Costimulatory cell engagers and methods of use thereof

Abstract

Compositions, e.g., compositions comprising fusion proteins, and methods of using such compositions for treating cancer are described.

Description

Attorney Docket No. 2012106-0161COSTIMULATORY CELL ENGAGERS AND METHODS OF USE THEREOFBACKGROUND OF THE INVENTION

[0001] Immune cell-based therapies are methods to treat cancer. However, there remains a need for improved methods for treating cancer using adoptive cell therapy.SUMMARY

[0002] The present disclosure provides methods and compositions useful for treatment of cancer and / or for initiating or modulating immune responses. In some embodiments, the present invention provides methods and compositions useful for initial treatment of cancers. In some embodiments, the present invention provides methods and compositions useful for treatment of cancers following relapse.

[0003] In some embodiments, the present disclosure provides fusion proteins for the activation of immune cells. In some embodiments, the present disclosure provides fusion proteins for the engagement and activation of CAR-T cells. In some embodiments, the present disclosure provides fusion proteins for the engagement and activation of TCR cells.

[0004] In some embodiments, a fusion protein of the present disclosure comprises (i) one or more accessory molecule(s) or a fragment thereof and (ii) one or more antigen-binding proteins or fragments that bind a tumor antigen. In some embodiments, a fusion protein of the present disclosure comprises (i) one or more accessory molecule(s) or a fragment thereof; (ii) one or more antigen-binding proteins or fragments that bind a tumor antigen; (iii) a target polypeptide for a cellular therapeutic. In some embodiments, an accessory molecule is or comprises a binding partner of an adhesion molecule, a costimulatory molecule, or a combination thereof. In some embodiments, an accessory molecule is or comprises an antigen binding protein or fragment that binds an adhesion molecule, a costimulatory molecule, or a combination thereof. In some embodiments, the accessory molecule is or is a binding partner of L-selectin, a4 integrin, lymphocyte function-associated antigen-1 (LFA-1), Intercellular- 1 - 13321680vlAttorney Docket No. 2012106-0161Adhesion Molecule 1 (ICAM-1) (CD54), CD2, CD3, CD28, B7-1 (CD80), B7-2 (CD86) or LFA-3 (CD58).

[0005] In some embodiments, a fusion protein comprises two or more accessory molecule(s) or a fragment thereof. In some embodiments, a fusion protein comprises two LFA-3 binding domains. In some embodiments, a fusion protein comprises B7-2 and one LFA-3 binding domain. In some embodiments, a fusion protein comprises, at least one accessory molecule that is an antigen binding protein or fragment that binds CD3.

[0006] In some embodiments, a tumor antigen is a tumor specific antigen (TSA) or a tumor associated antigen (TAA). In some embodiments, a tumor antigen is MART-l / MelanA (MART-I), gplOO (Pmel 17), tyrosinase, TRP-1, TRP-2, MAGE-1, MAGE-3, BAGE, GAGE-1, GAGE-2, pl 5, CEA, p53, Ras, HER-2 / neu, BCR-ABL, E2A-PRL, H4-RET, IGH-IGK, MYL-RAR, Epstein Barr virus antigens EBVA, human papillomavirus (HPV) antigen E6 or E7, TSP-180, MAGE-4, MAGE-5, MAGE-6, RAGE, NY-ESO, erbB, pl85erbB2, pl80erbB-3, c-met, nm-23Hl, PSA, TAG-72, CA 19-9, CA 72-4, CAM 17.1, NuMa, K-ras, beta-Catenin, CDK4, Mum-1, p 15, p 16, 43-9F, 5T4, 791Tgp72, alpha-fetoprotein, beta-HCG, BCA225, BTAA, CA 125, CA 15-3\CA 27.29\BCAA, CA 195, CA242, CA-50, CAM43, CD68\P1, CO-029, FGF-5, G250, Ga733\EpCAM, HTgp-175, M344, MA-50, MG7-Ag, M0V18, NB / 70K, NY-CO-1, RCAS1, SDCCAG16, TA-90\Mac-2 binding protein\cyclophilin C-associated protein, TAAL6, TAG72, TLP, MUC16, IL13Ra2, FRa, VEGFR2, Lewis Y, FAP, EphA2, CEACAM5, EGFR, CA6, CA9, GPNMB, EGP1, FOLR1, endothelial receptor, STEAP1, SLC44A4, Nectin-4, AGS-16, guanalyl cyclase C, MUC-1, CFC1B, integrin alpha 3 chain (of a3bl, a laminin receptor chain), TPS, CD19, CD20, CD22, CD30, CD72, CD180, CD171 (L1CAM), CD123, CD133, CD138, CD37, CD70, CD79a, CD79b, CD56, CD74, CD166, CD71, CLL-1 / CLEC12A, ROR1, Glypican 3 (GPC3), Mesothelin, CD33 / IL3Ra, c-Met, PSCA, PSMA, Glycolipid F77, EGFRvIII, BCMA, GD-2, MY-ESO-1, B7-H3 (CD276) or MAGE A3. In some embodiments, a tumor antigen is CD19, CD20, B7-H3, or IL13Ra2.

[0007] In some embodiments, an antigen-binding protein or fragment that binds a tumor antigen is an scFv, Fv, or VHH. In some embodiments, a fusion protein comprises two or more antigen-binding proteins or fragments that bind a tumor antigen. In some embodiments, a fusion - 2 - 13321680vlAttorney Docket No. 2012106-0161protein comprises an anti-B7-H3 VHH and an anti-IL13Ra2 VHH. In some embodiments, a fusion protein comprises an anti-B7-H3 VHH and an anti-HER2-scFv.

[0008] In some embodiments, a target polypeptide is a tumor antigen or fragment thereof. In some embodiments, a tumor antigen is or is a fragment of MART-l / MelanA (MART-I), gplOO (Pmel 17), tyrosinase, TRP-1, TRP-2, MAGE-1, MAGE-3, BAGE, GAGE-1, GAGE-2, pl 5, CEA, p53, Ras, HER-2 / neu, BCR-ABL, E2A-PRL, H4-RET, IGH-IGK, MYL-RAR, Epstein Barr virus antigens EBVA, human papillomavirus (HPV) antigen E6 or E7, TSP-180, MAGE-4, MAGE-5, MAGE-6, RAGE, NY-ESO, erbB, pl85erbB2, pl80erbB-3, c-met, nm-23H1, PSA, TAG-72, CA 19-9, CA 72-4, CAM 17.1, NuMa, K-ras, beta-Catenin, CDK4, Mum-1, p 15, p 16, 43-9F, 5T4, 791Tgp72, alpha-fetoprotein, beta-HCG, BCA225, BTAA, CA 125, CA 15-3\CA 27.29XBCAA, CA 195, CA 242, CA-50, CAM43, CD68\P1, CO-029, FGF-5, G250, Ga733\EpCAM, HTgp-175, M344, MA-50, MG7-Ag, M0V18, NB / 70K, NY-CO-1, RCAS1, SDCCAG16, TA-90\Mac-2 binding protein\cyclophilin C-associated protein, TAAL6, TAG72, TLP, MUC16, IL13Ra2, FRa, VEGFR2, Lewis Y, FAP, EphA2, CEACAM5, EGFR, CA6, CA9, GPNMB, EGP1, FOLR1, endothelial receptor, STEAP1, SLC44A4, Nectin-4, AGS-16, guanalyl cyclase C, MUC-1, CFC1B, integrin alpha 3 chain (of a3bl, a laminin receptor chain), TPS, CD19, CD20, CD22, CD30, CD72, CD180, CD171 (L1CAM), CD123, CD133, CD138, CD37, CD70, CD79a, CD79b, CD56, CD74, CD166, CD71, CLL-1 / CLEC12A, ROR1, Glypican 3 (GPC3), Mesothelin, CD33 / IL3Ra, c-Met, PSCA, PSMA, Glycolipid F77, EGFRvIII, BCMA, GD-2, MY-ESO-1, B7-H3 (CD276) or MAGE A3. In some embodiments, a tumor antigen is or is a fragment of CD20, CD 19, or BCMA.

[0009] In some embodiments, a cellular therapeutic is a CAR-T cell, CAR-NK cell, TCR-T cell, TIL cell, allogenic NK cell, or autologous NK cell.

[0010] In some embodiments, a fusion protein comprises a half-life extension polypeptide. In some embodiments, a half-life extension polypeptide is any one of a hyaluronan binding motif, PAS polypeptide, proline / alanine random coil polypeptide, and an antibody or antigen binding fragment thereof. In some embodiments, a half-life extension polypeptide is an antibody or antigen binding fragment thereof. In some embodiments, an anti-albumin antibody or antigen binding fragment thereof comprises an anti-albumin VHH.- 3 - 13321680vlAttorney Docket No. 2012106-0161

[0011] In some embodiments, a fusion protein comprises an immune suppression blockade domain. In some embodiments, an immune suppression blockade domain is a receptor for an immune suppression molecule. In some embodiments, an immune suppression blockade domain is TGFβ receptor 2 (TGFβr2).

[0012] In some embodiments, the present disclosure provides a nucleic acid encoding a fusion protein of the present disclosure. In some embodiments, the present disclosure provides a vector comprising a nucleic acid encoding a fusion protein of the present disclosure.

[0013] In some embodiments, the present disclosure provides an immune cell comprising an expression construct encoding a fusion protein comprising (i) one or more accessory molecule(s) or a fragment thereof; (ii) one or more antigen-binding proteins or fragments that bind a tumor antigen; and (iii) a target polypeptide for a cellular therapeutic. In some embodiments, a fusion protein described herein is secreted from the immune cell.

[0014] In some embodiments, the present disclosure provides a method of treating a subject having a tumor, comprising administering to the subject an effective amount of a fusion protein described herein, thereby treating the subject. In some embodiments, the present disclosure provides a method of treating a subject having a tumor, comprising administering to the subject an effective amount of an immune cell comprising an expression construct encoding a fusion protein comprising (i) one or more accessory molecule(s) or a fragment thereof; (ii) one or more antigen-binding proteins or fragments that bind a tumor antigen; and (iii) a target polypeptide for a cellular therapeutic. In some embodiments, a fusion protein described herein is secreted from the immune cell.BRIEF DESCRIPTION OF THE DRAWINGS

[0015] The foregoing and other objects of the present disclosure, the various features thereof, as well as the disclosure itself may be more fully understood from the following description, when read together with the accompanying drawings in which:- 4 - 13321680vlAttorney Docket No. 2012106-0161

[0016] Figure 1. Depicts Costimulatory CAR-T Engagers (“CTE”) organizing costimulatory proteins around the binding site on the CAR to mimic a canonical T cell immune synapse.

[0017] Figure 2. Shows binding of exemplary costimulatory CAR T engagers (e.g., CTE #607, CTE #686, and CTE #692) to soluble biotinylated CD2.

[0018] Figure 3. Shows binding of exemplary costimulatory T cell engagers to CD2 by ELISA.

[0019] Figure 4. Shows binding of exemplary costimulatory CAR T engagers with none, one, or two LFA3 binding domains to CD2.

[0020] Figures 5A-5C. Depict the distribution of key proteins across the APC-immune synapse and T cell immune synapse (FIG. 5A; Binder, et al., 2020), which, when bound by multi-antigen CTEs (FIG. 5B), organize co-stimulatory proteins around the binding site on the CAR to mimic the canonical T-cell immune synapse allowing for productive T cell signaling (FIG. 5C; van der Merwe, et al., 2000).

[0021] Figure 6A and 6B. Depict expression of LFA3 (FIG. 6A) and CD2 (FIG. 6B) on activated human T cells by flow cytometry staining.

[0022] Figure 7. Shows binding of exemplary costimulatory T cell engagers to activated T cells.

[0023] Figure 8. Shows exemplary costimulatory CAR T engagers dose-responsive binding to activated normal human T cells by flow cytometry.

[0024] Figure 9. Shows additional exemplary costimulatory T cell engagers having differential spacing of LFA3bds dose-responsive binding to activated normal human T cells by flow cytometry.

[0025] Figure 10. Shows CTE-directed, anti-CD19 CAR T mediated, cytotoxic activity against JeKo-1 lymphoma cells that are CD19 and CD58 deficient (JeKo-1 CD19 58 KO).

[0026] Figure 11. Shows evaluation of cytotoxicity directed by CTE #692 and anti-CD19 CAR T cells when the target lacks expression of CD19 and CD58.- 5 - 13321680vlAttorney Docket No. 2012106-0161

[0027] Figure 12. Shows anti-CD20 TCE binding of CD20 detected using biotinylated-CD20 and HRP-streptavidin.

[0028] Figure 13. Shows anti-CD20 TCE binding of CD2 detected using biotinylated-CD2 and HRP-streptavidin.

[0029] Figures 14A-14B. Show anti-CD20 TCE-mediated B cell depletion from Donor 67 (FIG. 14A) and Donor 22 (FIG. 14B).

[0030] Figure 15. Induction of NF AT -luciferase by TCE #754 and costimulatory-TCE #753 in the presence of A375 melanoma cells that express fL-13Roc2. CTE #573 served as the negative control engager protein.

[0031] Figure 16. Shows TCE-mediated cytotoxicity of A375 melanoma cells.

[0032] Figure 17. Shows detection of bridging proteins in cell culture supernatants by ELISA. Construct numbers are listed in Table 16.

[0033] Figures 18A-18B. Show quantification of bridging proteins in cell culture supernatants by ELISA. FIG 18A shows a standard curve using purified protein #357. FIG. 18B shows expression analysis at different cell culture dilutions. Two transfection dates are shown for #567; for #571 and #577 two amounts of DNA (2.5 and 5 pg) were used in transfections.

[0034] Figure 19. Shows expression of B7-H3, Her2 and CD19 on A375 melanoma cells by flow cytometry.

[0035] Figure 20. Shows binding of purified constructs #263, #567, #577, and #593 on A375 cells detected with FMC63-PE by flow cytometry.

[0036] Figures 21A-21B. Show dual-antigen bridging proteins binding using a B7-H3-capture plate and anti-His-tag detection antibody (FIG. 21A) and anti-Her2 detection antibody (FIG. 2 IB).

[0037] Figure 22. Shows dose response cytotoxicity against A375 cells mediated by bridging proteins in the presence of CAR-CD19 T cells.

[0038] Figure 23. Shows dose response cytotoxicity against A375 cells mediated by additional bridging proteins in the presence of CAR-CD19 T cells.- 6 - 13321680vlAttorney Docket No. 2012106-0161

[0039] Figure 24. Shows SKOV3 cytotoxicity data generated using bridging proteins and CAR-CD19 T cells (54.5% CAR-positive, 10:1 E: T ratio).

[0040] Figure 25. Shows exemplary bispecific anti-Her2 / anti-B7-H3 dual-antigen binding constructs mediated cytotoxicity using adjusted titers.

[0041] Figures 26A-26C. Shows the expression of CD 19 (FIG. 26 A), Her2 (FIG. 26B), and B7-H3 (FIG. 26C) on BT474 breast cancer cell line derived from ductal carcinoma.

[0042] Figure 27. Shows cytotoxicity of the dual-antigen binding constructs against BT474 ductal carcinoma cells.

[0043] Figure 28. Shows flow cytometry profiles of CTE binding to A375 cells.

[0044] Figure 29. Shows CTE-directed anti-CD19 CAR T-mediated cytotoxicity against A375 melanoma cells.

[0045] Figure 30. Shows exemplary bispecific dual-antigen-directed CTEs dose-responsive IL13Ra2 binding.

[0046] Figure 31. Shows exemplary bispecific dual-antigen-directed CTEs dose-responsive B7H3 binding.

[0047] Figures 32A-32C. Shows expression of B7-H3 (FIG. 32A) and IL-13Ra2 (FIG.32B) and CD19 antigens (FIG. 32C) on A375 melanoma cells by flow cytometry.

[0048] Figure 33. Shows exemplary bispecific dual-antigen-directed CTEs dose-responsive A375 cell binding.

[0049] Figure 34. Shows exemplary bispecific dual-antigen-directed CTE-mediated cytotoxicity.- 7 - 13321680vlAttorney Docket No. 2012106-0161

[0050] Figures 35A-35C. Shows expression of B7-H3 (FIG. 35A), and IL-13Ra2 (FIG.35B) and CD 19 antigens (FIG. 35C) on the U251MG cells by flow cytometry.

[0051] Figure 36. Shows exemplary bispecific dual-antigen-directed CTEs dose-responsive U251MG cell binding.

[0052] Figure 37. Shows exemplary bispecific dual-antigen-directed CTEs dose-responsive U251MG cell killing.

[0053] Figure 38. Shows U251MG cell killing mediated by exemplary single-antigen CTEs, bispecific dual-antigen directed CTEs, and CD19-directed CAR-T cells.

[0054] Figure 39. Shows the effect of adding multiple costimulatory domains to IL-13Ra2-directed CTEs dose-responsive binding to the anti-CD19 antibody FMC63 and to the IL-13Ra2 protein.

[0055] Figure 40. Shows IL13Ra2 binding by exemplary CTEs, containing 2 B7-2 binding domains and B7-H3 bispecifics with LFA3 or B7-2 by ELISA.

[0056] Figure 41. Shows B7-H3 binding activity for constructs #769 and #770 by ELISA.

[0057] Figure 42. Shows exemplary CTEs binding of anti-CD19 antibody FMC63 and CD2 protein by ELISA.

[0058] Figure 43. Shows CD2 binding by exemplary IL-13Rα2, B7-H3 bispecific dual-antigen directed constructs with LFA3.- 8 - 13321680vlAttorney Docket No. 2012106-0161

[0059] Figure 44. Shows CD28 binding by exemplary B7-2 containing constructs by ELISA.

[0060] Figure 45. Shows exemplary CTE-mediated cytotoxicity by CTEs listed in Table 35.

[0061] Figures 46A-46B Shows exemplary CTE-mediated induction of NFAT-luciferase expression in Jurkat-CAR19 cells by construct #607 in the presence of target 293-CD20 tumor cells (FIG 46A) and IL-13Ra2-directed CTEs in the presence IL-13Ra2-positive A375 melanoma cells (FIG. 46B).

[0062] Figure 47. Shows IL-13Roi2 x B7-H3 CTE binding to B7-H3-4Ig. The capture reagent was purified human serum albumin, and the detection reagent was biotinylated B7-H3-41g detected enzymatically.

[0063] Figure 48. Shows a CTE ELISA assay: The capture reagent was purified human serum albumin, and the detection reagent was biotinylated IL-13Roc2, detected enzymatically.

[0064] Figure 49. Shows induction of U251MG glioblastoma cell cytotoxicity using CD19-directed CAR T cells and CTEs containing the TGFβR2 ECD.

[0065] Figure 50. Shows dose response TGFP-induced SMAD2 phosphorylation by B7-H3 / IL-13Ra2 bi specific CTEs containing the TGFβR2 ECD.

[0066] Figure 51. Shows IL-13Ra2 / B7-H3 dual targeting CTE-mediated cytotoxicity in A375 melanoma cells.- 9 - 13321680vlAttorney Docket No. 2012106-0161

[0067] Figure 52. Shows CD2 binding by LFA3 and Her2-directed CTEs in the presence or absence of TGFp antagonist sequence.

[0068] Figure 53. Shows TGFp and Her2 targeting CTEs ability to bind to both the antiCD 19 antibody FMC63 and to the Her2 antigen by ELISA.

[0069] Figure 54. Shows TGFp and Her-2 targeting CTEs ability to bind to both the antiCD 19 antibody FMC63 and to the IL-13Ra2 antigen by ELISA.

[0070] Figure 55. Shows CTE binding to soluble TGFP by ELISA.

[0071] Figure 56. Shows a blockade of TGFP-mediated SMAD2 phosphorylation by CTE #727.

[0072] Figure 57. Shows induction of BT474 breast carcinoma cell cytotoxicity using CD19-directed CAR T cells and CTEs. The E: T ratio was 5:1 and the assay was run for 48 hours.

[0073] Figure 58. Shows induction of A375 Her2 KO melanoma cell cytotoxicity using CD19-directed CAR T cells and CTEs. The E: T ratio was 5:1 and the assay was run for 48 hours.

[0074] Figure 59. Shows induction of U251MG glioblastoma cell cytotoxicity using CD19-directed CAR T cells and CTEs. The E: T ratio was 5:1 and the assay was run for 48 hours.

[0075] Figure 60. Shows induction of BT474 CD58 KO ductal carcinoma cell cytotoxicity using CD19-directed CAR T cells and CTEs.- 10 - 13321680vlAttorney Docket No. 2012106-0161

[0076] Figures 61A-61C show confocal microscopy images of cell-cell contact between CD19-directed CAR T cells and Her2-expressing tumor cells without (Fig. 61A) or with the addition of CTE #782 (Fig. 61B) or CTE #786 (Fig. 61C).DEFINITIONS

[0077] In order for the present invention to be more readily understood, certain terms are first defined below. Additional definitions for the following terms and other terms are set forth throughout the specification.

[0078] Administration'. As used herein, the term “administration” refers to the administration of a composition to a subject or system. Administration to an animal subject (e.g., to a human) may be by any appropriate route. For example, in some embodiments, administration may be bronchial (including by bronchial instillation), buccal, enteral, interdermal, intra-arterial, intradermal, intragastric, intramedullary, intramuscular, intranasal, intraperitoneal, intrathecal, intravenous, intraventricular, within a specific organ (e.g., intrahepatic), mucosal, nasal, oral, rectal, subcutaneous, sublingual, topical, tracheal (including by intratracheal instillation), transdermal, vaginal and vitreal. In some embodiments, administration may be intratumoral or peritumoral. In some embodiments, administration may involve intermittent dosing. In some embodiments, administration may involve continuous dosing (e.g., perfusion) for at least a selected period of time.

[0079] Adoptive cell therapy. As used herein, “adoptive cell therapy” or “ACT” involves the transfer of immune cells with antitumour activity into cancer patients. In some embodiments, ACT is a treatment approach that involves the use of cells (e.g., myeloid or lymphoid) with antitumour activity, the in vitro expansion of these cells to large numbers and their infusion into a cancer-bearing host.

[0080] Agent: The term “agent” as used herein may refer to a compound or entity of any chemical class including, for example, polypeptides, nucleic acids, saccharides, lipids, small molecules, metals, or combinations thereof. As will be clear from context, in some- 11 - 13321680vlAttorney Docket No. 2012106-0161embodiments, an agent can be or comprise a cell or organism, or a fraction, extract, or component thereof. In some embodiments, an agent is or comprises a natural product in that it is found in and / or is obtained from nature. In some embodiments, an agent is or comprises one or more entities that is man-made in that it is designed, engineered, and / or produced through action of the hand of man and / or is not found in nature. In some embodiments, an agent may be utilized in isolated or pure form; in some embodiments, an agent may be utilized in crude form. In some embodiments, potential agents are provided as collections or libraries, for example that may be screened to identify or characterize active agents within them. Some particular embodiments of agents that may be utilized in accordance with the present invention include small molecules, antibodies, antibody fragments, aptamers, nucleic acids (e.g., siRNAs, shRNAs, DNA / RNA hybrids, antisense oligonucleotides, ribozymes), peptides, peptide mimetics, etc. In some embodiments, an agent is or comprises a polymer. In some embodiments, an agent is not a polymer and / or is substantially free of any polymer. In some embodiments, an agent contains at least one polymeric moiety. In some embodiments, an agent lacks or is substantially free of any polymeric moiety.

[0081] Amelioration'. As used herein, “amelioration” refers to prevention, reduction and / or palliation of a state, or improvement of the state of a subject. Amelioration includes, but does not require, complete recovery or complete prevention of a disease, disorder or condition.

[0082] Amino acid. As used herein, term “amino acid,” in its broadest sense, refers to any compound and / or substance that can be incorporated into a polypeptide chain. In some embodiments, an amino acid has the general structure H₂N–C(H)(R)–COOH. In some embodiments, an amino acid is a naturally occurring amino acid. In some embodiments, an amino acid is a synthetic amino acid; in some embodiments, an amino acid is a d-amino acid; in some embodiments, an amino acid is an 1-amino acid. “Standard amino acid” refers to any of the twenty standard 1-amino acids commonly found in naturally occurring peptides. “Nonstandard amino acid” refers to any amino acid, other than the standard amino acids, regardless of whether it is prepared synthetically or obtained from a natural source. As used herein, “synthetic amino acid” encompasses chemically modified amino acids, including but not limited to salts, amino acid derivatives (such as amides), and / or substitutions. Amino acids, including carboxy- and / or - 12 - 13321680vlAttorney Docket No. 2012106-0161amino-terminal amino acids in peptides, can be modified by methylation, amidation, acetylation, protecting groups, and / or substitution with other chemical groups that can change the peptide’s circulating half-life without adversely affecting their activity. Amino acids may participate in a disulfide bond. Amino acids may comprise one or posttranslational modifications, such as association with one or more chemical entities (e.g., methyl groups, acetate groups, acetyl groups, phosphate groups, formyl moieties, isoprenoid groups, sulfate groups, polyethylene glycol moieties, lipid moieties, carbohydrate moieties, biotin moieties, etc.). The term “amino acid” is used interchangeably with “amino acid residue,” and may refer to a free amino acid and / or to an amino acid residue of a peptide. It will be apparent from the context in which the term is used whether it refers to a free amino acid or a residue of a peptide.

[0083] Antibody. As used herein, the term “antibody” refers to a polypeptide that includes canonical immunoglobulin sequence elements sufficient to confer specific binding to a particular target antigen. As is known in the art, intact antibodies as produced in nature are approximately 150 kD tetrameric agents comprised of two identical heavy chain polypeptides (about 50 kD each) and two identical light chain polypeptides (about 25 kD each) that associate with each other into what is commonly referred to as a “Y-shaped” structure. Each heavy chain is comprised of at least four domains (each about 110 amino acids long)- an amino-terminal variable (VH) domain (located at the tips of the Y structure), followed by three constant domains: CHI, CH2, and the carboxy-terminal CH3 (located at the base of the Y’s stem). A short region, known as the “switch”, connects the heavy chain variable and constant regions. The “hinge” connects CH2 and CH3 domains to the rest of the antibody. Two disulfide bonds in this hinge region connect the two heavy chain polypeptides to one another in an intact antibody. Each light chain is comprised of two domains - an amino-terminal variable (VL) domain, followed by a carboxy-terminal constant (CL) domain, separated from one another by another “switch”. Intact antibody tetramers are composed of two heavy chain-light chain dimers in which the heavy and light chains are linked to one another by a single disulfide bond; two other disulfide bonds connect the heavy chain hinge regions to one another, so that the dimers are connected to one another and the tetramer is formed. Naturally-produced antibodies are also glycosylated, typically on the CH2 domain. Each domain in a natural antibody has a structure characterized by an “immunoglobulin fold” formed from two beta sheets (e.g., 3-, 4-, or 5- - 13 - 13321680vlAttorney Docket No. 2012106-0161stranded sheets) packed against each other in a compressed antiparallel beta barrel. Each variable domain contains three hypervariable loops known as “complement determining regions” (CDR1, CDR2, and CDR3) and four somewhat invariant “framework” regions (FR1, FR2, FR3, and FR4). When natural antibodies fold, the FR regions form the beta sheets that provide the structural framework for the domains, and the CDR loop regions from both the heavy and light chains are brought together in three-dimensional space so that they create a single hypervariable antigen binding site located at the tip of the Y structure. The Fc region of naturally-occurring antibodies binds to elements of the complement system, and also to receptors on effector cells, including for example effector cells that mediate cytotoxicity. As is known in the art, affinity and / or other binding attributes of Fc regions for Fc receptors can be modulated through glycosylation or other modification.

[0084] In some embodiments, antibody domains derived from camelids are used.Camelid single-domain antibodies (sdAbs), also known as VHH antibodies or nanobodies, are unique antigen-binding fragments derived from the heavy-chain antibodies naturally found in camelids such as camels, llamas, and alpacas. Unlike conventional antibodies composed of two heavy and two light chains, these heavy-chain antibodies lack light chains, and their antigenbinding region consists solely of a single variable domain referred to as VHH. The VHH domain, approximately 12-15 kDa in size, retains full antigen-binding capacity despite its smaller size compared to conventional antibodies (-150 kDa).

[0085] In some embodiments, antibodies produced and / or utilized in accordance with the present disclosure include glycosylated Fc domains, including Fc domains with modified or engineered such glycosylation. For purposes of the present disclosure, in certain embodiments, any polypeptide or complex of polypeptides that includes sufficient immunoglobulin domain sequences as found in natural antibodies can be referred to and / or used as an “antibody”, whether such polypeptide is naturally produced (e.g., generated by an organism reacting to an antigen), or produced by recombinant engineering, chemical synthesis, or other artificial system or methodology. In some embodiments, an antibody is polyclonal; in some embodiments, an antibody is monoclonal. In some embodiments, an antibody has constant region sequences that are characteristic of mouse, rabbit, primate, or human antibodies. In some embodiments,- 14 - 13321680vlAttorney Docket No. 2012106-0161antibody sequence elements are fully human, or are humanized, primatized, chimeric, etc, as is known in the art. Moreover, the term “antibody” as used herein, can refer in appropriate embodiments (unless otherwise stated or clear from context) to any of the art-known or developed constructs or formats for utilizing antibody structural and functional features in alternative presentation. For example, in some embodiments, an antibody utilized in accordance with the present disclosure is in a format selected from, but not limited to, intact IgG, IgE and IgM, bi- or multi- specific antibodies (e.g., Zybodies®, etc), single chain Fvs, polypeptide-Fc fusions, Fabs, cameloid antibodies, masked antibodies (e.g., Probodies®), Small Modular ImmunoPharmaceuticals (“SMIPsTM”), single chain or Tandem diabodies (TandAb®), VHHs, Anticalins®, Nanobodies®, minibodies, BiTE®s, ankyrin repeat proteins or DARPINs®, Avimers®, a DART, a TCR-like antibody, Adnectins®, Affilins®, Trans-bodies®, Affibodies®, a TrimerX®, MicroProteins, Fynomers®, Centyrins®, and a KALBITOR®. In some embodiments, an antibody may lack a covalent modification (e.g., attachment of a glycan) that it would have if produced naturally. In some embodiments, an antibody may contain a covalent modification (e.g., attachment of a glycan, a payload (e.g., a detectable moiety, a therapeutic moiety, a catalytic moiety, etc ), or other pendant group (e.g., poly-ethylene glycol, etc.)).

[0086] Antibody-Dependent Cellular Cytotoxicity. As used herein, the term “antibodydependent cellular cytotoxicity” or “ADCC” refers to a phenomenon in which target cells bound by antibody are killed by immune effector cells. Without wishing to be bound by any particular theory, ADCC is typically understood to involve Fc receptor (FcR)-bearing effector cells can recognizing and subsequently killing antibody-coated target cells (e g., cells that express on their surface specific antigens to which an antibody is bound). Effector cells that mediate ADCC can include immune cells, including but not limited to one or more of natural killer (NK) cells, macrophage, neutrophils, eosinophils.

[0087] Antibody Fragment. As used herein, an “antibody fragment” includes a portion of an intact antibody, such as, for example, the antigen-binding or variable region of an antibody. Examples of antibody fragments include Fab, Fab’, F(ab’)2, and Fv fragments; triabodies; tetrabodies; linear antibodies; single-chain antibody molecules (e.g., camelid heavy chain, VHH); and multi-specific antibodies formed from antibody fragments. For example, antibody - 15 - 13321680vlAttorney Docket No. 2012106-0161fragments include isolated fragments, “Fv” fragments (consisting of the variable regions of the heavy and light chains), recombinant single chain polypeptide molecules in which light and heavy chain variable regions are connected by a peptide linker (“ScFv proteins”), and minimal recognition units consisting of the amino acid residues that mimic the hypervariable region. In many embodiments, an antibody fragment contains sufficient sequence of the parent antibody of which it is a fragment that it binds to the same antigen as does the parent antibody; in some embodiments, a fragment binds to the antigen with a comparable affinity to that of the parent antibody and / or competes with the parent antibody for binding to the antigen. Examples of antigen binding fragments of an antibody include, but are not limited to, Fab fragment, Fab’ fragment, F(ab’)2 fragment, scFv fragment, Fv fragment, dsFv diabody, dAb fragment, Fd’ fragment, Fd fragment, and an isolated complementarity determining region (CDR) region. An antigen binding fragment of an antibody may be produced by any means. For example, an antigen binding fragment of an antibody may be enzymatically or chemically produced by fragmentation of an intact antibody and / or it may be recombinantly produced from a gene encoding the partial antibody sequence. Alternatively or additionally, antigen binding fragment of an antibody may be wholly or partially synthetically produced. An antigen binding fragment of an antibody may optionally comprise a single chain antibody fragment. Alternatively or additionally, an antigen binding fragment of an antibody may comprise multiple chains which are linked together, for example, by disulfide linkages. An antigen binding fragment of an antibody may optionally comprise a multimolecular complex. A functional antibody fragment typically comprises at least about 50 amino acids and more typically comprises at least about 200 amino acids.

[0088] Antigen: The term “antigen”, as used herein, refers to an agent that elicits an immune response; and / or an agent that binds to a T cell receptor (e.g., when presented by an MHC molecule) or to an antibody or antibody fragment. In some embodiments, an antigen elicits a humoral response (e.g., including production of antigen-specific antibodies); in some embodiments, an antigen elicits a cellular response (e.g., involving T-cells whose receptors specifically interact with the antigen). In some embodiments, an antigen binds to an antibody and may or may not induce a particular physiological response in an organism. In general, an antigen may be or include any chemical entity such as, for example, a small molecule, a nucleic - 16 - 13321680vlAttorney Docket No. 2012106-0161acid, a polypeptide, a carbohydrate, a lipid, a polymer (in some embodiments other than a biologic polymer (e.g., other than a nucleic acid or amino acid polymer)) etc. In some embodiments, an antigen is or comprises a polypeptide. In some embodiments, an antigen is or comprises a glycan. Those of ordinary skill in the art will appreciate that, in general, an antigen may be provided in isolated or pure form, or alternatively may be provided in crude form (e.g., together with other materials, for example in an extract such as a cellular extract or other relatively crude preparation of an antigen-containing source), or alternatively may exist on or in a cell. In some embodiments, an antigen is a recombinant antigen.

[0089] Antigen presenting cell'. The phrase “antigen presenting cell” or “APC,” as used herein, has its art understood meaning referring to cells that process and present antigens to T-cells. Exemplary APC include dendritic cells, macrophages, B cells, certain activated epithelial cells, and other cell types capable of TCR stimulation and appropriate T cell costimulation.

[0090] Approximately or about: As used herein, the term “approximately” or “about,” as applied to one or more values of interest, refers to a value that is similar to a stated reference value. In certain embodiments, the term “approximately” or “about” refers to a range of values that fall within 25%, 20%, 19%, 18%, 17%, 16%, 15%, 14%, 13%, 12%, 11%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, or less in either direction (greater than or less than) of the stated reference value unless otherwise stated or otherwise evident from the context (except where such number would exceed 100% of a possible value).

[0091] Binding. It will be understood that the term “binding”, as used herein, typically refers to a non-covalent association between or among two or more entities. “Direct” binding involves physical contact between entities or moieties; indirect binding involves physical interaction by way of physical contact with one or more intermediate entities. Binding between two or more entities can typically be assessed in any of a variety of contexts - including where interacting entities or moieties are studied in isolation or in the context of more complex systems (e.g., while covalently or otherwise associated with a carrier entity and / or in a biological system or cell).

[0092] Cancer'. The terms “cancer”, “malignancy”, “neoplasm”, “tumor”, and “carcinoma”, are used interchangeably herein to refer to cells that exhibit relatively abnormal, - 17 - 13321680vlAttorney Docket No. 2012106-0161uncontrolled, and / or autonomous growth, so that they exhibit an aberrant growth phenotype characterized by a significant loss of control of cell proliferation. In general, cells of interest for detection or treatment in the present application include precancerous (e.g., benign), malignant, pre-metastatic, metastatic, and non-metastatic cells. The teachings of the present disclosure may be relevant to any and all cancers. To give but a few, non-limiting examples, in some embodiments, teachings of the present disclosure are applied to one or more cancers such as, for example, hematopoietic cancers including leukemias, lymphomas (Hodgkins and nonHodgkins), myelomas and myeloproliferative disorders; sarcomas, melanomas, adenomas, carcinomas of solid tissue, squamous cell carcinomas of the mouth, throat, larynx, and lung, liver cancer, genitourinary cancers such as prostate, cervical, bladder, uterine, and endometrial cancer and renal cell carcinomas, bone cancer, pancreatic cancer, skin cancer, cutaneous or intraocular melanoma, cancer of the endocrine system, cancer of the thyroid gland, cancer of the parathyroid gland, head and neck cancers, breast cancer, gastro-intestinal cancers and nervous system cancers, benign lesions such as papillomas, and the like.

[0093] Chimeric antigen receptor: “Chimeric antigen receptor” or “CAR” or “CARs” as used herein refers to engineered receptors, which graft an antigen specificity onto cells (for example T cells such as naive T cells, central memory T cells, effector memory T cells or combination thereof). CARs are also known as artificial T-cell receptors, chimeric T-cell receptors or chimeric immunoreceptors. In some embodiments, CARs comprise an antigenspecific targeting regions, an extracellular domain, a transmembrane domain, one or more costimulatory domains, and an intracellular signaling domain.

[0094] Combination Therapy. As used herein, the term “combination therapy” refers to those situations in which a subject is simultaneously exposed to two or more therapeutic regimens (e.g., two or more therapeutic agents). In some embodiments, two or more agents may be administered simultaneously; in some embodiments, such agents may be administered sequentially; in some embodiments, such agents are administered in overlapping dosing regimens.

[0095] Domain. The term “domain” is used herein to refer to a section or portion of an entity. In some embodiments, a “domain” is associated with a particular structural and / or- 18 - 13321680vlAttorney Docket No. 2012106-0161functional feature of the entity so that, when the domain is physically separated from the rest of its parent entity, it substantially or entirely retains the particular structural and / or functional feature. Alternatively or additionally, a domain may be or include a portion of an entity that, when separated from that (parent) entity and linked with a different (recipient) entity, substantially retains and / or imparts on the recipient entity one or more structural and / or functional features that characterized it in the parent entity. In some embodiments, a domain is a section or portion of a molecular (e.g., a small molecule, carbohydrate, a lipid, a nucleic acid, or a polypeptide). In some embodiments, a domain is a section of a polypeptide; in some such embodiments, a domain is characterized by a particular structural element (e.g., a particular amino acid sequence or sequence motif, a-helix character, P-sheet character, coiled-coil character, random coil character, etc), and / or by a particular functional feature (e.g., binding activity, enzymatic activity, folding activity, signaling activity, etc).

[0096] Dosage form: As used herein, the terms “dosage form” and “unit dosage form” refer to a physically discrete unit of a therapeutic agent for the patient to be treated. Each unit contains a predetermined quantity of active material calculated to produce the desired therapeutic effect. It will be understood, however, that the total dosage of the composition will be decided by the attending physician within the scope of sound medical judgment.

[0097] Dosing regimen: As used herein, the term “dosing regimen” refers to a set of unit doses (typically more than one) that are administered individually to a subject, typically separated by periods of time. In some embodiments, a given therapeutic agent has a recommended dosing regimen, which may involve one or more doses. In some embodiments, a dosing regimen comprises a plurality of doses each of which are separated from one another by a time period of the same length; in some embodiments, a dosing regimen comprises a plurality of doses and at least two different time periods separating individual doses. In some embodiments, all doses within a dosing regimen are of the same unit dose amount. In some embodiments, different doses within a dosing regimen are of different amounts. In some embodiments, a dosing regimen comprises a first dose in a first dose amount, followed by one or more additional doses in a second dose amount different from the first dose amount. In some embodiments, a dosing regimen comprises a first dose in a first dose amount, followed by one or more additional - 19 - 13321680vlAttorney Docket No. 2012106-0161doses in a second dose amount same as the first dose amount. In some embodiments, a dosing regimen is correlated with a desired or beneficial outcome when administered across a relevant population (i.e., is a therapeutic dosing regimen).

[0098] Effector Function. As used herein, “effector function” refers a biochemical event that results from the interaction of an antibody Fc region with an Fc receptor or ligand. Effector functions include but are not limited to antibody-dependent cell-mediated cytotoxicity (ADCC), antibody-dependent cell-mediated phagocytosis (ADCP), and complement-mediated cytotoxicity (CMC). In some embodiments, an effector function is one that operates after the binding of an antigen, one that operates independent of antigen binding, or both.

[0099] Effector Cell'. As used herein, “effector cell” refers to a cell of the immune system that expresses one or more Fc receptors and mediates one or more effector functions. In some embodiments, effector cells may include, but may not be limited to, one or more of monocytes, macrophages, neutrophils, dendritic cells, eosinophils, mast cells, platelets, large granular lymphocytes, Langerhans' cells, natural killer (NK) cells, T-lymphocytes, B-lymphocytes and may be from any organism including but not limited to humans, mice, rats, rabbits, and monkeys.

[0100] Expression'. As used herein, “expression” of a nucleic acid sequence refers to one or more of the following events: (1) production of an RNA template from a DNA sequence (e.g., by transcription); (2) processing of an RNA transcript (e.g., by splicing, editing, 5’ cap formation, and / or 3’ end formation); (3) translation of an RNA into a polypeptide or protein; and / or (4) post-translational modification of a polypeptide or protein.

[0101] Fusion protein '. As used herein, the term “fusion protein” generally refers to a polypeptide including at least two segments, each of which shows a high degree of amino acid identity to a peptide moiety that (1) occurs in nature, and / or (2) represents a functional domain of a polypeptide. Typically, a polypeptide containing at least two such segments is considered to be a fusion protein if the two segments are moieties that (1) are not included in nature in the same peptide, and / or (2) have not previously been linked to one another in a single polypeptide, and / or (3) have been linked to one another through action of the hand of man.- 20 - 13321680vlAttorney Docket No. 2012106-0161

[0102] Gene: As used herein, the term “gene” has its meaning as understood in the art. It will be appreciated by those of ordinary skill in the art that the term “gene” may include gene regulatory sequences (e.g., promoters, enhancers, etc.) and / or intron sequences. It will further be appreciated that definitions of gene include references to nucleic acids that do not encode proteins but rather encode functional RNA molecules such as tRNAs, RNAi-inducing agents, etc. For the purpose of clarity we note that, as used in the present application, the term “gene” generally refers to a portion of a nucleic acid that encodes a protein; the term may optionally encompass regulatory sequences, as will be clear from context to those of ordinary skill in the art. This definition is not intended to exclude application of the term “gene” to non-proteincoding expression units but rather to clarify that, in most cases, the term as used in this document refers to a protein-coding nucleic acid.

[0103] Gene product or expression product. As used herein, the term “gene product” or “expression product” generally refers to an RNA transcribed from the gene (pre-and / or postprocessing) or a polypeptide (pre- and / or post-modification) encoded by an RNA transcribed from the gene.

[0104] Immune response: As used herein, the term “immune response” refers to a response elicited in an animal. An immune response may refer to cellular immunity, humoral immunity or may involve both. An immune response may also be limited to a part of the immune system. For example, in certain embodiments, an immunogenic composition may induce an increased IFNy response. In certain embodiments, an immunogenic composition may induce a mucosal IgA response (e.g., as measured in nasal and / or rectal washes). In certain embodiments, an immunogenic composition may induce a systemic IgG response (e.g., as measured in serum). In certain embodiments, an immunogenic composition may induce virusneutralizing antibodies or a neutralizing antibody response. In certain embodiments, an immunogenic composition may induce a cytolytic (CTL) response by T cells.

[0105] Immune Synapse. As used herein, the term “immune synapse” refers to a specialized junction between a T cell and an antigen-presenting cell (APC). This interface facilitates effective communication and activation of the T cell. Several accessory proteins are integral to the formation and function of the immune synapse. The CD3 complex is associated - 21 - 13321680vlAttorney Docket No. 2012106-0161non-covalently with the TCR, the CD3 complex consists of four invariant chains: CD3γ, CD3δ, CD3ε, and CD3ζ. These chains assemble into dimers (γε, δε, and ζζ) and play a pivotal role in signal transduction following antigen recognition. The co-receptors CD4 and CD8 are engaged to increase immune responses. CD4 is expressed on helper T cells and binds to MHC class II molecules on APCs to enhance the sensitivity of TCR-antigen recognition. CD8 is expressed by cytotoxic T cells and interacts with MHC class I molecules to facilitate the recognition of infected or malignant cells. Co-stimulatory proteins that modulate T cell activation and tolerance include CD28 that provides essential co-stimulatory signals upon binding to B7 molecules (CD80 / CD86) on APCs, promoting T cell activation and survival and CD2, that binds to LFA3 (CD58) to increase T cell sensitivity to antigen. The orchestrated interaction of the TCR with these accessory proteins at the immune synapse ensures precise antigen recognition and the initiation of appropriate T cell responses, which are vital for effective immunity.

[0106] Improve, increase, or reduce'. As used herein, the terms “improve,” “increase” or “reduce,” or grammatical equivalents, indicate values that are relative to a baseline measurement, such as a measurement in the same individual prior to initiation of the treatment described herein, or a measurement in a control individual (or multiple control individuals) in the absence of the treatment described herein.

[0107] Individual, subject, patient'. As used herein, the terms “subject,” “individual” or “patient” refer to a human or a non-human mammalian subject. The individual (also referred to as “patient” or “subject”) being treated is an individual (fetus, infant, child, adolescent, or adult) suffering from a disease, for example, cancer. In some embodiments, the subject is a human.

[0108] Linker: As used herein, the term “linker” refers to, e.g., in a fusion protein, an amino acid sequence of an appropriate length other than that appearing at a particular position in the natural protein and is generally designed to be flexible and / or to interpose a structure, such as an a-helix, between two protein moieties. In general, a linker allows two or more domains of a fusion protein to retain 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% or more of the biological activity of each of the domains. A linker may also be referred to as a spacer.

