abut each other in the first chimeric polypeptide.

5. The multi-chain chimeric polypeptide of any one of claims 1-3, wherein the first chimeric polypeptide further comprises a linker sequence between the soluble TR0C domain and the first domain of the pair of affinity domains in the first chimeric polypeptide.Attorney Docket No. 47039-0043W016. The multi-chain chimeric polypeptide of any one of claims 1-5, wherein the second domain of the pair of affinity domains and the second target-binding domain directly abut each other in the second chimeric polypeptide.

7. The multi-chain chimeric polypeptide of any one of claims 1-5, wherein the second chimeric polypeptide further comprises a linker sequence between the second domain of the pair of affinity domains and the second target-binding domain in the second chimeric polypeptide.

8. The multi-chain chimeric polypeptide of any one of claims 1-7, wherein the first chimeric polypeptide further comprises one or more additional target-binding domains.

9. The multi-chain chimeric polypeptide of claim 8, wherein the first domain of the pair of affinity domains and one of the one or more additional target-binding domains directly abut each other in the first chimeric polypeptide.

10. The multi-chain chimeric polypeptide of claim 8, wherein the first chimeric polypeptide further comprises a linker sequence between the first domain of the pair of affinity domains and one of the one or more additional target-binding domains in the first chimeric polypeptide.

11. The multi-chain chimeric polypeptide of any one of claims 1-10, wherein the second chimeric polypeptide further comprises one or more additional target-binding domains.

12. The multi-chain chimeric polypeptide of claim 11, wherein the second domain of the pair of affinity domains and one of the one or more additional target-binding domains directly abut each other in the second chimeric polypeptide.

13. The multi-chain chimeric polypeptide of claim 11, wherein the second chimeric polypeptide further comprises a linker sequence between the second domain of the pair of affinity domains and one of the one or more additional target-binding domains in the second chimeric polypeptide.Attorney Docket No. 47039-0043W0114. The multi-chain chimeric polypeptide of any one of claims 8-13, wherein the one or more additional target-binding domains bind specifically to CD3.

15. The multi-chain chimeric polypeptide of any one of claims 8-14, wherein the one or more additional target-binding domains comprises an anti-CD3 antibody.

16. The multi-chain chimeric polypeptide of any one of claims 1-15, wherein both the first target-binding domain and the second target-binding domain are an antigen-binding domain.

17. The multi-chain chimeric polypeptide of any one of claims 1-16, wherein the first target-binding domain or the second target-binding domain binds specifically to a target selected from the group consisting of Tissue Factor (CD142), 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, TNFa, CD26a, CD36, ULBP2, CD30, CD200, IGF-1R, MUC4AC, MUC5AC, Trop-2, CMET, EGFR, HER1, HER2, HER3, PSMA, CEA, B7H3, EPCAM, BCMA, P-cadherin, CEACAM5, uPAR, a UL 16-binding protein, HLA-DR, DLL4, TYR03, AXL, MER, CD122, CD155, PDGFDD, a ligand of TGF- receptor II (TGF-0RII), a ligand of TGF-PRIII, a ligand of DNAM1, a ligand of NKp46, a ligand of NKp44, a ligand of NKG2D, a ligand of NKp30, a ligand for a scMHCI, a ligand for a scMHCII, a ligand for a scTCR, a receptor for IL-1, a receptor for IL-2, a receptor for IL-3, a receptor for IL-4, a receptor for IL-7, a receptor for IL-8, a receptor for IL-10, a receptor for IL-12, a receptor for IL-15, a receptor for IL- 17, a receptor for IL- 18, a receptor for IL-21, a receptor for PDGF-DD, a receptor for stem cell factor (SCF), a receptor for stem cell-like tyrosine kinase 3 ligand (FLT3L), a receptor for MICA, a receptor for MICB, a receptor for a ULP16-binding protein, a receptor for CD155, a receptor for CD 122, and a receptor for CD28.

18. The multi-chain chimeric polypeptide of any one of claims 1-15, wherein one of the first target-binding domain and the second target-binding domain is a soluble interleukin or cytokine protein.Attorney Docket No. 47039-0043W0119. The multi-chain chimeric polypeptide of claim 18, wherein the soluble interleukin, cytokine, or ligand protein is selected from the group consisting of: IL-1, IL-2, IL-3, IL-4, IL-7, IL-8, IL-10, IL-12, IL-15, IL-17, IL-18, IL-21, PDGF-DD, SCF, and FLT3L.