[0109] Nucleic acid. As used herein, “nucleic acid”, in its broadest sense, refers to any compound and / or substance that is or can be incorporated into an oligonucleotide chain. In some - 22 - 13321680vlAttorney Docket No. 2012106-0161embodiments, a nucleic acid is a compound and / or substance that is or can be incorporated into an oligonucleotide chain via a phosphodiester linkage. As will be clear from context, in some embodiments, “nucleic acid” refers to individual nucleic acid residues (e.g., nucleotides and / or nucleosides); in some embodiments, “nucleic acid” refers to an oligonucleotide chain comprising individual nucleic acid residues. Tn some embodiments, a “nucleic acid” is or comprises RNA; in some embodiments, a “nucleic acid” is or comprises DNA. In some embodiments, a nucleic acid is, comprises, or consists of one or more natural nucleic acid residues. In some embodiments, a nucleic acid is, comprises, or consists of one or more nucleic acid analogs. In some embodiments, a nucleic acid analog differs from a nucleic acid in that it does not utilize a phosphodiester backbone. For example, in some embodiments, a nucleic acid is, comprises, or consists of one or more “peptide nucleic acids”, which are known in the art and have peptide bonds instead of phosphodiester bonds in the backbone, are considered within the scope of the present invention. Alternatively or additionally, in some embodiments, a nucleic acid has one or more phosphorothioate and / or 5'-N-phosphoramidite linkages rather than phosphodiester bonds. In some embodiments, a nucleic acid is, comprises, or consists of one or more natural nucleosides (e.g., adenosine, thymidine, guanosine, cytidine, uridine, deoxyadenosine, deoxythymidine, deoxy guanosine, and deoxycytidine). In some embodiments, a nucleic acid is, comprises, or consists of one or more nucleoside analogs (e.g., 2-aminoadenosine, 2-thiothymidine, inosine, pyrrolo-pyrimidine, 3 -methyl adenosine, 5 -methyl cytidine, C-5 propynyl-cytidine, C-5 propynyl-uridine, 2-aminoadenosine, C5-bromouridine, C5-fluorouridine, C 5 -iodouridine, C5-propynyl-uridine, C5 -propynyl-cytidine, C5-methylcytidine, 2-aminoadenosine, 7-deazaadenosine, 7-deazaguanosine, 8-oxoadenosine, 8-oxoguanosine, 0(6)-methylguanine, 2-thiocytidine, methylated bases, intercalated bases, and combinations thereof). In some embodiments, a nucleic acid comprises one or more modified sugars (e.g., 2'-fluororibose, ribose, 2'-deoxyribose, arabinose, and hexose) as compared with those in natural nucleic acids. In some embodiments, a nucleic acid has a nucleotide sequence that encodes a functional gene product such as an RNA or protein. In some embodiments, a nucleic acid includes one or more introns. In some embodiments, nucleic acids are prepared by one or more of isolation from a natural source, enzymatic synthesis by polymerization based on a complementary template (in vivo or in vitro), reproduction in a recombinant cell or system, and - 23 - 13321680vlAttorney Docket No. 2012106-0161chemical synthesis. In some embodiments, a nucleic acid is at least 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, 20, 225, 250, 275, 300, 325, 350, 375, 400, 425, 450, 475, 500, 600, 700, 800, 900, 1000, 1500, 2000, 2500, 3000, 3500, 4000, 4500, 5000 or more residues long. In some embodiments, a nucleic acid is single stranded; in some embodiments, a nucleic acid is double stranded. In some embodiments a nucleic acid has a nucleotide sequence comprising at least one element that encodes, or is the complement of a sequence that encodes, a polypeptide. In some embodiments, a nucleic acid has enzymatic activity.

[0110] Operably linked. As used herein, “operably linked” refers to a juxtaposition wherein the components described are in a relationship permitting them to function in their intended manner. A control sequence “operably linked” to a coding sequence is ligated in such a way that expression of the coding sequence is achieved under conditions compatible with the control sequences. “Operably linked” sequences include both expression control sequences that are contiguous with the gene of interest and expression control sequences that act in trans or at a distance to control the gene of interest. The term “expression control sequence” as used herein refers to polynucleotide sequences that are necessary to effect the expression and processing of coding sequences to which they are ligated. Expression control sequences include appropriate transcription initiation, termination, promoter and enhancer sequences; efficient RNA processing signals such as splicing and polyadenylation signals; sequences that stabilize cytoplasmic mRNA; sequences that enhance translation efficiency (i.e., Kozak consensus sequence); sequences that enhance protein stability; and when desired, sequences that enhance protein secretion. The nature of such control sequences differs depending upon the host organism. For example, in prokaryotes, such control sequences generally include promoter, ribosomal binding site, and transcription termination sequence, while in eukaryotes, typically, such control sequences include promoters and transcription termination sequence. The term “control sequences” is intended to include components whose presence is essential for expression and processing, and can also include additional components whose presence is advantageous, for example, leader sequences and fusion partner sequences.- 24 - 13321680vlAttorney Docket No. 2012106-0161

[0111] Patient. As used herein, the term “patient” refers to any organism to which a provided composition is or may be administered, e.g., for experimental, diagnostic, prophylactic, cosmetic, and / or therapeutic purposes. Typical patients include animals (e.g., mammals such as mice, rats, rabbits, non-human primates, and / or humans). In some embodiments, a patient is a human. In some embodiments, a patient is suffering from or susceptible to one or more disorders or conditions. In some embodiments, a patient displays one or more symptoms of a disorder or condition. In some embodiments, a patient has been diagnosed with one or more disorders or conditions. In some embodiments, the disorder or condition is or includes cancer, or presence of one or more tumors. In some embodiments, the patient is receiving or has received certain therapy to diagnose and / or to treat a disease, disorder, or condition.

[0112] Peptide-. The term “peptide” as used herein refers to a polypeptide that is typically relatively short, for example having a length of less than about 100 amino acids, less than about 50 amino acids, less than 20 amino acids, or less than 10 amino acids.

[0113] Pharmaceutically acceptable'. The term “pharmaceutically acceptable” as used herein, refers to substances that, within the scope of sound medical judgment, are suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit / risk ratio.

[0114] Polypeptide-. As used herein, a “polypeptide”, generally speaking, is a string of at least two amino acids attached to one another by a peptide bond. In some embodiments, a polypeptide may include at least 3-5 amino acids, each of which is attached to others by way of at least one peptide bond. Those of ordinary skill in the art will appreciate that polypeptides sometimes include “non-natural” amino acids or other entities that nonetheless are capable of integrating into a polypeptide chain, optionally.

[0115] Promoter-. As used herein, a “promoter” is a DNA sequence recognized by the synthetic machinery of the cell, or introduced synthetic machinery, required to initiate the specific transcription of a polynucleotide sequence. A “constitutive” promoter is a nucleotide sequence which, when operably linked with a polynucleotide that encodes or specifies a gene product, causes the gene product to be produced in a cell under most or all physiological - 25 - 13321680vlAttorney Docket No. 2012106-0161conditions of the cell. An “inducible” promoter is a nucleotide sequence that, when operably linked with a polynucleotide that encodes or specifies a gene product, causes the gene product to be produced in a cell substantially only when a promoter-specific inducer is present in the cell.

[0116] Protein: As used herein, the term “protein”, refers to a polypeptide (i.e., a string of at least two amino acids linked to one another by peptide bonds). Proteins may include moi eties other than amino acids (e.g., may be glycoproteins, proteoglycans, etc.) and / or may be otherwise processed or modified. Those of ordinary skill in the art will appreciate that a “protein” can be a complete polypeptide chain as produced by a cell (with or without a signal sequence), or can be a portion thereof. Those of ordinary skill will appreciate that a protein can sometimes include more than one polypeptide chain, for example linked by one or more disulfide bonds or associated by other means. Polypeptides may contain L-amino acids, D-amino acids, or both and may contain any of a variety of amino acid modifications or analogs known in the art. Useful modifications include, e.g., terminal acetylation, amidation, methylation, etc. In some embodiments, proteins may comprise natural amino acids, non-natural amino acids, synthetic amino acids, and combinations thereof.

[0117] Reference-. As used herein, “reference” describes a standard or control relative to which a comparison is performed. For example, in some embodiments, an agent, animal, individual, population, sample, sequence or value of interest is compared with a reference or control agent, animal, individual, population, sample, sequence or value. In some embodiments, a reference or control is tested and / or determined substantially simultaneously with the testing or determination of interest. In some embodiments, a reference or control is a historical reference or control, optionally embodied in a tangible medium. Typically, as would be understood by those skilled in the art, a reference or control is determined or characterized under comparable conditions or circumstances to those under assessment. Those skilled in the art will appreciate when sufficient similarities are present to justify reliance on and / or comparison to a particular possible reference or control.

[0118] Solid tumor-. As used herein, the term “solid tumor” refers to an abnormal mass of tissue that usually does not contain cysts or liquid areas. Solid tumors may be benign or malignant. Different types of solid tumors are named for the type of cells that form them.- 26 - 13321680vlAttorney Docket No. 2012106-0161Examples of solid tumors are sarcomas, carcinomas, lymphomas, mesothelioma, neuroblastoma, retinoblastoma, etc.

[0119] Stage of cancer'. As used herein, the term “stage of cancer” refers to a qualitative or quantitative assessment of the level of advancement of a cancer. Criteria used to determine the stage of a cancer include, but are not limited to, the size of the tumor and the extent of metastases (e.g., localized or distant).

[0120] Subject. By “subject” is meant a mammal (e.g., a human, in some embodiments including prenatal human forms). In some embodiments, a subject is suffering from a relevant disease, disorder or condition. In some embodiments, a subject is susceptible to a disease, disorder, or condition. In some embodiments, a subject displays one or more symptoms or characteristics of a disease, disorder or condition. In some embodiments, a subject does not display any symptom or characteristic of a disease, disorder, or condition. In some embodiments, a subject is someone with one or more features characteristic of susceptibility to or risk of a disease, disorder, or condition. In some embodiments, a subject is a patient. In some embodiments, a subject is an individual to whom diagnosis and / or therapy is and / or has been administered.

[0121] Suffering from: An individual who is “suffering from” a disease, disorder, or condition (e.g., cancer) has been diagnosed with and / or exhibits one or more symptoms of the disease, disorder, or condition.

[0122] Symptoms are reduced: According to the present invention, “symptoms are reduced” when one or more symptoms of a particular disease, disorder or condition is reduced in magnitude (e.g., intensity, severity, etc.) or frequency. For purposes of clarity, a delay in the onset of a particular symptom is considered one form of reducing the frequency of that symptom. It is not intended that the present invention be limited only to cases where the symptoms are eliminated. The present invention specifically contemplates treatment such that one or more symptoms is / are reduced (and the condition of the subject is thereby “improved”), albeit not completely eliminated.- 27 - 13321680vlAttorney Docket No. 2012106-0161

[0123] T cell receptor. As used herein, a “T cell receptor” or “TCR” refers to the antigen-recognition molecules present on the surface of T-cells. During normal T-cell development, each of the four TCR genes, a, P, y, and 8, can rearrange leading to highly diverse TCR proteins. During immune responses a TCR functions as a component of the adaptive immune system, enabling T cells to recognize and respond to specific antigens presented by other cells (e g., antigen presenting cells). The TCR complex is composed of several proteins that work together to initiate and propagate immune responses. In some embodiments, a TCR is a heterodimer consisting of two polypeptide chains: alpha (a) and beta (P). In some embodiments, a TCR is a heterodimer consisting of two polypeptide chains: gamma (y) and delta (8). In humans, approximately 95% of T cells express this aP TCR variant. A smaller subset, about 5%, expresses a gamma (y) and delta (8) chain combination, known as y6 T cells.

[0124] Therapeutic agent. As used herein, the phrase “therapeutic agent” in general refers to any agent that elicits a desired pharmacological effect when administered to an organism. In some embodiments, an agent is considered to be a therapeutic agent if it demonstrates a statistically significant effect across an appropriate population. In some embodiments, the appropriate population may be a population of model organisms. In some embodiments, an appropriate population may be defined by various criteria, such as a certain age group, gender, genetic background, preexisting clinical conditions, etc. In some embodiments, a therapeutic agent is a substance that can be used to alleviate, ameliorate, relieve, inhibit, prevent, delay onset of, reduce severity of, and / or reduce incidence of one or more symptoms or features of a disease, disorder, and / or condition. In some embodiments, a “therapeutic agent” is an agent that has been or is required to be approved by a government agency before it can be marketed for administration to humans. In some embodiments, a “therapeutic agent” is an agent for which a medical prescription is required for administration to humans.

[0125] Therapeutically effective amount: As used herein, the term “therapeutically effective amount” means an amount that is sufficient, when administered to a population suffering from or susceptible to a disease, disorder, and / or condition in accordance with a therapeutic dosing regimen, to treat the disease, disorder, and / or condition. In some embodiments, a therapeutically effective amount is one that reduces the incidence and / or severity - 28 - 13321680vlAttorney Docket No. 2012106-0161of, stabilizes one or more characteristics of, and / or delays onset of, one or more symptoms of the disease, disorder, and / or condition. Those of ordinary skill in the art will appreciate that the term “therapeutically effective amount” does not in fact require successful treatment be achieved in a particular individual. Rather, a therapeutically effective amount may be that amount that provides a particular desired pharmacological response in a significant number of subjects when administered to patients in need of such treatment. For example, in some embodiments, “therapeutically effective amount” refers to an amount which, when administered to an individual in need thereof in the context of inventive therapy, will block, stabilize, attenuate, or reverse a cancer-supportive process occurring in said individual, or will enhance or increase a cancer-suppressive process in said individual. In the context of cancer treatment, a “therapeutically effective amount” is an amount which, when administered to an individual diagnosed with a cancer, will prevent, stabilize, inhibit, or reduce the further development of cancer in the individual. A particularly preferred “therapeutically effective amount” of a composition described herein reverses (in a therapeutic treatment) the development of a malignancy such as a pancreatic carcinoma or helps achieve or prolong remission of a malignancy. A therapeutically effective amount administered to an individual to treat a cancer in that individual may be the same or different from a therapeutically effective amount administered to promote remission or inhibit metastasis. As with most cancer therapies, the therapeutic methods described herein are not to be interpreted as, restricted to, or otherwise limited to a “cure” for cancer; rather the methods of treatment are directed to the use of the described compositions to “treat” a cancer, i.e., to effect a desirable or beneficial change in the health of an individual who has cancer. Such benefits are recognized by skilled healthcare providers in the field of oncology and include, but are not limited to, a stabilization of patient condition, a decrease in tumor size (tumor regression), an improvement in vital functions (e.g., improved function of cancerous tissues or organs), a decrease or inhibition of further metastasis, a decrease in opportunistic infections, an increased survivability, a decrease in pain, improved motor function, improved cognitive function, improved feeling of energy (vitality, decreased malaise), improved feeling of well-being, restoration of normal appetite, restoration of healthy weight gain, and combinations thereof. In addition, regression of a particular tumor in an individual (e.g., as the result of treatments described herein) may also be assessed by taking - 29 - 13321680vlAttorney Docket No. 2012106-0161samples of cancer cells from the site of a tumor such as a pancreatic adenocarcinoma (e.g., over the course of treatment) and testing the cancer cells for the level of metabolic and signaling markers to monitor the status of the cancer cells to verify at the molecular level the regression of the cancer cells to a less malignant phenotype. For example, tumor regression induced by employing the methods of this invention would be indicated by finding a decrease in one or more pro-angiogenic markers, an increase in anti -angiogenic markers, the normalization (i.e., alteration toward a state found in normal individuals not suffering from cancer) of metabolic pathways, intercellular signaling pathways, or intracellular signaling pathways that exhibit abnormal activity in individuals diagnosed with cancer. Those of ordinary skill in the art will appreciate that, in some embodiments, a therapeutically effective amount may be formulated and / or administered in a single dose. In some embodiments, a therapeutically effective amount may be formulated and / or administered in a plurality of doses, for example, as part of a dosing regimen.

[0126] Transformation'. As used herein, “transformation” refers to any process by which exogenous DNA is introduced into a host cell. Transformation may occur under natural or artificial conditions using various methods well known in the art. Transformation may rely on any known method for the insertion of foreign nucleic acid sequences into a prokaryotic or eukaryotic host cell. In some embodiments, a particular transformation methodology is selected based on the host cell being transformed and may include, but is not limited to, viral infection, electroporation, mating, lipofection. In some embodiments, a “transformed” cell is stably transformed in that the inserted DNA is capable of replication either as an autonomously replicating plasmid or as part of the host chromosome. In some embodiments, a transformed cell transiently expresses introduced nucleic acid for limited periods of time.

[0127] Treatment. As used herein, the term “treatment” (also “treat” or “treating”) refers to any administration of a substance that partially or completely alleviates, ameliorates, relives, inhibits, delays onset of, reduces severity of, and / or reduces incidence of one or more symptoms, features, and / or causes of a particular disease, disorder, and / or condition (e g., cancer). Such treatment may be of a subject who does not exhibit signs of the relevant disease, disorder and / or condition and / or of a subject who exhibits only early signs of the disease, disorder, and / or - 30 - 13321680vlAttorney Docket No. 2012106-0161condition. Alternatively or additionally, such treatment may be of a subject who exhibits one or more established signs of the relevant disease, disorder and / or condition. In some embodiments, treatment may be of a subject who has been diagnosed as suffering from the relevant disease, disorder, and / or condition. In some embodiments, treatment may be of a subject known to have one or more susceptibility factors that are statistically correlated with increased risk of development of the relevant disease, disorder, and / or condition.

[0128] Tumor infiltrating lymphocyte: As used herein, the term “tumor-infiltrating lymphocytes” refers to white blood cells of a subject afflicted with a cancer (such as melanoma), that have left the blood stream and have migrated into a tumor. In some embodiments, tumorinfiltrating lymphocytes have tumor specificity.

[0129] Vector: As used herein, “vector” refers to a nucleic acid molecule capable of transporting another nucleic acid to which it is associated. In some embodiments, vectors are capable of extra-chromosomal replication and / or expression of nucleic acids to which they are linked in a host cell such as a eukaryotic and / or prokaryotic cell. Vectors capable of directing the expression of operatively linked genes are referred to herein as “expression vectors.”DETAILED DESCRIPTION

[0130] Among other things present therapies for cancer include immunotherapies.Immunotherapies can include chimeric antigen receptor therapy (CAR) and T-cell receptor therapy (TCR).

[0131] CAR therapy involves modification of an immune cell to encode and express a chimeric antigen receptor that can recognize an antigen (e.g., a tumor antigen) on a cell. TCR therapy involves modification of an immune cell to encode and express a T-cell receptor that can recognize an antigen (e.g., a tumor antigen) presented in a human leukocyte antigen (HLA). CAR and TCR therapies can be provide as adoptive cell therapy or in vivo cell therapy.

[0132] ACT is a treatment method in which cells are removed from a donor, cultured and / or manipulated in vitro, and then administered to a patient for the treatment of a disease. In vivo cell therapy is a treatment method in which cells are manipulated in vivo for treatment of - 31 - 13321680vlAttorney Docket No. 2012106-0161disease. In some embodiments, cells for ACT or in vivo cell therapy are transduced (e.g., ex vivo or in vivo) by viral particles or lipid nanoparticles comprising a nucleic acids, encapsulated RNA, and delivery of nucleic acid sequences. A variety of cell types have been used in immunotherapies (e.g., ACT or in vivo cell therapy) in an attempt to treat several classes of disorders. Cell types used in immunotherapies include lymphoid cells (e.g., T-cells, NK cells, etc.) and myeloid cells (e.g., monocytes, macrophages, etc.)..

[0133] Companion engager therapeutics are designed to bind to a therapeutic immune cell and can be used in conjunction with ACT and / or in vivo cell therapy. Interaction of an immune cell and a companion engager therapeutic, through the formation of an immune synapse, delivers signals to the immune cell, facilitating signal transduction and communication, enabling activation and effector functions of the therapeutic immune cell. However, there are known challenges which hinder the success of current companion engager strategies in facilitating the effector functions of immune cell therapeutics against cancers.

[0134] For example, some cancers (e.g., B cell lymphoma and leukemia), escape CAR19 treatment by reducing expression of CD 19, a well-known phenomenon referred to as “antigen escape”, whereby the cancer cells escape the therapy by downregulating expression of the cancer antigen that is being targeted. A second means of escape from CD19-directed CAR-T cell therapy is the downregulation of LFA3 expression. LFA3 binds to CD2 expressed by T cells, including CAR T cells. CD2 augments cytotoxic signaling by T cells, particularly when target antigen density is low (Bachmann, et al., 1999; Burton, et al., 2023). Reduced expression of LFA3 has been observed across various cancer types that are treated by immune-mediated therapeutics, including therapeutics that target the PD-1 / PD-L1 immune checkpoint pathway, CD3-based therapeutics that function as T cell engagers, and TCR-based therapeutics (Shen, et al., 2022; Ho, et al., 2023).

[0135] In contrast to the immune-cell derived cancers (e g., B cell lineage malignancies), cancers that constitute solid tumors are derived from diverse cell types. For example, solid tumors derived from epithelial cells can lead to carcinomas, from connective tissue cells can lead to sarcomas, from neuroendocrine cells can lead to neuroendocrine tumors, from germ cells can lead to teratomas, among others. In the nervous system, cancers derived from neural stem cells - 32 - 13321680vlAttorney Docket No. 2012106-0161can lead to tumors like gliomas or medulloblastomas. Cancers whose cellular origins are different often express different antigens on the cancer cell surface.

[0136] There are several known problems with therapies directed to antigens present on solid tumors that result in low response rates that are often short lived. For example, tumor cells that are engaged by cellular therapies (e.g., CAR T cells or TCEs) cells do not form a canonical immune relevant synapse with the T cell (Lindner, et al., 2020). The failure of an immune synapse to form between the targeted cancer cell and therapeutic T cells means that signaling cascades required to trigger productive T cell proliferation and differentiation are missing. The targeted tumor cell may still be killed, but the T cells are left with limited remaining proliferative, cytotoxic activity, and may show rapid signs of anergy, exhaustion and activation-induced cell death when infused into a patient because the correct antigen-specific immune synapses have not been formed. (Su et al., 2022).

[0137] Previous work has attempted to develop companion engager therapies into efficacious cancer therapies, for example, by circumventing T cell exhaustion. For example, several attempts at developing antibody-based companion engagers have been made. For example, Staufer, et al., characterized bi-specific antibody-based T-cell engager formats with antibody fragments or single chain variable fragments linked together in cis through immunoglobulin Ig hinge or in trans across the antibody constant fragment. US 5,928,643 Bl (“’643”) also described antibody -based T-cell engagers. The ‘643 patent described polypeptides that contained a portion of LFA3 containing a CD2-binding domain fused to the Fc region of an immunoglobulin. Nair et al., described more than twenty tri-specific antibody formats aimed at simultaneously engaging T cells in the tumor microenvironment by binding both CD3 and CD28 and a tumor associated antigen. Lu et al., 2022, developed an anti-CD19, anti-CD3, anti-CD2 IgG-like tri-specific antibody that simultaneously engages CD 19 on tumor cells, the T-cell receptor (TCR) signaling component CD3, and the T-cell costimulatory receptor CD2. It’s been reported that these tri-specific antibodies lead to redirected T-cell cytotoxicity towards CD 19-positive malignant B cells. However, results from these works resulted in differential and variable abilities of the engager therapy to bind the intended tumor target antigens and kill the targeted cancer cells. These works reported these variable results may be driven by the- 33 - 13321680vlAttorney Docket No. 2012106-0161formatting of the bi-specific and tri-specific antibodies, leading to suboptimal or variable contact formation between the engager therapy and the target cell. Furthermore, the ‘643 patent reported, unexpectedly, that the LFA3-Ig polypeptides inhibited T-cell activation.

[0138] CAR -T cells have been modified in a variety of ways in attempts to address the lack of productive synapse formation. For example, overexpressing cell surface molecules like CD2, or artificial integration of signaling domains that normally would be triggered by such cell surface molecules. However, these attempts are limited by the complexity of the multiple engineering steps required to alter the activity of the CAR -T cell and by the instability of the resulting protein expression by the CAR -T cell.

[0139] In some embodiments, the present disclosure provides fusion proteins useful for improvement of CAR-T therapy. In some embodiments, CAR-T cells are engaged and activated by fusion proteins of the present application (i.e., CAR-T engagers; CTE). In some embodiments, CTE fusion proteins form a bridge between CAR-T cells and cancer cells. In some embodiments, a bridge between a CAR-T cell and a cancer cell is formed by one portion of a fusion protein (e g., an antigen binding protein) binding a tumor antigen, one portion of a fusion protein (e.g., a target polypeptide) being bound by a CAR expressed on a CAR-T cell and one portion of a fusion protein (e.g., an accessory molecule) binding an accessory molecule binding partner or ligand on the CAR-T. Without wishing to be bound by a particular theory, this dual binding brings a CAR-T and a cancer cell into close proximity leading to CAR-T cell activation and tumor lysis.

[0140] In some embodiments, the present disclosure provides fusion proteins useful for improvement of TCR-directed therapy. In some embodiments, the present disclosure provides fusion proteins useful for activation of a subject’s T-cells. In some embodiments cells comprising a TCR (e g., cells for TCR therapy and / or patient T cells) are engaged and activated by fusion proteins of the present application (i.e., T cell engager proteins; TCE). In some embodiments, TCE fusion proteins form a bridge between cells comprising a TCR and cancer cells. In some embodiments, TCE fusion proteins form a bridge between a cell comprising a TCR and a cancer cell is formed by one portion of a fusion protein (e.g., an accessory molecule) binding to a cell comprising a TCR and one portion of a fusion protein (e.g., an antigen binding - 34 - 13321680vlAttorney Docket No. 2012106-0161protein) binding a tumor antigen on a cancer cell. Accessory molecules can be proteins found within, or recruited to, an immune synapse. Without wishing to be bound by a particular theory, this dual binding brings a cell comprising a TCR and a cancer cell into close proximity leading to activation of a cell comprising a TCR and tumor cell lysis. The interaction prompts T cells to release cytotoxic granules, resulting in the targeted destruction of the cancer cells.

[0141] The present disclosure provides solutions to prior unresolved problems with efficacy and suboptimal companion engager therapeutics. The present disclosure provides, among other things, technologies with several advantages to address the problem of cancer cells evading and escaping cancer therapeutics. Among other things, the present disclosure provides technologies that address suboptimal cancer- immune synapse formation.

[0142] In some embodiments, the present disclosure provides polypeptides useful in the treatment of cancer. In some embodiments, the present disclosure provides polypeptides useful in formation of a productive synapse between a therapeutic immune cell and a tumor cell. In some embodiments, the present disclosure provides a fusion protein comprising (i) one or more accessory molecules or a fragment thereof; (ii) one or more antigen-binding proteins or fragments thereof; and (iii) a target polypeptide for a cellular therapeutic.

[0143] In some embodiments, a fusion protein of the present disclosure is designed to be bound by a cellular therapeutic while simultaneously binding to one or more targeted tumor cell surface antigen(s). By providing accessory molecules that mimic the cell surface of an antigen presenting cell immune synapse fusion proteins of the present invention are useful in formation of a productive synapse between a therapeutic immune cell and a tumor cell. In some embodiments, formation of a productive synapse increases killing of target cells (e.g., tumor cells) by immune therapeutic cells that interact with the target tumor cells via accessory proteins on presently described fusion proteins is markedly enhanced, as depicted in Figure 1.Accessory Molecules

[0144] A fusion protein of the present disclosure contains one or more accessory molecule(s) or a fragment thereof. Accessory molecules (e g., costimulatory and adhesion- 35 - 13321680vlAttorney Docket No. 2012106-0161molecules) are described in, for example, Hodge et al., Frontiers in Bioscience 11, 788-803, January 1, 2006.

[0145] In some embodiments, an accessory molecule(s) can be a costimulatory molecule(s) or fragment thereof (e.g., CD28, CD2). In some embodiments, an accessory molecule or fragment thereof is a binding partner or ligand of a costimulatory molecule (e.g. LFA3, B7-1, B7-2). In some embodiments, an accessory molecule or fragment thereof is or comprises an antigen binding protein or fragment that binds an adhesion molecule, a costimulatory molecule, or a combination thereof. In some embodiments, an antigen binding protein includes an antibody or antigen-binding fragment thereof. In some embodiments, and antibody is an IgG, IgE and IgM, bi- or multi- specific antibodies (e.g., Zybodies®, etc). In some embodiments an antigen binding protein is an antibody fragment. In some embodiments, an antibody fragment includes a Fab fragment, Fab’ fragment, F(ab’)2 fragment, scFv fragment, Fv fragment, dsFv diabody, dAb fragment, Fd’ fragment, Fd fragment, an isolated complementarity determining region (CDR), a cameloid antibody, a masked antibody (e g., Probody®), a single chain or Tandem diabody (TandAb®), a VHH, an Anticalin®, a single-domain antibody (e.g., Nanobody®), an ankyrin repeat protein or DARPIN®, an Avimer®, an Adnectin®, an Affilin®, an Affibody®, a Fynomer®, or a Centyrin®). In some embodiments, an antigen binding protein is or includes a TCR or antigen-binding portion thereof. In some embodiments, an antigen binding protein is a pH sensitive domain (see, e.g., Schroter et al., MAbs 7:138-51 (2015)).

[0146] A costimulatory molecule can, in some embodiments, enhance the activation of immune cell therapy through engagement of one or more receptors on immune therapeutic cells. In some embodiments, a costimulatory molecule is CD2, CD3, CD28, B7-1 (CD80) or B7-2 (CD86), B7-HZB7H-RP-1, B7-H1 / PD-L1, B7-DC, ICAM-1, OX40L, 4-1BBL, CD40, CD70 / CD27L, SLAM-family proteins, CD226 (DNAM-1) or Light. In some embodiments, a costimulatory molecule is recruited to an immune synapse as a consequence of T cell activation.

[0147] In some embodiments, an accessory molecule(s) can be an adhesion molecule. In some embodiments, an accessory molecule is a binding partner or ligand of an adhesion molecule. In some embodiments and adhesion molecule is B7-1, B7-2, L-selectin, a4 integrin, lymphocyte function-associated antigen-1 (LFA-1), Intercellular Adhesion Molecule 1 (1CAM- - 36 - 13321680vlAttorney Docket No. 2012106-01611) (CD54), CD2, LFA-3 (CD58), CD28 / CTLA-4, ICOS, PD1, LFA1, 0X40 (CD134), 4-1BB (CD137), CD40L (CD154), CD27, SLAM-family proteins, CD155, CD112, T1G1T, orLight-R.

[0148] In some embodiments, a fusion protein of the present disclosure comprises one or more (e.g., 1, 2, 3, 4, 5 or more) accessory molecules. In some embodiments, a fusion protein of the present disclosure comprises one or more (e.g., 1, 2, 3, 4, 5 or more) costimulatory molecules. In some embodiments, a fusion protein of the present disclosure comprises one or more (e.g., 1, 2, 3, 4, 5 or more) adhesion molecules. In some embodiments, a fusion protein of the present disclosure comprises one or more (e.g., 1, 2, 3, 4, 5 or more) costimulatory molecules and / or one more (e.g., 1, 2, 3, 4, 5 or more) adhesion molecules.

[0149] In some embodiments, fusion proteins of the present disclosure contain the addition of multiple LFA3 binding domains (LFA3bd). In solution, LFA3 binds with low affinity to its cognate partner, CD2. However, CD2 is bound with higher affinity in the presence of multiple binding interactions with LFA3 when LFA3 and CD2 are expressed on targeted cancer cells and T cells, respectively, due to avidity effects. Thus, it is proposed that the presence of two or more domains will be required to provide the necessary avidity that triggers signaling through CD2.

[0150] In some embodiments fusion proteins of the present disclosure contain one or more B7-derived binding domains (B7bd) to enhance CTE-interacting CAR -T cell and TCE-interacting T cell activity.

[0151] In some embodiments fusion proteins of the present disclosure contain one or more B7 binding domains to provide additive or synergistic activity with LFA3bd when both are present.

[0152] In some embodiments fusion proteins of the present disclosure contain immune checkpoint inhibition domains. For example, as evidence by TGF R2 antagonism of TGFP-mediated SMAD phosphorylation. TGF[3 pathway antagonism will be synergistic with Costim-CTE and Costim-TCE activity.Antigen Binding Proteins- 37 - 13321680vlAttorney Docket No. 2012106-0161

[0153] In some embodiments, a fusion protein of the present disclosure comprises one or more antigen-binding proteins or fragments that bind a tumor antigen. In some embodiments, an antigen binding protein is a polypeptide that specifically binds to a target antigen, e.g., a tumor antigen described herein. In some embodiments, an antigen binding protein includes an antibody or antigen-binding fragment thereof. In some embodiments, and antibody is an IgG, IgE and IgM, bi- or multi- specific antibodies (e.g., Zybodies®, etc). In some embodiments an antigen binding protein is an antibody fragment. In some embodiments, an antibody fragment includes a Fab fragment, Fab’ fragment, F(ab’)2 fragment, scFv fragment, Fv fragment, dsFv diabody, dAb fragment, Fd’ fragment, Fd fragment, an isolated complementarity determining region (CDR), a cameloid antibody, a masked antibody (e.g., Probody®), a single chain or Tandem diabody (TandAb®), a VHH, an Anticalin®, a single-domain antibody (e.g., Nanobody®), an ankyrin repeat protein or DARPIN®, an Avimer®, an Adnectin®, an Affilin®, an Affibody®, a Fynomer®, or a Centyrin®). In some embodiments, an antigen binding protein is or includes a TCR or antigen-binding portion thereof. In some embodiments, an antigen binding protein is a pH sensitive domain (see, e.g., Schroter et al., MAbs 7:138-51 (2015)).

[0154] Antigen binding proteins can be selected based on, e.g., type and number of target antigens present on or near a surface of a target cell. For example, an antigen binding protein can be chosen to recognize an antigen that acts as a cell surface marker on a target cell associated with a particular disease state. In some embodiments, an antigen binding protein is selected to specifically bind to an antigen on a tumor cell. Selection of an antigen binding domain can depend on, e.g., a particular type of cancer to be treated.

[0155] In some embodiments, a tumor antigen described herein can be a tumor-specific antigen (TSA) or a tumor-associated antigen (TAA). A TSA is (or is believed to be) unique to tumor cells and does not occur on other cells in the body (e.g., does not occur to a significant extent on other cells). A TAA is not unique to a tumor cell and instead is also expressed on a normal cell (e.g., expressed under conditions that fail to induce a state of immunologic tolerance to the antigen). For example, TAAs can be antigens that are expressed on normal cells during fetal development when the immune system is immature and unable to respond, or they can be- 38 - 13321680vlAttorney Docket No. 2012106-0161antigens that are normally present at extremely low levels on normal cells but that are expressed at higher levels on tumor cells.

[0156] Non-limiting examples of tumor antigens can include tumor antigen is MART-1 / MelanA (MART-I), gplOO (Pmel 17), tyrosinase, TRP-1, TRP-2, MAGE-1, MAGE-3, BAGE, GAGE-1, GAGE-2, p!5, CEA, p53, Ras, HER-2 / neu, BCR-ABL, E2A-PRL, H4-RET, IGH-IGK, MYL-RAR, Epstein Barr virus antigens EBVA, human papillomavirus (HPV) antigen E6 orE7, TSP-180, MAGE-4, MAGE-5, MAGE-6, RAGE, NY-ESO, erbB, pl85erbB2, pl80erbB-3, c-met, nm-23Hl, PSA, TAG-72, CA 19-9, CA 72-4, CAM 17.1, NuMa, K-ras, beta-Catenin, CDK4, Mum-1, p 15, p 16, 43-9F, 5T4, 791Tgp72, alpha-fetoprotein, beta-HCG, BCA225, BTAA, CA 125, CA 15-3\CA 27.29\BCAA, CA 195, CA 242, CA-50, CAM43, CD68\P1, CO-029, FGF-5, G250, Ga733\EpCAM, HTgp-175, M344, MA-50, MG7-Ag, MOV18, NB / 70K, NY-CO-1, RCAS1, SDCCAG16, TA-90\Mac-2 binding protein\cyclophilin C-associated protein, TAAL6, TAG72, TLP, MUC16, IL13Ra2, FRa, VEGFR2, Lewis Y, FAP, EphA2, CEACAM5, EGFR, CA6, CA9, GPNMB, EGP1, FOLR1, endothelial receptor, STEAP1, SLC44A4, Nectin-4, AGS-16, guanalyl cyclase C, MUC-1, CFC1B, integrin alpha 3 chain (of a3bl, a laminin receptor chain), TPS, CD 19, CD20, CD22, CD30, CD72, CD 180, CD171 (L1CAM), CD123, CD133, CD138, CD37, CD70, CD79a, CD79b, CD56, CD74, CD166, CD71, CLL-1 / CLEC12A, ROR1, Glypican 3 (GPC3), Mesothelin, CD33 / IL3Ra, c-Met, PSCA, PSMA, Glycolipid F77, EGFRvIII, BCMA, GD-2, MY-ESO-1, B7-H3 (CD276) or MAGE A3. In some embodiments, a tumor antigen is CD19, CD20, Her2, B7-H3, or IL13Ra2.Target Polypeptide

[0157] In some embodiments, a fusion protein of the present disclosure comprises a target polypeptide for a cellular therapeutic. In some embodiments, a target polypeptide is bound by a cellular therapeutic. In some embodiments, a cellular therapeutic comprises a receptor that recognizes and / or binds a target polypeptide of a fusion protein described herein. In some embodiments, a target polypeptide is an antigen bound by an antigen binding protein (e.g., antibody or fragment thereof) on a cellular therapeutic.

[0158] In some embodiments, a target polypeptide comprises or consists of all or a portion of a tumor associated antigen (TAA) or tumor specific antigen (TSA). Non-limiting - 39 - 13321680vlAttorney Docket No. 2012106-0161examples of TSA or TAA antigens include differentiation antigens such as MART-l / MelanA (MART-1), gplOO (Pmel 17), tyrosinase, TRP-1, TRP-2 and tumor-specific multilineage antigens such as MAGE-1, MAGE-3, BAGE, GAGE-1, GAGE-2, pl5; overexpressed embryonic antigens such as CEA; overexpressed oncogenes and mutated tumor-suppressor genes such as p53, Ras, HER-2 / neu; unique tumor antigens resulting from chromosomal translocations such as BCR-ABL, E2A-PRL, H4-RET, IGH-IGK, MYL-RAR; and viral antigens, such as the Epstein Barr virus antigens EBVA and the human papillomavirus (HPV) antigens E6 and E7. Other tumor antigens include TSP- 180, MAGE-4, MAGE-5, MAGE-6, RAGE, NY-ESO, erbB, pl85erbB2, pl80erbB-3, c-met, nm-23Hl, PSA, TAG-72, CA 19-9, CA 72-4, CAM 17.1, NuMa, K-ras, beta-Catenin, CDK4, Mum-1, p 15, p 16, 43-9F, 5T4, 791Tgp72, alphafetoprotein, beta-HCG, BCA225, BTAA, CA 125, CA 15-3\CA 27.29\BCAA, CA 195, CA 242, CA-50, CAM43, CD68\P1, CO-029, FGF-5, G250, Ga733\EpCAM, HTgp-175, M344, MA-50, MG7-Ag, MOV 18, NB / 70K, NY-CO-1, RCAS1, SDCCAG16, TA-90\Mac-2 binding protein\cyclophilin C-associated protein, TAAL6, TAG72, TLP, MUC16, IL13Ra2, FRa, VEGFR2, Lewis Y, FAP, EphA2, CEACAM5, EGFR, CA6, CA9, GPNMB, EGP1, FOLR1, endothelial receptor, STEAP1, SLC44A4, Nectin-4, AGS-16, guanalyl cyclase C, MUC-1, CFC1B, integrin alpha 3 chain (of a3bl, a laminin receptor chain), and TPS.

[0159] In some embodiments, a target polypeptide comprises or consists of all or a portion of a tumor antigen selected from CD 19, CD20, CD22, CD30, CD72, CD 180, CD171 (L1CAM), CD123, CD133, CD138, CD37, CD70, CD79a, CD79b, CD56, CD74, CD166, CD71, CLL-1 / CLEC12A, ROR1, Glypican 3 (GPC3), Mesothelin, CD33 / IL3Ra, c-Met, PSCA, PSMA, Glycolipid F77, EGFRvIII, GD-2, MY-ESO-1, and MAGE A3.

[0160] In some embodiments, a target polypeptide comprises or consists of all or a portion of a B cell specific marker such as CD 19, CD20, CD21, CD22, CD23, CD24, CD40, CD72, CD180, ROR1, BCMA, HLA-DR10, CD1, CD5, CD21, CD25, CD27, CD30, CD38, CD78, CD80, CD86, CD 138, CD319, surface Ig, PD-1, PD-L1, PD-L2, TGFPR2, CD79a, and CD79b (see, e.g., LeBien et al., Blood 112:1570-1580 (2008).

[0161] The present disclosure provides, in part, multi-domain fusion protein accessory molecules designed to bind to a cellular therapeutic while simultaneously binding to one or more - 40 - 13321680vlAttorney Docket No. 2012106-0161targeted tumor cell antigen(s). These accessory molecules include costimulatory (e.g., CD28, CD2, or LFA-1) and / or adhesion (e.g., B7-1, B7-2, LFA3, or ICAM-1) domains that mimic the cell surface of an antigen presenting cell’s immune synapse. For example, in some embodiments, an accessory molecule includes one more (e g., 1, 2, 3, 4, 5 or more) costimulatory domains, one more (e.g., 1, 2, 3, 4, 5 or more) adhesion domains, and protein target domain for a CAR T cell (e.g., the CD 19 extracellular domain).Half-Life Extension

[0162] In some embodiments, a fusion protein of the disclosure includes an agent that increases the half-life of the fusion protein, e.g., relative to the fusion protein without the agent. In some embodiments, such an agent is a polypeptide, referred to herein as a “half-life extension polypeptide”.

[0163] In some embodiments, the half-life extension polypeptide is a transferrin polypeptide or a portion thereof. Transferrin is recycled by binding to a transferrin receptor (see, e.g., Widera et al., Adv. Drug Deliv. Rev. 55: 1439-66 (2003)). In some embodiments, the halflife extension polypeptide is albumin (e.g., bovine serum albumin (BSA), human serum albumin (HSA), or mouse serum albumin (MSA)) or a fragment thereof. In some embodiments, the halflife extension polypeptide is a polypeptide that binds a serum protein. In some embodiments, the half-life extension polypeptide is a serum albumin binder (e.g., a BSA, HSA, or MSA binder).

[0164] In some embodiments the serum albumin binder is an albumin binding peptide. Peptides that bind to albumin are described in WO200145746, W02002076489, W02008068280, WO2009127691, WO2011095545, and US Pat. Pub. Nos. 20040001827, 20080187517, and 20130316952. One of skill in the art is familiar with methods to link proteins and antibodies directly to albumin or domains of albumin as described, for example in Patterson et al., Bioconjugate Chem. 2016, 27, 10, 2271–2275; Bern et al., Sci. Trans. Med., 14 Oct 2020 • Vol 12, Issue 565.

[0165] In some embodiments, the half-life extension polypeptide is or comprises a hyaluronan binding domain. Hyaluronan (HA), also known as hyaluronic acid, is a glycosaminoglycan that is found in connective and other tissues and is abundant in synovial- 41 - 13321680vlAttorney Docket No. 2012106-0161fluid, skin, and the vitreous body. HA binds to a large number naturally occurring hyaluronan-binding proteins (HABPs). Some HABPs contain an HA binding domain referred to as a link module through which they bind to HA (Kohda, C. J et al., Cell 86 (1996) 767- 775.). Some HAPBs contain a linear 9-11 residue HA-binding motif containing multiple basic amino acids termed a B-X7-B motif (Yang B., et al. Identification of a common hyaluronan binding motif in the hyaluronan binding proteins RHAMM, CD44 and link protein. EMBO J., 13: 286-296, 1994).

[0166] In some embodiments, the half-life extension polypeptide is a PAS polypeptide. As used herein, a “PAS polypeptide” is a polypeptide characterized in that the sum of proline, alanine, and serine residues constitutes more than about 80%, or about 85%, or about 90%, or about 95%, or about 96%, or about 97%, or about 98%, or about 99%, or 100% of the total amino acid sequence of the half-life extension polypeptide. In general, a PAS polypeptide is characterized in that it adopts a random coil conformation under physiological conditions as described in US Pat. Pub. No. 20100292130. PAS polypeptides that may be used as half-life extension polypeptides in the fusion proteins of the present disclosure are further described in WO 2008 / 155134, US Pat. Pub. No. 20100292130, US Pat. No. 8,563,521 and / or US Pat. No. 9,260,494.

[0167] In some embodiments a half-life extension polypeptide consists solely of proline and alanine or consists predominantly of proline and alanine but can have up to 1%, 2%, 3%, 5%, or 10% other amino acid residues. Where other amino acids are present, they may all be the same, or multiple different amino acids may be present. Examples of polypeptides that are composed predominantly or entirely of proline and alanine and adopt a random coil conformation under physiological conditions, referred to as proline / alanine random coil polypeptides, are described in US Pat. Pub. No. 20130072420, US Pat. Nos. 9,221,882, and / or US Pat. No. 10,081,657.

[0168] In some embodiments a half-life extension polypeptide is characterized in that the sum of glycine, alanine, serine, threonine, glutamate, and proline residues constitutes more than about 80%, or about 85%, or about 90%, or about 95%, or about 96%, or about 97%, or about 98%, or about 99%, or 100% of the total amino acid sequence of the half-life extension- 42 - 13321680vlAttorney Docket No. 2012106-0161polypeptide, and the half-life extension polypeptide comprises at least 4 of these 6 different amino acids. Such a polypeptide may be referred to as being composed predominantly of amino acids selected from G, A, S, T, E, and P (see, e.g., US Pat. Pub. No. 20030228309; 9,926,351; 9,976,166; and 10,961,287).

[0169] In some embodiments, a half-life extension polypeptide is an antibody or fragment thereof. In some embodiments, a half-life extension polypeptide is an anti-albumin antibody or albumin-binding fragment thereof. In some embodiments, a half-life extension polypeptide is or comprises an anti-albumin single domain antibody. In some embodiments, a half-life extension polypeptide is or comprises an anti-albumin VUH. In some embodiments, an anti-albumin VHH comprises or consists of an amino acid sequence disclosed or described in U. S. Publ. No. 20070269422 the entire contents of which are incorporated herein by reference.Immune Suppression Blockade

[0170] Tumor microenvironments (TME) can be characterized by an increased level of immune suppression molecules. In some embodiments, tumor cells upregulate expression of immune suppression molecules. The present disclosure recognizes that removal, decrease, and or blockade of immune suppression molecules can be beneficial to cancer treatment.

[0171] In some embodiments, an immune suppression molecule is an immune checkpoint protein. In some embodiments, an immune suppression molecule is PD-L1, PD1, CTLA-4, VEGF, or TGFp. In some embodiments, fusion proteins of the present disclosure increase immune responses to cancer through blockade of immune suppression molecules.

[0172] In some embodiments, fusion proteins of the present disclosure comprise a domain that is a blockade of immune suppression molecules. In some embodiments, a domain that is a blockade of immune suppression molecules can be any polypeptide that binds to or is bound by an immune suppression molecule. In some embodiments, fusion proteins of the present disclosure comprise a receptor for immune suppression molecules. In some embodiments, fusion proteins of the present disclosure comprise a ligand for immune suppression molecules.

[0173] In some embodiments, fusion proteins of the present disclosure comprise a domain that is a blockade of TGFp. Exemplary blockades of TGFP are described in Kim et al.,- 43 - 13321680vlAttorney Docket No. 2012106-0161Journal of Hematology & Oncology volume 14, Article number: 55 (2021). In some embodiments, fusion proteins of the present disclosure comprise a receptor for TGFp. In some embodiments, fusion proteins of the present disclosure comprise TGFβ receptor 2 (TGFβr2). Tumors

[0174] The present disclosure provides technologies useful in the treatment of any tumor.

[0175] In some embodiments, a tumor is or comprises a hematologic malignancy, including but not limited to, acute lymphoblastic leukemia, acute myeloid leukemia, chronic lymphocytic leukemia, chronic myelogenous leukemia, hairy cell leukemia, AIDS-related lymphoma, Hodgkin lymphoma, non-Hodgkin lymphoma, Langerhans cell histiocytosis, multiple myeloma, or myeloproliferative neoplasms.

[0176] In some embodiments, a tumor is or comprises a solid tumor, including but not limited to breast carcinoma, a squamous cell carcinoma, a colon cancer, a head and neck cancer, ovarian cancer, a lung cancer, mesothelioma, a genitourinary cancer, a rectal cancer, a gastric cancer, or an esophageal cancer.

[0177] In some embodiments, a tumor is or comprises an advanced tumor, and / or a refractory tumor. In some embodiments, a tumor is characterized as advanced when certain pathologies are observed in a tumor (e.g., in a tissue sample, such as a biopsy sample, obtained from a tumor) and / or when cancer patients with such tumors are typically considered not to be candidates for conventional chemotherapy. In some embodiments, pathologies characterizing tumors as advanced can include tumor size, altered expression of genetic markers, invasion of adjacent organs and / or lymph nodes by tumor cells. In some embodiments, a tumor is characterized as refractory when patients having such a tumor are resistant to one or more known therapeutic modalities (e.g., one or more conventional chemotherapy regimens) and / or when a particular patient has demonstrated resistance (e.g., lack of responsiveness) to one or more such known therapeutic modalities.Cellular Therapeutics- 44 - 13321680vlAttorney Docket No. 2012106-0161

[0178] In some embodiments, a fusion protein described herein can be administered to a subject as a cellular therapeutic. For example, a nucleotide sequence encoding a fusion protein described herein can be introduced into a cell for administration to a subject as a cellular therapeutic. In some embodiments, a fusion protein of the present disclosure (e.g., a CTE or TCE) is expressed by a cellular therapeutic. In some embodiments, a fusion protein of the present disclosure (e.g., a CTE or TCE) is secreted from a cellular therapeutic.

[0179] In some embodiments, a cellular therapeutic can be produced from an immune cell, e.g., a cell useful in or capable of use in adoptive cell therapy. In some embodiments, a cellular therapeutic is produced from a cell type selected from a group consisting of TILs, T-cells, CD8+cells, CD4+cells, NK-cells, gamma-delta T-cells, regulatory T-cells, iNKT cells, Innate-like lymphocytes, monocytes, macrophages, IPSC-derived cells or peripheral blood mononuclear cells. As used herein “tumor-infiltrating lymphocytes” or TILs refer to white blood cells that have left the bloodstream and migrated into a tumor. Lymphocytes can be divided into three groups including B cells, T cells and natural killer cells. As used herein “T-cells” refers to CD3+cells, including CD4+helper cells, CD8+cytotoxic T-cells, and delta-gamma T cells.

[0180] In certain embodiments, a cellular therapeutic is produced by genetically modifying (e.g., transforming) a cell, e.g., an immune cell, with a nucleic acid encoding a fusion protein described herein. In some embodiments, such nucleic acid is included in a recombinant expression vector. The recombinant expression vector can comprise any type of nucleotides, including, but not limited to DNA and RNA, which can be single-stranded or double- stranded, synthesized or obtained in part from natural sources, and which can contain natural, non-natural or altered nucleotides. A recombinant expression vector can comprise naturally-occurring or non-naturally-occurring internucleotide linkages, or both types of linkages.

[0181] A recombinant expression vector can be any suitable recombinant expression vector. Suitable vectors include those designed for propagation and expansion or for expression or both, such as plasmids and viruses. For example, a vector can be selected from the pUC series (Fermentas Life Sciences, Glen Bumie, Md.), the pBluescript series (Stratagene, LaJolla, Calif.), the pET series (Novagen, Madison, Wis.), the pGEX series (Pharmacia Biotech, Uppsala, Sweden), and the pEX series (Clontech, Palo Alto, Calif). Bacteriophage vectors, such as - 45 - 13321680vlAttorney Docket No. 2012106-0161λGT10, λGTII, ZapII (Stratagene), λEMBL4, and NM1149, also can be used. Examples of plant expression vectors useful in the context of the disclosure include pBI101, pBI101.2, pBI101.3, pBI121 and pBIN19 (Clontech). Examples of animal expression vectors useful in the context of the disclosure include pcDNA, pEUK-Cl, pMAM, and pMAMneo (Clontech).

[0182] In some embodiments, a recombinant expression vector is a viral vector. Suitable viral vectors include, without limitation, retroviral vectors, alphaviral, vaccinial, adenoviral, adeno-associated viral, herpes viral, and fowl pox viral vectors, and preferably have a native or engineered capacity to transform an immune cell (e.g., T cell).

[0183] In ex-vivo applications such as cell therapy, gamma retroviral vectors, derived from murine leukemia virus (MLV), were developed first and are still used. Lentiviral vectors, based on human immunodeficiency virus (HIV), are widely used. The general strategy in designing lentiviral vectors is based on the deletion and alteration of the native viral sequences to prevent the generation of replication-competent viruses. Thus, the lentivirus components are segregated into three or four different plasmid constructs with the goal of preventing the possibility of complete recombination to a fully replication competent lentivirus (RCL). The viral vector genome contains at a minimum the transgene expression cassette, the long terminal repeats (LTRs), and the packaging signal. In most cases, three additional plasmids provide the factors required for virus production and packaging (e.g., gag, pol, env). The promoter-enhancer region from the 3’ LTR is also deleted, preventing transcription from this region and subsequent viral replication (termed a self-inactivating vector; SIN). The essential steps of ex-vivo cell transformation or transduction involve cell isolation and culture of the desired cell type to allow the selection, expansion, and differentiation either before or after the cell has been transduced with a viral vector. In the case of hematopoietic cells, most of these steps are performed in a closed system using single-use blood collection and processing bags. For CAR T cell therapy, patient blood cells are harvested, and the desired T cell populations are selected and grown to the required levels. They are then transduced with a viral vector carrying the desired gene cassette, followed by CAR T cell expansion to the billion-cell level. Lentiviral vectors have been shown to transduce T cells efficiently and are, therefore, the preferred vector for introducing CAR into patient target cells. Expanded cells are then reintroduced into the patient.- 46 - 13321680vlAttorney Docket No. 2012106-0161

[0184] In certain in-vivo applications, nucleic acids encoding fusion proteins described herein or vectors comprising nucleic acids encoding fusion proteins described herein are administered to an individual in need thereof. For example recombinant expression vectors comprising nucleic acids encoding fusion proteins described herein can be provided as described in, for example, Nawaz et al., Blood Cancer Journal volume 11, Article number: 119 (23 June 2021) Carbonaro-Sarracino et al., Molecular Therapy: Methods & Clinical Development Vol. 16 March 2020; Cantore and Naldini Haemophilia, Volume 27, Issue S3 p. 122-125; Gouze-Decaris, et al., Arthritis Res. 2001; 3(Suppl 1): P34; Breuer et al., Scientific Reports volume 10, Article number: 4544 (2020), Naldini et al., SCIENCE • 12 Apr 1996 • Vol 272, Issue 5259 • pp.263-267. Viral vector particles can be used to deliver nucleic acids directly in vivo. Examples of such viral vector particles include lentiviral, retroviral, AAV, HVS, vaccinia and many other viral types. Viral vector particles can be modified for optimized delivery, for example, to specific cell types including immune cells. See, for example, Yang et al., PNAS August 1, 2006 | 103 (31) 11479-11484; Schaffer et al., Annu Rev Biomed Eng. 2008; 10: 169-194; Lee et al., J. of Controlled Release Volume 334, 10 June 2021, Pages 106-113; Jang et al., Molecular Therapy Volume 19, Issue 8, August 2011, Pages 1407-1415.

[0185] Recombinant expression vectors can be prepared using standard recombinant DNA techniques described in, for example, Sambrook et al., Molecular Cloning: A Laboratory Manual, 3rd ed., Cold Spring Harbor Press, Cold Spring Harbor, N. Y. 2001; and Ausubel et al., Current Protocols in Molecular Biology, Greene Publishing Associates and John Wiley & Sons, NY, 1994. Constructs of expression vectors, which are circular or linear, can be prepared to contain a replication system functional in a prokaryotic or eukaryotic host cell. Replication systems can be derived, e.g., from ColEl, 2p plasmid, SV40, bovine papilloma virus, and the like.

[0186] A recombinant expression vector can include one or more marker genes, which allow for selection of transformed or transfected hosts. Marker genes include biocide resistance, e.g., resistance to antibiotics, heavy metals, etc., complementation in an auxotrophic host to provide prototrophy, and the like. Suitable marker genes for the recombinant expression vectors include, for instance, neomycin / G418 resistance genes, puromycin resistance genes, hygromycin - 47 - 13321680vlAttorney Docket No. 2012106-0161resistance genes, histidinol resistance genes, tetracycline resistance genes, and ampicillin resistance genes.

[0187] Vectors useful in the context of the disclosure can be “naked” nucleic acid vectors (i.e., vectors having little or no proteins, sugars, and / or lipids encapsulating them), or vectors complexed with other molecules. Other molecules that can be suitably combined with the vectors include without limitation viral coats, cationic lipids, liposomes, polyamines, gold particles, and targeting moieties such as ligands, receptors, or antibodies that target cellular molecules.

[0188] Vector DNA can be introduced into a cell, e.g., an immune cell (e.g., a T cell), via conventional transformation or transfection techniques. As used herein, the terms “transformation” and “transfection” and "transduction" are intended to refer to a variety of art-recognized techniques for introducing foreign nucleic acid (e.g., DNA) into a cell, including calcium phosphate or calcium chloride co-precipitation, DEAE-dextran-mediated transfection, lipofection, gene gun, or electroporation. Cell lines used to produce viral vector particles can themselves be modified to introduce useful characteristics including, without limitation, non-self shielding polypeptides, cell-type trophic polypeptides, anti-immunosuppression polypeptides and half-life extension polypeptides.Protein Therapeutics

[0189] In some embodiments, fusion proteins described herein can be produced and administered to a subject as protein therapeutics instead of, or in addition to, being produced by a cellular therapeutic described herein. Such polypeptides can be included in a composition, e.g., a pharmaceutical composition, and used as a protein therapeutic. For example, a protein therapeutic that includes a fusion protein described herein can be administered in combination with a cellular therapeutic, e.g., CAR-T cell or TCR.

[0190] A variety of methods of making polypeptides are known in the art and can be used to make a polypeptide to be included in a protein therapeutic. For example, a polypeptide can be recombinantly produced by utilizing a host cell system engineered to express a nucleic acid encoding the polypeptide. Recombinant expression of a gene can include construction of an- 48 - 13321680vlAttorney Docket No. 2012106-0161expression vector containing a polynucleotide that encodes the polypeptide. Once a polynucleotide has been obtained, a vector for the production of the polypeptide can be produced by recombinant DNA technology using techniques known in the art. Known methods can be used to construct expression vectors containing polypeptide coding sequences and appropriate transcriptional and translational control signals. These methods include, for example, in vitro recombinant DNA techniques, synthetic techniques, and in vivo genetic recombination.

[0191] An expression vector can be transferred to a host cell by conventional techniques, and transfected cells can then be cultured by conventional techniques to produce polypeptide.

[0192] A variety of host expression vector systems can be used (see, e.g., U. S. Pat. No.5,807,715). Such host-expression systems can be used to produce polypeptides and, where desired, subsequently purified. Such host expression systems include microorganisms such as bacteria (e.g., E. coli and B. subtilis) transformed with recombinant bacteriophage DNA, plasmid DNA, or cosmid DNA expression vectors containing polypeptide coding sequences; yeast (e.g., Saccharomyces and Pichia) transformed with recombinant yeast expression vectors containing polypeptide coding sequences; insect cell systems infected with recombinant virus expression vectors (e.g., baculovirus) containing polypeptide coding sequences; plant cell systems infected with recombinant virus expression vectors (e.g., cauliflower mosaic virus, CaMV; tobacco mosaic virus, TMV) or transformed with recombinant plasmid expression vectors (e.g., Ti plasmid) containing polypeptide coding sequences; or mammalian cell systems (e.g., COS, CHO, BHK, 293, NSO, and 3T3 cells) harboring recombinant expression constructs containing promoters derived from the genome of mammalian cells (e.g., metallothionein promoter) or from mammalian viruses (e g., the adenovirus late promoter; the vaccinia virus 7.5K promoter).

[0193] For bacterial systems, a number of expression vectors can be used, including, but not limited to, the E. coli expression vector pUR278 (Ruther et al., 1983, EMBO 12:1791); pIN vectors (Inouye & Inouye, 1985, Nucleic Acids Res. 13:3101-3109; Van Heeke & Schuster, 1989, J. Biol. Chem. 24:5503-5509); and the like. pGEX vectors can also be used to express foreign polypeptides as fusion proteins with glutathione 5-transferase (GST).

[0194] For expression in mammalian host cells, viral-based expression systems can be utilized (see, e.g., Logan & Shenk, 1984, Proc. Natl. Acad. Sci. USA 81:355-359). The - 49 - 13321680vlAttorney Docket No. 2012106-0161efficiency of expression can be enhanced by inclusion of appropriate transcription enhancer elements, transcription terminators, etc. (see, e.g., Bittner et al., 1987, Methods in Enzymol. 153:516-544).

[0195] In addition, a host cell strain can be chosen that modulates expression of inserted sequences, or modifies and processes the gene product in the specific fashion desired. Different host cells have characteristic and specific mechanisms for post-translational processing and modification of proteins and gene products. Appropriate cell lines or host systems can be chosen to ensure the correct modification and processing of the polypeptide expressed. Such cells include, for example, established mammalian cell lines and insect cell lines, animal cells, fungal cells, and yeast cells. Mammalian host cells include, e.g., BALB / c mouse myeloma line (NSO / 1, ECACC No: 85110503); human retinoblasts (PER. C6, CruCell, Leiden, The Netherlands); monkey kidney CV1 line transformed by SV40 (COS-7, ATCC CRL 1651); human embryonic kidney line (293 or 293 cells subcloned for growth in suspension culture, Graham et al., J. Gen Virol., 36:59,1977); human fibrosarcoma cell line (e.g., HT1080); baby hamster kidney cells (BHK, ATCC CCL 10); Chinese hamster ovary cells + / -DHFR (CHO, Urlaub and Chasin, Proc. Natl. Acad. Sci. USA, 77:4216, 1980); mouse sertoli cells (TM4, Mather, Biol. Reprod., 23:243-251, 1980); monkey kidney cells (CV1 ATCC CCL 70); African green monkey kidney cells (VERO-76, ATCC CRL-1 587); human cervical carcinoma cells (HeLa, ATCC CCL 2); canine kidney cells (MDCK, ATCC CCL 34); buffalo rat liver cells (BRL 3A, ATCC CRL 1442); human lung cells (W138, ATCC CCL 75); human liver cells (Hep G2, HB 8065); mouse mammary tumor (MMT 060562, ATCC CCL51); TRI cells (Mather et al., Annals N. Y. Acad. Sci., 383:44-68, 1982); MRC 5 cells; FS4 cells; and a human hepatoma line (Hep G2).

[0196] For long-term, high-yield production of recombinant proteins, host cells are engineered to stably express a polypeptide. Host cells can be transformed with DNA controlled by appropriate expression control elements known in the art, including promoter, enhancer, sequences, transcription terminators, polyadenylation sites, and selectable markers. Methods commonly known in the art of recombinant DNA technology can be used to select a desired recombinant clone.- 50 - 13321680vlAttorney Docket No. 2012106-0161

[0197] Once a protein described herein has been produced by recombinant expression, it may be purified by any method known in the art for purification, for example, by chromatography (e.g., ion exchange, affinity, and sizing column chromatography), centrifugation, differential solubility, or by any other standard technique for purification of proteins. For example, an antibody can be isolated and purified by appropriately selecting and combining affinity columns such as Protein A column with chromatography columns, filtration, ultra filtration, salting-out and dialysis procedures (see Antibodies: A Laboratory Manual, Ed Harlow, David Lane, Cold Spring Harbor Laboratory, 1988). Further, as described herein, a polypeptide can be fused to heterologous polypeptide sequences to facilitate purification.Alternatively or additionally, a polypeptide or fusion protein can be partially or fully prepared by chemical synthesis.Viral Delivery

[0198] In some embodiments, a nucleic acid encoding a fusion protein described herein can be introduced into a cell and / or administered to a subject as a viral vector. In some embodiments, such a viral vector can be used to introduce a fusion protein into a cell (e.g., an immune cell). Introduction of such fusion protein can increase susceptibility to a subject’s immune system and / or one or more additional therapeutic agents (see, e.g., WO2017 / 075533).Vector Design

[0199] A nucleic acid sequence encoding a fusion protein described herein can be introduced into a number of types of vectors. For example, a nucleic acid can be cloned into a plasmid, a phagemid, a phage derivative, an animal virus, and a cosmid. Other vectors can include expression vectors, replication vectors, probe generation vectors, sequencing vectors, and viral vectors. In other examples, the vector can be a foamy viral (FV) vector, a type of retroviral vector made from spumavirus. Viral vector design and technology is well known in the art as described in Sambrook et al., (Molecular Cloning: A Laboratory Manual, 2001), and in other virology and molecular biology manuals.Viral transduction- 51 - 13321680vlAttorney Docket No. 2012106-0161

[0200] Viruses are highly efficient at nucleic acid delivery to specific cell types, while often avoiding detection by the infected host immune system. These features make certain viruses attractive candidates as vehicles for introduction of nucleic acids (e.g., nucleic acids encoding fusion proteins described herein) into immune cells. A number of viral based systems have been developed for gene transfer into mammalian cells. Examples of viral vectors include, but are not limited to, retroviruses, adenoviruses, adeno-associated viruses, herpes viruses, lenti viruses, poxviruses, herpes simplex 1 virus, herpes virus, oncoviruses (e.g., murine leukemia viruses), and the like. In general, a suitable vector contains an origin of replication functional in at least one organism, a promoter sequence, convenient restriction endonuclease sites, and one or more selectable markers, (e.g., WO 01 / 96584; WO 01 / 29058; and U. S. Pat. No. 6,326,193).

[0201] Lentiviral and retroviral transduction can be enhanced by the addition of LentiBOOST (Mayflower Bioscience SBPLV10112 ) or TransDux (System Biosciences LV850 A-l) or polybrene (SantaCruz sc-134220; Millipore TR-1003-G; Sigma 107689), a cationic polymer (also known as hexadimethrine bromide) that is used to increase the efficiency of the lentiviral or retrovirus transduction.

[0202] For example, retroviruses provide a platform for gene delivery systems.Retroviruses are enveloped viruses that belong to the viral family Retroviridae. Once in a host’s cell, the virus replicates by using a viral reverse transcriptase enzyme to transcribe its RNA into DNA. The retroviral DNA replicates as part of the host genome, and is referred to as a provirus. A selected gene can be inserted into a vector and packaged in retroviral particles using techniques known in the art. The recombinant virus can then be isolated and delivered to cells of the subject in vivo. A number of retroviral systems are known in the art, (see, e.g., U. S. Pat Nos.5,994,136, 6,165, 782, and 6,428,953).

[0203] Retroviruses include the genus of Alpharetrovirus (e.g., avian leukosis virus), the genus of Betaretrovirus; (e.g., mouse mammary tumor virus), the genus of Deltaretrovirus (e.g., bovine leukemia virus and human T-lymphotropic virus), the genus of Epsilonretrovirus (e.g., Walleye dermal sarcoma virus), and the genus of Lentivirus. In some embodiments, a retrovirus is a lentivirus a genus of viruses of the Retroviridae family, e.g., characterized by a long incubation period. Lentiviruses are unique among the retroviruses in being able to infect non- - 52 - 13321680vlAttorney Docket No. 2012106-0161dividing cells; they can deliver a significant amount of genetic information into the DNA of the host cell, so can be used as an efficient gene delivery vector. In some examples, a lentivirus can be, but not limited to, human immunodeficiency viruses (HIV-1 and HIV-2), simian immunodeficiency virus (SIV), feline immunodeficiency virus (FIV), equine infections anemia (EIA), and visna virus. Vectors derived from lentiviruses offer the means to achieve significant levels of gene transfer in vivo.

[0204] In some embodiments, other methods of enhancing lentiviral and retroviral transduction include use of multiple promoters in lentiviral and retroviral vectors. In some embodiments incorporating multiple promoters within a single lentiviral vector enables independent expression of different genes. Each promoter drives the transcription of a specific gene, allowing for the simultaneous expression of multiple sequences. In another embodiment, a single promoter within a single lentiviral vector is capable of expressing multiple different genes integrated into the same viral vector. For example, in some embodiments a fusion protein (e.g., a costimulatory engager) is integrated into the same viral vector that is used to express a CAR. In some embodiments, a viral vector is engineered to express a fusion protein (e.g., a costimulatory engager) to be secreted from a cell. In some embodiments, a viral vector encodes an anti-CD19 CAR T sequence and a fusion protein with expression of both driven by a single promoter. In some embodiments a fusion protein (e.g., a costimulatory engager) is encoded upstream or downstream of an anti-CD19 CAR T sequence and has its expression driven by a single or multiple promoters. Methods to achieve secretion of multiple encoded sequences are known (see, for example, Labbé, 2021. doi: 10.3390 / v13081528). In some embodiments, only a CAR T cell viral vector is required in order to express the CAR domain, secrete the CTE, and achieve clearance of the tumor cells in vivo.

[0205] In some embodiments, a fusion protein (e.g., a costimulatory engager) and a CAR can be expressed at the same time (e.g., using the same or separate promoters), or can be expressed at different times. In some embodiments, a costimulatory engager construct includes a first promoter to express the costimulatory engager fusion protein, and includes a second promoter to express a second CAR. For example, a first promoter can mediate rapid expression- 53 - 13321680vlAttorney Docket No. 2012106-0161of the costimulatory engager fusion protein, and a second promoter can mediate delayed expression of the second CAR.

[0206] In some embodiments, a nucleic acid encoding a fusion protein and / or a CAR is operably linked to a promoter. Preferably, the promoter is functional in immune cells.Exemplary promoters include, e.g., CMV, E1F, VAV, TCRvbeta, MCSV, and PGK promoter. Operably linking of a nucleotide sequence with a promoter is within the skill of the artisan.

[0207] In some embodiments, other methods of enhancing lentiviral and retroviral transduction include the use of an Internal Ribosome Entry Site (IRES). In some embodiments, IRES elements allow for translation of multiple proteins from a single mRNA transcript. By placing an IRES between two coding sequences, a ribosome can initiate translation at both the 5' end and the IRES, producing two distinct proteins from one transcript.

[0208] In some embodiments, self-cleaving peptide sequences are used for enhancing transduction. In some embodiments, inserting self-cleaving peptide sequences, such as Thosea asigna virus 2A (T2A), between coding sequences enables the production of multiple proteins from a single open reading frame. During translation, a ribosome skips the formation of a peptide bond at the T2A site, resulting in separate proteins being produced from one mRNA. Additional methodologies are known in the art.

[0209] In some embodiments, a vector is an adenovirus vector. Adenoviruses are a large family of viruses containing double stranded DNA. They replicate the DNA of the host cell, while using a host’s cell machinery to synthesize viral RNA DNA and proteins. Adenoviruses are known in the art to affect both replicating and non-replicating cells, to accommodate large transgenes, and to code for proteins without integrating into the host cell genome.

[0210] In some embodiments, an AAVP vector is used. An AAVP vector is a hybrid of prokaryotic-eukaryotic vectors, which are chimeras of genetic cis-elements of recombinant adeno-associated virus (AAV) and phage. An AAVP combines selected elements of both phage and AAV vector systems, providing a vector that is simple to produce in bacteria and can exhibit little or no packaging limit, while allowing infection of mammalian cells combined with integration into the host chromosome. Vectors containing many of the appropriate elements are- 54 - 13321680vlAttorney Docket No. 2012106-0161commercially available, and can be further modified by standard methodologies to include the necessary sequences. Among other things, AAVPs do not require helper viruses or trans-acting factors. In addition, the native tropism of AAV for mammalian cells is eliminated since there is not AAV capsid formation. Other methods and details are in U. S. Patent 8,470,528 and Hajitou A. et al., Cell, 125: 358-398.

[0211] In some embodiments, a human papilloma (HPV) pseudovirus is used. DNA plasmids can be packaged into papillomavirus LI and L2 capsid protein to generate pseudovirion that can efficiently deliver DNA. The encapsulation can protect the DNA from nucleases and provides a targeted delivery with a high level of stability. Many of the safety concerns associated with the use of viral vectors can be mitigated with an HPV pseudovirus. Other methods and examples are in Hung, C., et al., Pios One, 7:7(e40983); 2012, U. S. Patent 8,394,411, and Kines, R., et al Int J of Cancer, 2015.

[0212] In some embodiments, an oncolytic virus is used. Oncolytic virus therapy can selectively replicate the virus in cancer cells, and can subsequently spread within a tumor, e.g., without affecting normal tissue. Alternatively, an oncolytic virus can preferentially infect and kill cells without causing damage to normal tissues. Oncolytic viruses can also effectively induce immune responses to themselves as well as to the infected tumor cell. Typically, oncolytic viruses fall into two classes: (I) viruses that naturally replicate preferentially in cancer cells and are nonpathogenic in humans. Exemplary class (I) oncolytic viruses include autonomous parvoviruses, myxoma virus (poxvirus), Newcastle disease virus (NDV; paramyxovirus), reovirus, and Seneca valley virus (picomavirus). A second class (II) includes viruses that are genetically manipulated for use as vaccine vectors, including measles virus (paramyxovims), poliovirus (picomavirus), and vaccinia vims (poxvirus). Additionally, oncolytic viruses may include those genetically engineered with mutations / deletions in genes required for replication in normal but not in cancer cells including adenovirus, herpes simplex virus, and vesicular stomatitis virus. Oncolytic viruses can be used as a viral transduction method due to their low probability of genetic resistance because they can target multiple pathways and replicate in a tumor-selective method. The viral dose within a tumor can increase- 55 - 13321680vlAttorney Docket No. 2012106-0161over time due to in situ viral amplification (as compared to small molecule therapies which decrease with time), and safety features can be built in (i.e., drug and immune sensitivity).Non-viral delivery

[0213] In some embodiments, non-viral methods are used to deliver and express nucleotides from cells. These methods are relative for both ex vivo and in vivo delivery. Non-viral methods for delivering chimeric antigen receptor (CAR) constructs into immune cells such as T cells include electroporation that uses electrical pulses to transiently permeabilize cell membranes, allowing the introduction of genetic material like DNA or RNA directly into the cells. Other non-viral methods include the use of Lipid Nanoparticles (LNPs) that can encapsulate nucleic acids, facilitating their delivery into cells by merging with the cell membrane. Another non-viral method uses Transposon Systems that are sequences that can change positions within the genome. Systems like Sleeping Beauty and piggyBac have been utilized to stably integrate CAR genes into the host genome, providing a non-viral means of achieving long-term expression. Another non-viral method uses CRISPR / Cas9-Mediated integration to introduce double-stranded breaks at specific genomic locations, facilitating the targeted insertion of CAR constructs. This method aims for precise integration, potentially reducing the risks associated with random insertion.Formulation and Administration

[0214] Certain embodiments of the disclosure include methods of administering to a subject a cellular therapeutic described herein (or a population thereof), a protein therapeutic described herein, a composition comprising a cellular therapeutic, and / or a composition comprising a protein therapeutic, e.g., in an amount effective to treat a subject. In some embodiments, the method effectively treats cancer in the subject.

[0215] In some embodiments a cellular therapeutic comprises an autologous cell that is administered into the same subject from which an immune cell was obtained. Alternatively, an immune cell is obtained from a subject and is transformed, e.g., transduced, with an expression construct described herein, to obtain a cellular therapeutic that is allogenically transferred into another subject.- 56 - 13321680vlAttorney Docket No. 2012106-0161

[0216] In some embodiments, a cellular therapeutic is autologous to a subject, and the subject can be immunologically naive, immunized, diseased, or in another condition prior to isolation of an immune cell from the subject.

[0217] In some embodiments, additional steps can be performed prior to administration to a subject. For instance, a cellular therapeutic can be expanded in vitro after contacting (e.g., transducing or transfecting) an immune cell with an expression construct described herein, but prior to the administration to a subject. In vitro expansion can proceed for 1 day or more, e g., 2 days or more, 3 days or more, 4 days or more, 6 days or more, or 8 days or more, prior to the administration to a subject. Alternatively, or in addition, in vitro expansion can proceed for 21 days or less, e.g., 18 days or less, 16 days or less, 14 days or less, 10 days or less, 7 days or less, or 5 days or less, prior to administration to a subject. For example, in vitro expansion can proceed for 1-7 days, 2-10 days, 3-5 days, or 8-14 days prior to the administration to a subject.

[0218] In some embodiments, during in vitro expansion, a cellular therapeutic can be stimulated with an antigen (e.g., a TCR antigen). Antigen specific expansion optionally can be supplemented with expansion under conditions that non-specifically stimulate lymphocyte proliferation such as, for example, anti-CD3 antibody, anti-Tac antibody, anti-CD28 antibody, or phytohemagglutinin (PHA). The expanded cellular therapeutic can be directly administered into a subject or can be frozen for future use, i.e., for subsequent administrations to a subject.

[0219] In some embodiments, a cellular therapeutic is treated ex vivo with interleukin-2 (IL-2) prior to infusion into a cancer patient, and the cancer patient is treated with IL-2 after infusion. Furthermore, in some embodiments, a cancer patient can undergo preparative lymphodepletion— the temporary ablation of the immune system— prior to administration of a cellular therapeutic. A combination of IL-2 treatment and preparative lymphodepletion can enhance persistence of a cellular therapeutic.

[0220] In some embodiments, a cellular therapeutic is transduced or transfected with a nucleic acid encoding a cytokine, which nucleic acid can be engineered to provide for constitutive, regulatable, or temporally-controlled expression of the cytokine. Suitable cytokines include, for example, cytokines which act to enhance the survival of T lymphocytes during the contraction phase, which can facilitate the formation and survival of memory T lymphocytes.- 57 - 13321680vlAttorney Docket No. 2012106-0161

[0221] In certain embodiments, a cellular therapeutic is administered prior to, substantially simultaneously with, or after the administration of another therapeutic agent, such as a cancer therapeutic agent. The cancer therapeutic agent can be, e.g., a chemotherapeutic agent, a biological agent, or radiation treatment. In some embodiments, a subject receiving a cellular therapeutic is not administered a treatment which is sufficient to cause a depletion of immune cells, such as lymphodepl eting chemotherapy or radiation therapy.

[0222] A cellular therapeutic described herein can be formed as a composition, e.g., a cellular therapeutic and a pharmaceutically acceptable carrier. In certain embodiments, a composition is a pharmaceutical composition comprising at least one cellular therapeutic described herein and a pharmaceutically acceptable carrier, diluent, and / or excipient.Pharmaceutically acceptable carriers described herein, for example, vehicles, adjuvants, excipients, and diluents, are well-known and readily available to those skilled in the art.Preferably, the pharmaceutically acceptable carrier is chemically inert to the active agent(s), e.g., a cellular therapeutic, and does not elicit any detrimental side effects or toxicity under the conditions of use.

[0223] A composition can be formulated for administration by any suitable route, such as, for example, intravenous, intratumoral, intraarterial, intramuscular, intraperitoneal, intrathecal, epidural, and / or subcutaneous administration routes. Preferably, the composition is formulated for a parenteral route of administration.

[0224] A composition suitable for parenteral administration can be an aqueous or nonaqueous, isotonic sterile injection solution, which can contain anti-oxidants, buffers, bacteriostats, and solutes, for example, that render the composition isotonic with the blood of the intended recipient. An aqueous or nonaqueous sterile suspension can contain one or more suspending agents, solubilizers, thickening agents, stabilizers, and preservatives.

[0225] Dosage administered to a subject, particularly a human, will vary with the particular embodiment, the composition employed, the method of administration, and the particular site and subject being treated. However, a dose should be sufficient to provide a therapeutic response. A clinician skilled in the art can determine the therapeutically effective amount of a composition to be administered to a human or other subject in order to treat or - 58 - 13321680vlAttorney Docket No. 2012106-0161prevent a particular medical condition. The precise amount of the composition required to be therapeutically effective will depend upon numerous factors, e.g., such as the specific activity of the cellular therapeutic, and the route of administration, the amount of available antigen or antigens on tumor cells (e.g., as a consequence of tumor bulk or extent of tumor burden) and / or on normal cells, in addition to many subject-specific considerations, which are within those of skill in the art. In some embodiments, the appropriate dose for a cellular therapeutic for a particular cancer indication or indications can be defined in a dose-escalation clinical trial.

[0226] Any suitable number of cellular therapeutic cells can be administered to a subject. While a single cellular therapeutic cell described herein is capable of expanding and providing a therapeutic benefit, in some embodiments, 102or more, e.g., 103or more, 104or more, 105or more, or 108or more, cellular therapeutic cells are administered. Alternatively, or additionally 1012or less, e.g., 1011or less, 109or less, 107or less, or 105or less, cellular therapeutic cells described herein are administered to a subject. In some embodiments, 102- 105, 104- 107, 103- 109, or 105-1010cellular therapeutic cells described herein are administered.

[0227] A dose of a cellular therapeutic described herein can be administered to a mammal at one time or in a series of subdoses administered over a suitable period of time, e.g., on a daily, semi-weekly, weekly, bi-weekly, semi-monthly, bi-monthly, semi-annual, or annual basis, as needed. A dosage unit comprising an effective amount of a cellular therapeutic may be administered in a single daily dose, or the total daily dosage may be administered in two, three, four, or more divided doses administered daily, as needed.

[0228] A polypeptide described herein can be incorporated into a pharmaceutical composition (e.g., for use as a protein therapeutic). Pharmaceutical compositions comprising a polypeptide can be formulated by methods known to those skilled in the art (see, e.g., Remington’s Pharmaceutical Sciences pp. 1447-1676 (Alfonso R. Gennaro, ed., 19th ed. 1995)). A pharmaceutical composition can be administered parenterally in the form of an injectable formulation comprising a sterile solution or suspension in water or another pharmaceutically acceptable liquid. For example, a pharmaceutical composition can be formulated by suitably combining a polypeptide with pharmaceutically acceptable vehicles or media, such as sterile water and physiological saline, vegetable oil, emulsifier, suspension agent, surfactant, stabilizer,- 59 - 13321680vlAttorney Docket No. 2012106-0161flavoring excipient, diluent, vehicle, preservative, binder, followed by mixing in a unit dose form required for generally accepted pharmaceutical practices. The amount of active ingredient included in pharmaceutical preparations is such that a suitable dose within the designated range is provided.

[0229] The sterile composition for injection can be formulated in accordance with conventional pharmaceutical practices using distilled water for injection as a vehicle. For example, physiological saline or an isotonic solution containing glucose and other supplements such as D-sorbitol, D-mannose, D-mannitol, and sodium chloride may be used as an aqueous solution for injection, optionally in combination with a suitable solubilizing agent, for example, alcohol such as ethanol and polyalcohol such as propylene glycol or polyethylene glycol, and a nonionic surfactant such as polysorbate 80™, HCO-50, and the like.

[0230] Nonlimiting examples of oily liquid include sesame oil and soybean oil, and it may be combined with benzyl benzoate or benzyl alcohol as a solubilizing agent. Other items that may be included are a buffer such as a phosphate buffer, or sodium acetate buffer, a soothing agent such as procaine hydrochloride, a stabilizer such as benzyl alcohol or phenol, and an antioxidant. The formulated injection can be packaged in a suitable ampule.

[0231] Protein half-life can be impacted by the degree of sialylation, which is the post-translational, covalent, addition of terminal sialic acid to glycosylated proteins (Hossler et al., Glycobiology. 2009; 19:936-49). Proteins with more sialyation may have longer half-lives in vivo (Flintegaard et al., Endocrinology. 2010; 151:5326-36; Bork et al., J Pharm Sci. 2009; 98:3499-508). In some embodiments, a fusion protein described herein is formulated as a sialylated protein therapeutic.

[0232] Route of administration can be parenteral, for example, administration by injection, transnasal administration, transpulmonary administration, or transcutaneous administration. Administration can be systemic or local by intravenous injection, intramuscular injection, intraperitoneal injection, subcutaneous injection. Biological CTEs are useful in with local or regional administration of CD19-targeting CAR T cells, for example, the treatment of glioblastoma in vivo. Glioblastoma is one example of a cancer in which antigens B7H3, Her2 and IL13Ra2 are expressed.- 60 - 13321680vlAttorney Docket No. 2012106-0161

[0233] A suitable means of administration can be selected based on the age and condition of the subject. A single dose of a pharmaceutical composition containing a polypeptide can be selected from a range of 0.001 to 1000 mg / kg of body weight. On the other hand, a dose can be selected in the range of 0.001 to 100000 mg / kg of body weight, but the present disclosure is not limited to such ranges. Dose and method of administration can vary depending on the weight, age, condition, and the like of the subject, and can be suitably selected as needed by those skilled in the art.

[0234] In some embodiments, a pharmaceutical composition containing a fusion protein is administered in combination with a cellular therapeutic, e.g., CAR-T cell or TCR, as described herein. In some embodiments, a pharmaceutical composition comprises a fusion protein described herein and a cellular therapeutic described herein. In some embodiments, a pharmaceutical composition containing a fusion protein is administered simultaneously, concomitantly, or sequentially with a cellular therapeutic described herein. In some embodiments, a pharmaceutical composition containing a fusion protein is administered about 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, or 7 days, prior to administration of a cellular therapeutic described herein. In some embodiments, a pharmaceutical composition containing a fusion protein is administered about 1 week, 2 weeks, 3 weeks, or 4 weeks prior to administration of a cellular therapeutic described herein. In some embodiments, a pharmaceutical composition containing a fusion protein is administered about 1 month, 2 months, 3 months, 4 months, or 5 months prior to administration of a cellular therapeutic described herein. In some embodiments, a pharmaceutical composition containing a fusion protein is administered about 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, or 7 days, subsequent to administration of a cellular therapeutic described herein. In some embodiments, a pharmaceutical composition containing a fusion protein is administered about 1 week, 2 weeks, 3 weeks, or 4 weeks subsequent to administration of a cellular therapeutic described herein. In some embodiments, a pharmaceutical composition containing a fusion protein is administered about 1 month, 2 months, 3 months, 4 months, or 5 months subsequent to administration of a cellular therapeutic described herein.

[0235] All publications, patent applications, patents, and other references mentioned herein, including GenBank Accession Numbers, are incorporated by reference in their entirety.- 61 - 13321680vlAttorney Docket No. 2012106-0161In addition, the materials, methods, and examples are illustrative only and not intended to be limiting. Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Although methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention, suitable methods and materials are described herein.Subject Identification

[0236] In some embodiments, a subject is identified and / or selected for administration of fusion protein as described herein. In some embodiments, a subject can be identified and / or selected for treatment based on diagnosis of refractory or resistant cancer. In some embodiments, a subject can be identified and / or selected for treatment based on the prescription to receive ACT therapy. In some embodiments, a subject can be identified and / or selected for treatment based on evidence of ACT therapy relapse. In some embodiments, a subject can be identified and / or selected for treatment based on one or more measured or observed sign of relapse in cancer (e.g., a non-beneficial response, loss or downregulation of the target antigen of a cell used in ACT or progressive disease). In some embodiments, the fusion protein is administered to the subject. In some embodiments, upon administration of the fusion protein therapy, the subject exhibits a positive clinical response to the ACT therapy, e.g., exhibits an improvement based on one or more clinical and / or objective criteria (e.g., exhibits a decrease in tumor burden, tumor size, and / or tumor stage).

[0237] Methods described herein can include preparing and / or providing a report, such as in electronic, web-based, or paper form. The report can include one or more outputs from a method described herein, e.g., tumor burden, tumor size, and / or tumor stage, stability of disease, loss or downregulation of target antigen. In some embodiments, a report is generated, such as in paper or electronic form, which identifies the presence or absence of one or more tumor antigens for a cancer patient, and optionally, a recommended course of cancer therapy. In some embodiments, the report includes an identifier for the cancer patient. In one embodiment, the report is in web-based form.- 62 - 13321680vlAttorney Docket No. 2012106-0161

[0238] In some embodiments, additionally or alternatively, a report includes information on prognosis, resistance, or potential or suggested therapeutic options. The report can include information on the likely effectiveness of a therapeutic option, the acceptability of a therapeutic option, or the advisability of applying the therapeutic option to a cancer patient, e.g., identified in the report. For example, the report can include information, or a recommendation, on the administration of a cancer therapy, e.g., the administration of a pre-selected dosage or in a preselected treatment regimen, e.g., in combination with one or more alternative cancer therapies, to the patient. The report can be delivered, e.g., to an entity described herein, within 7, 14, 21, 30, or 45 days from performing a method described herein. In some embodiments, the report is a personalized cancer treatment report.

[0239] In some embodiments, a report is generated to memorialize each time a cancer subject is tested using a method described herein. The cancer subject can be reevaluated at intervals, such as every month, every two months, every six months or every year, or more or less frequently, to monitor the subject for responsiveness to a cancer therapy and / or for an improvement in one or more cancer symptoms, e.g., described herein. In some embodiments, the report can record at least the treatment history of the cancer subject.

[0240] In one embodiment, the method further includes providing a report to another party. The other party can be, for example, the cancer subject, a caregiver, a physician, an oncologist, a hospital, clinic, third-party payor, insurance company or a government office.EXAMPLES

[0241] The following examples describe some of the preferred modes of making and practicing the present invention. However, it should be understood that these examples are for illustrative purposes only and are not meant to limit the scope of the invention. Furthermore, unless the description in an Example is presented in the past tense, the text, like the rest of the specification, is not intended to suggest that experiments were actually performed or data were actually obtained.Example 1: Generation of CD20-directed CTEs that contain LFA3 binding- 63 - 13321680vlAttorney Docket No. 2012106-0161

[0242] The present example describes CAR T engager fusion protein (CTE) expression constructs generated and used for characterizing relevant functional activity of CAR T engager fusion proteins described herein, including testing that augmenting LFA3 binding to CD2 expressed on the CAR T cell will improve CAR T functionality.

[0243] A lentiviral vector encoding a CD19-directed CAR sequence was used to transduce human donor T cells to create CAR T cells and a CAR-T engager fusion protein that binds CD20 and displays the CD 19 extracellular domain was cloned and expressed (see, for example, methods as described in Su et al. 2022, herein incorporated by reference in its entirety).

[0244] Some constructs were designed with and without accessory molecules (e.g., costimulatory domains). In some constructs the first 93 amino acids of the mature LFA3 protein were encoded to create an LFA3 binding domain (see, for example, Osborn et al., 1995). Flexible glycine / serine linkers were used between the domains (see, for example, van Rosmalen, et al., 2017). The CAR T engager fusion proteins listed in Table 1 were expressed and tested for their ability to improve CAR T cell anti-lymphoma activities including lymphoma cell binding and cytotoxicity. The domains are listed from the N terminus to the C terminus. The anti-CD20 VHH and anti-albumin VHH were derived from camelid single domain antibody sequences (sdAb; Lecocq et al. 2019, herein incorporated by reference in its entirety).Table 1. CD20-directed CTE constructs.Construct # Domains encoded607 anti-CD20 VHH - CD19ECD - anti -albumin VHH672 LFA3bd - anti-CD20 VHH - CD19ECD - anti-albumin VHH673 LFA3bd - anti-albumin VHH - anti-CD20 VHH - CD19ECD680 LFA3bd - anti-CD20 VHH - CD19ECD - anti-albumin VHH - LFA3bd681 LFA3bd - LFA3bd - anti-CD20 VHH - CD19ECD-anti-albumin VHH686 anti-CD20 VHH - CD19ECD - anti -albumin VHH - LFA3bd692 anti-CD20 VHH - LFA3 - CD19ECD - anti-albumin VHH - LFA3bd693 anti-CD20 VHH - CD19ECD-LFA3 - anti-albumin VHH - LFA3bd694 anti-CD20 VHH - CD19ECD - anti -albumin VHH - LFA3 - LFA3bd- 64 - 13321680vlAttorney Docket No. 2012106-0161

[0245] The LFA3 binding domains were tested in a variety of configurations relative to the CD 19 extracellular domain (ECD), anti-CD20, and anti-albumin domains. Initial experiments evaluated the binding of LFA3 binding domains containing CTEs to the CD2 protein in ELISA and cell binding formats.Example 2. Evaluation of the binding of LFA3 binding domains containing CTEs to the CD2 protein by ELISA

[0246] CTEs listed in Table 1 were screened for CD2 binding to LFA3 by ELISA.Briefly, 96-well ELISA assay plates were coated overnight with 1 pg / ml of the anti-CD19 antibody FMC63. The coded plates were incubated with blocking buffer for I hour at 37°C to reduce nonspecific binding. Blocking buffers were made in TBS and contained either 1% BSA or 0.3% nonfat dry milk. CTE proteins were added with a starting concentration of 750 ng / ml and titrated down using 2.5x dilutions in blocking buffer. Proteins were incubated with the plates for one hour at 37°C during which time the CD19ECD was able to bind to the anti-CD19 antibody. The plates were washed x3 with blocking buffer. Biotinylated CD2 (ACROBiosystems) was added at 1 pg / ml and allowed to incubate for one hour, followed by three washes with blocking buffer. Streptavidin-HRP (Thermo Fisher) was added for 30 minutes, followed by three more washes. The peroxidase enzymatic reaction was initiated to provide a colorimetric read out. The reaction was stopped with acidic buffer, and the plate was read in an ELISA plate reader using the 450nm wavelength.