20. The multi-chain chimeric polypeptide of claim 19, wherein the soluble interleukin, soluble cytokine protein, or soluble cell surface protein is IL-7.

21. The multi-chain chimeric polypeptide of any one of claims 1-15, wherein one or both of the first target-binding domain and the second target-binding domain is a soluble interleukin or cytokine receptor.

22. The multi-chain chimeric polypeptide of claim 21, wherein the soluble receptor is a soluble TGF-0 receptor II (TGF- RII), a soluble TGF- RIII, a soluble NKG2D, a soluble NKp30, a soluble NKp44, a soluble NKp46, a soluble DNAM-1, a scMHCI, a scMHCII, a scTCR, a soluble CD155, or a soluble CD28.

23. The multi-chain chimeric polypeptide of claim 22, wherein the soluble interleukin receptor, the soluble cytokine receptor, or the soluble cell surface receptor is a soluble TGF-βRII.

24. The multi-chain chimeric polypeptide of any one of claims 1-15, wherein the first target-binding domain is IL-7 and the second target-binding domain is the anti-mesothelin antigen-binding domain.

25. The multi-chain chimeric polypeptide of claim 24, wherein the second chimeric polypeptide further comprises an anti-CD3 antigen-binding domain.

26. The multi-chain chimeric polypeptide of any one of claims 1-15, wherein the first target-binding domain is a soluble TGF-βRII and the second target-binding domain comprises the anti-mesothelin antigen-binding domain.Attorney Docket No. 47039-0043W0127. The multi-chain chimeric polypeptide of claim 26, wherein the second target-binding domain further comprises an anti-tissue factor antigen-binding domain.

28. The multi-chain chimeric polypeptide of claim 27, wherein the second chimeric polypeptide further comprises an anti-CD3 antigen-binding domain.

29. The multi-chain chimeric polypeptide of any one of claims 1-15, wherein the first target-binding domain is a soluble TGF-βRII and the second target-binding domain is the anti-mesothelin antigen-binding domain.

30. The multi-chain chimeric polypeptide of claim 29, wherein the second chimeric polypeptide further comprises an anti-CD3 antigen-binding domain.

31. The multi-chain chimeric polypeptide of any one of claims 1-15, wherein one of the first target-binding domain and the second target-binding domain is an anti-tissue factor antigenbinding domain.

32. The multi-chain chimeric polypeptide of claim 31, wherein the first target-binding domain is the anti-mesothelin antigen-binding domain and the second target-binding domain is an anti -tissue factor antigen-binding domain.

33. The multi-chain chimeric polypeptide of claim 32, wherein the second chimeric polypeptide further comprises an anti-CD3 antigen-binding domain.

34. The multi-chain chimeric polypeptide of any one of claims 1-33, wherein the soluble TRpC domain is a soluble human TRpC domain.

35. The multi-chain chimeric polypeptide of any one of claims 1-34, wherein the soluble TRpC domain does not bind to an MHC protein with or without a loaded antigen.Attorney Docket No. 47039-0043W0136. The multi-chain chimeric polypeptide of any one of claims 1-35, wherein the soluble TRβC domain does not stimulate an immune response in a mammal.

37. The multi-chain chimeric polypeptide of any one of claims 1-36, wherein the soluble TRβC domain comprises or consists of a sequence from a wildtype soluble human TRβC.

38. The multi-chain chimeric polypeptide of any one of claims 1-37, wherein the pair of affinity domains is a sushi domain from an alpha chain of human IL- 15 receptor (IL15Ra) and a soluble IL-15.

39. The multi-chain chimeric polypeptide of any one of claims 1-37, wherein the pair of affinity domains is selected from the group consisting of: barnase and barstar, a PKA and an AKAP, adapter / docking tag modules based on mutated RNase I fragments, and SNARE modules based on interactions of the proteins syntaxin, synaptotagmin, synaptobrevin, and SNAP25.

40. The multi-chain chimeric polypeptide of any one of claims 1-39, wherein the first chimeric polypeptide and / or the second chimeric polypeptide further comprises a signal sequence at its N-terminal end.

41. A composition comprising any of the multi-chain chimeric polypeptides of claims 1-40.

42. The composition of claim 41, wherein the composition is a pharmaceutical composition.

43. A kit comprising at least one dose of the composition of claim 41 or 42.

44. A method of stimulating an immune cell, the method comprising:contacting an immune cell with an effective amount of any of the multi-chain chimeric polypeptides of claims 1-40 or the composition of claim 41 or 42.Attorney Docket No. 47039-0043W0145. A method of inducing or increasing proliferation of an immune cell, the method comprising:contacting an immune cell with an effective amount of any of the multi-chain chimeric polypeptides of claims 1-40 or the composition of claim 41 or 42.