[0247] Results shown in Figure 2 and Table 2 demonstrate that CTE #607, having no LFA3 domains, and CTE #686, having a single LFA3 domain, did not bind to CD2 in the ELISA assay. CTE #692 bound with an apparent affinity of 3 nM or -250 ng / ml. Without being bound by a theory, the binding avidity may have been gained by having multiple layers of protein binding, i.e. anti -CD 19 / CD 19 as the capture step, and LFA3 / CD2 the detection step, since this value is higher than the direct CD2 / LFA3 binding affinity demonstrated by surface plasma residence assays (see, for example, Ikemizu et al. 1999).Table 2. Binding affinity of CTEs for CD2 as measured by ELISA.- 65 - 13321680vlAttorney Docket No. 2012106-0161EC50EC50molecular weight(ng / ml) (n. M) (kDa)#607 57#686 - 69#692 249.7 3.1 81Example 3. Evaluation of immune synapse binding domain geometry for costimulation optimization by ELISA

[0248] CTE constructs are designed to engage the nascent immune synapse that forms when a therapeutic immune cell (e.g., CAR T) binds to its target tumor antigen. CD2 is expressed within the developing T cell immune synapse and, importantly, is also expressed at a high density in a ring encircling the immune synapse (i.e., the corolla) (see, Siokis, et al., 2021; Demetriou, et al., 2020). Engagement of the developing immune synapse with the appropriate geometry is useful in achieving optimal T cell co-stimulation. Therefore, the geometry (e.g., spacing and / or distance) of accessory molecules (e.g., LFA binding domain sequences) within a CTE was further investigated.

[0249] CTE binding was evaluated by ELISA according to similar methods as described in Example 2 with some modifications. Briefly, the capture reagent used was an anti-CD19 antibody (e.g, FMC63) and the detection reagents were a biotinylated CD2 and streptavidinperoxidase, whose binding was visualized enzymatically.

[0250] Results in Figure 3 demonstrated that the addition of 2 LFA3 binding domain sequences to the CD20-directed CTE allowed binding to CD2 in a manner that is dependent on LFA3 binding domain geometry (e.g., spacing and / or distance) within the CTE. The results show that adjacent LFA3bd do not support CD2 binding and that increasing the space between LFA3bd proteins is beneficial. The CTE binding ELISA was repeated to include additional CTEs having no (e.g., CTE #607) and one (e.g., CTE #686) LFA3 binding domain and the results are shown in Figure 4 and Tables 3-4. The pattern of binding shown in Figure 4 confirms the spacing preferences that were characterized in Figure 3.- 66 - 13321680vlAttorney Docket No. 2012106-0161Table 3. Binding affinities of CTEs for CD2 derived from dose response curves of Figure 3EC50EC50% Maximum Molecular weight (ng / ml) (nM) Binding (kDa) #680 315.5 3.9 95% 81#692 288.7 3.6 100% 81#693 279.1 3.5 47% 81#694 - - - 81Table 4. Binding affinities of CTEs for CD2 derived from dose response curves of Figure 4EC50EC50% Maximum Molecular weight (ng / ml) (nM) Binding (kDa) #607 4% 57#686 10% 69#692 253 3.1 100% 81#693 325 4 78% 81#694 - - 5% 81

[0251] Together, these data show that spatial constraints influence the functional positioning of 2 LFA3 binding domain sequences within the CD20-directed CTE. Specifically, adjacent LFA binding domains did not support CD2 binding (e.g., CTE #694), whereas the LFA3 binding domains separated by one or more other protein domains support CD2 binding (e.g., CTE #680). The effect appeared to be quantitative with saturable CD2 binding achieved when two protein domains intervened between the LFA3bds rather than one. These intervening domains are linked by flexible G4S linkers allowing for mobility within the CTE. These - 67 - 13321680vlAttorney Docket No. 2012106-0161intervening domains are small protein domains that occupy less than lOnm of the cell surface (Figure 5; the VHH domains are ~15kD and the CD19ECD is ~ 25kD). Further, the results obtained from CTEs with different LFA3 binding domain distribution suggests that the augmentation is due to supporting appropriate spacing of LFA3 such that CD2 is engaged in an immune synapse relevant manner (Figure 5A, 5B). Specifically, enhanced binding to CD2 was observed with the CTEs having LFA3bds separated by multiple other protein domains, thus: CTE #680 (LFA3bd - anti-CD20 VHH - CD19ECD - anti-albumin VHH - LFA3bd), and CTE# 692 (anti-CD20 VHH - LFA3bd - CD19ECD - anti-albumin VHH - LFA3bd). In this regard, the entire physical structure of the immune synapse that forms when protein pairs are bound between an APC and a T cell is needed to displace the very large phosphatases, including CD45, that act to reduce activated T cell signaling (Figure 5C).

[0252] The characterization of LFA3bd spacing within the CTE design is further demonstrated using more distant spacing between LFA3bd, by evaluating the impact of different protein domains placed between LFA3bd, by altering linker design, by altering linker length and by evaluating CTE with constrained architecture, e.g. by the placement of covalent linkages. Example 4. Evaluation of CTE binding to the surface of activated T cells by flow cytometry

[0253] CTEs listed in Table 1 were evaluated for binding to surface proteins on activated T cells using flow cytometry. Briefly, Human CD3+ T cells were isolated from PBMC samples using an EasySep human T cell isolation kit (STEMCELL Technologies). T cells were activated for 24 hours using anti-CD3 / anti-CD28 human T-activator Dynabeads (Thermo Fisher).Following activation, T cells were incubated for 1 hour on ice with CTEs or with antibodies that recognize human CD2 and CD3 proteins. Cells were then washed twice with cold FACs buffer (lx PBS without calcium and magnesium, 1% FBS, and 1mm EDTA). Bound CTEs were detected using an anti-antibody HIB19-PE that was incubated with the CTE samples for 30 minutes on ice. CD2, CD3, and LFA were detected using fluorescently labeled anti-CD2, anti-CD3, and anti-LFA3 antibodies (BioLegend) after two washes with cold FACs buffer. Samples were run on an Accuri C6 Plus flow cytometer and the data were analyzed using FlowJo software (BectonDickenson).- 68 - 13321680vlAttorney Docket No. 2012106-0161

[0254] Results in Figure 6 demonstrate that activated T cells brightly express LFA3 (Figure 6A) and CD2 (Figure 6B) on the cell surface. The abundant expression of LFA3 suggests that cis-interactions with CD2 may occur on these cells.

[0255] Results in Figure 7 and Table 5 demonstrate that CTE #607, having no LFA3 domains, and CTE #686, having a single LFA3 binding domain, did not bind in the flow cytometry assay. CTE #692 bound with an apparent affinity of 6.8 nM, similar to the affinity measured by ELISA (see Example 2). The substantial expression of CD2 coupled with the presence of two LFA3 domains in the CTE supports that avidity interactions that increase the apparent affinity. Together, these data show that the addition of two LFA3bd sequences to the CD20-directed CTE was sufficient to support binding to cell surface expressed CD2, consistent with the ELISA binding results showing binding to soluble CD2.Table 5. Binding affinity of CTEs for CD2 on activated T cells as measured by flow cytometry.EC50Molecular weightEC50(ng / ml) (nM) (kDa)#607 57#686 69#692 548.8 6.8 81

[0256] Results in Figure 8 and Table 6 further demonstrated that single LFA binding domain- containing CTEs (e.g., CTEs #672, #673) only bound at very high protein concentrations. Similarly, a CTE having adjacent LFA3 binding domains (e.g., CTE #681) only bound weakly to activated human T cells. The CTE with two LFA3 binding domains spaced apart (e.g., CTE #680), bound at ~1.4 nM.Table 6. CTE binding affinities to activated normal human T cells derived from dose response curves of Figure 8.- 69 - 13321680vlAttorney Docket No. 2012106-0161EC50EC50Molecularweightng / ml nM kDa#672 9695 140.6 69#673 11810 170.5 69#680 110 1.4 81#681 3556 44 81

[0257] Results in Figure 9 and Table 7 demonstrate additional CTE comparative binding data obtained via flow cytometry for CTEs #680, #692, #693, and #694 having differential spacing of LFA3 binding domains.Table 7. CTE binding affinities to activated normal human T cells derived from dose response curves of Figure 8.MolecularEC50EC50weight(ng / ml) (nM) (kDa)#680 97.9 1.2 81#692 91.5 1.1 81#693 441.5 5.5 81#694 1668 20.6 81

[0258] The patterns of CTE binding illustrated in Figure 9 and the affinity measurements derived from the binding curves in Table 7 are similar to those measured using the ELISA assay (see Example 3, Figures 3-4 and Tables 3-4). In the ELISA format, the LFA3 present in the CTEs gained an avidity advantage due to the mode of presentation. In the flow cytometry assay - 70 - 13321680vlAttorney Docket No. 2012106-0161format, cell surface expressed CD2 gained an avidity advantage likely due to high levels of cell surface expression. Furthermore, the calculated affinities observed in the present Example are similar to published affinities for binding of LFA3 to CD2 in the context of added avidity (Tibaldi et al., 2002; Zhu et al., 2006).

[0259] Together these results demonstrate using CD20-directed CTEs yielded surprising results since LFA3 binding is already abundantly expressed on lymphoma cells. These data demonstrate that augmentation of LFA3 binding to CD2 will be useful to enhance anti-CD19 CAR T cell treatment of B cell lymphoma. In support of these data, is had been shown that B cell lymphoma cells actively downregulate LFA3 as a mechanism of escape from immune directed therapeutics (e.g., anti-CD19 CAR T cell therapy; Younes et al., 2023; Yan et al., 2022).Example 5. Evaluation of CTE binding to CD20 and CD2 by ELISA and flow cytometry

[0260] It is known that the interaction of LFA3 and CD2 is most immunologically relevant in the context of low antigen expression to the T cell. Therefore, it is also important to characterize the binding of CTE to the targeted antigen, alongside of CD2 binding. Therefore, a selection of CTEs listed in Table 1 were evaluated for their ability to bind CD20 and CD2 by ELISA and flow cytometry according to similar methods as described in Examples 2-4.

[0261] Results shown in Table 8 demonstrated that in both ELISA and flow cytometry experiments, CTE #680 and CTE #692 that showed low nM affinity binding to CD2 and low nM affinity binding to CD20.Table 8. Summary of CTE CD20 and CD2 binding data by ELISA and flow cytometryBinding data expressed as average EC50 derived, in nMCD2 CD2 CD20 CD20 (ELISA) (flow cytometry) (ELISA) (flow cytometry)- 71 - 13321680vlAttorney Docket No. 2012106-0161#607 - - 0.7 0.2#680 3.9 1.3 2 9.5#692 3.3 4 0.4 0.5#693 3.8 5.5 0.7 ndExample 6. Evaluation of CTE-directed, anti-CD19 CAR T mediated, cytotoxic activity.

[0262] To evaluate the contribution of CD2 binding to cytotoxicity, CTEs listed in Table 1 were screened in cytotoxic activity assays using JeKo-1 lymphoma cells that are CD19 and CD58 deficient (JeKo-1 CD19 CD58 KO). For the cytotoxicity assay, a 96 well round bottom plate was seeded with 50 pL of target cells (JeKo-1 CD19 CD58 KO) carrying the luciferase gene at 1 x 104cells / well in RPMI 1640 medium containing 10% FBS without antibiotics (RPMI / FBS). Four-fold dilutions of test proteins which was titered using ELISA were made in 25 pL RPMEFBS and added to the cells. The CAR19 T cells, primary T cells transduced with lentiviral particles that encode an anti-CD19 CAR-T construct (A254), were thawed and washed once with RPMEFBS, collected via centrifugation at 550 RCF for 10 minutes and added to the wells in a volume of 25 pL to give a 5: 1 CAR:target (E: T) cell ratio. The plates were incubated at 37°C for 48 hours. The plate was centrifuged at 500 RCF for 5 minutes, the pellet was rinsed with PBS and spun again. Then, 20 pL of 1 x lysis buffer (Promega) was added to the pellet, and the lysate was transferred into a 96 well opaque tissue culture plate (Fisher Scientific). The plates were read in a luminometer with an injector to dispense the substrate (Promega). The percent killing was calculated based upon the average loss of luminescence of the experimental condition vs the control (target cell plus CAR19 only).

[0263] Results shown in Figure 10 and Table 9 demonstrated that CTE #680, which incorporates LFA3 binding domain sequences at a distance from each other, improved antiCD 19 CAR-T -mediated cytotoxicity by >65% versus the control CTE #607 that lacks LFA3 domains. In contrast, CTE #681, which incorporates adjacent LF A3 binding domain sequences - 72 - 13321680vlAttorney Docket No. 2012106-0161that do not bind CD2 with high affinity / avidity, supported anti-CD19 CAR-T-mediated % cytotoxicity at a level very similar to the #607 control.Table 9. IC50 values for CTE-directed, anti-CD19 CAR-T-mediated cytotoxicity.IC50 IC50 Molecular weight(ng / ml) (pM) (kDa)#607 0.28 4.9 57#680 0.13 1.6 81#681 0.33 4.1 81

[0264] The cytotoxic activity assay was repeated to evaluate additional CTEs (e g., CTE #692 and CTE #607) and the results shown in Figure 11 and Table 10 demonstrated that CTE #692, which incorporates spatially discrete LFA3bds while returning the anti-CD20 VHH to the N-terminus, supported potent CTE-directed, anti-CD19 CAR T-mediated cytotoxicity against lymphoma cells lacking expression of both CD 19 and CD58. The potency was improved by over 70% when compared to the control CTE # 607.

[0265] These accumulated results demonstrate that adding LFA3bd sequences specifically distanced from each other enables CD2 binding, does not interfere with CTE activity, and improves cytotoxicity in the absence of CD58 expression.Table 10. Cytotoxic activity assay IC50 values comparing CTE #607 and CTE #692.IC50 IC50 Molecular weight(ng / ml) (pM) (kDa)#607 0.54 9.5 57#692 0.22 2.7 81- 73 - 13321680vlAttorney Docket No. 2012106-0161Example 7. Novel use of LFA3 binding domains to augment T cell engagers activity against Non-Hodgkin Lymphoma (NHL) cancer cells.

[0266] The present Example describes CD20-directed T cell engager fusion proteins (TCEs) developed and used for characterizing relevant functional activities of costimulatory T cell engagers described herein, including binding, cell activation, and cytotoxicity.

[0267] TCEs #656, 657, 722, and 752 directed to CD20 listed in Table 11 were developed by linking an anti-CD20 VHH domain to an anti-CD3 scFv (see, for example, methods as described in (Bargou R, et al., 2008); and in some constructs an anti-human serum albumin domain and LFA3 costimulatory domain were also included.

[0268] To establish the functional characteristics of costimulatory TCEs, the constructs designed included LFA3 binding domains (LFA3bd) placed within the TCEs, either adjacent to each other or separated by other protein domains. Using other domains to separate the LFA3 domains allowed for investigation of the spacing needed to signal effectively to CD2 on T cells. The TCEs listed in Table 11 all contain an anti-CD20 VHH, the same anti-albumin VHH, and the same anti-CD3 scFV sequences. Construct #656 also contained a C terminal VHH to human serum albumin. TCEs #656 and #657 use the same anti-CD20 VHH, TCEs #722 and #752 use the same anti-CD20 VHH and anti-human serum albumin VHH domains. TCEs #722 and #752 differ by the presence or absence of LFA3bds.Table 11. CD20-directed TCE constructs.Construct # Domains Encoded656 N - anti-CD20 VHH1 - anti-CD3 scFv - anti-albumin - C657 N - anti-CD20 VHH1 - anti-CD3 scFv - C722 N - anti-CD20 VHH2 - LFA3 - anti-CD3 scFv - anti-albumin VHH - LFA3 - C 752 N - anti-CD20 VHH2 - anti-CD3 scFv - anti-albumin VHH - CXXX N - anti-CD20 VHH2 - LFA3 - anti-CD3 scFv - LFA3 - anti-albumin VHH - C yyy N - anti-CD20 VHH2 - LFA3 - LFA3 - anti-CD3 scFv - anti-albumin VHH - C- 74 - 13321680vlAttorney Docket No. 2012106-0161

[0269] Cell binding was assessed by two ELISA assays that were developed using human serum albumin (HSA) as the capture reagent. Briefly, 96-well plates were coated with HSA followed by the CTE. During the first ELISA (Figure 12), the bound CTE was detected using biotinylated-CD20 and HRP-streptavidin for detection. During the second ELISA (Figure 13), the bound CTE was detected using biotinylated-CD2 and HRP-streptavidin for detection. Results shown in Figures 12 and 13 and Tables 12 and 13 demonstrated that CTE #722 and CTE #752 proteins bound to CD20 with approximately 2.1 and 0.2nM affinity, respectively. These data additionally demonstrate that CTE #722 binds to biotinylated CD2 with an approximate affinity of 15.5nM.Table 12. CTE CD20 binding affinities derived from Figure 12EC50EC50Molecular weight(ng / ml) (nM) (kDa)#722 166.8 2.1 78.8#752 12.8 0.2 54.7Table 13. CTE CD2 binding affinities derived from Figure 13MolecularEC50(ng / ml) EC50(nM) Molecular weight(kDa)#722 1.2 15.5 78.8

[0270] Cytotoxicity activity of the TCEs was assessed using a PBMC depletion assay. Briefly, 2x105donor PBMCs were incubated with 3-fold serial dilutions of TCE proteins and incubated for 48 hours. TCEs bind to both T cells and B cells in the PBMC preparation leading to B cell cytotoxicity. Post-incubation, PBMC cellularity was evaluated by flow cytometry. Anti-CD79b-APC was used to detect B cells and anti-CD4-FITC plus anti-CD8-FITC were used to - 75 - 13321680vlAttorney Docket No. 2012106-0161detect T cells. PBMCs from 2 different donors were used. The dose response curves are shown in Figure 14 A and B. TCEs #722 and #752 demonstrated an ability to deplete B cells and 1C50 values are shown in Table 14.Table 14. Anti-CD20 TCE B cell depletion EC50s derived from Figure 14Donor # #752 #722 #693EC50 (ng / ml) 67 0.29 6.04 -EC50 (pM) 67 5.4 76.6 0.0MW (kDa) 67 54.69 78.82 81Donor # #752 #722 #693EC50 (ng / ml) 22 1.55 8.27 -EC50 (pM) 22 28.4 104.9 0.0MW (kDa) 22 54.69 78.82 81

[0271] Additional methods of evaluating cytotoxicity for TCEs are known in the art. For example, cytotoxicity can be assessed by a luciferase reporter cell line assay. Briefly, 1x10e4 of JeKo-1 CD19 KO target cells (T) that stably express luciferase are added to each well of a 96-well round bottom plate. Primary T effector cells (E) are transduced with transduced with lentiviral particles that encode an anti-CD19 CAR-T construct (A254). An effector target ratio (E: T) of 5 T cells: 1 target cell is calculated. Each TCE is represented by triplicates (n = 3). Once a TCE concentration of 30 ng / ml is achieved, for example, it is followed by 3x serial dilutions to establish a titration. The final volume per well is 150 pl media containing 50 pl target cells and 50 pl of 3x-serial diluted TCE and 50 pl T cells. The assay components, all contained RPMI media containing 10% fetal bovine serum with no antibiotics. After a 48-hour incubation, the cells in each well are lysed and subjected to a standard luciferase assay.- 76 - 13321680vlAttorney Docket No. 2012106-0161

[0272] Additional constructs (e.g., xxx and yyy, shown in Table 11) are examples of varying LFAbd placement within CD20-directed TCE constructs that can be evaluated. For example, a construct that contains a N - anti-CD20 VHH2 - LFA3 - anti-CD3 scFv - LFA3 -anti-albumin VHH - C is useful in evaluating placement of LFA3bd on either side of the CD3 binder. In contrast, a construct that contains N - anti-CD20 VHH2 - LFA3 - LFA3 - anti-CD3 scFv - anti -albumin VHH - C, where placement of LFA3bds is adjacent to each other, would not be expected to have improved binding to CD2 nor improved cytotoxic efficacy.Example 8. Induction of T cell cytotoxic activity by IL13-R«2-directed T Cell Engagers.

[0273] The present example describes TCE expression constructs generated and used for characterizing relevant functional activity in a solid tumor setting. TCEs containing IL-13Ra2-directed VHHs were designed. Construct #753 also contains 2 LFA3 binding domains. The TCE constructs listed in Table 15 were expressed and evaluated.Table 15. Exemplary TCE constructs containing LFA3 binding domains.Construct # Domains encoded753 anti-IL-13Ra2 VHH - LFA3bd - anti-CD3 scFv- anti-albumin VHH - LFA3bd754 anti-IL-13Roc2 VHH - anti-CD3 scFv - anti-albumin VHH574 anti-IL-13Ra2 VHH - CD19ECD

[0274] The TCEs listed in Table 15, which contain LFA3 binding domains, were evaluated for their ability to activate T cells in the presence of IL- 13Ra2 -positive tumor cells (A375 melanoma cells) using an NF AT -luciferase reporter assay. Briefly, 50 pl Jurkat-CAR19 cells, containing the NFAT-luciferase reporter gene, were added to 50 pl 5 x 10e4 of A375 cells in RPMI assay media containing 10% FBS. The E. T. ratio was 1: 1 with a cell concentration of 5 x 10e4 cells each. 50 pl of the TCE was added from a 3-fold serial dilution, where the starting concentration was 10 ng / ml. The assay was run for 5 hours at 37°C. Samples were collected for the luciferase assay. Fold induction of NF AT -Luc was calculated by comparing relative luminescence units (RLU) of each TCE treatment with the RLU of the Jurkat cells only.- 77 - 13321680vlAttorney Docket No. 2012106-0161

[0275] Results shown in Figure 15 and Table 16 demonstrated a > 10-fold improvement in T cell activation when LFA3bds were added to the TCE.Table 16. TCE binding affinities derived from the induction of T cell activation / signaling derived from dose response curves in Figure 15.EC50 EC50 Molecular weight(ng / ml) (pM) (kDa)TCE #7530.065 14.7 44+A375TCE #75420.64 134 79+A375CTE #574- 0.0 55+A375

[0276] The results shown in Table 16 were extended to a cell-based cytotoxicity assay as described for TCEs, using the A375 cell line as the source of antigen. The E: T ratio used for the assay was 2.5:1 and toxicity was measured after 48 hours. Results shown in Figure 16 and Table 17 confirm results from the prior assay showing an approximately eight-fold difference in cytotoxicity potential produced by adding appropriately spaced LFA3bds to the TCE.Table 17. Calculation of IC50 values from Figure 16.574 753 i 754IC50 (ng / ml) 0.18 0.87IC50 (PM) 0.0 2.24 15.83MW (kDa) 79 55Example 9. Evaluation of additional costimulatory domains to TCEs.

[0277] The present example describes the evaluation of (1) placement of LFA3bds to TCEs, (2) the addition of one or more B7-2 binding domains, (3) the addition of both LFA3bds and B7-2bds to TCEs, and (4) the addition of TGFp antagonist to costimulatory TCE.- 78 - 13321680vlAttorney Docket No. 2012106-0161

[0278] Evaluation consists of assays for the following characteristics: binding to tumor antigen or antigens, CD2 binding, CD28 binding, antigen-positive tumor cytotoxicity, in vitro and in vivo. Evidentiary tools used to assist in characterization are tumor cell lines, expressing one or more tumor antigens, tumor cell lines that express CD58, tumor cell lines that are engineered not to express CD58, tumor cell lines known to express TGFp, and additional tools as are known. Evidentiary assays are binding assays, T cell signaling assays, cytotoxicity assays, cytokine secretion assays and additional assays as are known, including those that are performed as in vitro assays, and those that are performed as an analysis of in vivo activity.Example 10. Generation of multi-targeting CTE for the treatment of solid tumors.

[0279] The present example describes the construction and characterization of B7-H3 bridging proteins and evaluated the activity of dual antigen CTEs to target tumor antigens (e.g., B7-H3 and Her2).

[0280] CTEs generated are listed in Table 18. All anti-B7-H3 monospecific CTEs contain a unique anti-B7-H3 scFv or VHH sequence followed by a GGGGS (G4S) linker sequences repeated 4 times (G4Sx4) and then a stabilized C terminal CD 19 sequence (Klesmith, Molecular Pharmaceutics 201916 (8), 3544-3558) and a C terminal 6 Histidine tag (6x His). They were chemically synthesized and cloned into the expression vector pcDNA3.1 (+) hygro, by GenScript. The anti-Her2 scFv (trastuzumab)-CD19-anti-B7-H3 bispecific CTEs were generated by synthesizing the anti-B7-H3 component and cloning it into a backbone plasmid which contains the anti-Her2 (Trastuzumab scFv) and the stabilized C terminal CD19 sequence. The constructs contained G4S x 4 linkers between the domains and contained a C terminal 6x His tag.Table 18. Construct numbers and CTEs sequence.Construct # Description of the CTEs protein#263 anti-HER2 scFv, CD19ECD sequence#567 anti-B7-H3 VHH (ProSci Clone 1G4), CD19ECD sequence- 79 - 13321680vlAttorney Docket No. 2012106-0161#568 anti-B7-H3 scFv sequence (Doth; Ab 376.96, US20180371053A1)anti-B7-H3 scFv sequence (Mackall; “SEQ ID NO 12”, as listed in #569US2018 / 0346544A1), CD19ECD sequenceanti-B7-H3 scFv sequence (GenbankAVC01686; “SEQ ID NO. 45” as #570listed in US9790282)anti-HER2 scFv, CD19ECD, and anti-B7-H3 scFv sequence (Doth; Ab #571376.96, US20180371053A1)anti-HER2 scFv, CD19ECD and anti-B7-H3 scFv sequence (Mackall;#572“SEQ ID NO. 12”, as listed in US2018 / 0346544A1)anti-HER2 scFv, CD19ECD, and anti-B7-H3 scFv sequence (Genbank;#573Seq ID 45, AVC01686)anti-HER2 scFv, CD19ECD, and anti-B7-H3 VHH sequence (ProSci#577Clone 1G4)#593 B7-H3 ProSci Clone 1G4, CD19 ECD and anti-HER2 scFv sequence

[0281] Expression constructs were transfected into 293T cells. Briefly, constructs #263, #567, #577 and #593 were expressed in 293T cells in 6 well plates. Approximately 1.6 x 106cells were plated and left to settle for about 1 hour. Per well, 2 pg of a DNA construct and 10 pl of Lipofectamine 2000 (Invitrogen) was used to transfect the cells following the manufacturer’s protocol. The supernatants were harvested 72 hours post transfection by spinning at 12,000 RCF for 4 minutes at 4°C and used in assays or frozen at -20° C

[0282] The expression of each CTEs in the cell culture supernatants was detected in an ELISA assay in which anti-CD19 antibody FMC63 was coated on a plate and an anti -His tag antibody was used for detection. Briefly, plates were coated overnight at 4°C with 1.0 pg / mL anti-CD19 mAb FMC63 (NOVUS) in 0.1 M carbonate, pH 9.5. The plate was blocked with 0.3% non-fat milk in TBS for 1 hour at room temperature. After washing in TBST 3 times, the cell culture supernatants containing B7-H3 VHH CTEs was diluted 10 fold, or the purified CTEs were diluted to the required concentration, in TBS / 1% BSA, and then added to the wells in 3-- 80 - 13321680vlAttorney Docket No. 2012106-0161fold serial dilutions; the plate was incubated for 1 hour. The plates were then washed 3 times, followed by a 1 hour incubation with a 1:2000 dilution of HRP-anti-His (BioLegend). Then, 1-Step Ultra TMB-ELISA solution (Thermo Fisher) was added to develop the peroxidase signal, and the plate was read at 405 nm. IL-13RO.2 protein and CD33 protein were used as a specificity controls.

[0283] Results shown in Figure 17 demonstrated that the CTE constructs expressed levels of CTE protein as to be detectable in this ELISA format. Additional analysis of the constructs to evaluate protein concentration was performed. Here, a purified CTE protein #357 was used as a means of setting the standard curve. The #357 CTE protein is a highly purified CAR-T engager protein produced by Wuxi Biologies and consists of three domains: anti-CLEC12A ScFv - anti-CLEC12A VHH 2H3 - CD 19 ECD. The ELISA plates were coated with anti-CD19 antibody FMC63 and an anti -His tag antibody was used for detection. A standard curve (Figure 18A) was used to calculate the concentration of the expressed bridging proteins (Figure 18B) and calculated concentrations are shown in Table 19. These binding assays showed that the CTEs were able to bind to anti-CD19 antibody FMC63 and retained the C-terminal His-tag.Table 19. Concentrations calculated of expressed bridging proteins.CTE µg / ml#567 (4 / 17) 24#567 (6 / 1) 31.9#568 (6 / 1) 24.7#571 (5) 7.0#571 (2.5) 6.4#577 (5) 4.8#577 (2.5) 5.1- 81 - 13321680vlAttorney Docket No. 2012106-0161

[0284] The expression of B7-H3 and Her2 on A375 melanoma cells was evaluated via flow cytometry. Results shown in Figure 19 demonstrated that A375 cells express both B7-H3 and Her2.

[0285] Additional binding analysis of constructs listed in Table 18 was performed by flow cytometry. Briefly, 50 pl of Fc blocked A375 cells (5xl05) were incubated with 50 pl of a 3x serial diluted CTE for 30 minutes at 4°C. The samples were then spun and washed twice. A fluorescent labeled anti-CD19 FMC63 antibody was used for detection. Results of the binding assay are shown in Figure 20 and Table 20.Table 20. Binding affinities of purified constructs on A375 cells detected with FMC63-PE#263 #567 #577 #593IC50 (ng / ml) 224.3 112.7 22.35 12IC50 (nM) 3.939 2.531 0.310 0.167MW (kDa) 56.95 44.52 72.07 72.07Example 11. Evaluation of dual-antigen CTEs binding activity to target tumor antigens Her2 and B7-H3.

[0286] The present example describes methods used to evaluate the ability of the CTEs listed in Table 18 (see, Example 10) for their ability to bind to target tumor antigens Her2 and B7-H3.

[0287] A bispecific-binding ELISA was used to evaluate the CTEs, Briefly, plates were coated overnight at 4°C with 1.0 pg / mL B7-H3-His-4 Ig (ACROBiosystems) or Her2-Fc (ACROBiosystems) in 0.1 M carbonate, pH 9.5. The plate was blocked with 0.3% non-fat milk in TBS for 1 hour at room temperature. After washing in TBST 3 times, the supernatant of #571, 572 and 577 were titrated with 3-fold dilution in TBS / 1% BSA and incubated 1 hour. Then, 100 pl at 0.5 pg / ml concentration of biotinylated Her2-His (ACROBiosystems) if plate was coated- 82 - 13321680vlAttorney Docket No. 2012106-0161with B7-H3-4 Ig or biotinylated B7-H3-4 Ig VHH protein (labeled in house) if plate was coated with Her2-Fc were added into plate for 1 hr. The plates were then washed 3 times, followed by a 1-hour incubation with a 1:2000 dilution of HRP-streptavidin (Thermo Fisher). 1-Step Ultra TMB-ELISA solution was added and the plate was read at 405 nm.

[0288] Results, shown in Figure 21 demonstrated that constructs #571 and #577 retain binding when captured with B7-H3 whether the detection is with anti-His tag antibody or with biotinylated Her2. Construct #572 appeared in this case to lose some degree of binding when detected with biotinylated Her2.Example 12. Evaluation of CTE mediated cytotoxicity by CAR-CD19 T cells.

[0289] The present example describes methods to evaluate the efficacy of CTE mediated cytotoxicity by CAR-CD19 T cells. First, CTEs were assayed in a dose-response study with CAR-CD19 T cells using a Her2-positive and B7-H3 -positive A375 cell line. The CAR T cell preparation was 54.5% CAR-positive and was used at a ratio of T cells to target cells of 10. Briefly, adherent cells (Her2-positive and B7-H3-positive A375 cells) were removed from the flask with Accutase (Corning) and washed with FACS buffer. A 96 well flat bottom plate was seeded with 50 pL of A375 cells carrying the luciferase gene at 1 x 104cells / well in RPMI 1640 medium + 10% FBS without antibiotics (RPMI+FBS).

[0290] Dilutions of the bridging proteins were made in 50 pL RPMI + FBS and added to the cells. The CAR-CD19 T cells (CAR-254, see Report No. Aleta-CD 19-001) were thawed and washed once with RPMI + FBS via centrifugation at 550 RCF for 10 minutes. The CAR T cells were added to the wells right after the bridging protein dilution at a volume of 50 pL to give a CAR:target cell ratio of 10: 1. The plates were incubated at 37°C for 48 hours. The plate was centrifuged at 550 RCF for 5 minutes, the pellet was rinsed with PBS, and spun again. Then, 20 pL of 1 x lysis buffer (Promega) was added to the pellet. The plates were read in a luminometer with an injector to dispense the substrate (Promega). The percent killing was calculated based upon the average loss of luminescence of the experimental condition vs the control (target cell plus CAR-CD19 T cells).- 83 - 13321680vlAttorney Docket No. 2012106-0161

[0291] Results shown in Figure 22 demonstrated that A375 cells can be killed by CAR-CD19 T cells either through Her2 antigen binding (#263, anti-Her2-CD19 ECD CTE) or through B7-H3 antigen binding (#576 and #568, anti-B7-H3-CD19 ECD CTEs). All of the dual-antigen binding constructs support anti-CD19 CAR-T cell-mediated cytotoxicity, and do so at lower concentration than do the single antigen CTEs (Table 21).Table 21. Cytotoxicity data derived from Figure 22.Construct EC50 EC50Description of the CTE protein(ng / ml) (pM) #567 anti-B7-H3 1G4 VHH - CD 19 ECD 8.7 195#568 anti-B7-H3 scFv (Doth) - CD 19 ECD 5.7 101.3 #569 anti-B7-H3 scFv (Mackall) - CD 19 ECD - - anti-HER2 - CD 19 ECD- anti-B7-H3 scFv 3.9 #571 0.33(Dotti)#577 anti-HER2 - CD 19 ECD - anti-B7-H3 1G4 VHH 0.31 4.3anti-HER2 - CD 19 ECD - anti-B7-H3 scFv 27.7 #572 2.3(Mackall)#263 anti-HER2 - CD 19 ECD 6.9 120.5

[0292] To further explore the relationship between single antigen and dual antigen potency a second assay was performed to compare CTEs. Results shown in Figure 23 and the derived cytotoxicity EC50 values in Table 22, are very similar to the prior experiment, with #571 showing a 30-fold increase in potency relative to #568 and #577 showing a 7-fold improvement in potency.Table 22. Cytotoxicity data derived from Figure 23.13321680vlAttorney Docket No. 2012106-0161Construct # Description EC50 EC50 (ng / ml) (pM) 567 Anti-B7-H3 1G4 VHH - CD 19 ECD 9 202568 Anti-B7-H3 scFv (Dotti) - CD 19 ECD 6 106571 Anti-Her2 - CD 19 ECD - anti-B7-H3 scFv (Dotti) 0.2 2.4577 Anti-Her2 - CD 19 ECD - anti-B7-H3 1G4 VHH 0.19 2.6

[0293] Figure 24 shows the results of dual-antigen binding constructs mediating anti-CD19 CAR-T cell cytotoxicity using an additional dual anti -positive cell line. Here, Skov3 cells express both Her2 and B7-H3 were used. As seen previously, #577 was more potent than #567. However, #263 (Her2 -targeting only) was as potent as #577 on this cell line, possibly due to the very high level of Her2 expression (a 4-log shift over control staining by flow cytometry which can correspond to up to 2 million copies of Her2).

[0294] Figure 25 and Table 23 shows the results of dual-antigen binding constructs mediate cytotoxicity using adjusted titers. These results demonstrate that the dual-antigen directed bispecific constructs, #593 and #577, are more potent than the monospecific constructs when evaluated using the A375 cell line.Table 23. Cytotoxicity data derived from Figure 25EC50 ng / ml EC50 pM MW kDa#263 3.5 61.5 56.95#577 0.39 5.4 72#593 0.44 6.1 72#567 2.3 51.7 44.5- 85 - 13321680vlAttorney Docket No. 2012106-0161Figure 26 and Table 24 shows the expression of CD19 (FIG. 26A), Her2 (FIG. 26B), and B7-H3 (FIG. 26C) on BT474 breast cancer cell line derived from ductal carcinoma.Table. 24. BT474 cell line expression dataCount % of This | Mean FL2-AHOI BT474 unstained 85 1.4% 7,553.7H02 BT474 +mslgGl-PE 84 1.4% 9,499.1H03 BT474+ FMC63-PE 119 1.9% 7,973.6H04 BT474 + a-Her2-PE 5,010 99.9% 2,182,926.3H05 BT474 + a-B7H3-PE 6,257 100.0% 294,465.0Using the BT474 cell line, cytotoxicity of the dual-antigen binding constructs was evaluated. Results shown in Figure 27 and Table 25 demonstrated that the bispecific constructs are not more potent than the anti-Her2 monospecific construct (#263) in the BT474 cell line.Table 25. Cytotoxicity data derived from Figure 27EC50 ng / ml EC50 pM MW kDa#263 0.23 4.04 56.95#577 0.16 2.2 72#593 0.18 2.5 72#567 2.29 51.46 44.5

[0295] Here we present proof-of-concept data that a dual antigen targeting CTE can mediate binding to both antigens and induce cytotoxicity against a dual-antigen-positive cell line, A375, that appears to be superior to single antigen targeting.

[0296] The relative contribution of the two binding domains in the B7-H3 / Her2 dual targeting CTE proteins are evaluated by flow cytometry analyses of target cell lines, derivation - 86 - 13321680vlAttorney Docket No. 2012106-0161of antigen-specific cell lines (Her2-only, B7-H3-only), analysis of activity at different antigen densities, measurement of corresponding cytotoxic activities, and with novel forms of the bridging proteins.Example 13. Generation of CTEs comprising anti- IL-13Ra2 VHH sequences.

[0297] The present example describes CTE expression constructs that incorporate anti-IL-13Ra2 VHH sequences that were generated and used for characterizing relevant functional activity of CAR T engager fusion proteins described herein. Methods used to incorporate the CD 19 extracellular domain and anti-albumin VHH into CTE constructs are similar to those described in Su et al 2022 ( 10.1080 / 2162402X.2022.2111904) and Ambrose et al 2021 (10.1371 / joumal.pone.0247701) while also using the anti-IL-13Roc2 VHH described herein.

[0298] The CTE fusion proteins listed in Table 26 were expressed and tested for their ability to bind cell surface expressed IL-13Ra2 by flow cytometry. A375 cells were incubated with culture supernatants containing expressed CTEs; the concentration was estimated by ELISA. Three CTEs bound to A375 in a dose response manner and presented a bright mean fluorescent intensity by flow cytometry (Figure 28). These results show that the expressed CTEs bound to IL-13Ra2on the surface of A375 cells and displayed the CD19 ECD.Table 26. CTEs containing VHH sequences, directed to IL - 13 Ra2.CTE VHH used#574 anti-IL-13Ra2 VHH IF 10 - CD19 ECD#575 anti-IL-13Ra2 VHH 2C1 - CD19 ECD#576 anti-IL-13Ra2 VHH 2C 10 CD 19 ECD

[0299] The ability of CTEs to direct an anti -CD 19 CAR T cell cytotoxicity against the A375 melanoma cells was tested using a cytotoxicity assay according to similar methods as described in Example 6. Dose responsive cytotoxicity results shown in Figure 29 and the resulting IC50 values (Table 27) demonstrated that CTEs created with VHH IF 10, 2C1 and 2C10 produced very similar cytotoxicity IC50 values.Table 27. IC50 values calculated from the cytotoxicity curves.IC50 (ng / mL) IC50 (pM) MW (kDa)- 87 - 13321680vlAttorney Docket No. 2012106-0161#574 0.33 7.42 44#575 0.17 3.87 44#576 0.46 10.75 43Example 14. Generation and characterization of dual antigen binding CTE directed to IL-13Ra2 and B7-H3.

[0300] The present example describes CTE expression constructs designed to improve CAR T cell activity against IL- 13Ra2 -expressing tumor cells and IL-13Ra2 / B7-H3-expressing tumor cells. Further, two costimulatory pathways, LFA3 / CD2 and B7 / CD28, were investigated using a series of constructs directed to solid tumors by targeting several different tumor-associated antigens (TAA).

[0301] The dual antigen binding CTE fusion proteins listed in Table 28 were expressed and tested for their binding characteristics by ELISA and flow cytometry.Table 28. Dual antigen binding CTE directed to IL13Ra2 and B7H3.Construct # Encoded domains748 anti-IL-13Ra2 VHH - anti-B7-H3 VHH - CD 19 ECD749 anti-B7-H3 VHH - anti-IL-13Ra2 VHH - CD 19 ECD750 anti-B7-H3 VHH - CD 19 ECD - anti-IL-13Ra2 VHH751 anti-IL-13Ra2 VHH - CD 19 ECD - anti-B7-H3 VHH

[0302] Dual-antigen binding CTE fusion proteins listed in Table 28 were tested for the binding activity to IL-13Ra2 using similar methods as described in Examples 2 and 5 with some modifications. Briefly, 96-well plates were coated with the anti-CD19 antibody FMC63. The CTEs were added to the wells and allowed to incubate. Bound CTEs were detected with biotinylated IL-13Ra2 protein that was in turn detected by peroxidase-coupled streptavidin using an enzymatic reaction.- 88 - 13321680vlAttorney Docket No. 2012106-0161

[0303] Dose response binding shown in Figure 30 and the apparent EC50s for IL-13Ra2 binding (Table 29) show that the preferred design for the bispecific CTE binding to IL-13Ra2 protein was #750 having the anti-IL-13Ra2 VHH at the N-terminus (anti-IL-13Ra2 VHH - CD19 ECD - anti-B7-H3 VHH) with 50 pM binding affinity.Table 29. CTE binding affinities for IL-13Ra2 by ELISA.EC5O EC5O Molecular weight(ng / ml) (nM) (kDa)#574 17.4 0.4 44#748 9.8 0.2 60#749 10.1 0.2 60#750 2.9 0.05 60#751 10.9 0.2 60

[0304] Additionally, dual-antigen binding CTE fusion proteins listed in Table 28 were also tested for the binding activity to B7-H3. Here, 96-well plates were coated anti-CD19 antibody FMC63. The CTEs were added to the wells and allowed to incubate. Bound CTEs were detected with biotinylated B7-H3 protein that was in turn detected by peroxidase-coupled streptavidin and an enzymatic reaction. Resulting dose response binding curves are shown in Figure 31 and apparent EC50s for binding of CTEs to B7-H3 protein were calculated from the ELISA dose response curves (Table 30).Table 30. Dose response EC50 for binding of CTEs to B7-H3.3inding to B7-H ’ 41gMolecularEC50ng / mlnM weight kDa#567 12.8 0.3 4513321680vlAttorney Docket No. 2012106-0161#748 20 0.3 60#749 14.9 0.25 60#750 4.8 0.08 60#751 4.5 0.08 60

[0305] These data show that the dual-antigen bispecific CTE binding to B7-H3 was optimized in CTEs #750 and #751. These CTEs feature a design in which the 2 anti-antigen VHHs are separated by the CD 19 ECD. This suggests complexity in the CTE design that requires consideration of antigen access by the different VHH. In contrast to data collected in the anti-IL-13Ra2 ELISA format there was no difference in having anti-B7-H3 VHH located at the N-terminus versus the C-terminus.

[0306] Evaluation of CTE binding affinities was extended to cell surface binding using antigen-expressing cell lines and flow cytometry analyses. Flow cytometry was used to show the uniform expression of B7-H3 and IL-13Ra2 expression on the surface of A375 cells (Figure 32).Dual-antigen bispecific CTEs were incubated with A375 cells and detected with the commercial anti-CD19 antibody HIB19-PE. Dose response curves for cell surface antigen binding are shown in Figure 33. The data demonstrated that IL-13Ra2 x B7-H3 CTEs bound to A375 cells better than the single antigen CTEs #567 and #574; CTE #750 bound with the lowest EC50 of 140 pM (Table 31)Table 31. Apparent affinity for CTEs binding to a A375 melanoma cells.B inding to A375 cellsMolecularEC50ng / ml EC50nMi weight, kl)a#567 54 1.2 45#574 18 0.4 44- 90 - 13321680vlAttorney Docket No. 2012106-0161#748 12.8 0.2 60#749 10.6 0.18 60#750 3.4 0.06 60#751 8.4 0.14 60

[0307] Cytotoxicity activity of the dual-antigen bispecific CTEs was evaluated using similar methods as described in Examples 6 and 15 with some modifications. Here, the dualantigen positive melanoma cell line A375 was targeted for CD19-CAR-T mediated killing via the CD 19 ECD contained within the CTEs. The cytotoxicity dose response curves shown in Figure 34 and IC50 values show that the CTE designs #574 and #750 produce the lowest concentration IC50 values for inducing CD19-CAR-T mediated A375 melanoma cell death (Table 32). This suggests that on the A375 cell line, having the anti-IL-13Ra2 domain present on the N terminus may be favorable. Targeting B7-H3 alone via CTE #567 was less potent in the cytotoxicity assay.Table 32. IC50 values derived from A375 cytotoxicity curves generated in the presence of CTEs and CD19-directed CAR T cells.CytotoxicityMolecular weightIC50ng / ml IC50pMkDa#567 10.9 244.2 45#574 3 67.8 44#748 5.8 97 60#749 5.4 90.5 60#750 2.8 47.4 60#751 6.8 113 6013321680vlAttorney Docket No. 2012106-0161

[0308] Dual-antigen bispecific CTE binding and cytotoxic activity was further evaluated using the glioblastoma U251MG cell line. Flow cytometry was used to show the uniform expression of B7-H3 and IL-13Ra2 expression on the surface of U251MG cells (Figure 35). Dose response curves for cell surface antigen binding are shown in Figure 36. The data demonstrated that all IL-13Ra2 x B7-H3 CTEs bound to U25 IMG cells better than the single antigen CTEs; Aleta #750 had the lowest EC50 (Table 33). These results demonstrate synergistic binding to dual antigen-positive tumor cells.Table 33. Apparent binding affinities for U251MG cells derived from the CTE dose responsive binding curves.Binding to U251MG cellsMolecularEC50ng / ml EC50nMweight kDa#567 19.9 0.4 45#574 38.6 0.9 44#748 11.7 0.2 60#749 10.6 0.2 60#750 3.1 0.05 60#751 8.8 0.15 60

[0309] These data demonstrate that the bispecific CTEs all bound to the dual-antigenpositive glioblastoma cells better than the single antigen CTEs (#567 and #574); CTE #750 bound with the lowest EC50 of 50 pM and CTE #751 was similar at 150 pM. Therefore, successfully incorporating two distinct VHH directed to two distinct TAA created synergistic binding.- 92 - 13321680vlAttorney Docket No. 2012106-0161

[0310] Cytotoxicity activity of the dual-antigen bispecific CTEs was evaluated to determine the impact of differential binding on killing of U251MG glioblastoma cells as mediated by CD19-CAR-T cells through the CTEs. Results shown in Figure 37 and the calculated IC50 values (Table 34) showed that cytotoxicity curves did not reach a maximal killing. Therefore, this assay was repeated with the single-antigen CTEs and the dual antigen CTEs #750 and #751. The assay was performed at a 5: 1 E: T ratio and allowed to proceed for 48 hours.Table 34. Cytotoxicity IC50 values for U251MG cells targeted with CTEs and CD19-directed CAR T cells.CytotoxicityIC50IC50Molecular(ng / ml) (pM) weight(kDa)#567 3.3 73.8 45#574 5.1 114.9 44#748 4.8 81.2 60#749 4.6 77.1 60#750 1.1 17.6 60#751 2.4 40.7 60

[0311] Dose response curves shown in Figure 38 and calculated IC50s (Table 35) demonstrated that the dual antigen targeting CTEs (#750 and #751) were more potent than single antigen CTEs. In agreement with prior results CTEs #750 and #751 enhanced the potency of anti-CD19 CAR-T cells against the glioblastoma cells compared to single antigen CTEs.Table 35. Cytotoxicity IC50 values for U251MG cells targeted with single antigen CTEs (#567, #574) and dual antigen CTEs (#750, #751) added with anti-CD19 CAR-T cells.- 93 - 13321680vlAttorney Docket No. 2012106-0161U251MG cytotoxicity assayMolecular weightIC50ng / ml IC50pMkDa#567 23.9 540 45#574 36.2 820 44#750 3.7 62 60#751 9.9 166 60Example 15. Generation and characterization of CTEs targeting solid tumors antigens and containing costimulation domains.

[0312] The present example describes CTE expression constructs with a CD28 binding domain of B7-2 that were generated and used for characterizing relevant functional activity of CTE fusion proteins described herein. Some constructs were generated by cloning an anti-IL-13Ra2 VHH in frame with LFA3bds and / or a single B7-2 domain known to mediate binding to CD28 (B7-2bd; Rennert 1997. doi: 10.1093 / intimm / 9.6.805.). The CTE fusion proteins listed in Table 36 were expressed and tested for their ability to mediate CAR T cell costimulation alongside inducing anti-tumor antigen specific CAR T cell cytotoxicity.Table 36. CTE targeting solid tumor, antigens, and containing costimulation domains.Construct # Domains encoded758 IL-13Ra2 VHH - LFA3bd - CD 19 ECD - LFA3bd759 IL-13Ra2 VHH - LFA3bd - CD 19 ECD - LFA3bd - B7-2bd760 IL-13Ra2 VHH - LFA3bd - CD19 ECD - B7-2bd - LFA3761 IL-13Ra2 VHH - CD 19 ECD - B7-2bd- 94 - 13321680vlAttorney Docket No. 2012106-0161766 IL-13Ra2 VHH - CD19 ECD - B7-2bd - B7-2bd767 B7-2bd - IL-13Ra2 VHH - CD19 ECD - B7-2bd768 IL-13Ra2 VHH - B7-2bd - CD19 ECD - B7-2bd769 IL-13Ra2 VHH - LFA3bd - CD 19 ECD - B7-H3 VHH - LFA3bd770 IL-13Ra2 VHH - CD 19 ECD - B7-H3 VHH - B7-2bd771 IL-13Ra2 VHH - B7-2bd - CD 19 ECD

[0313] The effect of adding multiple costimulatory domains to IL-13Ra2-directed CTEs was investigated using ELISA binding assays. The ability to bind the anti-CD19 antibody FMC63 and IL-13Ra2 protein was evaluated by ELISA where bound CTEs were detected with biotinylated IL-13Ra2 that was visualized enzymatically. Results shown in Figure 39 showed that the measured binding events had EC50 values within a tight range of roughly 2 to 9 pM (Table 37)Table 37. EC50 values calculated from the ELISA assay.MolecularEC50ng / ml EC50pM weight,kDa#758 0.6 8.6 69#759 0.1 1.7 83#760 0.6 7.4 83#761 0.1 2.2 58

[0314] The binding assay was repeated, where the anti-CD19 antibody FMC63 and IL-13Ra2 protein was evaluated by ELISA where bound CTEs, containing 2 B7-2 binding domains- 95 - 13321680vlAttorney Docket No. 2012106-0161and B7-H3 bispecifics with LFA3 or B7-2, were detected with biotinylated IL-13Ra2 that was visualized enzymatically and results are shown in Figure 40.! L-13Ra2 binding affinities (Table 38) were derived from the dose response curves in Figure 40.Table 38. IL-13Rα2 binding affinities derived from Figure 40.Binding to IL-13Rα2 EC50ng / ml EC50nM MW kDa#766 2.25 0.03 72.5#767 3.17 0.04 72.5#768 3.32 0.04 72.5#769 3.01 0.03 83.7#770 3.21 0.04 73.6#771 2.93 0.05 58.5

[0315] B7-H3 binding activity for constructs #769 and #770 was evaluated by ELISA. Briefly, an anti-FMC63 was used to coat the plate, followed by biotinylated B7-H3 41g, and detection was done using HRP-SA. Dose response curves and the derived binding affinities are shown in Figure 41 and Table 39.Table 39. B7-H3 4Ig binding affinities derived from Figure 41.Binding to B7-H3 4Ig EC50ng / ml EC50MW kDanM#567 8.91 0.2 44.51#769 5.48 0.07 83.7- 96 - 13321680vlAttorney Docket No. 2012106-0161#770 3.13 0.04 73.6

[0316] Additional binding analysis was performed to evaluate the ability of the CTEs to bind the anti-CD19 antibody FMC63 and CD2 protein by ELISA where CTEs were captured onto the anti-CD19 antibody-coated plate and bound CTEs were detected with biotinylated CD2, streptavidin-HRP and the peroxidase reaction. Here, the ability to bind CD2 requires the presence and appropriate spacing of LFA3bds (see, for example, Example 3).

[0317] Results shown in Figure 42 demonstrated that CTEs lacking LFA3bd were unable to bind to CD2. CTE #759 appeared to have more robust binding than the similar CTE #758 and #760. It appears that the positioning of the B7-2bd at the C-terminus may confer favorable geometry with respect to the LFA3bd and CD19ECD proteins, as evidenced by the binding affinity data (Table 40).Table 40. CTE binding affinities for CD2 by ELISA.EC50Molecular weightEC50ng / mlnM (kDa)#574 - - 44#758 16 0.2 69#759 2.9 0.04 83#760 14.4 0.2 83#761 - - 58

[0318] An evaluation of CD2 binding for IL-13Ra2 x B7-H3 bispecific with LFA3 was performed by ELISA. Here, constructs #764 and #769 were evaluated. Results from this binding assay and CD2 binding affinities derived from dose response curves are shown in Figure 43 and Table 41Table 41. CD2 binding affinities by constructs #764 and # 769 derived from Figure 43.13321680vlAttorney Docket No. 2012106-0161Binding to CD2 EC50ng / ml ECso MW kDanM#764 67.3 0.83 81.2#769 83.3 1.0 83.7

[0319] An evaluation of CD28 binding for B7-2 containing constructs was performed by ELISA. Here, constructs #766, #767, #768, #770, and #771 were evaluated. Briefly, an anti-FMC63 was used to coat the plate, followed by biotinylated CD28, and detection was done using HRP-SA. Dose response curves and the derived binding affinities are shown in Figure 44 and Table 42Table 42. CD28 binding affinities of by constructs #766, #767, #768, #770, and #771 derived from Figure 44.Binding to CD28 EC50ng / ml ECso nM MW kDa#766 69.8 0.96 72.5#767 131.2 1.81 72.5#768 161.2 2.22 72.5#770 - - 73.6#771 - - 58.5

[0320] Cytotoxicity was be assessed by a luciferase reporter cell line assay. Briefly, antiCD 19 CAR T (CAR19) effector cells (E) were transduced with “A254” lentiviral particles. An effector target ratio (E: T) of 5 CAR-positive cells: 1 target cell (A375 cells) was calculated. After a 48-hour incubation, the cells in each well were lysed and subjected to a standard luciferase assay. Here constructs #574, #758, #759, #760, and #763 were evaluated. Percent13321680vlAttorney Docket No. 2012106-0161killing cytotoxicity dose response curved are shown in Figure 45. Derived IC50 values are shown in Table 43.Table 43. Cytotoxicity IC50 values derived from Figure 45.A375 IC50ng / ml IC50pM MW kDa#574 9.2 207 44.4#758 0.22 3.21 68.55#759 0.04 0.48 82.59#760 0.05 0.61 82.59#761 0.13 2.22 58.47

[0321] An additional assay was established to evaluate CTE activation of CAR-T cells in the presence of targeted antigen-expressing cells. Jurkat cells expressing an anti-CD19 CAR domain were further engineered to express an NF AT promoter / luciferase reporter construct. NF AT activity is correlated with T cell activation. To align the NF AT assay with cytotoxicity assays the CAR-CD19-NFAT-luc cells were incubated with HEK293 cells expressing CD20 with or without CTE #607 (anti-CD20 VHH - CD 19 ECD - anti-albumin VHH). CTE #607 binding to CD20 on the target cell and the anti-CD19 domain on the reporter cell was sufficient to trigger CAR T cell activation. Prior work to had demonstrated CTE #607 dependent cytotoxicity against 293-CD20 and against CD20 positive lymphoma cell lines (see, for example, Su, et al., 2022).

[0322] CTE-mediated induction of NF AT -luciferase expression on Jurkat-CAR19 cells in the presence of target 293-CD20 tumor cells is shown in Figure 46A. The effective concentration for half-maximum induction of NF AT luciferase is shown in Table 44 and EC50 for the activity of CTE #607 was 5.4 pM, or 0.3 ng / ml. These results demonstrated a three-log dynamic range. The EC50 for NF AT induction is very similar to the IC50 of 0.12 ng / ml derived - 99 - 13321680vlAttorney Docket No. 2012106-0161from multiple experiments assessing CD20-directed anti-CD19 CAR T cell-mediated cytotoxicity (see, for example, Su, et al., 2022).Table 44. EC50 values calculated from CTE-mediated induction of NFAT-luciferase expression.MolecularEC50(ng / ml) EC50(pM)weight kDa#607 0.3No CTEnacontrolNo CARnacontrol

[0323] The NFAT-luciferase expression assay was then used to evaluate the IL-13Roc2- directed CTE and the addition of cost stimulation domains to those CTEs. Here, induction of NFAT-luciferase expression was evaluated in Jurkat-CAR19 cells that were incubated with IL- 13Ra2 -positive A375 melanoma cells and IL-13Ra2-directed CTEs. TheE: T ratio was 1:1 and the incubation period was five hours. Results shown in Figure 46B and Table 45 demonstrated that CTEs showed the same pattern of enhanced activity in the presence of two LFA3bds.Adding the B7-2bd to CTEs containing anti-IL-13Ra2similarly improved CAR T cell activation in the NFAT-luciferase activation assay. Furthermore, the combination of the B7-2bd domain with two LFA3bds, separated by at least the CD19ECD, extended this enhancement even further.Table 45. Impact of adding costimulation domains to CTEs.Half maximal induction ofluciferaseMolecular weightEC50ng / ml ECso pMkDa#574 2.7 60.8 44- 100 - 13321680vlAttorney Docket No. 2012106-0161#758 0.15 2.2 69#759 0.01 0.1 83#761 0.05 0.9 59

[0324] Striking and novel results were obtained with a CTE designed using the combination of LFA3bds and a B7-2bd (#759) that mediated T cell activation with a EC50 (100 fM) that was > 600-fold more potent than the control CTE (#574) lacking the costimulatory domains.

[0325] Without wishing to be bound by a theory, the extent of improvement may be due to two different phenomena with respect to the A375 melanoma cell line. First the A375 melanoma cells do not express the CD28 ligands B7-1 and B7-2. Second, LFA3 expression A375 melanoma cells is not organized in a manner that has evolved to support binding to a T cell immune synapse.Example 16. Use of a monomeric to TGFp receptor 2 domain as a TGFp TRAP for immune suppression blockade.

[0326] Solid tumor environments are often dominated by the activity of the immunosuppressive cytokine TGFP (Shi 2022. doi: 10.1186 / sl3045-022-01349-6). TGFP is capable of blunting T cell activation and immune function. In many cancers, TGFp acts as a master control signal for the tumor microenvironment (TME) and it has been shown that activated T cells and other immune cells that migrate into TME can be shut down completely by TGFp, preventing the secretion of IFNy and other proinflammatory cytokines that mediate antitumor immunity.

[0327] A monomeric TGFp receptor 2 sequence was encoded as a decoy for TGFp (Bedi et al. US 8,993,524 B2). The sequence encoding a monomeric receptor TRAP domain was cloned with B7-H3 and IL-13Ra2 VHH to create a multi-targeting CD19-engaging CTE (Table 46)- 101 - 13321680vlAttorney Docket No. 2012106-0161Table 46. Components encoded in CTEs #755, #756, #757.CTE# Encoded domains#755 anti-B7-H3 VHH - CD19 ECD - anti-IL-13Ra2 VHH - TGFβR2 ECD#756 TGFβR2 ECD - anti-B7-H3 VHH - CD19 ECD - anti-IL-13Ra2 VHH#757 anti-B7-H3 VHH - CD 19 ECD - TGF0R2ECD - anti-IL-13Ra2 VHH

[0328] The CTE were evaluated for binding activity. Dose response curves of IL-13Ra2 x B7-H3 CTE binding to B7-H3-4Ig are shown in Figure 47. The capture reagent was purified human serum albumin, and the detection reagent was biotinylated B7-H3-4Ig, detected enzymatically. Binding affinities (Table 47) demonstrated equivalent binding by CTEs irrespective of the presence or placement of the TGFβR2 ECD, as detected by B7-H3 in the ELISA format.Table 47. The placement of a TGFBR2 ECD within CTEs does not interfere with binding to tumor antigens.Binding to B7-H3 EC50ng / ml EC50nM Molecular41g weight, kDa#755 3.6 0.05 76#756 2.8 0.04 76#757 2.6 0.03 76#750 3.2 0.05 60

[0329] A second binding assay was run using purified human serum albumin as the capture agent and biotinylated IL-13Ra2 as the detection agent (Figure 48). The derived binding affinities are shown in Table 48.- 102 - 13321680vlAttorney Docket No. 2012106-0161Table 48. Apparent binding affinity EC50 values derived from the ELISA assay.Binding to IL- EC50ng / ml EC Molecula13 « 3.2 0„ nM rR weight, kDa#755 2.1 0.03 76#756 2.1 0.03 76#757 1.9 0.03 76#750 3.3 0.06 60

[0330] Since the presence of the TGF R2 ECD within CTEs had no impact on binding to antigen (Table 48), the ability of these CTE to mediate anti-CD19 CAR T cell cytotoxicity was evaluated by flow cytometry. Induction of U251MG glioblastoma cell cytotoxicity by CD19-directed CAR T cells and CTEs containing the TGFβR2 ECD was measured (Figure 49). The E: T ratio was 5:1 and the assay was run for 48 hours. These data suggest that the activity of the TGFPR2 ECD-containing IL-13Ra.2 x B7-H3 bispecific CTEs are similar to the CTEs without TGFPR2 ECD on U251MG cells. The calculated IC50 values are shown in Table 49, which demonstrated that TGFPR2 ECD-containing CTEs mediated cytotoxicity in the range of 3 - 6 ng / ml.Table 49. Affinities of CTEs binding U251MG cells as measured by flow cytometry.ic molecularU251MG IC50 ng / ml50pM weight kDa#750 3 50.6 60#755 4 52.1 76#756 5.2 69.0 76#757 5.6 73.2 76- 103 - 13321680vlAttorney Docket No. 2012106-0161

[0331] As the CTE activities of binding and mediating cytotoxicity were unaffected, the impact of the TGFβR2 ECD on TGFP activity was measured. The phosphorylation of SMAD2 in response to TGFp incubation was measured in Ramos cell lysates using an ELISA assay. Figure 50 shows dose response TGF -induced SMAD2 phosphorylation by B7-H3 / IL-13Ra2 bispecific CTEs containing the TGF[3R2 ECD. These data show that B7-H3 / ZL-13RO.2 bispecific CTEs containing the TGFPR2 ECD inhibited TGFp-induced SMAD2 phosphorylation. A single antigen CTE targeting Her2 (#628) also blocked TGF beta activation of SMAD2. The derived IC50 values are shown in Table 50. These data indicate that multi-antigen targeting CTEs successfully blocked TGFp signaling to Ramos cells. Furthermore, the utility of the TGFPR2 ECD to antagonize TGFp was confirmed using a second CTE that targeted Her2 (#628).Table 50. ICso values for the inhibition of SMAD2 phosphorylation by CTEs containing the TGFBR2 ECD.Molecular weight IC50 ng / ml IC50 nMkDa #628 20.4 0.3 73.4 #755 103 1.4 76#756 88.7 1.2 76#757 181.2 2.4 76

[0332] Additional cytotoxicity activity was evaluated for IL-13Ra2 / B7-H3 dual targeting CTEs with the addition of a TGFPR2 ECD domain. Results shown in Figure 51 demonstrated that the addition of the TGFPR2 ECD domain to the IL-13Ra2 / B7-H3 dual targeting CTEs (#755 and #756) does not alter cytotoxicity against A375 melanoma cells. The dual antigen targeting CTEs (#755, #756) are shown again to be more potent than the IL-13Ra2 CTE (#574), and inserting the TGFPR2 ECD does not inhibit cytotoxicity (Table 51). Together, these data demonstrated that a TGFP antagonist was successfully added to the CTE format.Table 51. IC50 values for the induction of CTE-mediated A375 cell cytotoxicity by anti-CD19 CAR T cells as mediated by CTEs.- 104 - 13321680vlAttorney Docket No. 2012106-0161A375 cellIC50IC50 Molecularcytotoxicity ng / ml pM weight, kDa#574 10.7 238.3 44#750 2.4 39.7 60#755 2.3 29.7 76#756 2.6 34 76Example 17. Use of a monomeric TGFp receptor 2 domain ECD as a TGFp TRAP with an additional antigen.

[0333] Evaluation of TGFp receptor 2 ECD monomeric sequence was extended to include a second antigen, Her2. Further, the sequence encoding a monomeric receptor TRAP domain was cloned with Her2 scFv and IL-13Ra2 VHH to create a multi -targeting CD19-engaging CTE (Table 52). The use of LFA3bds to augment binding of a CTE directed to a second antigen, Her2, was evaluated in the presence or absence of TGF R2 antagonist sequence.Table 52. CTE created and characterized to evaluate the influence of TGFPR2 antagonist sequence on LFA3bd-mediated binding.CTE # Encoded Domains#674 LFA3bd anti-Her2 scFv - CD 19 ECD#762 anti-Her2 scFv - LFA3bd - CD 19 ECD - LFA3bd - TGFPR2 ECD#763 anti-Her2 scFv -CD 19 ECD - LFA3bd - TGFpR2 ECD - LFA3bd#764 anti-Her2 scFv - LFA3bd - CD 19 ECD - LFA3bd#765 anti-Her2 scFv - CD 19 ECD - LFA3bd - LFA3bd#628 anti-Her2 scFv - CD 19 ECD - TGFPR2 ECD13321680vlAttorney Docket No. 2012106-0161

[0334] CD2 binding by LFA3 and Her2-directed CTEs in the presence or absence of TGF0R2 antagonist sequence was evaluated by ELISA. Dose response curves for CD2 binding are shown in Figure 52. Binding affinities were calculated (Table 53). These results confirmed that the spacing of LFA3bds is critical for CD2 binding. The result showed that the presence of the TGFPR2 antagonist sequence did not interfere with CD2 binding.Table 53. CD2 binding affinities derived from Figure 43.EC50 ng / ml EC50nM MWkDa#674 1062 15.4 69#762 130.5 1.3 97#763 131.8 1.4 97#764 115.1 1.4 81#765 - - 83#628 - - 73Table 54. Her-2 targeting CTE constructs, based on CTE #597, that incorporate a TGFPR2 antagonist sequence and an anti-IL13Ra2 VHH.CTE # Encoded domains#597 anti-IL-13Ra2 VHH - anti-Her2 scFv - CD19 ECD#727 anti-IL-13Ra2 VHH - anti-HER2 scFv - CD19 ECD - TGFpR2 ECD #728 anti-IL-13Ra2 VHH - anti-HER2 scFv- TGFPR2 ECD - CD 19 ECD

[0335] Additional CTEs shown in Table 54 were evaluated for their ability to bind to both the anti-CD19 antibody FMC63 and to the Her2 antigen (Figure 53). The results demonstrated that CTE #728 was not capable of binding to one or the other or both of the capture and detection reagents. In contrast, CTE #727 had binding characteristics very similar to the control CTE #597. The apparent affinities are shown in Table 55.- 106 - 13321680vlAttorney Docket No. 2012106-0161Table 55. EC50 values from the ELISA dose response curves of Figure 53.Molecular weight ECso (ng / ml) ECso(nM)(kDa) #597 8.3 0.12 72#727 3.2 0.04 88#728 91.5 - 88

[0336] Figure 54 shows additional binding analysis performed to evaluate the ability of the CTEs listed in Table 54 to a second antigen, IL-13Ra2. The resulting data (Table 56) show that the CTE #728 is compromised in its binding ability to bind a second antigen, IL-13Ra2. This result illustrates the difficulty in deriving a CTE format that is multifunctional.Table 56. EC50 values from the ELISA dose response curves.ECso (ng / ml)#597 4.2#727 1.4#728 86

[0337] A further analysis of binding to soluble TGFp (Figure 55) showed that the TGFβR2 ECD was functional. It was therefore concluded that it was the TGFβR2 ECD itself that was interfering with the additional functions within the CTE (Table 57).Table 57. Derived EC50 values from Figure 55.Molecular weight ECso (ng / ml) ECso(nM)(kDa) #727 20.8 0.24 88#728 78 0.88 88- 107 - 13321680vlAttorney Docket No. 2012106-0161

[0338] Since the CTE #727 incorporated a functional TGFpR.2 ECD, the ability of the CTE to block TGF0 signaling was examined (Figure 56). The 1C50 value derived from the curve for CTE # 727 was 7.8 ng / ml.

[0339] To demonstrate CTE #727-directed anti-CD19 CAR T-mediated cytotoxicity, the breast cancer cell line BT474 was used. Here, CD19-directed CAR T cells and CTE-mediated induction of BT474 breast carcinoma cell cytotoxicity was measured. The E: T ratio was 5:1 and the assay was run for 48 hours. Results are shown in Figure 57 and Table 58.Table 58. IC50 values derived from the cytotoxicity dose response curve of Figure 57.IC50 ICso Molecular weight (ng / ml) (nM) (kDa) #574 - 44#597 0.6 8.2 72#727 0.5 5.7 88

[0340] Cytotoxic activity was next demonstrated against the A375 melanoma cell line. Here, induction of A375 Her2 KO melanoma cell cytotoxicity using CD19-directed CAR T cells and CTEs was measured. The E: T ratio was 5:1 and the assay was run for 48 hours. Results are shown in Figure 58 and Table 59.Table 59. IC50 values derived from the cytotoxicity dose response curve of Figure 58.IC50 ng / ml IC50 nM Molecular weight kDa #574 6.8 153 44#597 2 27 72#727 0.9 10 88

[0341] Cytotoxic activity was next demonstrated against the U25 IMG glioblastoma cell line. Here, induction of U251MG glioblastoma cell cytotoxicity using CD19-directed CAR T cells and CTEs was measured. The E: T ratio was 5:1 and the assay was run for 48 hours. Results are shown in Figure 59 and Table 60.- 108 - 13321680vlAttorney Docket No. 2012106-0161Table 60. IC50 values derived from the cytotoxicity dose response curve of Figure 59.Molecular weight ICso ng / ml IC50 nMkDa #574 5.7 128.7 44#597 1.4 19.5 72#727 1.4 15.8 88

[0342] Further cytotoxic activity was demonstrated against the BT474 CD58 KO ductal carcinoma cell line. Here, induction of BT474 CD58 KO ductal carcinoma cell cytotoxicity using CD19-directed CAR T cells and CTEs was measured. The E: T ratio was 5:1 and the assay was run for 48 hours. Results are shown in Figure 60 and Table 61.Table 61. IC50 values derived from the cytotoxicity dose response curve of Figure 60.exp 968_Cytotox IC50 on BT474 ( Her2+) usi ng A254 (D 6008) at E: T = 5:1F r F y£: T 5: 1 628 674 762 763 764 765EC50 (ng / ml) 0.1373 0.1957 0.4564 0.1787 0.4613 0.4121 IC50(PM) 1.871 2.836 4.681 1.833 5.681 4.969MW (Da) 73400 69010 97500 97500 81200 82940 Example 18. Costimulatory Engagers for use as biologies for administration and as cell-secreted.

[0343] The present example describes methods for delivery of costimulatory engagers. Additionally, the present example describes exemplary models for evaluating methods of administration of costimulatory engagers described herein. Additional models for evaluating administration of costimulatory engagers are known in the art.

[0344] Purified fusion proteins (e.g., a costimulatory engager) for use as a biologic alongside a cellular therapeutic (e.g., CD19-targeting CAR T cells ) to be administered to a patient can be prepared using methods known in the art. Administration of biological costimulatory engagers (e.g., CTEs and TCEs) can be evaluated using a variety of models known in the art (e.g., in vitro models, in vivo models, ex vivo models).- 109 - 13321680vlAttorney Docket No. 2012106-0161

[0345] Routes of administration are evaluated. For example, CTE engagers are administered locally or regionally alongside administration of CD19-targeting CAR T cells using an in vivo glioblastoma cancer model in which the antigens B7H3 and IL13Ra2 are expressed.

[0346] Additional models known in the art are used for evaluating administration of costimulatory engagers (e.g., CTEs or TCEs) as a biologic include models of tumor growth. For example, models include orthotopic models in which tumor cells are placed in the tissues or organs from which they were derived, e.g., glioblastoma cells are placed orthopedically within the central nervous system. Additional models are used for many indications and appropriate models (e.g., in vitro, ex vivo, in vivo) are known in the art depending on the indication of interest. For example, ovarian carcinoma models make use of intraperitoneal injection, colorectal carcinoma models make use of placement within the mucosa of the gut, breast cancer models use of placement in or adjacent to the breast, for example, in the breast fat pad.

[0347] In the context of administration of an anti-CD19 CAR T cells the model organism, for example, an immune-deficient mouse model, is infused with CAR T cells. CAR T cells circulate throughout the organism until the tumor is encountered. A costimulatory engager (e.g., CTE) injected locally at the site of tumor growth directs a CAR T cell(s) to attack a tumor via a CTE..

[0348] Methods for evaluating CAR-T cell activity using in vitro and in vivo models are translatable to the treatment of cancer patients.

[0349] Methods described for delivery and / or administration of are applicable to the delivery of TCEs and costimulatory TCEs described herein. TCEs can be delivered to test animals as biologicals that are purified, using methods widely known in the art. Such animals can be implanted with cells express one or more targeted antigens and TCE injected to induce cytotoxicity in the presence of human immune cells. Methods for placing human immune cells in immune-deficient animals are widely known and include the injection of PBMC into immunodeficient mice such as NSG or NOG mice. Other methods include injection of CD34 positive stem cells into immunodeficient mice (e.g., NSG or NOG mice (Guil-Luna, 2021.10.1146 / annurev-cancerbio-050520100526)). TCEs and Costimulatory TCEs are readily secreted from T cells, including CAR T cells, using ex vivo and in vivo viral and non-viral transduction - 110 - 13321680vlAttorney Docket No. 2012106-0161methods. Without being bound by a theory, TCEs are evaluated using in vivo models of cancer and are shown to be more effective in killing cancer cells than are non-costimulatory TCE.

[0350] The methods evaluating TCE using in vitro and in vivo models are translatable to the treatment of cancer patients.Example 19. Demonstration of Immune Synapse FormationThe present example demonstrates the ability of CTEs described herein to effectively form an immune synapse.Methods.

[0351] The MB-MDA-468 cell line used in the present example was derived from a metastatic breast cancer biopsy and is widely used in studies with Her2 -targeting therapeutics. Fluorescent-labeled CD19-targeting CAR T cells were co-cultured with Her2-expressing MB-MDA-468 tumor cells with or without CAR T Engager (CTE) proteins described herein. Lattice light-sheet confocal microscopy was used to visualize cell-cell contact.Table 62 Her-2 targeting CTE constructsCTE # Encoded domains#782 anti-HER2 VHH-LFA3bd-CD19ECD-LFA3bd-anti-albumin VHH #786 anti-HER2 VHH-CD19ECD-anti-albumin VHHResults:

[0352] Fluorescent signals allowed visualization of CAR-T cells (pink) and tumor cells (brown). The intensity of the signal is relative to the plane of the cell relative to the light source.

[0353] No cell-cell contact was observed when CD19-CAR T cells were co-cultured alone with MB-MDA-468 tumor cells (Fig. 61A). In contrast, when Her-2 directed CTE proteins were added to the co-culture, cell-cell contact was observed (Figs. 6 IB, 61C).

[0354] When CTE #782 was added to the co-culture, large and persistent contact was observed in a ring pattern that closely resembled immune synapse formation (Fig. 61B). CTE - Ill - 13321680vlAttorney Docket No. 2012106-0161#782 contains 2 LFA3 binding domains to costimulate CD2 on the CAR-T surface. When CTE #786 was added to the coculture, small and focal contact was observed (C) suggesting a sub-optimal immune synapse was formed. These results are consistent with cytotoxicity and IFNv secretion data that show that the inclusion of LFA3 binding domains in the CTE optimizes CAR-T activity upon binding.LISTING OF SEQUENCES

[0355] Key for the amino acid domain assignments:Domain Identifying FormatSignal sequence dotted underlineCD19 extracellular domain solid underlineAnti-albumin domain italic fontLFA3 domain bold and solid underlineFirst VHH or scFv in a sequence bold + italicSecond VHH or scFv in the sequence italic + solid underlineTGF0R2 domain boldAnti-CD3 scFv solid underline + bold + italicB7-2 binding domain dashed underlineG4S linkers no formattingHis tag no formatting13321680vlAttorney Docket No. 2012106-0161Aleta #263Nucleotide sequence SEQ ID NO: 1

[0356] ATGGAGTTTGGGCTGAGCTGGGTTTTCCTCGTTGCTCTTTTTAGAGGTG TCCAGTGTGAGGTGCAGCTGGTGGAGTCTGGTGGTGGTCTTGTTCAACCTGGTGGTT CTCTTCGTCTTTCTTGTGCTGCTTCTGGTTTTAATATTAAAGATACTTATATTCATTGG GTTCGTCAAGCTCCTGGTAAAGGTCTTGAATGGGTTGCTCGTATTTATCCTACTAATG GTTATACTCGTTATGCTGATTCTGTTAAAGGTCGTTTTACTATTTCTGCTGATACTTCT AAAAATACTGCTTATCTTCAAATGAACTCTCTTCGTGCTGAAGATACTGCTGTTTATT ATTGTTCTCGTTGGGGTGGTGATGGTTTTTATGCTATGGATTATTGGGGTCAAGGTAC TCTTGTCACCGTCTCCTCAGCTAGCACCGGGGGAGGTGGGTCTGGAGGTGGAGGATC TGGTGGAGGTGGGTCTGACATCCAGATGACCCAGTCTCCTTCTTCTCTTTCTGCTTCT GTTGGTGATCGTGTTACTATTACTTGTCGTGCTTCTCAAGATGTTAATACTGCTGTTG CTTGGTATCAACAAAAACCTGGTAAAGCTCCTAAACTTCTTATTTATTCTGCTTCTTT TCTTTATTCTGGTGTTCCTTCTCGTTTTTCTGGTTCTCGTTCTGGTACTGATTTTACTCT TACTATTTCTTCTCTTCAACCTGAAGATTTTGCTACTTATTATTGTCAACAACATTAT ACTACTCCTCCTACTTTTGGTCAAGGTACCAAGGTGGAGATCAAACGTACGGGAGGA GGTGGGTCTGGAGGTGGAGGATCTGGTGGAGGTGGGTCTGGAGGAGGTGGATCCCC CGAGGAACCTCTAGTGGTGAAGGTGGAAGAGGGAGATACCGCTGCCCTGTGGTGCC TCAAGGGGACCTCAGATGGCCCCACTCAGCAGCTGACCTGGTCTCGGGAGTCCCCG CTTAAACCCTTCTTAAAATACAGCCTGGGGGTGCCAGGCCTGGGAGTGCACGTGAG GCCCGACGCCATCAGCGTGGTTATCCGGAACGTCTCTCAACAGATGGGGGGCTTCTA CCTGTGCCAGCCGGGGCCCCCCTCTGAGAAGGCCTGGCAGCCTGGCTGGACAGTCA ATGTGGAGGGCAGCGGGGAGCTGTTCCGGTGGAATGTTTCGGACCTAGGTGGCCTG GGCTGTGGCCTGAAGAACAGGTCCTCAGAGGGCCCCAGCTCCCCTTCCGGGAAGCT CATGAGCCCCAAGCTGTATGTGTGGGCCAAAGACCGCCCTGAGATCTGGGAGGGAG AGCCTCCGTGTCTCCCACCGAGGGACAGCCTGAACCAGAGCCTCAGCCGGGACCTC ACCGTTGCCCCTGGCTCCACACTCTGGCTGTCCTGTGGGGTACCCCCTGACTCTGTGT CCAGGGGCCCCCTCTCCTGGACCCATGTGCACCCCAAGGGGCCTAAGTCATTGCTGA GCCTAGAGCTGAAGGACGATCGCCCGGCCAGAGATATGTGGGTAATGGGCACGAGC- 113 - 13321680vlAttorney Docket No. 2012106-0161CTGATGTTGCCCCGGGCCACAGCTCAAGACGCTGGAAAGTGGTATTGTCACCGTGGC AACCTGACCATGTCATTCCACCTGGAGATCACTGCTCGGCCATCTAGACATCATCAC CATCACCATAmino acid sequence SEQ ID NO: 2

[0357] MEFGLSWVFLVALFRGVQCEVQLVESGGGLVQPGGSLRLSCAASGFNIKDTYIHWVRQAPGKGLEWVARIYPTNGYTRYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCSRWGGDGFYAMDYWGQGTLVTVSSASTGGGGSGGGGSGGGGSDIQMTQSPSSLSASVGDRVTITCRASQDVNTAVAWYQQKPGKAPKLLIYSASFLYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQHYTTPPTFGQGTKVEIКRTGGGGSGGGGSGGGGSGGGSPEEPLVVKVEEGDTAALWCLKGTSDGPTQQLTWSRESPLKPFLKYSLGVPGLGVHVRPDAISVVIRNVSQQMGGFYLCQPGPPSEKAWQPGWTVNVEGSGELFRWNVSDLGGLGCGLKNRSSEGPSSPSGKLMSPKLYVWAKDRPEIWEGEPPCLPPRDSLNQSLSRDLTVAPGSTLWLSCGVPPDSVSRGPLSWTHVHPKGPKSLLSLELKDDRPARDMWVMGTSLMLPRATAQDAGKWYCHRGNLTMSFHLEITARPSRHHHHHHAleta #567Nucleotide sequence SEQ ID NO: 3

[0358] ATGGAGTTTGGGCTGAGCTGGGTTTTCCTCGTTGCTCTTTTTAGAGGTG TCCAGTGTCAGGTGCAGTTGCAGCAGTCTGGGGGAGGATTGGTGCAGGCTGGGGGC TCTCTGAGACTCTCCTGTGCAGCCTCTGGACGCACCTTCACCCGCCTTCCCATGGGCT GGTTCCGCCAGGCTCCAGGAAAGGAGCGAGAGTTTGTAGCAGCTCTTAGCTGGAGT GGTGGCAACACAGACTATGCAAATACTGCAGACTCCGTGAAGGGCCGATTCACCAT GTCCAGAGACAGCGCCAAGAACACGGTGTATCTGTGGATGAACAACCTGAAACCTG AGGACACGGCCGTTTATTACTGTGCAGCCCGACAGAGTGGTTGGGGTACCGCCTTCA CTGATGACTATGACTACTGGGGCCAGGGGACCCAGGTCACCGTCTCCTCAGGAGGA GGTGGGTCTGGAGGTGGAGGATCTGGTGGAGGTGGGTCTGGAGGAGGTGGATCCCC CGAGGAACCTCTAGTGGTGAAGGTGGAAGAGGGAGATACCGCTGCCCTGTGGTGCC- 114 - 13321680vlAttorney Docket No. 2012106-0161TCAAGGGGACCTCAGATGGCCCCACTCAGCAGCTGACCTGGTCTCGGGAGTCCCCG CTTAAACCCTTCTTAAAATACAGCCTGGGGGTGCCAGGCCTGGGAGTGCACGTGAG GCCCGACGCCATCAGCGTGGTTATCCGGAACGTCTCTCAACAGATGGGGGGCTTCTA CCTGTGCCAGCCGGGGCCCCCCTCTGAGAAGGCCTGGCAGCCTGGCTGGACAGTCA ATGTGGAGGGCAGCGGGGAGCTGTTCCGGTGGAATGTTTCGGACCTAGGTGGCCTG GGCTGTGGCCTGAAGAACAGGTCCTCAGAGGGCCCCAGCTCCCCTTCCGGGAAGCT CATGAGCCCCAAGCTGTATGTGTGGGCCAAAGACCGCCCTGAGATCTGGGAGGGAG AGCCTCCGTGTCTCCCACCGAGGGACAGCCTGAACCAGAGCCTCAGCCGGGACCTC ACCGTTGCCCCTGGCTCCACACTCTGGCTGTCCTGTGGGGTACCCCCTGACTCTGTGT CCAGGGGCCCCCTCTCCTGGACCCATGTGCACCCCAAGGGGCCTAAGTCATTGCTGA GCCTAGAGCTGAAGGACGATCGCCCGGCCAGAGATATGTGGGTAATGGGCACGAGC CTGATGTTGCCCCGGGCCACAGCTCAAGACGCTGGAAAGTGGTATTGTCACCGTGGC AACCTGACCATGTCATTCCACCTGGAGATCACTGCTCGGCCATCTAGACATCATCAC CATCACCATAmino acid sequence SEQ ID NO: 4

[0359] MEFGLSWVFLVALFRGVQCQVLQQSGGGLVQAGGSLRLSCAASGRTFTRLPMGWFRQAPGKEREFVAALSWSGGNTDYANTADSVKGRFTMSRDSAKNTVYLWMNNLKPEDTAVYYCAARQSGWGTAFTDDYDYWGQGTQVTVSSGGGGSGGGGSGGGGSGGGSPEEPLVVKVEEGDTAALWCLKGTSDGPTQQLTWSRESPLKPFLKYSLGVPGLGVHVRPDAISVVIRNVSQQMGGFYLCQPGPPSEKAWQPGWTVNVEGSGELFRWNVSDLGGLGCGLKNRSSEGPSSPSGKLMSPKLYVWAKDRPEIWEGEPPCLPPRDSLNQSLSRDLTVAPGSTLWLSCGVPPDSVSRGPLSWTHVHPKGPKSLLSLELKDDRPARDMWVMGTSLMLPRATAQDAGKWYCHRGNLTMSFHLEITARPSRHHHHHHAleta #568Nucleotide sequence SEQ ID NO: 5- 115 - 13321680vlAttorney Docket No. 2012106-0161

[0360] ATGGAGTTTGGGCTGAGCTGGGTTTTCCTCGTTGCTCTTTTTAGAGGTG TCCAGTGTGATATCGTGATGACCCAGAGCCACAAGTTCATGAGCACCAGCATCGGC GCCAGAGTGTCCATCACCTGCAAGGCCAGCCAGGACGTGCGGACAGCTGTGGCCTG GTACCAGCAGAAACCCGGCCAAAGCCCTAAGCTGCTGATCTACAGCGCCAGCTATA GATACACAGGCGTGCCAGATAGATTCACAGGCAGCGGATCCGGCACCGACTTCACC TTCACCATCTCTTCTGTGCAGGCCGAGGACCTGGCCGTGTACTATTGCCAGCAGCAC TACGGCACACCTCCTTGGACATTTGGCGGAGGAACCAAGCTGGAAATCAAGGGCGG CGGCGGCTCTGGCGGCGGAGGCAGCGGCGGTGGCGGCAGCGAGGTGCAGCTGGTCG AGAGCGGAGGCGGCCTGGTTAAGCCTGGCGGATCCCTGAAACTCAGCTGCGAGGCC TCCAGATTCACCTTCAGCAGCTACGCCATGTCTTGGGTGCGGCAGACCCCTGAGAAG CGGCTGGAATGGGTCGCCGCTATTTCTGGCGGAGGCAGATACACCTACTACCCCGAC AGCATGAAAGGCCGCTTTACAATCAGCCGGGACAACGCCAAGAACTTCCTGTACCT GCAGATGAGCAGCCTGAGAAGCGAGGACACCGCCATGTACTACTGCGCCAGACACT ACGACGGCTACCTTGATTACTGGGGCCAGGGCACAACCCTGACCGTGTCCAGCGGA GGAGGTGGGTCTGGAGGTGGAGGATCTGGTGGAGGTGGGTCTGGAGGAGGTGGATC CCCCGAGGAACCTCTAGTGGTGAAGGTGGAAGAGGGAGATACCGCTGCCCTGTGGT GCCTCAAGGGGACCTCAGATGGCCCCACTCAGCAGCTGACCTGGTCTCGGGAGTCC CCGCTTAAACCCTTCTTAAAATACAGCCTGGGGGTGCCAGGCCTGGGAGTGCACGTG AGGCCCGACGCCATCAGCGTGGTTATCCGGAACGTCTCTCAACAGATGGGGGGCTT CTACCTGTGCCAGCCGGGGCCCCCCTCTGAGAAGGCCTGGCAGCCTGGCTGGACAG TCAATGTGGAGGGCAGCGGGGAGCTGTTCCGGTGGAATGTTTCGGACCTAGGTGGC CTGGGCTGTGGCCTGAAGAACAGGTCCTCAGAGGGCCCCAGCTCCCCTTCCGGGAA GCTCATGAGCCCCAAGCTGTATGTGTGGGCCAAAGACCGCCCTGAGATCTGGGAGG GAGAGCCTCCGTGTCTCCCACCGAGGGACAGCCTGAACCAGAGCCTCAGCCGGGAC CTCACCGTTGCCCCTGGCTCCACACTCTGGCTGTCCTGTGGGGTACCCCCTGACTCTG TGTCCAGGGGCCCCCTCTCCTGGACCCATGTGCACCCCAAGGGGCCTAAGTCATTGC TGAGCCTAGAGCTGAAGGACGATCGCCCGGCCAGAGATATGTGGGTAATGGGCACG AGCCTGATGTTGCCCCGGGCCACAGCTCAAGACGCTGGAAAGTGGTATTGTCACCGT GGCAACCTGACCATGTCATTCCACCTGGAGATCACTGCTCGGCCATCTAGACATCAT CACCATCACCAT- 116 - 13321680vlAttorney Docket No. 2012106-0161Amino acid sequence SEQ ID NO: 6

[0361] MEFGLSWVFLVALFRGVQCDIVMTQSHKFMSTSIGARVSITCKASQDVRTAVAWYQQKPGQSPKLLIYSASYRYTGVPDRFTGSGSGTDFTFTTSSVQAEDLAVYYCQQHYGTPPWTFGGGTKLEIKGGGGSGGGGSGGGGSEVQLVESGGGLVKPGGSLKLSCEASRFTFSSYAMSWVRQTPEKRLEWVAAISGGGRYTYYPDSMKGRFTISRDNAKNFLYLQMSSLRSEDTAMYYCARHYDGYLDYWGQGTTLTVSSGGGGSGGGGSGGGGSGGGGSPEEPLVVKVEEGDTAALWCLKGTSDGPTQQLTWSRESPLKPFLKYSLGVPGLGVHVRPDAISVVIRNVSQQMGGFYLCQPGPPSEKAWQPGWTVNVEGSGELFRWNVSDLGGLGCGLKNRSSEGPSSPSGKLMSPKLYVWAKDRPEIWEGEPPCLPPRDSLNQSLSRDLTVAPGSTLWLSCGVPPDSVSRGPLSWTHVHPKGPKSLLSLELKDDRPARDMWVMGTSLMLPRATAQDAGKWYCHRGNLTMSFHLEITARPSRHHHHHHAleta #569Nucleotide sequence SEQ ID NO: 7