46. A method of inducing differentiation of an immune cell into a memory or memorylike immune cell, the method comprising:contacting an immune cell with an effective amount of any of the multi-chain chimeric polypeptides of claims 1-40 or the composition of claim 41 or 42.

47. The method of any one of claims 44-46, wherein the immune cell is contacted in vitro.

48. The method of any one of claims 44-46, wherein the immune cell is contacted in vivo.

49. The method of any one of claims 44-48, wherein the immune cell is selected from the group consisting of: an immature thymocyte, a peripheral blood lymphocyte, a naive T cell, a pluripotent Th cell precursor, a lymphoid progenitor cell, a Treg cell, a Thl7 cell, a Th22 cell, a Th9 cell, a Th2 cell, a Thl cell, a Th3 cell, y8 T cell, an o.p T cell, a tumor-infiltrating T cell, a CD8+ T cell, a CD4+ T cell, a natural killer T cell, a mast cell, a macrophage, a neutrophil, a dendritic cell, a basophil, an eosinophil, and a natural killer cell.

50. The method of claim 49, wherein the CD8+T cell is a progenitor exhausted CD8+T (Tpex) cell, an effector memory CD8+T (Tern) cell, a central memory CD8+T (Tcm) cell, a memory CD8+stem (Tscm) cell, a virtual memory CD8+T (Tvm) cell, an antigen-experienced CD8+T cell, or a terminally-exhausted CD8+T (Tex) cell.

51. The method of any one of claims 44-50, wherein the immune cell has previously been genetically modified to express a chimeric antigen receptor or a recombinant T-cell receptor.Attorney Docket No. 47039-0043W0152. A method of killing a cancer cell, an infected cell, or a senescent cell in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of any of the multi-chain chimeric polypeptides claims 1-40 or the composition of claim 41 or 42.

53. A method of treating a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of any of the multi-chain chimeric polypeptides of claims 1-40 or the composition of claim 41 or 42.

54. The method of claim 52 or 53, wherein the subject has been identified or diagnosed as having a cancer, an aging-related disease or condition, or an infectious disease.

55. The method of claim 54, wherein the cancer is selected from the group consisting of: solid tumor, hematological tumor, sarcoma, osteosarcoma, 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 lymphocytic leukemia (ALL), myelodysplastic syndromes (MDS), cutaneous T-cell lymphoma, retinoblastoma, stomach cancer, urothelial carcinoma, lung cancer, renal cell carcinoma, gastic and esophageal cancer, pancreatic cancer, prostate cancer, breast cancer, colorectal cancer, ovarian cancer, non-small cell lung carcinoma, squamous cell head and neck carcinoma, endometrial cancer, cervical cancer, liver cancer, biliary cancer, and hepatocellular carcinoma.

56. The method of claim 54, wherein the cancer is selected from lung cancer, ovarian cancer, colon cancer, pancreatic cancer, cholangiocarcinoma, glioblastoma, thymic cancer, gastric cancer, endometrial cancer, cervical cancer, breast cancer, acute myeloid leukemia, liver cancer, a head and neck cancer, biliary cancer, and mesothelioma.

57. The method of claim 54, wherein the cancer is selected from triple-negative breast cancer (TNBC), malignant mesothelioma, lung adenocarcinoma, non-small cell lung cancer,Attorney Docket No. 47039-0043W01pancreatic ductal adenocarcinoma, platinum-resistant ovarian cancer, colorectal cancer, uterine serous carcinoma, epithelial ovarian carcinoma.

58. The method of claim 54, wherein the aging-related disease or condition is selected from the group consisting of: a senescence-associated skin condition, atopic dermatitis, wrinkles, senile lentigo, age spots, photoaged skin, radiation-induced dermatitis, radiation-induced skin fibrosis / injury, cutaneous wound healing, Alzheimer’s disease, aneurysm, cystic fibrosis, fibrosis in pancreatitis, glaucoma, hypertension, idiopathic pulmonary fibrosis, inflammatory bowel disease, intervertebral disc degeneration, macular degeneration, osteoarthritis, type 2 diabetes mellitus, adipose atrophy, lipodystrophy, atherosclerosis, cataracts, COPD, idiopathic pulmonary fibrosis, kidney transplant failure, liver fibrosis, loss of bone mass, myocardial infarction, sarcopenia, wound healing, alopecia, cardiomyocyte hypertrophy, osteoarthritis, Parkinson’s disease, age-associated loss of lung tissue elasticity, macular degeneration, cachexia, glomerulosclerosis, liver cirrhosis, NAFLD, osteoporosis, amyotrophic lateral sclerosis, Huntington’s disease, spinocerebellar ataxia, multiple sclerosis, and renal dysfunction.