[0362] ATGGAGTTTGGGCTGAGCTGGGTTTTCCTCGTTGCTCTTTTTAGAGGTG TCCAGTGTGAGGTGCAGCTGGTCGAGAGCGGCGGCGGACTGGTGCAACCTGGCGGC AGCCTGAGACTGAGCTGCGCCGCATCTGGCTTCACATTTAGCAGCTTCGGCATGCAC TGGGTTAGACAGGCCCCTGGAAAAGGCCTGGAATGGGTGGCCTACATCAGCTCTGA CAGCAGCGCTATCTACTACGCCGACACAGTGAAGGGCAGATTCACCATCTCTAGAG ATAACGCCAAGAATAGCCTGTACCTGCAGATGAACAGCCTCAGGGACGAGGACACC GCCGTGTACTACTGTGGCCGGGGCAGAGAGAACATCTATTACGGCTCCCGCCTGGA CTACTGGGGACAGGGCACAACCGTGACCGTGTCCTCCGGCGGAGGAGGCAGCGGCG GCGGCGGCTCTGGCGGCGGCGGCAGCGATATCCAGCTGACACAGAGCCCTTCTTTTC TGAGCGCCAGCGTGGGGGATAGAGTGACCATTACATGCAAGGCTTCACAGAATGTG GACACCAACGTGGCTTGGTACCAGCAGAAACCAGGCAAGGCCCCTAAGGCCCTGAT CTACAGCGCCAGCTACCGGTACTCTGGCGTCCCCAGCCGGTTCAGCGGATCCGGCTC CGGCACCGATTTTACCCTGACCATCAGCAGCCTGCAGCCTGAGGACTTCGCCACATA- 117 - 13321680vlAttorney Docket No. 2012106-0161CTATTGCCAGCAATACAACAACTACCCCTTCACCTTCGGCCAGGGAACCAAGCTGGA AATCAAGGGAGGAGGTGGGTCTGGAGGTGGAGGATCTGGTGGAGGTGGGTCTGGAG GAGGTGGATCCCCCGAGGAACCTCTAGTGGTGAAGGTGGAAGAGGGAGATACCGCT GCCCTGTGGTGCCTCAAGGGGACCTCAGATGGCCCCACTCAGCAGCTGACCTGGTCT CGGGAGTCCCCGCTTAAACCCTTCTTAAAATACAGCCTGGGGGTGCCAGGCCTGGG AGTGCACGTGAGGCCCGACGCCATCAGCGTGGTTATCCGGAACGTCTCTCAACAGA TGGGGGGCTTCTACCTGTGCCAGCCGGGGCCCCCCTCTGAGAAGGCCTGGCAGCCTG GCTGGACAGTCAATGTGGAGGGCAGCGGGGAGCTGTTCCGGTGGAATGTTTCGGAC CTAGGTGGCCTGGGCTGTGGCCTGAAGAACAGGTCCTCAGAGGGCCCCAGCTCCCC TTCCGGGAAGCTCATGAGCCCCAAGCTGTATGTGTGGGCCAAAGACCGCCCTGAGA TCTGGGAGGGAGAGCCTCCGTGTCTCCCACCGAGGGACAGCCTGAACCAGAGCCTC AGCCGGGACCTCACCGTTGCCCCTGGCTCCACACTCTGGCTGTCCTGTGGGGTACCC CCTGACTCTGTGTCCAGGGGCCCCCTCTCCTGGACCCATGTGCACCCCAAGGGGCCT AAGTCATTGCTGAGCCTAGAGCTGAAGGACGATCGCCCGGCCAGAGATATGTGGGT AATGGGCACGAGCCTGATGTTGCCCCGGGCCACAGCTCAAGACGCTGGAAAGTGGT ATTGTCACCGTGGCAACCTGACCATGTCATTCCACCTGGAGATCACTGCTCGGCCAT CTAGACATCATCACCATCACCATAmino acid sequence SEQ ID NO: 8

[0363] MEFGLSWVFLVALFRGVQCEVQLVESGGGLVQPGGSLRLSCAASGFTFSSFGMHWVRQAPGKGLEWVAYISSDSSAIYYADTVKGRFTISRDNAKNSLYLQMNSLRDEDTAVYYCGRGRENIYYGSRLDYWGQGTTVTVSSGGGGSGGGGSGGGGSDIQLTQSPSFLS ASVGDRVTITCKASQNVDTNVAWYQQKPGKAPKALIYSASYRYSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQYNNYPFTFGQGTKLEIKGGGGSGGGGSGGGGSGGGGSP EEPLVVKVEEGDTAALWCLKGTSDGPTQQLTWSRESPLKPFLKYSLGVPGLGVHVRPDAISVVIRNVSQQMGGFYLCQPGPPSEKAWQPGWTVNVEGSGELFRWNVSDLGGLGCGLKNRSSEGPSSPSGKLMSPKLYVWAKDRPEIWEGEPPCLPPRDSLNQSLSRDLTVAPGSTLWLSCGVPPDSVSRGPLSWTHVHPKGPKSLLSLELKDDRPARDMWVMGTSLMLPRATAQDAGKWYCHRGNLTMSFHLEITARPSRHHHHHH- 118 - 13321680vlAttorney Docket No. 2012106-0161Aleta #570Nucleotide sequence SEQ ID NO: 9

[0364] ATGGAGTTTGGGCTGAGCTGGGTTTTCCTCGTTGCTCTTTTTAGAGGTG TCCAGTGTCAGGTGCAGCTGGTGCAGTCTGGCGCCGAGGTGAAAAAGCCCGGCAGC TCTGTCAAGGTGTCCTGCAAGGCCAGCGGAGGTACATTTAGCAGCTACGCCATCAGC TGGGTGCGGCAGGCCCCAGGCCAGGGCCTGGAGTGGATGGGCGGAATCATCCCTAT CCTGGGCACAGCCAACTACGCCCAGAAATTCCAGGGCAGAGTGACAATCACCGCCG ACGAGAGCACCAGCACAGCCTACATGGAACTGTCCAGCCTGAGAAGCGAGGATACC GCTGTGTATTACTGCGCCAGGTGGGGCGGCGGAGCCTTCGACATCTGGGGCCAAGG CACAATGGTGACCGTGTCTTCTGGCGGCGGAGGGAGCGGCGGCGGAGGCAGCGGCG GCGGAGGAAGCGAGATCGTGCTGACCCAGAGCCCCGGCACACTGTCTCTGAGCCCT GGCGAGCGGGCCACCCTGTCCTGCAGAGCTAGCCAGAGCGTGGGCGGCTACCTGGC CTGGTACCAGCAGAAGCCTGGACAAGCTCCTAGACTGCTGATCTACGACGCCTCTAA TAGAGCAACCGGCATCCCCGCTAGATTCAGCGGCTCTGGCTCCGGAACCGATTTTAC CCTCACCATCAGCAGCCTGGAACCTGAGGACTTCGCCGTCTACTACTGCCAGCAGCG GAACAACTGGCCTCCAATGTACACCTTCGGCCAGGGCACCAAGCTGGAAATCAAGG GAGGAGGTGGGTCTGGAGGTGGAGGATCTGGTGGAGGTGGGTCTGGAGGAGGTGG ATCCCCCGAGGAACCTCTAGTGGTGAAGGTGGAAGAGGGAGATACCGCTGCCCTGT GGTGCCTCAAGGGGACCTCAGATGGCCCCACTCAGCAGCTGACCTGGTCTCGGGAG TCCCCGCTTAAACCCTTCTTAAAATACAGCCTGGGGGTGCCAGGCCTGGGAGTGCAC GTGAGGCCCGACGCCATCAGCGTGGTTATCCGGAACGTCTCTCAACAGATGGGGGG CTTCTACCTGTGCCAGCCGGGGCCCCCCTCTGAGAAGGCCTGGCAGCCTGGCTGGAC AGTCAATGTGGAGGGCAGCGGGGAGCTGTTCCGGTGGAATGTTTCGGACCTAGGTG GCCTGGGCTGTGGCCTGAAGAACAGGTCCTCAGAGGGCCCCAGCTCCCCTTCCGGG AAGCTCATGAGCCCCAAGCTGTATGTGTGGGCCAAAGACCGCCCTGAGATCTGGGA GGGAGAGCCTCCGTGTCTCCCACCGAGGGACAGCCTGAACCAGAGCCTCAGCCGGG ACCTCACCGTTGCCCCTGGCTCCACACTCTGGCTGTCCTGTGGGGTACCCCCTGACTC TGTGTCCAGGGGCCCCCTCTCCTGGACCCATGTGCACCCCAAGGGGCCTAAGTCATT- 119 - 13321680vlAttorney Docket No. 2012106-0161GCTGAGCCTAGAGCTGAAGGACGATCGCCCGGCCAGAGATATGTGGGTAATGGGCA CGAGCCTGATGTTGCCCCGGGCCACAGCTCAAGACGCTGGAAAGTGGTATTGTCAC CGTGGCAACCTGACCATGTCATTCCACCTGGAGATCACTGCTCGGCCATCTAGACAT CATCACCATCACCATAmino acid sequence SEQ ID NO: 10

[0365] MEFGLSWVFLVALFRGVQCQVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYAISWVRQAPGQGLEWMGGIIPILGTANYAQKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCARWGGGAFDIWGQGTMVTVSSGGGGSGGGGSGGGGSEIVLTQSPGTLSLSPGERATLSCRASQSVGGYLAWYQQKPGQAPRLLIYDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRNNWPPMYTFGQGTKLEIKGGGGSGGGGSGGGGSGGGGSPEE PLVVKVEEGDTAALWCLKGTSDGPTQQLTWSRESPLKPFLKYSLGVPGLGVHVRPDAISVVIRNVSQQMGGFYLCQPGPPSEKAWQPGWTVNVEGSGELFRWNVSDLGGLGCGLKNRSSEGPSSPSGKLMSPKLYVWAKDRPEIWEGEPPCLPPRDSLNQSLSRDLTVAPGSTLWLSCGVPPDSVSRGPLSWTHVHPKGPKSLLSLELKDDRPARDMWVMGTSLMLPRATAQDAGKWYCHRGNLTMSFHLEITARPSRHHHHHHAleta #571Nucleotide sequence SEQ ID NO: 11

[0366] ATGGAGTTTGGGCTGAGCTGGGTTTTCCTCGTTGCTCTTTTTAGAGGTG TCCAGTGTGAGGTGCAGCTGGTGGAGTCTGGTGGTGGTCTTGTTCAACCTGGTGGTT CTCTTCGTCTTTCTTGTGCTGCTTCTGGTTTTAATATTAAAGATACTTATATTCATTGG GTTCGTCAAGCTCCTGGTAAAGGTCTTGAATGGGTTGCTCGTATTTATCCTACTAATG GTTATACTCGTTATGCTGATTCTGTTAAAGGTCGTTTTACTATTTCTGCTGATACTTCT AAAAATACTGCTTATCTTCAAATGAACTCTCTTCGTGCTGAAGATACTGCTGTTTATT ATTGTTCTCGTTGGGGTGGTGATGGTTTTTATGCTATGGATTATTGGGGTCAAGGTAC TCTTGTCACCGTCTCCTCAGCTAGCACCGGGGGAGGTGGGTCTGGAGGTGGAGGATC TGGTGGAGGTGGGTCTGACATCCAGATGACCCAGTCTCCTTCTTCTCTTTCTGCTTCT- 120 - 13321680vlAttorney Docket No. 2012106-0161GTTGGTGATCGTGTTACTATTACTTGTCGTGCTTCTCAAGATGTTAATACTGCTGTTG CTTGGTATCAACAAAAACCTGGTAAAGCTCCTAAACTTCTTATTTATTCTGCTTCTTT TCTTTATTCTGGTGTTCCTTCTCGTTTTTCTGGTTCTCGTTCTGGTACTGATTTTACTCT TACTATTTCTTCTCTTCAACCTGAAGATTTTGCTACTTATTATTGTCAACAACATTAT ACTACTCCTCCTACTTTTGGTCAAGGTACCAAGGTGGAGATCAAACGTACGGGAGGA GGTGGGTCTGGAGGTGGAGGATCTGGTGGAGGTGGGTCTGGAGGAGGTGGATCCCC CGAGGAACCTCTAGTGGTGAAGGTGGAAGAGGGAGATACCGCTGCCCTGTGGTGCC TCAAGGGGACCTCAGATGGCCCCACTCAGCAGCTGACCTGGTCTCGGGAGTCCCCG CTTAAACCCTTCTTAAAATACAGCCTGGGGGTGCCAGGCCTGGGAGTGCACGTGAG GCCCGACGCCATCAGCGTGGTTATCCGGAACGTCTCTCAACAGATGGGGGGCTTCTA CCTGTGCCAGCCGGGGCCCCCCTCTGAGAAGGCCTGGCAGCCTGGCTGGACAGTCA ATGTGGAGGGCAGCGGGGAGCTGTTCCGGTGGAATGTTTCGGACCTAGGTGGCCTG GGCTGTGGCCTGAAGAACAGGTCCTCAGAGGGCCCCAGCTCCCCTTCCGGGAAGCT CATGAGCCCCAAGCTGTATGTGTGGGCCAAAGACCGCCCTGAGATCTGGGAGGGAG AGCCTCCGTGTCTCCCACCGAGGGACAGCCTGAACCAGAGCCTCAGCCGGGACCTC ACCGTTGCCCCTGGCTCCACACTCTGGCTGTCCTGTGGGGTACCCCCTGACTCTGTGT CCAGGGGCCCCCTCTCCTGGACCCATGTGCACCCCAAGGGGCCTAAGTCATTGCTGA GCCTAGAGCTGAAGGACGATCGCCCGGCCAGAGATATGTGGGTAATGGGCACGAGC CTGATGTTGCCCCGGGCCACAGCTCAAGACGCTGGAAAGTGGTATTGTCACCGTGGC AACCTGACCATGTCATTCCACCTGGAGATCACTGCTCGGCCATCTAGAGGAGGAGGT GGGTCTGGAGGTGGAGGATCTGGTGGAGGTGGGTCTGGAGGAGGTGGGTCTGATAT CGTGATGACCCAGAGCCACAAGTTCATGAGCACCAGCATCGGCGCCAGAGTGTCCA TCACCTGCAAGGCCAGCCAGGACGTGCGGACAGCTGTGGCCTGGTACCAGCAGAAA CCCGGCCAAAGCCCTAAGCTGCTGATCTACAGCGCCAGCTATAGATACACAGGCGT GCCAGATAGATTCACAGGCAGCGGATCCGGCACCGACTTCACCTTCACCATCTCTTC TGTGCAGGCCGAGGACCTGGCCGTGTACTATTGCCAGCAGCACTACGGCACACCTCC TTGGACATTTGGCGGAGGAACCAAGCTGGAAATCAAGGGCGGCGGCGGCTCTGGCG GCGGAGGCAGCGGCGGTGGCGGCAGCGAGGTGCAGCTGGTCGAGAGCGGAGGCGG CCTGGTTAAGCCTGGCGGATCCCTGAAACTCAGCTGCGAGGCCTCCAGATTCACCTT CAGCAGCTACGCCATGTCTTGGGTGCGGCAGACCCCTGAGAAGCGGCTGGAATGGG- 121 - 13321680vlAttorney Docket No. 2012106-0161TCGCCGCTATTTCTGGCGGAGGCAGATACACCTACTACCCCGACAGCATGAAAGGC CGCTTTACAATCAGCCGGGACAACGCCAAGAACTTCCTGTACCTGCAGATGAGCAG CCTGAGAAGCGAGGACACCGCCATGTACTACTGCGCCAGACACTACGACGGCTACC TTGATTACTGGGGCCAGGGCACAACCCTGACCGTGTCCAGCCATCATCACCATCACC ATAmino acid sequence SEQ ID NO: 12

[0367] MEFGLSWVFLyALFRGy. QCEVQLVESGGGLVQPGGSLRLSCAASGFNIK DTYIHWVRQAPGKGLEWVARIYPTNGYTRYADSVKGRFTISADTSKNTAYLQMNSLRA EDTAVYYCSRWGGDGFYAMDYWGQGTLVTVSSASTGGGGSGGGGSGGGGSD1QMTQ SPSSLSASVGDRVTITCRASQDVNTAVAWYQQKPGKAPKLLIYSASFLYSGVPSRFSGSR SGTDFTLTISSLQPEDFATYYCQQHYTTPPTFGQGTKVEIKRTGGGGSGGGGSGGGGSG GGGSPEEPLVVKVEEGDTAALWCLKGTSDGPTOQLTWSRESPLKPFLKYSLGVPGLGV HVRPDAISVVIRNVSOOMGGFYLCOPGPPSEKAWOPGWTVNVEGSGELFRWNVSDLGG LGC GLKNRS SEGP S SP S GKLM SPKL Y VW AKDRPEIWEGEPPCLPPRD SLNQ SL SRDLT VA PGSTLWLSCGVPPDSVSRGPLSWTHVHPKGPKSLLSLELKDDRPARDMWVMGTSLMLP RATAQDAGKWYCHRGNLTMSFHLEITARPSRGGGGSGGGGSGGGGSGGGGSDIVMTQ SHKFMSTSIGARVSITCKASODVRTAVAWYOQKPGOSPKLLIYSASYRYTGVPDRFTGS GSGTDFTFTISSVQAEDLAVYYCQQHYGTPPWTFGGGTKLEIKGGGGSGGGGSGGGGSE VOLVESGGGLVKPGGSLKLSCEASRFTFSSYAMSWVRQTPEKRLEWVAAISGGGRYTY YPDSMKGRFTISRDNAKNFLYLOMSSLRSEDTAMYYCARHYDGYLDYWGOGTTLTVS SHHHHHHAleta #572Nucleotide sequence SEQ ID NO: 13

[0368] ATGGAGTTTGGGCTGAGCTGGGTTTTCCTCGTTGCTCTTTTTAGAGGTG TCCAGTGTGAGGTGCAGCTGGTGGAGTCTGGTGGTGGTCTTGTTCAACCTGGTGGTT C TC TTCGTC TTTC TTGTGCTGC TTC TGGTTTT AATATTAAAGAT AC TT ATATTC ATTGG- 122 - 13321680vlAttorney Docket No. 2012106-0161GTTCGTCAAGCTCCTGGTAAAGGTCTTGAATGGGTTGCTCGTATTTATCCTACTAATG GTTATACTCGTTATGCTGATTCTGTTAAAGGTCGTTTTACTATTTCTGCTGATACTTCT AAAAATACTGCTTATCTTCAAATGAACTCTCTTCGTGCTGAAGATACTGCTGTTTATT ATTGTTCTCGTTGGGGTGGTGATGGTTTTTATGCTATGGATTATTGGGGTCAAGGTAC TCTTGTCACCGTCTCCTCAGCTAGCACCGGGGGAGGTGGGTCTGGAGGTGGAGGATC TGGTGGAGGTGGGTCTGACATCCAGATGACCCAGTCTCCTTCTTCTCTTTCTGCTTCT GTTGGTGATCGTGTTACTATTACTTGTCGTGCTTCTCAAGATGTTAATACTGCTGTTG CTTGGTATCAACAAAAACCTGGTAAAGCTCCTAAACTTCTTATTTATTCTGCTTCTTT TCTTTATTCTGGTGTTCCTTCTCGTTTTTCTGGTTCTCGTTCTGGTACTGATTTTACTCT TACTATTTCTTCTCTTCAACCTGAAGATTTTGCTACTTATTATTGTCAACAACATTAT ACTACTCCTCCTACTTTTGGTCAAGGTACCAAGGTGGAGATCAAACGTACGGGAGGA GGTGGGTCTGGAGGTGGAGGATCTGGTGGAGGTGGGTCTGGAGGAGGTGGATCCCC CGAGGAACCTCTAGTGGTGAAGGTGGAAGAGGGAGATACCGCTGCCCTGTGGTGCC TCAAGGGGACCTCAGATGGCCCCACTCAGCAGCTGACCTGGTCTCGGGAGTCCCCG CTTAAACCCTTCTTAAAATACAGCCTGGGGGTGCCAGGCCTGGGAGTGCACGTGAG GCCCGACGCCATCAGCGTGGTTATCCGGAACGTCTCTCAACAGATGGGGGGCTTCTA CCTGTGCCAGCCGGGGCCCCCCTCTGAGAAGGCCTGGCAGCCTGGCTGGACAGTCA ATGTGGAGGGCAGCGGGGAGCTGTTCCGGTGGAATGTTTCGGACCTAGGTGGCCTG GGCTGTGGCCTGAAGAACAGGTCCTCAGAGGGCCCCAGCTCCCCTTCCGGGAAGCT CATGAGCCCCAAGCTGTATGTGTGGGCCAAAGACCGCCCTGAGATCTGGGAGGGAG AGCCTCCGTGTCTCCCACCGAGGGACAGCCTGAACCAGAGCCTCAGCCGGGACCTC ACCGTTGCCCCTGGCTCCACACTCTGGCTGTCCTGTGGGGTACCCCCTGACTCTGTGT CCAGGGGCCCCCTCTCCTGGACCCATGTGCACCCCAAGGGGCCTAAGTCATTGCTGA GCCTAGAGCTGAAGGACGATCGCCCGGCCAGAGATATGTGGGTAATGGGCACGAGC CTGATGTTGCCCCGGGCCACAGCTCAAGACGCTGGAAAGTGGTATTGTCACCGTGGC AACCTGACCATGTCATTCCACCTGGAGATCACTGCTCGGCCATCTAGAGGAGGAGGT GGGTCTGGAGGTGGAGGATCTGGTGGAGGTGGGTCTGGAGGAGGTGGGTCTGAGGT GCAGCTGGTCGAGAGCGGCGGCGGACTGGTGCAACCTGGCGGCAGCCTGAGACTGA GCTGCGCCGCATCTGGCTTCACATTTAGCAGCTTCGGCATGCACTGGGTTAGACAGG CCCCTGGAAAAGGCCTGGAATGGGTGGCCTACATCAGCTCTGACAGCAGCGCTATC- 123 - 13321680vlAttorney Docket No. 2012106-0161TACTACGCCGACACAGTGAAGGGCAGATTCACCATCTCTAGAGATAACGCCAAGAA TAGCCTGTACCTGCAGATGAACAGCCTCAGGGACGAGGACACCGCCGTGTACTACT GTGGCCGGGGCAGAGAGAACATCTATTACGGCTCCCGCCTGGACTACTGGGGACAG GGCACAACCGTGACCGTGTCCTCCGGCGGAGGAGGCAGCGGCGGCGGCGGCTCTGG CGGCGGCGGCAGCGATATCCAGCTGACACAGAGCCCTTCTTTTCTGAGCGCCAGCGT GGGGGATAGAGTGACCATTACATGCAAGGCTTCACAGAATGTGGACACCAACGTGG CTTGGTACCAGCAGAAACCAGGCAAGGCCCCTAAGGCCCTGATCTACAGCGCCAGC TACCGGTACTCTGGCGTCCCCAGCCGGTTCAGCGGATCCGGCTCCGGCACCGATTTT ACCCTGACCATCAGCAGCCTGCAGCCTGAGGACTTCGCCACATACTATTGCCAGCAA TACAACAACTACCCCTTCACCTTCGGCCAGGGAACCAAGCTGGAAATCAAGCATCA TCACCATCACCATAmino acid sequence SEQ ID NO: 14

[0369] MEFGLSWVFLVALFRGY. QCEVQLVESGGGLVQPGGSLRLSCAASGFNIK DTYIHWVRQAPGKGLEWVARIYPTNGYTRYADSVKGRFTISADTSKNTAYLQMNSLRA EDTAVYYCSRWGGDGFYAMDYWGQGTLVTVSSASTGGGGSGGGGSGGGGSDIQMTQ SPSSLSASVGDRVTITCRASQDVNTAVAWYQQKPGKAPKLLIYSASFLYSGVPSRFSGSR SGTDFTLTISSLQPEDFATYYCQQHYTTPPTFGQGTKVEIKRTGGGGSGGGGSGGGGSG GGGSPEEPLVVKVEEGDTAALWCLKGTSDGPTQQLTWSRESPLKPFLKYSLGVPGLGVHVRPDAISVVIRNVSQQMGGFYLCQPGPPSEKAWQPGWTVNVEGSGELFRWNVSDLGG LGCGLKNRSSEGPSSPSGKLMSPKLYVWAKDRPEIWEGEPPCLPPRDSLNQSLSRDLTVA PGSTLWLSCGVPPDSVSRGPLSWTHVHPKGPKSLLSLELKDDRPARDMWVMGTSLMLP RATAQDAGKWYCHRGNLTMSFHLEITARPSRGGGGSGGGGSGGGGSGGGGSEVQLVE SGGGLVQPGGSLRLSCAASGFTFSSFGMHWVROAPGKGLEWVAYISSDSSAIYYADTV KGRFTISRDNAKNSLYLQMNSLRDEDTAVYYCGRGRENIYYGSRLDYWGOGTTVTVSS GGGGSGGGGSGGGGSDIQLTOSPSFLSASVGDRVTITCKASONVDTNVAWYOQKPGKA PKALIYSASYRYSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCOOYNNYPFTFGOGTKL EIKHHHHHH- 124 - 13321680vlAttorney Docket No. 2012106-0161Aleta #573Nucleotide sequence SEQ ID NO: 15

[0370] ATGGAGTTTGGGCTGAGCTGGGTTTTCCTCGTTGCTCTTTTTAGAGGTG TCCAGTGTGAGGTGCAGCTGGTGGAGTCTGGTGGTGGTCTTGTTCAACCTGGTGGTT CTCTTCGTCTTTCTTGTGCTGCTTCTGGTTTTAATATTAAAGATACTTATATTCATTGG GTTCGTCAAGCTCCTGGTAAAGGTCTTGAATGGGTTGCTCGTATTTATCCTACTAATG GTTATACTCGTTATGCTGATTCTGTTAAAGGTCGTTTTACTATTTCTGCTGATACTTCT AAAAATACTGCTTATCTTCAAATGAACTCTCTTCGTGCTGAAGATACTGCTGTTTATT ATTGTTCTCGTTGGGGTGGTGATGGTTTTTATGCTATGGATTATTGGGGTCAAGGTAC TCTTGTCACCGTCTCCTCAGCTAGCACCGGGGGAGGTGGGTCTGGAGGTGGAGGATC TGGTGGAGGTGGGTCTGACATCCAGATGACCCAGTCTCCTTCTTCTCTTTCTGCTTCT GTTGGTGATCGTGTTACTATTACTTGTCGTGCTTCTCAAGATGTTAATACTGCTGTTG CTTGGTATCAACAAAAACCTGGTAAAGCTCCTAAACTTCTTATTTATTCTGCTTCTTT TCTTTATTCTGGTGTTCCTTCTCGTTTTTCTGGTTCTCGTTCTGGTACTGATTTTACTCT TACTATTTCTTCTCTTCAACCTGAAGATTTTGCTACTTATTATTGTCAACAACATTAT ACTACTCCTCCTACTTTTGGTCAAGGTACCAAGGTGGAGATCAAACGTACGGGAGGA GGTGGGTCTGGAGGTGGAGGATCTGGTGGAGGTGGGTCTGGAGGAGGTGGATCCCC CGAGGAACCTCTAGTGGTGAAGGTGGAAGAGGGAGATACCGCTGCCCTGTGGTGCC TCAAGGGGACCTCAGATGGCCCCACTCAGCAGCTGACCTGGTCTCGGGAGTCCCCG CTTAAACCCTTCTTAAAATACAGCCTGGGGGTGCCAGGCCTGGGAGTGCACGTGAG GCCCGACGCCATCAGCGTGGTTATCCGGAACGTCTCTCAACAGATGGGGGGCTTCTA CCTGTGCCAGCCGGGGCCCCCCTCTGAGAAGGCCTGGCAGCCTGGCTGGACAGTCA ATGTGGAGGGCAGCGGGGAGCTGTTCCGGTGGAATGTTTCGGACCTAGGTGGCCTG GGCTGTGGCCTGAAGAACAGGTCCTCAGAGGGCCCCAGCTCCCCTTCCGGGAAGCT CATGAGCCCCAAGCTGTATGTGTGGGCCAAAGACCGCCCTGAGATCTGGGAGGGAG AGCCTCCGTGTCTCCCACCGAGGGACAGCCTGAACCAGAGCCTCAGCCGGGACCTC ACCGTTGCCCCTGGCTCCACACTCTGGCTGTCCTGTGGGGTACCCCCTGACTCTGTGT CCAGGGGCCCCCTCTCCTGGACCCATGTGCACCCCAAGGGGCCTAAGTCATTGCTGA GCCTAGAGCTGAAGGACGATCGCCCGGCCAGAGATATGTGGGTAATGGGCACGAGC- 125 - 13321680vlAttorney Docket No. 2012106-0161CTGATGTTGCCCCGGGCCACAGCTCAAGACGCTGGAAAGTGGTATTGTCACCGTGGC AACCTGACCATGTCATTCCACCTGGAGATCACTGCTCGGCCATCTAGAGGAGGAGGT GGGTCTGGAGGTGGAGGATCTGGTGGAGGTGGGTCTGGAGGAGGTGGGTCTCAGGT GCAGCTGGTGCAGTCTGGCGCCGAGGTGAAAAAGCCCGGCAGCTCTGTCAAGGTGT CCTGCAAGGCCAGCGGAGGTACATTTAGCAGCTACGCCATCAGCTGGGTGCGGCAG GCCCCAGGCCAGGGCCTGGAGTGGATGGGCGGAATCATCCCTATCCTGGGCACAGC CAACTACGCCCAGAAATTCCAGGGCAGAGTGACAATCACCGCCGACGAGAGCACCA GCACAGCCTACATGGAACTGTCCAGCCTGAGAAGCGAGGATACCGCTGTGTATTACT GCGCCAGGTGGGGCGGCGGAGCCTTCGACATCTGGGGCCAAGGCACAATGGTGACC GTGTCTTCTGGCGGCGGAGGGAGCGGCGGCGGAGGCAGCGGCGGCGGAGGAAGCG AGATCGTGCTGACCCAGAGCCCCGGCACACTGTCTCTGAGCCCTGGCGAGCGGGCC ACCCTGTCCTGCAGAGCTAGCCAGAGCGTGGGCGGCTACCTGGCCTGGTACCAGCA GAAGCCTGGACAAGCTCCTAGACTGCTGATCTACGACGCCTCTAATAGAGCAACCG GCATCCCCGCTAGATTCAGCGGCTCTGGCTCCGGAACCGATTTTACCCTCACCATCA GCAGCCTGGAACCTGAGGACTTCGCCGTCTACTACTGCCAGCAGCGGAACAACTGG CCTCCAATGTACACCTTCGGCCAGGGCACCAAGCTGGAAATCAAGCATCATCACCAT CACCATAmino acid sequence SEQ ID NO: 16

[0371] MEFGLSWVFLVALFRGY. QCEVQLVESGGGLVQPGGSLRLSCAASGFNIK DTYIHWVRQAPGKGLEWVARIYPTNGYTRYADSVKGRFTISADTSKNTAYLQMNSLRA EDTAVYYC SRWGGDGFYAMDYWGQGTLVTVS S ASTGGGGSGGGGSGGGGSDIQMTQ SPSSLSASVGDRVTITCRASQDVNTAVAWYQQKPGKAPKLLIYSASFLYSGVPSRFSGSR SGTDFTLTISSLQPEDFATYYCQQHYTTPPTFGQGTKVEIKRTGGGGSGGGGSGGGGSG GGGSPEEPLVVKVEEGDTAALWCLKGTSDGPTQQLTWSRESPLKPFLKYSLGVPGLGVHVRPDAISVVIRNVSQQMGGFYLCQPGPPSEKAWQPGWTVNVEGSGELFRWNVSDLGG LGC GLKNRS SEGP S SP S GKLM SPKL YVW AKDRPEIWEGEPPCLPPRD SLNQ SL SRDLT V A PGSTLWLSCGVPPDSVSRGPLSWTHVHPKGPKSLLSLELKDDRPARDMWVMGTSLMLP RATAQDAGKWYCHRGNLTMSFHLEITARPSRGGGGSGGGGSGGGGSGGGGSQVQLVQ - 126 - 13321680vlAttorney Docket No. 2012106-0161SGAEVKKPGSSVKVSCKASGGTFSSYNAISWVRQAPGQGLEWMGGIIPILGTANYAQKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCARWGGGAFDIWGQGTMVTVSSGGGGSGGGGSGGGGSEIVLTQSPGTLSLSPGERATLSCRASQSVGGYLAWYQQKPGQAPRLLIYDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRNNWPPMYTFGQGTKLEIKHHHHHHAleta #574Nucleotide sequence SEQ ID NO: 17

[0372] ATGGAGTTTGGGCTGAGCTGGGTTTTCCTCGTTGCTCTTTTTAGAGGTG TCCAGTGTCAGGTGCAGCTGCAGGAGTCTGGGGGAGGCTTGGTGCAGGCTGGGGGC TCTCTGAAACTCTCCTGTGGAGTCTCTGGACGCACCCTCAGTGACTATGCCATGGGC TGGTTCCGCCAGCCTCCAGGGAAGGAGCGTGAGTTTGTAGCAACTATTAGGTGGAGT GGTGGTTACACATACTCTGCAGACTCCGTGAAGGGCCGATTCACCATCTCCAGAGAC GCCGCCAAGAACACGGTGTATCTGCAAATGAACAGCCTGAAACCTGAGGACACGGC CGTTTATCACTGTGCCGCCTGCAGACTATACAATGGTAGACGGTTAGGCGACCCGAG TGAGTATGACTACTGGGGCCAGGGGACCCTGGTCACCGTCTCCTCAGGAGGAGGTG GGTCTGGAGGTGGAGGATCTGGTGGAGGTGGGTCTGGAGGAGGTGGATCCCCCGAG GAACCTCTAGTGGTGAAGGTGGAAGAGGGAGATACCGCTGCCCTGTGGTGCCTCAA GGGGACCTCAGATGGCCCCACTCAGCAGCTGACCTGGTCTCGGGAGTCCCCGCTTAA ACCCTTCTTAAAATACAGCCTGGGGGTGCCAGGCCTGGGAGTGCACGTGAGGCCCG ACGCCATCAGCGTGGTTATCCGGAACGTCTCTCAACAGATGGGGGGCTTCTACCTGT GCCAGCCGGGGCCCCCCTCTGAGAAGGCCTGGCAGCCTGGCTGGACAGTCAATGTG GAGGGCAGCGGGGAGCTGTTCCGGTGGAATGTTTCGGACCTAGGTGGCCTGGGCTG TGGCCTGAAGAACAGGTCCTCAGAGGGCCCCAGCTCCCCTTCCGGGAAGCTCATGA GCCCCAAGCTGTATGTGTGGGCCAAAGACCGCCCTGAGATCTGGGAGGGAGAGCCT CCGTGTCTCCCACCGAGGGACAGCCTGAACCAGAGCCTCAGCCGGGACCTCACCGT TGCCCCTGGCTCCACACTCTGGCTGTCCTGTGGGGTACCCCCTGACTCTGTGTCCAG GGGCCCCCTCTCCTGGACCCATGTGCACCCCAAGGGGCCTAAGTCATTGCTGAGCCT AGAGCTGAAGGACGATCGCCCGGCCAGAGATATGTGGGTAATGGGCACGAGCCTGA- 127 - 13321680vlAttorney Docket No. 2012106-0161TGTTGCCCCGGGCCACAGCTCAAGACGCTGGAAAGTGGTATTGTCACCGTGGCAAC CTGACCATGTCATTCCACCTGGAGATCACTGCTCGGCCATCTAGACATCATCACCAT CACCATAmino acid sequence SEQ ID NO: 18

[0373] VIHI Gl. SW^ D YAMGWFRQPPGKEREFVA TIR WSGGYTYSADSVKGRFTISRDAAKNTVYLQMNSLKP EDTAVYHCAACRLYNGRRLGDPSEYDYWGQGTLVTVSSGGGGSGGGGSGGGGSGGGG SPEEPLVVKVEEGDTAALWCLKGTSDGPTQQLTWSRESPLKPFLKYSLGVPGLGVHVRPDAISVVIRNVSQQMGGFYLCQPGPPSEKAWQPGWTVNVEGSGELFRWNVSDLGGLGCGLKNRSSEGPSSPSGKLMSPKLYVWAKDRPEIWEGEPPCLPPRDSLNQSLSRDLTVAPGSTLWLSCGVPPDSVSRGPLSWTHVHPKGPKSLLSLELKDDRPARDMWVMGTSLMLPRATAQDAGKWYCHRGNLTMSFHLEITARPSRHHHHHHAleta #575Nucleotide sequence SEQ ID NO: 19

[0374] ATGGAGTTTGGGCTGAGCTGGGTTTTCCTCGTTGCTCTTTTTAGAGGTG TCCAGTGTCAGGTGCAGTTGCAGGCGTTTGGGGGAGGCTTGGTGCAGGCTGGGGGG TCTCTGAGACTCTCCTGTGCAGCCCCTGGGATTATCTTCAGTACCATTGCCATGGGCT GGTACCGCCAGGCTCCAGGGAAGGGGCGCGAGTTGGAGCGCGAGTTGATCGCAGCC ATTGCTACTGATGGTAGCACAGACTATGCAGACTCCGTGAAGGGCCGATTCACCATC TACAGAGACAACGCCAAGAACACGGCATATCTGCAAATGAACAGCCTGAAACCTGA GGACACAGGCGTCTATTACTGTAATGCACGTCGGGGCGCTTCCGTACTATGGGGACA GGGGACCCTGGTCACCGTTTCCTCAGGAGGAGGTGGGTCTGGAGGTGGAGGATCTG GTGGAGGTGGGTCTGGAGGAGGTGGATCCCCCGAGGAACCTCTAGTGGTGAAGGTG GAAGAGGGAGATACCGCTGCCCTGTGGTGCCTCAAGGGGACCTCAGATGGCCCCAC TCAGCAGCTGACCTGGTCTCGGGAGTCCCCGCTTAAACCCTTCTTAAAATACAGCCT GGGGGTGCCAGGCCTGGGAGTGCACGTGAGGCCCGACGCCATCAGCGTGGTTATCC- 128 - 13321680vlAttorney Docket No. 2012106-0161GGAACGTCTCTCAACAGATGGGGGGCTTCTACCTGTGCCAGCCGGGGCCCCCCTCTG AGAAGGCCTGGCAGCCTGGCTGGACAGTCAATGTGGAGGGCAGCGGGGAGCTGTTC CGGTGGAATGTTTCGGACCTAGGTGGCCTGGGCTGTGGCCTGAAGAACAGGTCCTCA GAGGGCCCCAGCTCCCCTTCCGGGAAGCTCATGAGCCCCAAGCTGTATGTGTGGGCC AAAGACCGCCCTGAGATCTGGGAGGGAGAGCCTCCGTGTCTCCCACCGAGGGACAG CCTGAACCAGAGCCTCAGCCGGGACCTCACCGTTGCCCCTGGCTCCACACTCTGGCT GTCCTGTGGGGTACCCCCTGACTCTGTGTCCAGGGGCCCCCTCTCCTGGACCCATGT GCACCCCAAGGGGCCTAAGTCATTGCTGAGCCTAGAGCTGAAGGACGATCGCCCGG CCAGAGATATGTGGGTAATGGGCACGAGCCTGATGTTGCCCCGGGCCACAGCTCAA GACGCTGGAAAGTGGTATTGTCACCGTGGCAACCTGACCATGTCATTCCACCTGGAG ATCACTGCTCGGCCATCTAGACATCATCACCATCACCATAmino acid sequence SEQ ID NO: 20

[0375] XnJ GLSWV IAMGWYRQAPGKGRELERELIAAIATDGSTDYADSVKGRFTIYRDNAKNTAYLQMNSLK PEDTGVYYCNARRGASVLWGQGTLVTVSSGGGGSGGGG GGGGSGGGGSP^PINN^ EEGDTAALWCLKGTSDGPTQQLTWSRESPLKPFLKYSLGVPGLGVHVRPDAISVVIRNVSQQMGGFYLCQPGPPSEKAWQPGWTVNVEGSGELFRWNVSDLGGLGCGLKNRSSEGPSSPSGKLMSPKLYVWAKDRPEIWEGEPPCLPPRDSLNQSLSRDLTVAPGSTLWLSCGVPPDSVSRGPLSWTHVHPKGPKSLLSLELKDDRPARDMWVMGTSLMLPRATAQDAGKWYCHRGNLTMSFHLEITARPSRHHHHHHAleta #576Nucleotide sequence SEQ ID NO: 21

[0376] ATGGAGTTTGGGCTGAGCTGGGTTTTCCTCGTTGCTCTTTTTAGAGGTG TCCAGTGTCAGGTGCAGCTGCAGGAGTCTGGGGGAGGCTTGGTGCAGCCTGGGGGG TCTCTGAGACTCTCCTGTGCAGCCTCTGGAAGTATCTTCAGTATCAGTTCCTTGGGGT GGTTCCGCCAGGCTCCAGGGAAGCAGCGCGAGTTGGTCGCAGACATTACTAGTGGT- 129 - 13321680vlAttorney Docket No. 2012106-0161GGTAGCACAAACTATGCAGACTCCGTGAAGGGCCGATTCACCATCTCCAGAGACAA CGCCAAGAGCACGGTGTATCTGCAAATGAACAGCCTGAAACCTGACGACACGGCCG TCTATTATTGTAATGCCCGAAGGGGCGATACGATCCTGTGGGCCGAGGGGACCCAG GTCACCGTCTCCTCAGGAGGAGGTGGGTCTGGAGGTGGAGGATCTGGTGGAGGTGG GTCTGGAGGAGGTGGATCCCCCGAGGAACCTCTAGTGGTGAAGGTGGAAGAGGGAG ATACCGCTGCCCTGTGGTGCCTCAAGGGGACCTCAGATGGCCCCACTCAGCAGCTGA CCTGGTCTCGGGAGTCCCCGCTTAAACCCTTCTTAAAATACAGCCTGGGGGTGCCAG GCCTGGGAGTGCACGTGAGGCCCGACGCCATCAGCGTGGTTATCCGGAACGTCTCTC AACAGATGGGGGGCTTCTACCTGTGCCAGCCGGGGCCCCCCTCTGAGAAGGCCTGG CAGCCTGGCTGGACAGTCAATGTGGAGGGCAGCGGGGAGCTGTTCCGGTGGAATGT TTCGGACCTAGGTGGCCTGGGCTGTGGCCTGAAGAACAGGTCCTCAGAGGGCCCCA GCTCCCCTTCCGGGAAGCTCATGAGCCCCAAGCTGTATGTGTGGGCCAAAGACCGCC CTGAGATCTGGGAGGGAGAGCCTCCGTGTCTCCCACCGAGGGACAGCCTGAACCAG AGCCTCAGCCGGGACCTCACCGTTGCCCCTGGCTCCACACTCTGGCTGTCCTGTGGG GTACCCCCTGACTCTGTGTCCAGGGGCCCCCTCTCCTGGACCCATGTGCACCCCAAG GGGCCTAAGTCATTGCTGAGCCTAGAGCTGAAGGACGATCGCCCGGCCAGAGATAT GTGGGTAATGGGCACGAGCCTGATGTTGCCCCGGGCCACAGCTCAAGACGCTGGAA AGTGGTATTGTCACCGTGGCAACCTGACCATGTCATTCCACCTGGAGATCACTGCTC GGCCATCTAGACATCATCACCATCACCATAmino acid sequence SEQ ID NO: 22

[0377] MEFGLSWVTLVALFMSSLGWFRQAPGKQRELVADITSGGSTNYADSVKGRFTISRDNAKSTVYLQMNSLKPDDT A VYYCNARRGDTIL WAEGTOVTVSSGGGGSGGGGSGGGGSGGGGSPEEPLNVKVEEGD TAALWCLKGTSDGPTOQLTWSRESPLKPFLKYSLGVPGLGVHVRPDAISVVIRNVSOOM GGFYLCQPGPPSEKAWQPGWTVNVEGSGELFRWNVSDLGGLGCGLKNRSSEGPSSPSG KLMSPKLYVWAKDRPEIWEGEPPCLPPRDSLNOSLSRDLTVAPGSTLWLSCGVPPDSVS RGPLSWTHVHPKGPKSLLSLELKDDRPARDMWVMGTSLMLPRATAQDAGKWYCHRG NLTMSFHLE1TARPSRHHHHHH- 130 - 13321680vlAttorney Docket No. 2012106-0161Aleta #577Nucleotide sequence SEQ ID NO: 23