59. The method of claim 54, wherein 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.

60. Nucleic acid encoding any of the multi-chain chimeric polypeptides of any one of claims 1-40.

61. A vector comprising the nucleic acid of claim 60.

62. A cell comprising the nucleic acid of claim 60 or the vector of claim 61.

63. A method of producing a multi-chain chimeric polypeptide, the method comprising: culturing the cell of claim 62 in a culture medium under conditions sufficient to result in the production of the multi-chain chimeric polypeptide; andAttorney Docket No. 47039-0043W01recovering the multi-chain chimeric polypeptide from the cell and / or the culture medium.

64. A multi-chain chimeric polypeptide produced by the method of claim 63.

65. A single-chain chimeric polypeptide comprising:(i) a first target-binding domain;(ii) a soluble T cell receptor 0-chain constant region (TRpC) domain; and (iii) a second target-binding domain,wherein the first-target binding domain and / or the second target binding domain is an anti-mesothelin antigen-binding domain.

66. The single-chain chimeric polypeptide of claim 65, wherein the first target-binding domain and the soluble TRpC domain directly abut each other.

67. The single-chain chimeric polypeptide of claim 65, wherein the single-chain chimeric polypeptide further comprises a linker sequence between the first target-binding domain and the soluble TRPC domain.

68. The single-chain chimeric polypeptide of any one of claims 65-67, wherein the soluble TRpC domain and the second target-binding domain directly abut each other.

69. The single-chain chimeric polypeptide of any one of claims 65-67, wherein the single-chain chimeric polypeptide further comprises a linker sequence between the soluble TRpC domain and the second target-binding domain.

70. The single-chain chimeric polypeptide of any one of claims 65-69, wherein the first target-binding domain and the second target-binding domain bind specifically to the same antigen.

71. The single-chain chimeric polypeptide of claim 70, wherein the first target-binding domain and the second target-binding domain bind specifically to the same epitope.Attorney Docket No. 47039-0043W0172. The single-chain chimeric polypeptide of claim 71, wherein the first target-binding domain and the second target-binding domain comprise the same amino acid sequence.

73. The single-chain chimeric polypeptide of any one of claims 65-69, wherein the first target-binding domain and the second target-binding domain bind specifically to different antigens.

74. The single-chain chimeric polypeptide of any one of claims 65-73, wherein both of the first target-binding domain and the second target-binding domain are an antigen-binding domain.

75. The single-chain chimeric polypeptide of any one of claims 65-74, wherein one of the first target-binding domain and the second target-binding domain bind to a target selected from the group consisting of: tissue factor (CD142), 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, TNFa, CD26, CD26a, CD36, ULBP2, CD30, CD200, IGF-1R, MUC4AC, MUC5AC, Trop-2, CMET, EGFR, HER1, HER2, HER3, PSMA, CEA, B7H3, EPCAM, BCMA, P-cadherin, CEACAM5, uPAR, aUL16-binding protein, HLA-DR, DLL4, TYR03, AXL, MER, CD122, CD155, PDGFDD, a ligand of TGF- receptor II (TGF-0RII), a ligand of TGF-PRIII, a ligand of DNAM1, a ligand of NKp46, a ligand of NKp44, a ligand of NKG2D, a ligand of NKp30, a ligand for a scMHCI, a ligand for a scMHCII, a ligand for a scTCR, a receptor for IL-1, a receptor for IL-2, a receptor for IL-3, a receptor for IL-4, a receptor for IL-7, a receptor for IL-8, a receptor for IL-10, a receptor for IL-12, a receptor for IL-15, a receptor for IL- 17, a receptor for IL- 18, a receptor for IL-21, a receptor for PDGF-DD, a receptor for stem cell factor (SCF), a receptor for stem cell-like tyrosine kinase 3 ligand (FLT3L), a receptor for MICA, a receptor for MICB, a receptor for a ULP16-binding protein, a receptor for CD155, a receptor for CD 122, and a receptor for CD28.