[0378] ATGGAGTTTGGGCTGAGCTGGGTTTTCCTCGTTGCTCTTTTTAGAGGTG TCCAGTGTGAGGTGCAGCTGGTGGAGTCTGGTGGTGGTCTTGTTCAACCTGGTGGTT CTCTTCGTCTTTCTTGTGCTGCTTCTGGTTTTAATATTAAAGATACTTATATTCATTGG GTTCGTCAAGCTCCTGGTAAAGGTCTTGAATGGGTTGCTCGTATTTATCCTACTAATG GTTATACTCGTTATGCTGATTCTGTTAAAGGTCGTTTTACTATTTCTGCTGATACTTCT AAAAATACTGCTTATCTTCAAATGAACTCTCTTCGTGCTGAAGATACTGCTGTTTATT ATTGTTCTCGTTGGGGTGGTGATGGTTTTTATGCTATGGATTATTGGGGTCAAGGTAC TCTTGTCACCGTCTCCTCAGCTAGCACCGGGGGAGGTGGGTCTGGAGGTGGAGGATC TGGTGGAGGTGGGTCTGACATCCAGATGACCCAGTCTCCTTCTTCTCTTTCTGCTTCT GTTGGTGATCGTGTTACTATTACTTGTCGTGCTTCTCAAGATGTTAATACTGCTGTTG CTTGGTATCAACAAAAACCTGGTAAAGCTCCTAAACTTCTTATTTATTCTGCTTCTTT TCTTTATTCTGGTGTTCCTTCTCGTTTTTCTGGTTCTCGTTCTGGTACTGATTTTACTCT TACTATTTCTTCTCTTCAACCTGAAGATTTTGCTACTTATTATTGTCAACAACATTAT ACTACTCCTCCTACTTTTGGTCAAGGTACCAAGGTGGAGATCAAACGTACGGGAGGA GGTGGGTCTGGAGGTGGAGGATCTGGTGGAGGTGGGTCTGGAGGAGGTGGATCCCC CGAGGAACCTCTAGTGGTGAAGGTGGAAGAGGGAGATACCGCTGCCCTGTGGTGCC TCAAGGGGACCTCAGATGGCCCCACTCAGCAGCTGACCTGGTCTCGGGAGTCCCCG CTTAAACCCTTCTTAAAATACAGCCTGGGGGTGCCAGGCCTGGGAGTGCACGTGAG GCCCGACGCCATCAGCGTGGTTATCCGGAACGTCTCTCAACAGATGGGGGGCTTCTA CCTGTGCCAGCCGGGGCCCCCCTCTGAGAAGGCCTGGCAGCCTGGCTGGACAGTCA ATGTGGAGGGCAGCGGGGAGCTGTTCCGGTGGAATGTTTCGGACCTAGGTGGCCTG GGCTGTGGCCTGAAGAACAGGTCCTCAGAGGGCCCCAGCTCCCCTTCCGGGAAGCT CATGAGCCCCAAGCTGTATGTGTGGGCCAAAGACCGCCCTGAGATCTGGGAGGGAG AGCCTCCGTGTCTCCCACCGAGGGACAGCCTGAACCAGAGCCTCAGCCGGGACCTC ACCGTTGCCCCTGGCTCCACACTCTGGCTGTCCTGTGGGGTACCCCCTGACTCTGTGT CCAGGGGCCCCCTCTCCTGGACCCATGTGCACCCCAAGGGGCCTAAGTCATTGCTGA- 131 - 13321680vlAttorney Docket No. 2012106-0161GCCTAGAGCTGAAGGACGATCGCCCGGCCAGAGATATGTGGGTAATGGGCACGAGC CTGATGTTGCCCCGGGCCACAGCTCAAGACGCTGGAAAGTGGTATTGTCACCGTGGC AACCTGACCATGTCATTCCACCTGGAGATCACTGCTCGGCCATCTAGAGGAGGAGGT GGGTCTGGAGGTGGAGGATCTGGTGGAGGTGGGTCTGGAGGAGGTGGGTCTCAGGT GCAGTTGCAGCAGTCTGGGGGAGGATTGGTGCAGGCTGGGGGCTCTCTGAGACTCT CCTGTGCAGCCTCTGGACGCACCTTCACCCGCCTTCCCATGGGCTGGTTCCGCCAGG CTCCAGGAAAGGAGCGAGAGTTTGTAGCAGCTCTTAGCTGGAGTGGTGGCAACACA GACTATGCAAATACTGCAGACTCCGTGAAGGGCCGATTCACCATGTCCAGAGACAG CGCCAAGAACACGGTGTATCTGTGGATGAACAACCTGAAACCTGAGGACACGGCCG TTTATTACTGTGCAGCCCGACAGAGTGGTTGGGGTACCGCCTTCACTGATGACTATG ACTACTGGGGCCAGGGGACCCAGGTCACCGTCTCCTCACATCATCACCATCACCATAmino acid sequence SEQ ID NO: 24

[0379] MEFGLSWVFLVALFRGY. QCEVQLVESGGGLVQPGGSLRLSCAASGFNIK DTYIHWVRQAPGKGLEWVARIYPTNGYTRYADSVKGRFTISADTSKNTAYLQMNSLRA EDTAVYYCSRWGGDGFYAMDYWGQGTLVTVSSASTGGGGSGGGGSGGGGSDIQMTQ SPSSLSASVGDRVTITCRASQDVNTAVAWYQQKPGKAPKLLIYSASFLYSGVPSRFSGSR SGTDFTLTISSLQPEDFATYYCQQHYTTPPTFGQGTKVEIKRTGGGGSGGGGSGGGGSG GGGSPEEPLVVKVEEGDTAALWCLKGTSDGPTQQLTWSRESPLKPFLKYSLGVPGLGVHVRPDAISVVIRNVSQQMGGFYLCQPGPPSEKAWQPGWTVNVEGSGELFRWNVSDLGG LGCGLKNRSSEGPSSPSGKLMSPKLYVWAKDRPEIWEGEPPCLPPRDSLNQSLSRDLTVA PGSTLWLSCGVPPDSVSRGPLSWTHVHPKGPKSLLSLELKDDRPARDMWVMGTSLMLP RATAQDAGKWYCHRGNLTMSFHLEITARPSRGGGGSGGGGSGGGGSGGGGSOVOLQO SGGGLVQAGGSLRLSCAASGRTFTRLPMGWFRQAPGKEREFVAALSWSGGNTDYANT ADSVKGRFTMSRDSAKNTVYLWMNNLKPEDTAVYYCAARQSGWGTAFTDDYDYWG QGTQ VT VS SHHHHHHAleta #578- 132 - 13321680vlAttorney Docket No. 2012106-0161Nucleotide sequence SEQ ID NO: 25

[0380] ATGGAGTTTGGGCTGAGCTGGGTTTTCCTCGTTGCTCTTTTTAGAGGTG TCCAGTGTCAGGTGCAGTTGCAGGAGTCTGGGGGAGGATTGGTGCAGCCTGGGGGG TCTCTGAGACTCTCCTGTGCAGCCTCTGGAACCATCTTCCCTATCAGCACCATGGGCT GGTTCCGCCAGGCTCCAGGGAAGCAGCGCGAGTTGGTCGCAACTAGTACTGAGGAT GGTAGTACAACCTACGCAGACTCCGTGAAGGGCCGATTCACCGCCTCCAGAGACAA CGCCAAGAATACGGTGTTTCTGCAAATGAACAGCCTGAAACCTGAGGACACGGCCG TCTATTTGTGTTATGCAGAACACAGACGGCGGACCACCGACTTCTATCGGACCTATT GGGGCCAGGGGACCCAGGTCACCGTTTCCTCAGGAGGAGGTGGGTCTGGAGGTGGA GGATCTGGTGGAGGTGGGTCTGGAGGAGGTGGATCCCCCGAGGAACCTCTAGTGGT GAAGGTGGAAGAGGGAGATACCGCTGCCCTGTGGTGCCTCAAGGGGACCTCAGATG GCCCCACTCAGCAGCTGACCTGGTCTCGGGAGTCCCCGCTTAAACCCTTCTTAAAAT ACAGCCTGGGGGTGCCAGGCCTGGGAGTGCACGTGAGGCCCGACGCCATCAGCGTG GTTATCCGGAACGTCTCTCAACAGATGGGGGGCTTCTACCTGTGCCAGCCGGGGCCC CCCTCTGAGAAGGCCTGGCAGCCTGGCTGGACAGTCAATGTGGAGGGCAGCGGGGA GCTGTTCCGGTGGAATGTTTCGGACCTAGGTGGCCTGGGCTGTGGCCTGAAGAACAG GTCCTCAGAGGGCCCCAGCTCCCCTTCCGGGAAGCTCATGAGCCCCAAGCTGTATGT GTGGGCCAAAGACCGCCCTGAGATCTGGGAGGGAGAGCCTCCGTGTCTCCCACCGA GGGACAGCCTGAACCAGAGCCTCAGCCGGGACCTCACCGTTGCCCCTGGCTCCACA CTCTGGCTGTCCTGTGGGGTACCCCCTGACTCTGTGTCCAGGGGCCCCCTCTCCTGG ACCCATGTGCACCCCAAGGGGCCTAAGTCATTGCTGAGCCTAGAGCTGAAGGACGA TCGCCCGGCCAGAGATATGTGGGTAATGGGCACGAGCCTGATGTTGCCCCGGGCCA CAGCTCAAGACGCTGGAAAGTGGTATTGTCACCGTGGCAACCTGACCATGTCATTCC ACCTGGAGATCACTGCTCGGCCATCTAGACATCATCACCATCACCATAmino acid sequence SEQ ID NO: 26

[0381] MEFGLS^yTLNALFRGyQCQVQLQESGGGLVQPGGSLRLSCAASGTIFPI STMGWFRQAPGKQREL VA TSTEDGSTTYADSVKGRFTASRDNAKNTVFLQMNSLKPED TAVYLCYAEHRRRTTDFYRTYWGQGTQVTVSSGGGG GGGG GGGG GGGGSPEEYAN- 133 - 13321680vlAttorney Docket No. 2012106-0161VKVEEGDTAALWCLKGTSDGPTOQLTWSRESPLKPFLKYSLGVPGLGVHVRPDAISVVI RNVSOOMGGFYLCOPGPPSEKAWOPGWTVNVEGSGELFRWNVSDLGGLGCGLKNRSS EGPSSPSGKLMSPKLYVWAKDRPEIWEGEPPCLPPRDSLNQSLSRDLTVAPGSTLWLSCG VPPD S VSRGPLS WTHVHPKGPKSLL SLELKDDRP ARDMWVMGT SLMLPRATAQD AGK WYCHRGNI. TMSFHI. F. ITARPSRHHHHHHAleta #579Nucleotide sequence SEQ ID NO: 27

[0382] ATGGAGTTTGGGCTGAGCTGGGTTTTCCTCGTTGCTCTTTTTAGAGGTG TCCAGTGTCAGGTGCAGCTGCAGCAGTCTGGGGGAGGATTGGTGCAGGCTGGGGGC TCTCTGAGACTCTCCTGTACAGCCTCTGGACGCACCAAGACTACCACTTACATGGGC TGGTTCCGCCAGGTTCCAGGGAAGGAGCGTGAGTTTGTAGCCACTATTGGCTGGACT GGTAGAAGCGCATACTATGCAGACTCCGTAGAGGGCCGATTCACCATCTCCAGGGA CAACGCCAAGAACACGGTGTATCTACAAATGAACAACCTGAGTCCTGACGACACGG CCGTTTATTACTGTGCAACGGACCCGTCGAGTAGTAGTTGGCACCATCTCCGCAACC CCGATCACTATGTCTACTGGGGCCAGGGGACCCAGGTCACCGTTTCCTCAGGAGGA GGTGGGTCTGGAGGTGGAGGATCTGGTGGAGGTGGGTCTGGAGGAGGTGGATCCCC CGAGGAACCTCTAGTGGTGAAGGTGGAAGAGGGAGATACCGCTGCCCTGTGGTGCC TCAAGGGGACCTCAGATGGCCCCACTCAGCAGCTGACCTGGTCTCGGGAGTCCCCG CTTAAACCCTTCTTAAAATACAGCCTGGGGGTGCCAGGCCTGGGAGTGCACGTGAG GCCCGACGCCATCAGCGTGGTTATCCGGAACGTCTCTCAACAGATGGGGGGCTTCTA CCTGTGCCAGCCGGGGCCCCCCTCTGAGAAGGCCTGGCAGCCTGGCTGGACAGTCA ATGTGGAGGGCAGCGGGGAGCTGTTCCGGTGGAATGTTTCGGACCTAGGTGGCCTG GGCTGTGGCCTGAAGAACAGGTCCTCAGAGGGCCCCAGCTCCCCTTCCGGGAAGCT CATGAGCCCCAAGCTGTATGTGTGGGCCAAAGACCGCCCTGAGATCTGGGAGGGAG AGCCTCCGTGTCTCCCACCGAGGGACAGCCTGAACCAGAGCCTCAGCCGGGACCTC ACCGTTGCCCCTGGCTCCACACTCTGGCTGTCCTGTGGGGTACCCCCTGACTCTGTGT CCAGGGGCCCCCTCTCCTGGACCCATGTGCACCCCAAGGGGCCTAAGTCATTGCTGA GCCTAGAGCTGAAGGACGATCGCCCGGCCAGAGATATGTGGGTAATGGGCACGAGC- 134 - 13321680vlAttorney Docket No. 2012106-0161CTGATGTTGCCCCGGGCCACAGCTCAAGACGCTGGAAAGTGGTATTGTCACCGTGGC AACCTGACCATGTCATTCCACCTGGAGATCACTGCTCGGCCATCTAGACATCATCAC CATCACCATAmino acid sequence SEQ ID NO: 28

[0383] MEEGLS yYLNALERGyQCQVQLQQSGGGLVQAGGSLRLSCTASGRTKT TTYMGWFRQVPGKEREFVATIGWTGRSAYYADSVEGRFTISRDNAKNTVYLQMNNLSP DDTA VYYCA TDPSSSSWHHLRNPDHYVYWGQGTQVTVSSGGGGSGGGGSGGGGSGGG GSPEEPLVVKVEEGDTAALWCLKGTSDGPTOOLTWSRESPLKPFLKYSLGVPGLGVHV RPDA1SVV1RNVSOOMGGFYLCOPGPPSEKAWOPGWTVNVEGSGELFRWNVSDLGGLG CGLKNRS SEGP S SP SGKLMSPKLYVWAKDRPEIWEGEPPCLPPRD SLNQ SLSRDLT VAPG STLWLSCGVPPDSVSRGPLSWTHVHPKGPKSLLSLELKDDRPARDMWVMGTSLMLPRA TAQDAGKWYCFiRGNLTMSFFiLEITARPSRHHHHHHAleta #580Nucleotide sequence SEQ ID NO: 29

[0384] ATGGAGTTTGGGCTGAGCTGGGTTTTCCTCGTTGCTCTTTTTAGAGGTG TCCAGTGTCAGGTGCAGCTGCAGCAGTTTGGGGGAGGATTGGTGCAGGCTGACGGC TCTCTGAGACTCTCCTGTGCAGCCTCTGGACGCTCCTTCACTCATACTTACATGGGCT GGTTCCGCCAGGTTCCAGGGAAGGAGCGTGAGTTTGTAGCCACTGTTGGCTGGACTG GTAATAGTGCATATTATGCAGACTCCGTGAAGGGCCGATTCACCATCTCCAGAGACA ACGCCAAGAACACGGTGTATCTACAAATGAACAACCTGAAACCTGAGGACACGGCC CTTTATTACTGTGCATCGTACCCGTCGAGTAGTAGTTGGCACCATCTCCGCGACCCC AATCACTATGCCTACTGGGGCCAGGGGACCCAGGTCACCGTTTCCTCAGGAGGAGG TGGGTCTGGAGGTGGAGGATCTGGTGGAGGTGGGTCTGGAGGAGGTGGATCCCCCG AGGAACCTCTAGTGGTGAAGGTGGAAGAGGGAGATACCGCTGCCCTGTGGTGCCTC AAGGGGACCTCAGATGGCCCCACTCAGCAGCTGACCTGGTCTCGGGAGTCCCCGCTT AAACCCTTCTTAAAATACAGCCTGGGGGTGCCAGGCCTGGGAGTGCACGTGAGGCC- 135 - 13321680vlAttorney Docket No. 2012106-0161CGACGCCATCAGCGTGGTTATCCGGAACGTCTCTCAACAGATGGGGGGCTTCTACCT GTGCCAGCCGGGGCCCCCCTCTGAGAAGGCCTGGCAGCCTGGCTGGACAGTCAATG TGGAGGGCAGCGGGGAGCTGTTCCGGTGGAATGTTTCGGACCTAGGTGGCCTGGGC TGTGGCCTGAAGAACAGGTCCTCAGAGGGCCCCAGCTCCCCTTCCGGGAAGCTCAT GAGCCCCAAGCTGTATGTGTGGGCCAAAGACCGCCCTGAGATCTGGGAGGGAGAGC CTCCGTGTCTCCCACCGAGGGACAGCCTGAACCAGAGCCTCAGCCGGGACCTCACC GTTGCCCCTGGCTCCACACTCTGGCTGTCCTGTGGGGTACCCCCTGACTCTGTGTCCA GGGGCCCCCTCTCCTGGACCCATGTGCACCCCAAGGGGCCTAAGTCATTGCTGAGCC TAGAGCTGAAGGACGATCGCCCGGCCAGAGATATGTGGGTAATGGGCACGAGCCTG ATGTTGCCCCGGGCCACAGCTCAAGACGCTGGAAAGTGGTATTGTCACCGTGGCAA CCTGACCATGTCATTCCACCTGGAGATCACTGCTCGGCCATCTAGACATCATCACCA TCACCATAmino acid sequence SEQ ID NO: 30

[0385] MEFGLSWyYLNALFRG QCQVQLQQFGGGLVQADGSLRLSCAASGRSFT HTYMGWFRQVPGKEREFVA TVGWTGNSA YYADSVKGRFTISRDNAKNTVYLQMNNLK PEDTALYYCASYPSSSSWHHLRDPNHYAYWGQGTQVTVSSGGGGSGGGGSGGGGSGGG GSPEEPLVVKVEEGDTAALWCLKGTSDGPTOOLTWSRESPLKPFLKYSLGVPGLGVHV RPDAISVVIRNVSQQMGGFYLCQPGPPSEKAWOPGWTVNVEGSGELFRWNVSDLGGLG CGLKNRS SEGP S SP SGKLMSPKLYVWAKDRPEIWEGEPPCLPPRD SLNQ SLSRDLT VAPG STLWLSCGVPPDSVSRGPLSWTHVHPKGPKSLLSLELKDDRPARDMWVMGTSLMLPRA TAQDAGKWYCHRGNLTMSFHLEITARPSRHHHHHHAleta #593Nucleotide sequence SEQ ID NO: 31

[0386] ATGGAGTTTGGGCTGAGCTGGGTTTTCCTCGTTGCTCTTTTTAGAGGTG TCCAGTGTCAGGTGCAGTTGCAGCAGTCTGGGGGAGGATTGGTGCAGGCTGGGGGC TCTCTGAGACTCTCCTGTGCAGCCTCTGGACGCACCTTCACCCGCCTTCCCATGGGCT- 136 - 13321680vlAttorney Docket No. 2012106-0161GGTTCCGCCAGGCTCCAGGAAAGGAGCGAGAGTTTGTAGCAGCTCTTAGCTGGAGT GGTGGCAACACAGACTATGCAAATACTGCAGACTCCGTGAAGGGCCGATTCACCAT GTCCAGAGACAGCGCCAAGAACACGGTGTATCTGTGGATGAACAACCTGAAACCTG AGGACACGGCCGTTTATTACTGTGCAGCCCGACAGAGTGGTTGGGGTACCGCCTTCA CTGATGACTATGACTACTGGGGCCAGGGGACCCAGGTCACCGTCTCCTCAGGAGGA GGTGGGTCTGGAGGTGGAGGATCTGGTGGAGGTGGGTCTGGAGGAGGTGGATCCCC CGAGGAACCTCTAGTGGTGAAGGTGGAAGAGGGAGATACCGCTGCCCTGTGGTGCC TCAAGGGGACCTCAGATGGCCCCACTCAGCAGCTGACCTGGTCTCGGGAGTCCCCG CTTAAACCCTTCTTAAAATACAGCCTGGGGGTGCCAGGCCTGGGAGTGCACGTGAG GCCCGACGCCATCAGCGTGGTTATCCGGAACGTCTCTCAACAGATGGGGGGCTTCTA CCTGTGCCAGCCGGGGCCCCCCTCTGAGAAGGCCTGGCAGCCTGGCTGGACAGTCA ATGTGGAGGGCAGCGGGGAGCTGTTCCGGTGGAATGTTTCGGACCTAGGTGGCCTG GGCTGTGGCCTGAAGAACAGGTCCTCAGAGGGCCCCAGCTCCCCTTCCGGGAAGCT CATGAGCCCCAAGCTGTATGTGTGGGCCAAAGACCGCCCTGAGATCTGGGAGGGAG AGCCTCCGTGTCTCCCACCGAGGGACAGCCTGAACCAGAGCCTCAGCCGGGACCTC ACCGTTGCCCCTGGCTCCACACTCTGGCTGTCCTGTGGGGTACCCCCTGACTCTGTGT CCAGGGGCCCCCTCTCCTGGACCCATGTGCACCCCAAGGGGCCTAAGTCATTGCTGA GCCTAGAGCTGAAGGACGATCGCCCGGCCAGAGATATGTGGGTAATGGGCACGAGC CTGATGTTGCCCCGGGCCACAGCTCAAGACGCTGGAAAGTGGTATTGTCACCGTGGC AACCTGACCATGTCATTCCACCTGGAGATCACTGCTCGGCCATCTAGAGGAGGAGGT GGGTCTGGAGGTGGAGGATCTGGTGGAGGTGGGTCTGGAGGAGGTGGGTCTGAGGT GCAGCTGGTGGAGTCTGGTGGTGGTCTTGTTCAACCTGGTGGTTCTCTTCGTCTTTCT TGTGCTGCTTCTGGTTTTAATATTAAAGATACTTATATTCATTGGGTTCGTCAAGCTC CTGGTAAAGGTCTTGAATGGGTTGCTCGTATTTATCCTACTAATGGTTATACTCGTTA TGCTGATTCTGTTAAAGGTCGTTTTACTATTTCTGCTGATACTTCTAAAAATACTGCT TATCTTCAAATGAACTCTCTTCGTGCTGAAGATACTGCTGTTTATTATTGTTCTCGTT GGGGTGGTGATGGTTTTTATGCTATGGATTATTGGGGTCAAGGTACTCTTGTCACCG TCTCCTCAGCTAGCACCGGGGGAGGTGGGTCTGGAGGTGGAGGATCTGGTGGAGGT GGGTCTGACATCCAGATGACCCAGTCTCCTTCTTCTCTTTCTGCTTCTGTTGGTGATC GTGTTACTATTACTTGTCGTGCTTCTCAAGATGTTAATACTGCTGTTGCTTGGTATCA- 137 - 13321680vlAttorney Docket No. 2012106-0161ACAAAAACCTGGTAAAGCTCCTAAACTTCTTATTTATTCTGCTTCTTTTCTTTATTCTG GTGTTCCTTCTCGTTTTTCTGGTTCTCGTTCTGGTACTGATTTTACTCTTACTATTTCTT CTCTTCAACCTGAAGATTTTGCTACTTATTATTGTCAACAACATTATACTACTCCTCC TACTTTTGGTCAAGGTACCAAGGTGGAGATCAAACGTACGCATCATCACCATCACCA TAmino acid sequence SEQ ID NO: 32

[0387] MEYGLSNyYYNALYRG QCQVQLQQSGGGLVQAGGSLRLSCAASGRTFT RLPMGWFRQAPGKEREFVAALSWSGGNTDYANTADSVKGRFTMSRDSAKNTVYLWMN NLKPEDTAVYYCAARQSGWGTAFTDDYDYWGQGTQVTVSSGGGGSGGGGSGGGGSGG GGSPEEPLVVKVEEGDTAALWCLKGTSDGPTQQLTWSRESPLKPFLKYSLGVPGLGVH VRPDAISVVIRNVSOOMGGFYLCOPGPPSEKAWOPGWTVNVEGSGELFRWNVSDLGGL GCGLKNRSSEGPSSPSGKLMSPKLYVWAKDRPEIWEGEPPCLPPRDSLNQSLSRDLTVAP GSTLWLSCGVPPDSVSRGPLSWTHVHPKGPKSLLSLELKDDRPARDMWVMGTSLMLPR ATAODAGI< WYCHRGNLTMSFHLE1TARPSRGGGGSGGGGSGGGGSGGGGSA1'G / J7AG GGLVQPGGSLRLSCAASGFNIKDTYIHmrRQAPGKGLErARIYPTNGYTRYADSVKGRFTIS ADTSKNTAYLQMNSLRAEDTAVYYCSRWGGDGFYAMDYWGQGTLVTVSSASTGGGGSGGGG SGGGGSDIQMTQSPSSLSASVGDRVTITCRASQDVNTAVAWYQOKPGKAPKLLIYSASFLYSGV PSRFSGSRSGTDFTLTISSLOPEDFATYYCOOHYTTPPTFGOGTKVEIKRTEEAEEAEEAAleta #597Nucleotide sequence SEQ ID NO: 33

[0388] ATGGAGTTTGGGCTGAGCTGGGTTTTCCTCGTTGCTCTTTTTAGAGGTG TCCAGTGTCAGGTGCAGCTGCAGGAGTCTGGGGGAGGCTTGGTGCAGGCTGGGGGC TCTCTGAAACTCTCCTGTGGAGTCTCTGGACGCACCCTCAGTGACTATGCCATGGGC TGGTTCCGCCAGCCTCCAGGGAAGGAGCGTGAGTTTGTAGCAACTATTAGGTGGAGT GGTGGTTACACATACTCTGCAGACTCCGTGAAGGGCCGATTCACCATCTCCAGAGAC GCCGCCAAGAACACGGTGTATCTGCAAATGAACAGCCTGAAACCTGAGGACACGGC- 138 - 13321680vlAttorney Docket No. 2012106-0161CGTTTATCACTGTGCCGCCTGCAGACTATACAATGGTAGACGGTTAGGCGACCCGAG TGAGTATGACTACTGGGGCCAGGGGACCCTGGTCACCGTCTCCTCAGGAGGAGGTG GGTCTGGAGGTGGAGGATCTGGTGGAGGTGGGTCTGGAGGAGGTGGGTCTGAGGTG CAGCTGGTGGAGTCTGGTGGTGGTCTTGTTCAACCTGGTGGTTCTCTTCGTCTTTCTT GTGCTGCTTCTGGTTTTAATATTAAAGATACTTATATTCATTGGGTTCGTCAAGCTCC TGGTAAAGGTCTTGAATGGGTTGCTCGTATTTATCCTACTAATGGTTATACTCGTTAT GCTGATTCTGTTAAAGGTCGTTTTACTATTTCTGCTGATACTTCTAAAAATACTGCTT ATCTTCAAATGAACTCTCTTCGTGCTGAAGATACTGCTGTTTATTATTGTTCTCGTTG GGGTGGTGATGGTTTTTATGCTATGGATTATTGGGGTCAAGGTACTCTTGTCACCGTC TCCTCAGCTAGCACCGGGGGAGGTGGGTCTGGAGGTGGAGGATCTGGTGGAGGTGG GTCTGACATCCAGATGACCCAGTCTCCTTCTTCTCTTTCTGCTTCTGTTGGTGATCGT GTTACTATTACTTGTCGTGCTTCTCAAGATGTTAATACTGCTGTTGCTTGGTATCAAC AAAAACCTGGTAAAGCTCCTAAACTTCTTATTTATTCTGCTTCTTTTCTTTATTCTGGT GTTCCTTCTCGTTTTTCTGGTTCTCGTTCTGGTACTGATTTTACTCTTACTATTTCTTCT CTTCAACCTGAAGATTTTGCTACTTATTATTGTCAACAACATTATACTACTCCTCCTA CTTTTGGTCAAGGTACCAAGGTGGAGATCAAACGTACGGGAGGAGGTGGGTCTGGA GGTGGAGGATCTGGTGGAGGTGGGTCTGGAGGAGGTGGATCCCCCGAGGAACCTCT AGTGGTGAAGGTGGAAGAGGGAGATACCGCTGCCCTGTGGTGCCTCAAGGGGACCT CAGATGGCCCCACTCAGCAGCTGACCTGGTCTCGGGAGTCCCCGCTTAAACCCTTCT TAAAATACAGCCTGGGGGTGCCAGGCCTGGGAGTGCACGTGAGGCCCGACGCCATC AGCGTGGTTATCCGGAACGTCTCTCAACAGATGGGGGGCTTCTACCTGTGCCAGCCG GGGCCCCCCTCTGAGAAGGCCTGGCAGCCTGGCTGGACAGTCAATGTGGAGGGCAG CGGGGAGCTGTTCCGGTGGAATGTTTCGGACCTAGGTGGCCTGGGCTGTGGCCTGAA GAACAGGTCCTCAGAGGGCCCCAGCTCCCCTTCCGGGAAGCTCATGAGCCCCAAGC TGTATGTGTGGGCCAAAGACCGCCCTGAGATCTGGGAGGGAGAGCCTCCGTGTCTCC CACCGAGGGACAGCCTGAACCAGAGCCTCAGCCGGGACCTCACCGTTGCCCCTGGC TCCACACTCTGGCTGTCCTGTGGGGTACCCCCTGACTCTGTGTCCAGGGGCCCCCTCT CCTGGACCCATGTGCACCCCAAGGGGCCTAAGTCATTGCTGAGCCTAGAGCTGAAG GACGATCGCCCGGCCAGAGATATGTGGGTAATGGGCACGAGCCTGATGTTGCCCCG- 139 - 13321680vlAttorney Docket No. 2012106-0161GGCCACAGCTCAAGACGCTGGAAAGTGGTATTGTCACCGTGGCAACCTGACCATGT CATTCCACCTGGAGATCACTGCTCGGCCATCTAGACATCATCACCATCACCATAmino acid sequence SEQ ID NO: 34

[0389] MEFGLSWVFLVALFRGVQCQVQLQESGGGLVQAGGSLKLSCGVSGRT LSDYAMGWFRQPPGKEREFVATIRWSGGYTYSADSVKGRFTISRDAAKNTVYLQM NSLKPEDTAVYHCAACRLYNGRRLGDPSEYDYWGQGTLVTVSSGGGGSGGGGSGG GGSGGGGSEVQLVESGGGLVQPGGSLRLSCAASGFNIKDTYIHWVROAPGKGLEWVAR IYPTNGYTRYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCSRWGGDGFYAM DYWGOGTLVTVSSASTGGGGSGGGGSGGGGSD1OMTOSPSSLSASVGDRVT1TCRASO DVNTAVAWYOOKPGKAPKLLIYSASFLYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYY COQHYTTPPTFGQGTKVEIKRTGGGGSGGGGSGGGGSGGGGSPEEPLVVKVEEGDTAA LWCLKGTSDGPTQOLTWSRESPLKPFLKYSLGVPGLGVHVRPDAISVVTRNVSOQMGGF YLCOPGPPSEKAWQPGWTVNVEGSGELFRWNVSDLGGLGCGLKNRSSEGPSSPSGKLM SPKLYVWAKDRPEIWEGEPPCLPPRDSLNQSLSRDLTVAPGSTLWLSCGVPPDSVSRGPL SWTHVHPKGPKSLLSLELKDDRPARDMWVMGTSLMLPRATAQDAGKWYCHRGNLTM SFHLEIT ARP SRHHHHHHAleta #607Nucleotide sequence SEQ ID NO: 35

[0390] ATGGAGTTTGGGCTGAGCTGGGTTTTCCTCGTTGCTCTTTTTAGAGGTG TCCAGTGTGAGGTGCAGCTGGTGGAGTCCGGAGGAGGCCTGGTGCAGCCAGGAGGC TCTCTGAGGCTGAGCTGCACCTTCTCCGGCGGCACCTTCAGCAGCTACACAATGGGC TGGTTCAGGCAGGCACCAGGCAAGGAGAGAGAGTTTGTGGCAGAGGTGAGGTGGG GAGGAGTGACCACATACTCCAACTCTCTGAAGGACCGCTTCAGCATCTCCGAGGATT CTGTGAAGAACGCCGTGTATCTGCAGATGAATAGCCTGAAGCCCGAGGACACAGCC GTGTACTATTGTGCCGCCGTGCGGCAGATGTACATGACCGTGGTGCCTGATTATTGG GGCCAGGGCACCCTGGTGACAGTGTCTAGCGGAGGAGGTGGGTCTGGAGGTGGAGG- 140 - 13321680vlAttorney Docket No. 2012106-0161ATCTGGTGGAGGTGGGTCTGGAGGAGGTGGATCCCCCGAGGAACCTCTAGTGGTGA AGGTGGAAGAGGGAGATACCGCTGCCCTGTGGTGCCTCAAGGGGACCTCAGATGGC CCCACTCAGCAGCTGACCTGGTCTCGGGAGTCCCCGCTTAAACCCTTCTTAAAATAC AGCCTGGGGGTGCCAGGCCTGGGAGTGCACGTGAGGCCCGACGCCATCAGCGTGGT TATCCGGAACGTCTCTCAACAGATGGGGGGCTTCTACCTGTGCCAGCCGGGGCCCCC CTCTGAGAAGGCCTGGCAGCCTGGCTGGACAGTCAATGTGGAGGGCAGCGGGGAGC TGTTCCGGTGGAATGTTTCGGACCTAGGTGGCCTGGGCTGTGGCCTGAAGAACAGGT CCTCAGAGGGCCCCAGCTCCCCTTCCGGGAAGCTCATGAGCCCCAAGCTGTATGTGT GGGCCAAAGACCGCCCTGAGATCTGGGAGGGAGAGCCTCCGTGTCTCCCACCGAGG GACAGCCTGAACCAGAGCCTCAGCCGGGACCTCACCGTTGCCCCTGGCTCCACACTC TGGCTGTCCTGTGGGGTACCCCCTGACTCTGTGTCCAGGGGCCCCCTCTCCTGGACC CATGTGCACCCCAAGGGGCCTAAGTCATTGCTGAGCCTAGAGCTGAAGGACGATCG CCCGGCCAGAGATATGTGGGTAATGGGCACGAGCCTGATGTTGCCCCGGGCCACAG CTCAAGACGCTGGAAAGTGGTATTGTCACCGTGGCAACCTGACCATGTCATTCCACC TGGAGATCACTGCTCGGCCATCTAGAGGAGGCGGAGGAAGTGGAGGAGGAGGGAG CGGGGGGGGGGGAAGTGAAGTTCAGTTGGTCGAATCTGGAGGGGGACTGGTACAGC CTGGCAACTCTCTGCGATTGTCTTGTGCTGCTAGTGGTTTTACATTTTCTTCATTTGGC ATGAGTTGGGTCCGACAAGCCCCAGGAAAAGGGTTGGAATGGGTGAGCAGCATTTC TGGAAGCGGTTCTGATACACTCTACGCCGACAGCGTAAAAGGCCGGTTCACTATATC TCGCGATAATGCAAAAACAACACTTTATCTGCAAATGAATAGTCTGCGACCTGAAG ATACAGCAGTATATTATTGTACAATAGGAGGTAGTCTCAGTAGGTCATCACAAGGA ACACTGGTAACCGTGAGTTCCCATCATCACCATCACCATAmino acid sequence SEQ ID NO: 36

[0391] MEFGLSWVFLVALFRGVQCEVQLVESGGGLVQPGGSLRLSCTFSGGTFSSYTMGWFRQAPGKEREFVAEVRWGGVTTYSNSLKDRFSISEDSVKNAVYLQMNSLKPEDTAVYYCAAVRQMYMTWPDYWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSPEEPL VVKVEEGDTAALWCLKGTSDGPTQQLTWSRESPLKPFLKYSLGVPGLGVHVRPDAISVVIRNVSQQMGGFYLCQPGPPSEKAWQPGWTVNVEGSGELFRWNVSDLGGLGCGLKNR - 141 - 13321680vlAttorney Docket No. 2012106-0161SSEGPSSPS GKLMSPKL YVW AKDRPEIWEGEPPCLPPRD SLNQ SL SRDLT VAPGS TL WLS CGVPPDSVSRGPLSWTHVHPKGPKSLLSLELKDDRPARDMWVMGTSLMLPRATAODA GKWYCHRGNLTMSFHLEITARPSRGGGGSGGGGSGGGGSEFQZ VESGGGL VQPGNSLRL SCAASGFTFSSFGMSWVRQAPGKGLEWVSSISGSGSDTLYADSVKGRFTISRDNAKTTLYLQM NSLRPEDTAVYYCTIGGSLSRSSQGTLVTVSSAAAAAAAleta #628Nucleotide sequence SEQ ID NO: 37

[0392] ATGGAGTTTGGGCTGAGCTGGGTTTTCCTCGTTGCTCTTTTTAGAGGTG TCCAGTGTGAGGTGCAGCTGGTGGAGTCTGGTGGTGGTCTTGTTCAACCTGGTGGTT CTCTTCGTCTTTCTTGTGCTGCTTCTGGTTTTAATATTAAAGATACTTATATTCATTGG GTTCGTCAAGCTCCTGGTAAAGGTCTTGAATGGGTTGCTCGTATTTATCCTACTAATG GTTATACTCGTTATGCTGATTCTGTTAAAGGTCGTTTTACTATTTCTGCTGATACTTCT AAAAATACTGCTTATCTTCAAATGAACTCTCTTCGTGCTGAAGATACTGCTGTTTATT ATTGTTCTCGTTGGGGTGGTGATGGTTTTTATGCTATGGATTATTGGGGTCAAGGTAC TCTTGTCACCGTCTCCTCAGCTAGCACCGGGGGAGGTGGGTCTGGAGGTGGAGGATC TGGTGGAGGTGGGTCTGACATCCAGATGACCCAGTCTCCTTCTTCTCTTTCTGCTTCT GTTGGTGATCGTGTTACTATTACTTGTCGTGCTTCTCAAGATGTTAATACTGCTGTTG CTTGGTATCAACAAAAACCTGGTAAAGCTCCTAAACTTCTTATTTATTCTGCTTCTTT TCTTTATTCTGGTGTTCCTTCTCGTTTTTCTGGTTCTCGTTCTGGTACTGATTTTACTCT TACTATTTCTTCTCTTCAACCTGAAGATTTTGCTACTTATTATTGTCAACAACATTAT ACTACTCCTCCTACTTTTGGTCAAGGTACCAAGGTGGAGATCAAACGTACGGGAGGA GGTGGGTCTGGAGGTGGAGGATCTGGTGGAGGTGGGTCTGGAGGAGGTGGATCCCC CGAGGAACCTCTAGTGGTGAAGGTGGAAGAGGGAGATACCGCTGCCCTGTGGTGCC TCAAGGGGACCTCAGATGGCCCCACTCAGCAGCTGACCTGGTCTCGGGAGTCCCCG CTTAAACCCTTCTTAAAATACAGCCTGGGGGTGCCAGGCCTGGGAGTGCACGTGAG GCCCGACGCCATCAGCGTGGTTATCCGGAACGTCTCTCAACAGATGGGGGGCTTCTA CCTGTGCCAGCCGGGGCCCCCCTCTGAGAAGGCCTGGCAGCCTGGCTGGACAGTCA ATGTGGAGGGCAGCGGGGAGCTGTTCCGGTGGAATGTTTCGGACCTAGGTGGCCTG- 142 - 13321680vlAttorney Docket No. 2012106-0161GGCTGTGGCCTGAAGAACAGGTCCTCAGAGGGCCCCAGCTCCCCTTCCGGGAAGCT CATGAGCCCCAAGCTGTATGTGTGGGCCAAAGACCGCCCTGAGATCTGGGAGGGAG AGCCTCCGTGTCTCCCACCGAGGGACAGCCTGAACCAGAGCCTCAGCCGGGACCTC ACCGTTGCCCCTGGCTCCACACTCTGGCTGTCCTGTGGGGTACCCCCTGACTCTGTGT CCAGGGGCCCCCTCTCCTGGACCCATGTGCACCCCAAGGGGCCTAAGTCATTGCTGA GCCTAGAGCTGAAGGACGATCGCCCGGCCAGAGATATGTGGGTAATGGGCACGAGC CTGATGTTGCCCCGGGCCACAGCTCAAGACGCTGGAAAGTGGTATTGTCACCGTGGC AACCTGACCATGTCATTCCACCTGGAGATCACTGCTCGGCCATCTAGAGGCGGAGGC GGCAGCGGAGGCGGCGGCAGCGGCGGCGGCGGAAGCACCATCCCCCCCCACGTGC AGAAAAGCGTGAACAATGACATGATCGTGACCGACAACAACGGCGCCGTCAAGTTT CCTCAACTGTGCAAGTTCTGCGACGTGCGGTTCAGCACATGTGATAACCAGAAGTCC TGTATGAGCAACTGCTCCATCACCAGCATCTGCGAGAAACCTCAGGAGGTGTGCGTG GCCGTGTGGAGAAAGAACGACGAGAACATTACACTGGAAACCGTGTGTCACGATCC TAAGCTGCCTTACCACGACTTCATCCTGGAAGATGCCGCTTCTCCAAAGTGCATCAT GAAGGAAAAGAAAAAGCCCGGCGAGACATTTTTCATGTGCTCTTGTAGCAGCGACG AGTGCAACGATAATATCATCTTCAGCGAGGAATACAACACCTCTAATCCTGACCATC ATCACCATCACCATAmino acid sequence SEQ ID NO: 38

[0393] MEFGLSWVFLVALFRGY. QCEVQLVESGGGLVQPGGSLRLSCAASGFNIK DTYIHWVRQAPGKGLEWVARIYPTNGYTRYADSVKGRFTISADTSKNTAYLQMNSLRA EDTAVYYC SRWGGDGFYAMDYWGQGTLVTVS S ASTGGGGSGGGGSGGGGSDIQMTQ SPSSLSASVGDRVTITCRASQDVNTAVAWYQQKPGKAPKLLIYSASFLYSGVPSRFSGSR SGTDFTLTISSLQPEDFATYYCQQHYTTPPTFGQGTKVEIKRTGGGGSGGGGSGGGGSG GGGSPEEPLVVKVEEGDTAALWCLKGTSDGPTQQLTWSRESPLKPFLKYSLGVPGLGVHVRPDAISVVIRNVSQQMGGFYLCQPGPPSEKAWQPGWTVNVEGSGELFRWNVSDLGG LGC GLKNRS SEGP S SP S GKLM SPKL YVW AKDRPEIWEGEPPCLPPRD SLNQ SL SRDLT V A PGSTLWLSCGVPPDSVSRGPLSWTHVHPKGPKSLLSLELKDDRPARDMWVMGTSLMLP RATAODAGKWYCHRGNLTMSFHLE1TARPSRGGGGSGGGGSGGGGST1PPHVOKSVNN - 143 - 13321680vlAttorney Docket No. 2012106-0161DMIVTDNNGAVKFPQLCKFCDVRFSTCDNQKSCMSNCSITSICEKPQEVCVAVWRKNDENITLETVCHDPKLPYHDFILEDAASPKCIMKEKKKPGETFFMCSCSSDECNDNIIFSEEY NTSNPDHHHHHHAleta #656Nucleotide sequence SEQ ID NO: 39

[0394] ATGGAGTTTGGGCTGAGCTGGGTTTTCCTCGTTGCTCTTTTTAGAGGTG TCCAGTGTCAGGTGCAGCTGCAGGAGTCTGGAGGAGGCCTGGCACAGGCAGGAGGC AGCCTGCGGCTGTCCTGCGCCGCCTCTGGCAGGACCTTCAGCATGGGCTGGTTTAGG CAGGCACCAGGCAAGGAGAGGGAGTTCGTGGCCGCCATCACATACTCCGGCGGCTC TCCCTACTATGCCAGCTCCGTGCGGGGCCGGTTCACCATCAGCCGGGACAACGCCAA GAATACAGTGTACCTGCAGATGAACTCCCTGAAGCCCGAGGACACCGCCGTGTACT ATTGTGCCGCCAATCCTACATATGGCTCCGATTGGAACGCCGAGAATTGGGGCCAGG GCACCCAGGTGACAGTGTCTAGCGGAGGAGGTGGGTCTGGAGGTGGAGGATCTGGT GGAGGTGGGTCTGGAGGAGGTGGAAGCGACATCAAGCTGCAGCAGAGCGGCGCCG AGCTGGCCCGGCCCGGAGCCAGTGTGAAAATGAGCTGCAAGACCTCTGGCTACACC TTTACCAGATACACCATGCACTGGGTCAAGCAGCGGCCTGGCCAGGGCCTGGAGTG GATCGGCTACATCAACCCCAGCAGAGGATACACAAACTACAATCAGAAGTTCAAGG ATAAGGCCACACTGACCACAGATAAGAGCAGCAGCACCGCCTATATGCAGCTGAGC AGCCTGACAAGCGAGGACAGCGCCGTGTACTACTGCGCCAGATACTATGATGACCA CTACTGTCTGGACTACTGGGGACAAGGCACAACCCTCACAGTGTCTAGCGTGGAAG GCGGCTCTGGTGGCAGCGGCGGCTCTGGCGGCTCCGGCGGAGTGGACGACATTCAG CTGACCCAGTCCCCTGCTATCATGTCTGCCTCTCCTGGCGAGAAGGTGACCATGACC TGTAGAGCCAGCAGCAGCGTGAGCTACATGAACTGGTACCAGCAGAAATCTGGCAC CTCTCCAAAGCGGTGGATCTACGACACCTCCAAAGTGGCCAGCGGCGTGCCTTATAG ATTCAGCGGCAGCGGAAGCGGCACCTCCTACAGCCTGACCATCAGCAGCATGGAAG CCGAGGATGCCGCTACATACTACTGCCAGCAATGGTCCAGCAACCCTCTGACCTTCG GCGCTGGCACAAAGCTGGAACTGAAGGGAGGCGGAGGAAGTGGAGGAGGAGGGAG CGGGGGGGGGGGAAGTGAAGTTCAGTTGGTCGAATCTGGAGGGGGACTGGTACAGC- 144 - 13321680vlAttorney Docket No. 2012106-0161CTGGCAACTCTCTGCGATTGTCTTGTGCTGCTAGTGGTTTTACATTTTCTTCATTTGGC ATGAGTTGGGTCCGACAAGCCCCAGGAAAAGGGTTGGAATGGGTGAGCAGCATTTC TGGAAGCGGTTCTGATACACTCTACGCCGACAGCGTAAAAGGCCGGTTCACTATATC TCGCGATAATGCAAAAACAACACTTTATCTGCAAATGAATAGTCTGCGACCTGAAG ATACAGCAGTATATTATTGTACAATAGGAGGTAGTCTCAGTAGGTCATCACAAGGA ACACTGGTAACCGTGAGTTCCCATCATCACCATCACCATAmino acid sequence SEQ ID NO: 40

[0395] MEFGLSWVFLVALFRGY. QCQVQLQESGGGLAQAGGSLRLSCAASGRTFS MGWFRQAPGKEREFVAA1TYSGGSPYYASSVRGRFT1SRDNAKNTVYLQMNSLKPEDT AVYYCAANPTYGSDWNAENWGQGTQVTVSSGGGGSGGGGSGGGGSGGGGSDIKLQQ SGAELARPGASVKMSCKTSGYTFTRYTMHWVKQRPGOGLEWIGYINPSRGYTNYNOKF KDKATLTTDKSSSTAYMQLSSLTSEDSAVYYCARYYDDHYCLDYWGOGTTLTVSSVEG GSGGSGGSGGSGGVDDIQLTQSPAIMSASPGEKVTMTCRASSSVSYMNWYOOKSGTSP KRWIYDTSKVASGVPYRFSGSGSGTSYSLTISSMEAEDAATYYCOQWSSNPLTFGAGTK LELKGGGGSGGGGSGGGGSEVQLVESGGGLVQPGNSLRLSCAASGFTFSSFGMSWVRQ APGKGLEWVSSISGSGSDTLYADSVKGRFTISRDNAKTTLYLQMNSLRPEDTAVYYCTI GGSLSRS SQGTL VTVS SHHHHHHAleta #657Nucleotide sequence SEQ ID NO: 41