76. The single-chain chimeric polypeptide of any one of claims 65-69, wherein the first target-binding domain or the second target-binding domain is a soluble interleukin, a soluble cytokine protein, or a soluble cell surface protein.Attorney Docket No. 47039-0043W0177. The single-chain chimeric polypeptide of claim 76, wherein 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-4, IL-7, IL-8, IL-10, IL-12, IL-15, IL-17, IL-18, IL-21, PDGF-D, SCF, FLT3L, MICA, MICB, and a ULP16-binding protein.

78. The single-chain chimeric polypeptide of claim 77, wherein the soluble interleukin, soluble cytokine protein, or soluble cell surface protein is IL-7.

79. The single-chain chimeric polypeptide of any one of claims 65-69, wherein the first target-binding domain or the second target-binding domain is a soluble interleukin receptor, a soluble cytokine receptor, or a soluble cell surface receptor.

80. The single-chain chimeric polypeptide of claim 79, wherein the soluble interleukin receptor, the soluble cytokine receptor, or the soluble cell surface receptor is a soluble TGF-P receptor II (TGF-PRII), a soluble TGF-PRIII, a soluble NKG2D, a soluble NKp30, a soluble NKp44, a soluble NKp46, a soluble DNAM-1, a scMHCI, a scMHCII, a scTCR, a soluble CD155, or a soluble CD28.

81. The single-chain chimeric polypeptide of claim 80, wherein the soluble interleukin receptor, the soluble cytokine receptor, or the soluble cell surface receptor is a soluble TGF-βRII.

82. The single -chain chimeric polypeptide of any one of claims 65-69, wherein the first target-binding domain is IL-7 and the second target-binding domain is the anti-mesothelin antigen-binding domain.

83. The single -chain chimeric polypeptide of any one of claims 65-69, wherein the first target-binding domain is a soluble TGF-βRII and the second target-binding domain is the anti-mesothelin antigen-binding domain.Attorney Docket No. 47039-0043W0184. The single-chain chimeric polypeptide of any one of claims 65-83, wherein the single-chain chimeric polypeptide further comprises one or more additional target-binding domains at its N- and / or C-terminus.

85. The single-chain chimeric polypeptide of any one of claims 65-84, wherein the soluble TR0C domain is a soluble human TR0C domain.

86. The single-chain chimeric polypeptide of any one of claims 65-85, wherein the soluble TR0C domain does not bind to an MHC protein with or without a loaded antigen.

87. The single-chain chimeric polypeptide of any one of claims 65-86, wherein the soluble TR0C domain does not stimulate an immune response in a mammal.

88. The single-chain chimeric polypeptide of any one of claims 65-87, wherein the soluble TR0C domain comprises or consists of a sequence from a wildtype soluble human TR0C.

89. A composition comprising any of the single-chain chimeric polypeptides of claims 65-88.

90. The composition of claim 89, wherein the composition is a pharmaceutical composition.

91. A kit comprising at least one dose of the composition of claim 89 or 90.

92. A method of stimulating an immune cell, the method comprising:contacting an immune cell with an effective amount of any of the single-chain chimeric polypeptides of claims 65-88 or the composition of claim 89 or 90.

93. A method of inducing or increasing proliferation of an immune cell, the method comprising:Attorney Docket No. 47039-0043W01contacting an immune cell with an effective amount of any of the single-chain chimeric polypeptides of claims 65-88 or the composition of claim 89 or 90.

94. A method of inducing differentiation of an immune cell into a memory or memorylike immune cell, the method comprising:contacting an immune cell with an effective amount of any of the single-chain chimeric polypeptides of claims 65-88 or the composition of claim 89 or 90.

95. The method of any one of claims 92-94, wherein the immune cell is contacted in vitro.

96. The method of any one of claims 92-94, wherein the immune cell is contacted in vivo.

97. The method of any one of claims 92-96, wherein the immune cell is selected from the group consisting of: an immature thymocyte, a peripheral blood lymphocyte, a naive T cell, a pluripotent Th cell precursor, a lymphoid progenitor cell, a Treg cell, a Thl7 cell, a Th22 cell, a Th9 cell, a Th2 cell, a Thl cell, a Th3 cell, y8 T cell, an aP T cell, a tumor-infiltrating T cell, a CD8+ T cell, a CD4+ T cell, a natural killer T cell, a mast cell, a macrophage, a neutrophil, a dendritic cell, a basophil, an eosinophil, and a natural killer cell.

98. The method of claim 97, wherein the CD8+T cell is a progenitor exhausted CD8+T (Tpex) cell, an effector memory CD8+T (Tern) cell, a central memory CD8+T (Tcm) cell, a memory CD8+stem (Tscm) cell, a virtual memory CD8+T (Tvm) cell, an antigen-experienced CD8+T cell, or a terminally-exhausted CD8+T (Tex) cell.

99. The method of any one of claims 92-98, wherein the immune cell has previously been genetically modified to express a chimeric antigen receptor or a recombinant T-cell receptor.

100. The method of any one of claims 92-99, wherein the method further comprises administering the immune cell to a subject in need thereof.Attorney Docket No. 47039-0043W01101. A method of killing a cancer cell, an infected cell, or a senescent cell in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of any of the single-chain chimeric polypeptides of claims 65-88 or the composition of claim 89 or 90.

102. A method of treating a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of any of the single-chain chimeric polypeptides of claims 65-88 or the composition of claim 89 or 90.

103. The method of claim 101 or 102, wherein the subject has been identified or diagnosed as having a cancer, an aging-related disease or condition, or an infectious disease.

104. The method of claim 103, wherein the cancer is selected from the group consisting of: solid tumor, hematological tumor, sarcoma, osteosarcoma, 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 lymphocytic leukemia (ALL), myelodysplastic syndromes (MDS), cutaneous T-cell lymphoma, retinoblastoma, stomach cancer, urothelial carcinoma, lung cancer, renal cell carcinoma, gastic and esophageal cancer, pancreatic cancer, prostate cancer, breast cancer, colorectal cancer, ovarian cancer, non-small cell lung carcinoma, squamous cell head and neck carcinoma, endometrial cancer, cervical cancer, liver cancer, biliary cancer, and hepatocellular carcinoma.

105. The method of claim 103, wherein the cancer is selected from lung cancer, ovarian cancer, colon cancer, pancreatic cancer, cholangiocarcinoma, glioblastoma, thymic cancer, gastric cancer, endometrial cancer, cervical cancer, breast cancer, acute myeloid leukemia, liver cancer, a head and neck cancer, biliary cancer, and mesothelioma.

106. The method of claim 103, wherein the cancer is selected from triple-negative breast cancer (TNBC), malignant mesothelioma, lung adenocarcinoma, non-small cell lung cancer,Attorney Docket No. 47039-0043W01pancreatic ductal adenocarcinoma, platinum-resistant ovarian cancer, colorectal cancer, uterine serous carcinoma, and epithelial ovarian carcinoma107. The method of claim 103, wherein the aging-related disease or condition is selected from the group consisting of: a senescence-associated skin condition, atopic dermatitis, wrinkles, senile lentigo, age spots, photoaged skin, radiation-induced dermatitis, radiation-induced skin fibrosis / injury, cutaneous wound healing, Alzheimer’s disease, aneurysm, cystic fibrosis, fibrosis in pancreatitis, glaucoma, hypertension, idiopathic pulmonary fibrosis, inflammatory bowel disease, intervertebral disc degeneration, macular degeneration, osteoarthritis, type 2 diabetes mellitus, adipose atrophy, lipodystrophy, atherosclerosis, cataracts, COPD, idiopathic pulmonary fibrosis, kidney transplant failure, liver fibrosis, loss of bone mass, myocardial infarction, sarcopenia, wound healing, alopecia, cardiomyocyte hypertrophy, osteoarthritis, Parkinson’s disease, age-associated loss of lung tissue elasticity, macular degeneration, cachexia, glomerulosclerosis, liver cirrhosis, NAFLD, osteoporosis, amyotrophic lateral sclerosis, Huntington’s disease, spinocerebellar ataxia, multiple sclerosis, and renal dysfunction.

108. The method of claim 103, wherein 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.

109. Nucleic acid encoding any of the single-chain chimeric polypeptides of any one of claims 65-88.

110. A vector comprising the nucleic acid of claim 109.

111. A cell comprising the nucleic acid of claim 109 or the vector of claim 110.

112. A method of producing a single-chain chimeric polypeptide, the method comprising:culturing the cell of claim 111 in a culture medium under conditions sufficient to result in the production of the single-chain chimeric polypeptide; andAttorney Docket No. 47039-0043W01recovering the single-chain chimeric polypeptide from the cell and / or the culture medium.

113. A single-chain chimeric polypeptide produced by the method of claim 112.

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