[0396] ATGGAGTTTGGGCTGAGCTGGGTTTTCCTCGTTGCTCTTTTTAGAGGTG TCCAGTGTCAGGTGCAGCTGCAGGAGTCTGGAGGAGGCCTGGCACAGGCAGGAGGC AGCCTGCGGCTGTCCTGCGCCGCCTCTGGCAGGACCTTCAGCATGGGCTGGTTTAGG CAGGCACCAGGCAAGGAGAGGGAGTTCGTGGCCGCCATCACATACTCCGGCGGCTC TCCCTACTATGCCAGCTCCGTGCGGGGCCGGTTCACCATCAGCCGGGACAACGCCAA GAATACAGTGTACCTGCAGATGAACTCCCTGAAGCCCGAGGACACCGCCGTGTACT ATTGTGCCGCCAATCCTACATATGGCTCCGATTGGAACGCCGAGAATTGGGGCCAGG- 145 - 13321680vlAttorney Docket No. 2012106-0161GCACCCAGGTGACAGTGTCTAGCGGAGGAGGTGGGTCTGGAGGTGGAGGATCTGGT GGAGGTGGGTCTGGAGGAGGTGGAAGCGACATCAAGCTGCAGCAGAGCGGCGCCG AGCTGGCCCGGCCCGGAGCCAGTGTGAAAATGAGCTGCAAGACCTCTGGCTACACC TTTACCAGATACACCATGCACTGGGTCAAGCAGCGGCCTGGCCAGGGCCTGGAGTG GATCGGCTACATCAACCCCAGCAGAGGATACACAAACTACAATCAGAAGTTCAAGG ATAAGGCCACACTGACCACAGATAAGAGCAGCAGCACCGCCTATATGCAGCTGAGC AGCCTGACAAGCGAGGACAGCGCCGTGTACTACTGCGCCAGATACTATGATGACCA CTACTGTCTGGACTACTGGGGACAAGGCACAACCCTCACAGTGTCTAGCGTGGAAG GCGGCTCTGGTGGCAGCGGCGGCTCTGGCGGCTCCGGCGGAGTGGACGACATTCAG CTGACCCAGTCCCCTGCTATCATGTCTGCCTCTCCTGGCGAGAAGGTGACCATGACC TGTAGAGCCAGCAGCAGCGTGAGCTACATGAACTGGTACCAGCAGAAATCTGGCAC CTCTCCAAAGCGGTGGATCTACGACACCTCCAAAGTGGCCAGCGGCGTGCCTTATAG ATTCAGCGGCAGCGGAAGCGGCACCTCCTACAGCCTGACCATCAGCAGCATGGAAG CCGAGGATGCCGCTACATACTACTGCCAGCAATGGTCCAGCAACCCTCTGACCTTCG GCGCTGGCACAAAGCTGGAACTGAAGCATCATCACCATCACCATAmino acid sequence SEQ ID NO: 42

[0397] MEFGLSWVFLVALFRGVQCQVQLQESGGGLAQAGGSLRLSCAASGRTFSMGWFRQAPGKEREFVAAITYSGGSPYYASSVRGRFTISRDNAKNTVYLQMNSLKPEDTAVYYCAANPTYGSDWNAENWGQGTQVTVSSGGGGSGGGGSGGGGSGGGGSDIKLQQSGAELARPGASVKMSCKTSGYTFTRYTMHWVKQRPGQGLEWIGYINPSRGYTNYNQKFKDKATLTTDKSSSTAYMQLSSLTSEDSAVYYCARYYDDHYCLDYWGQGTTLTVSSVEG GSGGSGGSGGSGGVDDIQLTQSPAIMSASPGEKVTMTCRASSSVSYMNWYQQKSGTSPKRWIYDTSKVASGVPYRFSGSGSGTSYSLTISSMEAEDAATYYCOQWSSNPLTFGAGTKLELKHHHHHHAleta #672Nucleotide sequence SEQ ID NO: 43- 146 - 13321680vlAttorney Docket No. 2012106-0161

[0398] ATGGAGTTTGGGCTGAGCTGGGTTTTCCTCGTTGCTCTTTTTAGAGGTG TCCAGTGTTTTAGCCAGCAGATCTATGGCGTGGTGTACGGCAACGTGACCTTCCACG TGCCTAGCAACGTCCCCCTGAAAGAGGTGCTCTGGAAGAAGCAGAAGGACAAGGTG GCCGAGCTGGAAAACAGCGAGTTCCGGGCCTTCAGCAGCTTTAAGAACAGAGTGTA CCTGGACACAGTGTCCGGCAGCCTGACAATCTACAACCTGACCAGCTCTGATGAAG ATGAGTACGAGATGGAAAGCCCTAATATCACCGACACCATGAAATTCTTCCTGTACG TGCTGGGCGGAGGCGGCAGCGGCGGCGGCGGCTCCGGCGGAGGCGGATCTGGCGG AGGCGGCTCTGAGGTGCAGCTGGTGGAGTCCGGAGGAGGCCTGGTGCAGCCAGGAG GCTCTCTGAGGCTGAGCTGCACCTTCTCCGGCGGCACCTTCAGCAGCTACACAATGG GCTGGTTCAGGCAGGCACCAGGCAAGGAGAGAGAGTTTGTGGCAGAGGTGAGGTGG GGAGGAGTGACCACATACTCCAACTCTCTGAAGGACCGCTTCAGCATCTCCGAGGA TTCTGTGAAGAACGCCGTGTATCTGCAGATGAATAGCCTGAAGCCCGAGGACACAG CCGTGTACTATTGTGCCGCCGTGCGGCAGATGTACATGACCGTGGTGCCTGATTATT GGGGCCAGGGCACCCTGGTGACAGTGTCTAGCGGAGGAGGTGGGTCTGGAGGTGGA GGATCTGGTGGAGGTGGGTCTGGAGGAGGTGGATCCCCCGAGGAACCTCTAGTGGT GAAGGTGGAAGAGGGAGATACCGCTGCCCTGTGGTGCCTCAAGGGGACCTCAGATG GCCCCACTCAGCAGCTGACCTGGTCTCGGGAGTCCCCGCTTAAACCCTTCTTAAAAT ACAGCCTGGGGGTGCCAGGCCTGGGAGTGCACGTGAGGCCCGACGCCATCAGCGTG GTTATCCGGAACGTCTCTCAACAGATGGGGGGCTTCTACCTGTGCCAGCCGGGGCCC CCCTCTGAGAAGGCCTGGCAGCCTGGCTGGACAGTCAATGTGGAGGGCAGCGGGGA GCTGTTCCGGTGGAATGTTTCGGACCTAGGTGGCCTGGGCTGTGGCCTGAAGAACAG GTCCTCAGAGGGCCCCAGCTCCCCTTCCGGGAAGCTCATGAGCCCCAAGCTGTATGT GTGGGCCAAAGACCGCCCTGAGATCTGGGAGGGAGAGCCTCCGTGTCTCCCACCGA GGGACAGCCTGAACCAGAGCCTCAGCCGGGACCTCACCGTTGCCCCTGGCTCCACA CTCTGGCTGTCCTGTGGGGTACCCCCTGACTCTGTGTCCAGGGGCCCCCTCTCCTGG ACCCATGTGCACCCCAAGGGGCCTAAGTCATTGCTGAGCCTAGAGCTGAAGGACGA TCGCCCGGCCAGAGATATGTGGGTAATGGGCACGAGCCTGATGTTGCCCCGGGCCA CAGCTCAAGACGCTGGAAAGTGGTATTGTCACCGTGGCAACCTGACCATGTCATTCC ACCTGGAGATCACTGCTCGGCCATCTAGAGGAGGCGGAGGAAGTGGAGGAGGAGG GAGCGGGGGGGGGGGAAGTGAAGTTCAGTTGGTCGAATCTGGAGGGGGACTGGTAC- 147 - 13321680vlAttorney Docket No. 2012106-0161AGCCTGGCAACTCTCTGCGATTGTCTTGTGCTGCTAGTGGTTTTACATTTTCTTCATTT GGCATGAGTTGGGTCCGACAAGCCCCAGGAAAAGGGTTGGAATGGGTGAGCAGCAT TTCTGGAAGCGGTTCTGATACACTCTACGCCGACAGCGTAAAAGGCCGGTTCACTAT ATCTCGCGATAATGCAAAAACAACACTTTATCTGCAAATGAATAGTCTGCGACCTGA AGATACAGCAGTATATTATTGTACAATAGGAGGTAGTCTCAGTAGGTCATCACAAG GAACACTGGTAACCGTGAGTTCCCATCATCACCATCACCATAmino acid sequence SEQ ID NO: 44

[0399] MEFGLSWVFLVALFRGVQCFSQQIYGVVYGNVTFHVPSNVPLKEVLWKKQKDKVAELENSEFRAFSSFKNRVYLDTVSGSLTIYNLTSSDEDEYEMESPNITDTMKFFLYVLGGGGSGGGGSGGGGSGGGGSEVQLVESGGGLVQPGGSLRLSCTFSGGTFSSYTMGWFRQAPGKEREFVAEVRWGGVTTYSNSLKDRFSISEDSVKNAVYLQMNSLKPEDTAVYYCAAVRQMYMTWPDYWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSPEEPLVVKVEEGDTAALWCLKGTSDGPTQQLTWSRESPLKPFLKYSLGVPGLGVHVRPDAISVVIRNVSQQMGGFYLCQPGPPSEKAWQPGWTVNVEGSGELFRWNVSDLGGLGCGLKNRSSEGPSSPSGKLMSPKLYVWAKDRPEIWEGEPPCLPPRDSLNQSLSRDLTVAPGSTLWLSCGVPPDSVSRGPLSWTHVHPKGPKSLLSLELKDDRPARDMWVMGTSLMLPRATAQDAGKWYCHRGNLTMSFHLEITARPSRGGGGSGGGGSGGGGSEFQLVESGGGLVQPGNSLRLSCAASGFTFSSFGMSWVRQAPGKGLEWVSSISGSGSDTLYADSVKGRFTISRDNAKTTLYLQMNSLRPEDTAVYYCTIGGSLSRSSQGTLVTVSSHHHHHHAleta #673Nucleotide sequence SEQ ID NO: 45

[0400] ATGGAGTTTGGGCTGAGCTGGGTTTTCCTCGTTGCTCTTTTTAGAGGTG TCCAGTGTTTTAGCCAGCAGATCTATGGCGTGGTGTACGGCAACGTGACCTTCCACG TGCCTAGCAACGTCCCCCTGAAAGAGGTGCTCTGGAAGAAGCAGAAGGACAAGGTG GCCGAGCTGGAAAACAGCGAGTTCCGGGCCTTCAGCAGCTTTAAGAACAGAGTGTA CCTGGACACAGTGTCCGGCAGCCTGACAATCTACAACCTGACCAGCTCTGATGAAG- 148 - 13321680vlAttorney Docket No. 2012106-0161ATGAGTACGAGATGGAAAGCCCTAATATCACCGACACCATGAAATTCTTCCTGTACG TGCTGGGCGGAGGCGGCAGCGGCGGCGGCGGCTCCGGCGGAGGCGGATCTGGCGG AGGCGGCTCTGAAGTTCAGTTGGTCGAATCTGGAGGGGGACTGGTACAGCCTGGCA ACTCTCTGCGATTGTCTTGTGCTGCTAGTGGTTTTACATTTTCTTCATTTGGCATGAGT TGGGTCCGACAAGCCCCAGGAAAAGGGTTGGAATGGGTGAGCAGCATTTCTGGAAG CGGTTCTGATACACTCTACGCCGACAGCGTAAAAGGCCGGTTCACTATATCTCGCGA TAATGCAAAAACAACACTTTATCTGCAAATGAATAGTCTGCGACCTGAAGATACAG CAGTATATTATTGTACAATAGGAGGTAGTCTCAGTAGGTCATCACAAGGAACACTGG TAACCGTGAGTTCCGGAGGCGGAGGAAGTGGAGGAGGAGGGAGCGGGGGGGGGGG AAGTGGAGGTGGAGGATCTGAGGTGCAGCTGGTGGAGTCCGGAGGAGGCCTGGTGC AGCCAGGAGGCTCTCTGAGGCTGAGCTGCACCTTCTCCGGCGGCACCTTCAGCAGCT ACACAATGGGCTGGTTCAGGCAGGCACCAGGCAAGGAGAGAGAGTTTGTGGCAGAG GTGAGGTGGGGAGGAGTGACCACATACTCCAACTCTCTGAAGGACCGCTTCAGCAT CTCCGAGGATTCTGTGAAGAACGCCGTGTATCTGCAGATGAATAGCCTGAAGCCCG AGGACACAGCCGTGTACTATTGTGCCGCCGTGCGGCAGATGTACATGACCGTGGTGC CTGATTATTGGGGCCAGGGCACCCTGGTGACAGTGTCTAGCGGAGGAGGTGGGTCT GGAGGTGGAGGATCTGGTGGAGGTGGGTCTGGAGGAGGTGGATCCCCCGAGGAACC TCTAGTGGTGAAGGTGGAAGAGGGAGATACCGCTGCCCTGTGGTGCCTCAAGGGGA CCTCAGATGGCCCCACTCAGCAGCTGACCTGGTCTCGGGAGTCCCCGCTTAAACCCT TCTTAAAATACAGCCTGGGGGTGCCAGGCCTGGGAGTGCACGTGAGGCCCGACGCC ATCAGCGTGGTTATCCGGAACGTCTCTCAACAGATGGGGGGCTTCTACCTGTGCCAG CCGGGGCCCCCCTCTGAGAAGGCCTGGCAGCCTGGCTGGACAGTCAATGTGGAGGG CAGCGGGGAGCTGTTCCGGTGGAATGTTTCGGACCTAGGTGGCCTGGGCTGTGGCCT GAAGAACAGGTCCTCAGAGGGCCCCAGCTCCCCTTCCGGGAAGCTCATGAGCCCCA AGCTGTATGTGTGGGCCAAAGACCGCCCTGAGATCTGGGAGGGAGAGCCTCCGTGT CTCCCACCGAGGGACAGCCTGAACCAGAGCCTCAGCCGGGACCTCACCGTTGCCCC TGGCTCCACACTCTGGCTGTCCTGTGGGGTACCCCCTGACTCTGTGTCCAGGGGCCC CCTCTCCTGGACCCATGTGCACCCCAAGGGGCCTAAGTCATTGCTGAGCCTAGAGCT GAAGGACGATCGCCCGGCCAGAGATATGTGGGTAATGGGCACGAGCCTGATGTTGC- 149 - 13321680vlAttorney Docket No. 2012106-0161CCCGGGCCACAGCTCAAGACGCTGGAAAGTGGTATTGTCACCGTGGCAACCTGACC ATGTCATTCCACCTGGAGATCACTGCTCGGCCATCTAGACATCATCACCATCACCATAmino acid sequence SEQ ID NO: 46

[0401] MEFGLSWVFLVALFRGVQCFSQQIYGVVYGNVTFHVPSNVPLKEVLWKKQKDKVAELENSEFRAFSSFKNRVYLDTVSGSLTIYNLTSSDEDEYEMESPNITDTMKFFLYVLGGGGSGGGGSGGGGSGGGGSEFQLVESGGGLVQPGNSLRLSCAASGFTFSSFGMSWVRQAPGKGLEWVSSISGSGSDTLYADSVKGRFTISRDNAKTTLYLQMNSLRPEDTAVYYCTIGGSLSRSSQGTLVTVSSGGGGSGGGGSGGGGSGGGGSEVQLVESGGGLVQPGGSLRLSCTFSGGTFSSYTMGWFRQAPGKEREFVAEVRWGGVTTYSNSLKDRFSISEDSVKNAVYLQMNSLKPEDTAVYYCAAVRQMYMTWPDYWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSPEEPLVVKVEEGDTAALWCLKGTSDGPTQQLTWSRESPLKPFLKYSLGVPGLGVHVRPDAISVVIRNVSQQMGGFYLCQPGPPSEKAWQPGWTVNVEGSGELFRWNVSDLGGLGCGLKNRSSEGPSSPSGKLMSPKLYVWAKDRPEIWEGEPPCLPPRDSLNQSLSRDLTVAPGSTLWLSCGVPPDSVSRGPLSWTHVHPKGPKSLLSLELKDDRPARDMWVMGTSLMLPRATAQDAGKWYCHRGNLTMSFHLEITARPSRHHHHHHAleta #674Nucleotide sequence SEQ ID NO: 47

[0402] ATGGAGTTTGGGCTGAGCTGGGTTTTCCTCGTTGCTCTTTTTAGAGGTG TCCAGTGTTTTAGCCAGCAGATCTATGGCGTGGTGTACGGCAACGTGACCTTCCACG TGCCTAGCAACGTCCCCCTGAAAGAGGTGCTCTGGAAGAAGCAGAAGGACAAGGTG GCCGAGCTGGAAAACAGCGAGTTCCGGGCCTTCAGCAGCTTTAAGAACAGAGTGTA CCTGGACACAGTGTCCGGCAGCCTGACAATCTACAACCTGACCAGCTCTGATGAAG ATGAGTACGAGATGGAAAGCCCTAATATCACCGACACCATGAAATTCTTCCTGTACG TGCTGGGCGGAGGCGGCAGCGGCGGCGGCGGCTCCGGCGGAGGCGGATCTGGCGG AGGCGGCTCTGAGGTGCAGCTGGTGGAGTCTGGTGGTGGTCTTGTTCAACCTGGTGG TTCTCTTCGTCTTTCTTGTGCTGCTTCTGGTTTTAATATTAAAGATACTTATATTCATT- 150 - 13321680vlAttorney Docket No. 2012106-0161GGGTTCGTCAAGCTCCTGGTAAAGGTCTTGAATGGGTTGCTCGTATTTATCCTACTA ATGGTTATACTCGTTATGCTGATTCTGTTAAAGGTCGTTTTACTATTTCTGCTGATAC TTCTAAAAATACTGCTTATCTTCAAATGAACTCTCTTCGTGCTGAAGATACTGCTGTT TATTATTGTTCTCGTTGGGGTGGTGATGGTTTTTATGCTATGGATTATTGGGGTCAAG GTACTCTTGTCACCGTCTCCTCAGCTAGCACCGGGGGAGGTGGGTCTGGAGGTGGAG GATCTGGTGGAGGTGGGTCTGACATCCAGATGACCCAGTCTCCTTCTTCTCTTTCTGC TTCTGTTGGTGATCGTGTTACTATTACTTGTCGTGCTTCTCAAGATGTTAATACTGCT GTTGCTTGGTATCAACAAAAACCTGGTAAAGCTCCTAAACTTCTTATTTATTCTGCTT CTTTTCTTTATTCTGGTGTTCCTTCTCGTTTTTCTGGTTCTCGTTCTGGTACTGATTTTA CTCTTACTATTTCTTCTCTTCAACCTGAAGATTTTGCTACTTATTATTGTCAACAACAT TATACTACTCCTCCTACTTTTGGTCAAGGTACCAAGGTGGAGATCAAACGTACGGGA GGAGGTGGGTCTGGAGGTGGAGGATCTGGTGGAGGTGGGTCTGGAGGAGGTGGATC CCCCGAGGAACCTCTAGTGGTGAAGGTGGAAGAGGGAGATACCGCTGCCCTGTGGT GCCTCAAGGGGACCTCAGATGGCCCCACTCAGCAGCTGACCTGGTCTCGGGAGTCC CCGCTTAAACCCTTCTTAAAATACAGCCTGGGGGTGCCAGGCCTGGGAGTGCACGTG AGGCCCGACGCCATCAGCGTGGTTATCCGGAACGTCTCTCAACAGATGGGGGGCTT CTACCTGTGCCAGCCGGGGCCCCCCTCTGAGAAGGCCTGGCAGCCTGGCTGGACAG TCAATGTGGAGGGCAGCGGGGAGCTGTTCCGGTGGAATGTTTCGGACCTAGGTGGC CTGGGCTGTGGCCTGAAGAACAGGTCCTCAGAGGGCCCCAGCTCCCCTTCCGGGAA GCTCATGAGCCCCAAGCTGTATGTGTGGGCCAAAGACCGCCCTGAGATCTGGGAGG GAGAGCCTCCGTGTCTCCCACCGAGGGACAGCCTGAACCAGAGCCTCAGCCGGGAC CTCACCGTTGCCCCTGGCTCCACACTCTGGCTGTCCTGTGGGGTACCCCCTGACTCTG TGTCCAGGGGCCCCCTCTCCTGGACCCATGTGCACCCCAAGGGGCCTAAGTCATTGC TGAGCCTAGAGCTGAAGGACGATCGCCCGGCCAGAGATATGTGGGTAATGGGCACG AGCCTGATGTTGCCCCGGGCCACAGCTCAAGACGCTGGAAAGTGGTATTGTCACCGT GGCAACCTGACCATGTCATTCCACCTGGAGATCACTGCTCGGCCATCTAGACATCAT CACCATCACCATAmino acid sequence SEQ ID NO: 48- 151 - 13321680vlAttorney Docket No. 2012106-0161

[0403] MEFGLSWVFLVALFRGVOCFSOQIYGVVYGNVTFHVPSNVPLKEVLWKK QKDKVAELENSEFRAFSSFKNRVYLDTVSGSLT1YNLTSSDEDEYEMESPN1TDTMKFFL VLGGGGSGGGGSGGGGSGGGGSEVQLVESGGGLVQPGGSLRLSCAASGFNIKDTYIHWV RQAPGKGEEWVAR1YPTNGYTRYADSFKGRFTISADTSKNTA YLQMNSLRAEDTA VYYCSRWG GDGFYAMDYWGQGTLVTVSSASTGGGGSGGGGSGGGGSDIQMTQSPSSLSASVGDRVT1TCR ASQDVNTA VA WYQQKPGKAPKLLIYSASFLYSGVPSRFSGSRSGTDFTLTISSLQPEDFA TYYC OOHYTTPPTFGQGTKVEIKRTGGGGSGGGGSGGGGSGGGGSYEEPENVEVEEGDEAAE CLKGTSDGPTOQLTWSRESPLKPFLKYSLGVPGLGVHVRPDAISVVIRNVSOOMGGFYL COPGPPSEKAWQPGWTVNVEGSGELFRWNVSDLGGLGCGLKNRSSEGPSSPSGKLMSP KLYVWAKDRPEIWEGEPPCLPPRDSLNQSLSRDLTVAPGSTLWLSCGVPPDSVSRGPLS WTHVHPKGPKSLLSLELKDDRPARDMWVMGTSLMLPRATAQDAGKWYCHRGNLTMS FHLEIT ARP SRHHHHHHAleta #680Nucleotide sequence SEQ ID NO: 49

[0404] ATGGAGTTTGGGCTGAGCTGGGTTTTCCTCGTTGCTCTTTTTAGAGGTG TCCAGTGTTTTAGCCAGCAGATCTATGGCGTGGTGTACGGCAACGTGACCTTCCACG TGCCTAGCAACGTCCCCCTGAAAGAGGTGCTCTGGAAGAAGCAGAAGGACAAGGTG GCCGAGCTGGAAAACAGCGAGTTCCGGGCCTTCAGCAGCTTTAAGAACAGAGTGTA CCTGGACACAGTGTCCGGCAGCCTGACAATCTACAACCTGACCAGCTCTGATGAAG ATGAGTACGAGATGGAAAGCCCTAATATCACCGACACCATGAAATTCTTCCTGTACG TGCTGGGCGGAGGCGGCAGCGGCGGCGGCGGCTCCGGCGGAGGCGGATCTGGCGG AGGCGGCTCTGAGGTGCAGCTGGTGGAGTCCGGAGGAGGCCTGGTGCAGCCAGGAG GCTCTCTGAGGCTGAGCTGCACCTTCTCCGGCGGCACCTTCAGCAGCTACACAATGG GCTGGTTCAGGCAGGCACCAGGCAAGGAGAGAGAGTTTGTGGCAGAGGTGAGGTGG GGAGGAGTGACCACATACTCCAACTCTCTGAAGGACCGCTTCAGCATCTCCGAGGA TTCTGTGAAGAACGCCGTGTATCTGCAGATGAATAGCCTGAAGCCCGAGGACACAG CCGTGTACTATTGTGCCGCCGTGCGGCAGATGTACATGACCGTGGTGCCTGATTATT GGGGCCAGGGCACCCTGGTGACAGTGTCTAGCGGAGGAGGTGGGTCTGGAGGTGGA- 152 - 13321680vlAttorney Docket No. 2012106-0161GGATCTGGTGGAGGTGGGTCTGGAGGAGGTGGATCCCCCGAGGAACCTCTAGTGGT GAAGGTGGAAGAGGGAGATACCGCTGCCCTGTGGTGCCTCAAGGGGACCTCAGATG GCCCCACTCAGCAGCTGACCTGGTCTCGGGAGTCCCCGCTTAAACCCTTCTTAAAAT ACAGCCTGGGGGTGCCAGGCCTGGGAGTGCACGTGAGGCCCGACGCCATCAGCGTG GTTATCCGGAACGTCTCTCAACAGATGGGGGGCTTCTACCTGTGCCAGCCGGGGCCC CCCTCTGAGAAGGCCTGGCAGCCTGGCTGGACAGTCAATGTGGAGGGCAGCGGGGA GCTGTTCCGGTGGAATGTTTCGGACCTAGGTGGCCTGGGCTGTGGCCTGAAGAACAG GTCCTCAGAGGGCCCCAGCTCCCCTTCCGGGAAGCTCATGAGCCCCAAGCTGTATGT GTGGGCCAAAGACCGCCCTGAGATCTGGGAGGGAGAGCCTCCGTGTCTCCCACCGA GGGACAGCCTGAACCAGAGCCTCAGCCGGGACCTCACCGTTGCCCCTGGCTCCACA CTCTGGCTGTCCTGTGGGGTACCCCCTGACTCTGTGTCCAGGGGCCCCCTCTCCTGG ACCCATGTGCACCCCAAGGGGCCTAAGTCATTGCTGAGCCTAGAGCTGAAGGACGA TCGCCCGGCCAGAGATATGTGGGTAATGGGCACGAGCCTGATGTTGCCCCGGGCCA CAGCTCAAGACGCTGGAAAGTGGTATTGTCACCGTGGCAACCTGACCATGTCATTCC ACCTGGAGATCACTGCTCGGCCATCTAGAGGAGGCGGAGGAAGTGGAGGAGGAGG GAGCGGGGGGGGGGGAAGTGAAGTTCAGTTGGTCGAATCTGGAGGGGGACTGGTAC AGCCTGGCAACTCTCTGCGATTGTCTTGTGCTGCTAGTGGTTTTACATTTTCTTCATTT GGCATGAGTTGGGTCCGACAAGCCCCAGGAAAAGGGTTGGAATGGGTGAGCAGCAT TTCTGGAAGCGGTTCTGATACACTCTACGCCGACAGCGTAAAAGGCCGGTTCACTAT ATCTCGCGATAATGCAAAAACAACACTTTATCTGCAAATGAATAGTCTGCGACCTGA AGATACAGCAGTATATTATTGTACAATAGGAGGTAGTCTCAGTAGGTCATCACAAG GAACACTGGTAACCGTGAGTTCCGGCGGAGGCGGCTCCGGCGGCGGCGGCAGCGGA GGCGGCGGCTCTGGCGGAGGCGGATCTTTTAGCCAGCAGATCTATGGCGTGGTCTAC GGCAACGTGACCTTCCACGTGCCCAGCAACGTGCCTCTGAAGGAAGTGCTCTGGAA GAAACAGAAGGACAAGGTGGCCGAGCTGGAAAACAGCGAGTTCCGGGCCTTCAGC AGCTTTAAGAACAGAGTGTACCTGGACACCGTGTCCGGCAGCCTGACAATCTACAA CCTGACCAGCTCTGATGAGGACGAGTACGAGATGGAAAGCCCTAATATCACCGATA CAATGAAATTCTTCCTGTACGTGCTGCATCATCACCATCACCAT- 153 - 13321680vlAttorney Docket No. 2012106-0161Amino acid sequence SEQ ID NO: 50

[0405] MEFGLSWVFLVALFRGVOCFSOQIYGVVYGNVTFHVPSNVPLKEVLWKK OKDKVAELENSEFRAFSSFKNRVYLDTVSGSLTIYNLTSSDEDEYEMESPNITDTMKFFL VLGGGGSGGGGSGGGGSGGGGSEVQLFESGGGLVQPGGSLRLSCTFSGGTFSSYTMGWF RQAPGKEREFVAEVRWGGVTTYSNSLKDRFSISEDSVKNA VYLQMNSLKPEDTA VYYCAA VRQ M TOFEFGOGZZF YGGGGSGGGGSGGGGSGGGGSPEEPLVVKVEEGDTAALW CLKGTSDGPTOQLTWSRESPLKPFLKYSLGVPGLGVHVRPDAISVVIRNVSOOMGGFYL CQPGPPSEKAWQPGWTVNVEGSGELFRWNVSDLGGLGCGLKNRSSEGPSSPSGKLMSP KLYVWAKDRPEIWEGEPPCLPPRDSLNQSLSRDLTVAPGSTLWLSCGVPPDSVSRGPLS WTHVHPKGPKSLLSLELKDDRPARDMWVMGTSLMLPRATAQDAGKWYCHRGNLTMS YYEYANRPSYiGGGGSGGGGSGGGGSEVOLVESGGGLVOPGNSLRLSCAASGFTFSSFGMS WVRQAPGKGLEWVSSISGSGSDTLYADSVKGRFTISRDNAKTTLYLQMNSLRPEDTAVYYCTIGG5Z5A55GGTEJTFA5GGGGSGGGGSGGGGSGGGGSFSOOIYGVVYGNVTFHVPSNVPLK EVLWKKQKDKVAELENSEFRAFSSFKNRVYLDTVSGSLTIYNLTSSDEDEYEMESPNITD TMKFFLYVLHHHHHHAleta #681Nucleotide sequence SEQ ID NO: 51

[0406] ATGGAGTTTGGGCTGAGCTGGGTTTTCCTCGTTGCTCTTTTTAGAGGTG TCCAGTGTTTTAGCCAGCAGATCTATGGCGTGGTGTACGGCAACGTGACCTTCCACG TGCCTAGCAACGTCCCCCTGAAAGAGGTGCTCTGGAAGAAGCAGAAGGACAAGGTG GCCGAGCTGGAAAACAGCGAGTTCCGGGCCTTCAGCAGCTTTAAGAACAGAGTGTA CCTGGACACAGTGTCCGGCAGCCTGACAATCTACAACCTGACCAGCTCTGATGAAG ATGAGTACGAGATGGAAAGCCCTAATATCACCGACACCATGAAATTCTTCCTGTACG TGCTGGGCGGAGGCGGCTCCGGCGGCGGCGGCAGCGGAGGCGGCGGCTCTGGCGG AGGCGGATCTTTTAGCCAGCAGATCTATGGCGTGGTCTACGGCAACGTGACCTTCCA CGTGCCCAGCAACGTGCCTCTGAAGGAAGTGCTCTGGAAGAAACAGAAGGACAAGG TGGCCGAGCTGGAAAACAGCGAGTTCCGGGCCTTCAGCAGCTTTAAGAACAGAGTG- 154 - 13321680vlAttorney Docket No. 2012106-0161TACCTGGACACCGTGTCCGGCAGCCTGACAATCTACAACCTGACCAGCTCTGATGAG GACGAGTACGAGATGGAAAGCCCTAATATCACCGATACAATGAAATTCTTCCTGTAC GTGCTGGGCGGAGGCGGCAGCGGCGGCGGCGGCTCCGGCGGAGGCGGATCTGGCG GAGGCGGCTCTGAGGTGCAGCTGGTGGAGTCCGGAGGAGGCCTGGTGCAGCCAGGA GGCTCTCTGAGGCTGAGCTGCACCTTCTCCGGCGGCACCTTCAGCAGCTACACAATG GGCTGGTTCAGGCAGGCACCAGGCAAGGAGAGAGAGTTTGTGGCAGAGGTGAGGTG GGGAGGAGTGACCACATACTCCAACTCTCTGAAGGACCGCTTCAGCATCTCCGAGG ATTCTGTGAAGAACGCCGTGTATCTGCAGATGAATAGCCTGAAGCCCGAGGACACA GCCGTGTACTATTGTGCCGCCGTGCGGCAGATGTACATGACCGTGGTGCCTGATTAT TGGGGCCAGGGCACCCTGGTGACAGTGTCTAGCGGAGGAGGTGGGTCTGGAGGTGG AGGATCTGGTGGAGGTGGGTCTGGAGGAGGTGGATCCCCCGAGGAACCTCTAGTGG TGAAGGTGGAAGAGGGAGATACCGCTGCCCTGTGGTGCCTCAAGGGGACCTCAGAT GGCCCCACTCAGCAGCTGACCTGGTCTCGGGAGTCCCCGCTTAAACCCTTCTTAAAA TACAGCCTGGGGGTGCCAGGCCTGGGAGTGCACGTGAGGCCCGACGCCATCAGCGT GGTTATCCGGAACGTCTCTCAACAGATGGGGGGCTTCTACCTGTGCCAGCCGGGGCC CCCCTCTGAGAAGGCCTGGCAGCCTGGCTGGACAGTCAATGTGGAGGGCAGCGGGG AGCTGTTCCGGTGGAATGTTTCGGACCTAGGTGGCCTGGGCTGTGGCCTGAAGAACA GGTCCTCAGAGGGCCCCAGCTCCCCTTCCGGGAAGCTCATGAGCCCCAAGCTGTATG TGTGGGCCAAAGACCGCCCTGAGATCTGGGAGGGAGAGCCTCCGTGTCTCCCACCG AGGGACAGCCTGAACCAGAGCCTCAGCCGGGACCTCACCGTTGCCCCTGGCTCCAC ACTCTGGCTGTCCTGTGGGGTACCCCCTGACTCTGTGTCCAGGGGCCCCCTCTCCTG GACCCATGTGCACCCCAAGGGGCCTAAGTCATTGCTGAGCCTAGAGCTGAAGGACG ATCGCCCGGCCAGAGATATGTGGGTAATGGGCACGAGCCTGATGTTGCCCCGGGCC ACAGCTCAAGACGCTGGAAAGTGGTATTGTCACCGTGGCAACCTGACCATGTCATTC CACCTGGAGATCACTGCTCGGCCATCTAGAGGAGGCGGAGGAAGTGGAGGAGGAG GGAGCGGGGGGGGGGGAAGTGAAGTTCAGTTGGTCGAATCTGGAGGGGGACTGGT ACAGCCTGGCAACTCTCTGCGATTGTCTTGTGCTGCTAGTGGTTTTACATTTTCTTCA TTTGGCATGAGTTGGGTCCGACAAGCCCCAGGAAAAGGGTTGGAATGGGTGAGCAG CATTTCTGGAAGCGGTTCTGATACACTCTACGCCGACAGCGTAAAAGGCCGGTTCAC TATATCTCGCGATAATGCAAAAACAACACTTTATCTGCAAATGAATAGTCTGCGACC- 155 - 13321680vlAttorney Docket No. 2012106-0161TGAAGATACAGCAGTATATTATTGTACAATAGGAGGTAGTCTCAGTAGGTCATCACA AGGAACACTGGTAACCGTGAGTTCCCATCATCACCATCACCATAmino acid sequence SEQ ID NO: 52

[0407] MEFGLSWVFLVALFRGy. QCFSQQIYGVVYGNVTFHVPSNVPLKEVLWKK QKDKVAELENSEFRAFSSFKNRVYLDTVSGSLTIYNLTSSDEDEYEMESPNITDTMKFFL YVLGGGGSGGGGSGGGGSGGGGSFSQQIYGVVYGNVTFHVPSNVPLKEVLWKKQKDK VAELENSEFRAFSSFKNRVYLDTVSGSLTIYNLTSSDEDEYEMESPNITDTMKFFLYVLG GGGSGGGGSGGGGSGGGGSEVQLVESGGGLVQPGGSLRLSCTFSGGTFSSYTMGWFRQAP GKEREFVAEVRWGGVTTYSNSLKDRFSISEDSVKNA VYLQMNSLKPEDTA VYYCAA VRQMYM TOF gG YyS GGGGSGGGGSGGGGSGGGGSPEEPLVVKVEEGDTAALWCLK GTSDGPTQQLTWSRESPLKPFLKYSLGVPGLGVHVRPDAISVVIRNVSQQMGGFYLCQP GPPSEKAWQPGWTVNVEGSGELFRWNVSDLGGLGCGLKNRSSEGPSSPSGKLMSPKLY VWAKDRPEIWEGEPPCLPPRD SLNQ SLSRDLT VAPGSTLWLSC GVPPD S VSRGPL S WTH VHPKGPKSLLSLELKDDRPARDMWVMGTSLMLPRATAQDAGKWYCHRGNLTMSFHL EITARPSRGGGGSGGGGSGGGGSEFgZ VESGGGL VQPGNSLRLSCAASGFTFSSFGMSWVR QAPGKGLEm7SSISGSGSDTLYADSVKGRFTISRDNAKTTLYLQMNSLRPEDTAmCTIGGSL SRSSQGTL E7TYSHHHHHHAleta #686Nucleotide sequence SEQ ID NO: 53

[0408] ATGGAGTTTGGGCTGAGCTGGGTTTTCCTCGTTGCTCTTTTTAGAGGTG TCCAGTGTGAGGTGCAGCTGGTGGAGTCCGGAGGAGGCCTGGTGCAGCCAGGAGGC TCTCTGAGGCTGAGCTGCACCTTCTCCGGCGGCACCTTCAGCAGCTACACAATGGGC TGGTTCAGGCAGGCACCAGGCAAGGAGAGAGAGTTTGTGGCAGAGGTGAGGTGGG GAGGAGTGACCACATACTCCAACTCTCTGAAGGACCGCTTCAGCATCTCCGAGGATT CTGTGAAGAACGCCGTGTATCTGCAGATGAATAGCCTGAAGCCCGAGGACACAGCC GTGTACTATTGTGCCGCCGTGCGGCAGATGTACATGACCGTGGTGCCTGATTATTGG- 156 - 13321680vlAttorney Docket No. 2012106-0161GGCCAGGGCACCCTGGTGACAGTGTCTAGCGGAGGAGGTGGGTCTGGAGGTGGAGG ATCTGGTGGAGGTGGGTCTGGAGGAGGTGGATCCCCCGAGGAACCTCTAGTGGTGA AGGTGGAAGAGGGAGATACCGCTGCCCTGTGGTGCCTCAAGGGGACCTCAGATGGC CCCACTCAGCAGCTGACCTGGTCTCGGGAGTCCCCGCTTAAACCCTTCTTAAAATAC AGCCTGGGGGTGCCAGGCCTGGGAGTGCACGTGAGGCCCGACGCCATCAGCGTGGT TATCCGGAACGTCTCTCAACAGATGGGGGGCTTCTACCTGTGCCAGCCGGGGCCCCC CTCTGAGAAGGCCTGGCAGCCTGGCTGGACAGTCAATGTGGAGGGCAGCGGGGAGC TGTTCCGGTGGAATGTTTCGGACCTAGGTGGCCTGGGCTGTGGCCTGAAGAACAGGT CCTCAGAGGGCCCCAGCTCCCCTTCCGGGAAGCTCATGAGCCCCAAGCTGTATGTGT GGGCCAAAGACCGCCCTGAGATCTGGGAGGGAGAGCCTCCGTGTCTCCCACCGAGG GACAGCCTGAACCAGAGCCTCAGCCGGGACCTCACCGTTGCCCCTGGCTCCACACTC TGGCTGTCCTGTGGGGTACCCCCTGACTCTGTGTCCAGGGGCCCCCTCTCCTGGACC CATGTGCACCCCAAGGGGCCTAAGTCATTGCTGAGCCTAGAGCTGAAGGACGATCG CCCGGCCAGAGATATGTGGGTAATGGGCACGAGCCTGATGTTGCCCCGGGCCACAG CTCAAGACGCTGGAAAGTGGTATTGTCACCGTGGCAACCTGACCATGTCATTCCACC TGGAGATCACTGCTCGGCCATCTAGAGGAGGCGGAGGAAGTGGAGGAGGAGGGAG CGGGGGGGGGGGAAGTGAAGTTCAGTTGGTCGAATCTGGAGGGGGACTGGTACAGC CTGGCAACTCTCTGCGATTGTCTTGTGCTGCTAGTGGTTTTACATTTTCTTCATTTGGC ATGAGTTGGGTCCGACAAGCCCCAGGAAAAGGGTTGGAATGGGTGAGCAGCATTTC TGGAAGCGGTTCTGATACACTCTACGCCGACAGCGTAAAAGGCCGGTTCACTATATC TCGCGATAATGCAAAAACAACACTTTATCTGCAAATGAATAGTCTGCGACCTGAAG ATACAGCAGTATATTATTGTACAATAGGAGGTAGTCTCAGTAGGTCATCACAAGGA ACACTGGTAACCGTGAGTTCCGGCGGAGGCGGCTCCGGCGGCGGCGGCAGCGGAGG CGGCGGCTCTGGCGGAGGCGGATCTTTTAGCCAGCAGATCTATGGCGTGGTCTACGG CAACGTGACCTTCCACGTGCCCAGCAACGTGCCTCTGAAGGAAGTGCTCTGGAAGA AACAGAAGGACAAGGTGGCCGAGCTGGAAAACAGCGAGTTCCGGGCCTTCAGCAG CTTTAAGAACAGAGTGTACCTGGACACCGTGTCCGGCAGCCTGACAATCTACAACCT GACCAGCTCTGATGAGGACGAGTACGAGATGGAAAGCCCTAATATCACCGATACAA TGAAATTCTTCCTGTACGTGCTGCATCATCACCATCACCAT- 157 - 13321680vlAttorney Docket No. 2012106-0161Amino acid sequence SEQ ID NO: 54

[0409] MEYGLS yYLy AFRGyQCEVQLVESGGGLVQPGGSLRLSCTFSGGTFS SYTMGWFRQAPGKEREFVAEVRWGGVTTYSNSLKDRFSISEDSVKNAVYLQMNSLKPE DTA VYYCAA VRQMYMTWPDYWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSPEEPE VVKVEEGDTAALWCLKGTSDGPTOQLTWSRESPLKPFLKYSLGVPGLGVHVRPDAISV VIRNVSOOMGGFYLCOPGPPSEKAWQPGWTVNVEGSGELFRWNVSDLGGLGCGLKNR S SEGP S SP SGKLMSPKL YVWAKDRPEIWEGEPPCLPPRD SLNQ SLSRDLT VAPGSTLWLS CGVPPDSVSRGPLSWTHVHPKGPKSLLSLELKDDRPARDMWVMGTSLMLPRATAODA GKWYCHRGNLTMSFHLEITARPSRGGGGSGGGGSGGGGSEFQZ VESGGGL VQPGNSLRL SCAASGFTFSSFGMSWVRQAPGKGLEWVSSISGSGSDTLYADSVKGRFTISRDNAKTTLYLQM NSLRPEDTAVYYCnGGSLSRSSOGTL JVSSGGGGSGGGGSGGGGSGGGGSYSOOTYGVV YGNVTFHVPSNVPLKEVLWKKQKDKVAELENSEFRAFSSFKNRVYLDTVSGSLTIY NLTSSDEDEYEMESPNITDTMKFFLYVLHHHHHHAleta #692Nucleotide sequence SEQ ID NO: 55

[0410] ATGGAGTTTGGGCTGAGCTGGGTTTTCCTCGTTGCTCTTTTTAGAGGTG TCCAGTGTGAGGTGCAGCTGGTGGAGTCCGGAGGAGGCCTGGTGCAGCCAGGAGGC TCTCTGAGGCTGAGCTGCACCTTCTCCGGCGGCACCTTCAGCAGCTACACAATGGGC TGGTTCAGGCAGGCACCAGGCAAGGAGAGAGAGTTTGTGGCAGAGGTGAGGTGGG GAGGAGTGACCACATACTCCAACTCTCTGAAGGACCGCTTCAGCATCTCCGAGGATT CTGTGAAGAACGCCGTGTATCTGCAGATGAATAGCCTGAAGCCCGAGGACACAGCC GTGTACTATTGTGCCGCCGTGCGGCAGATGTACATGACCGTGGTGCCTGATTATTGG GGCCAGGGCACCCTGGTGACAGTGTCTAGCGGCGGAGGTGGCTCTGGAGGCGGTGG ATCTGGTGGTGGCGGATCAGGTGGAGGCGGGAGCTTTTCTCAGCAGATCTATGGCGT GGTCTACGGCAACGTTACCTTCCACGTGCCTAGCAACGTGCCTCTGAAGGAAGTGCT GTGGAAGAAGCAGAAAGACAAGGTGGCCGAGCTGGAAAACAGCGAGTTCCGGGCC TTCAGCAGCTTTAAGAATAGAGTGTACCTGGACACCGTGTCCGGAAGCCTGACAATC- 158 - 13321680vlAttorney Docket No. 2012106-0161TACAACCTGACAAGCAGCGACGAGGATGAGTACGAGATGGAAAGCCCCAACATCAC CGATACCATGAAATTCTTCCTGTACGTGCTCGGAGGAGGTGGGTCTGGAGGTGGAG GATCTGGTGGAGGTGGGTCTGGAGGAGGTGGATCCCCCGAGGAACCTCTAGTGGTG AAGGTGGAAGAGGGAGATACCGCTGCCCTGTGGTGCCTCAAGGGGACCTCAGATGG CCCCACTCAGCAGCTGACCTGGTCTCGGGAGTCCCCGCTTAAACCCTTCTTAAAATA CAGCCTGGGGGTGCCAGGCCTGGGAGTGCACGTGAGGCCCGACGCCATCAGCGTGG TTATCCGGAACGTCTCTCAACAGATGGGGGGCTTCTACCTGTGCCAGCCGGGGCCCC CCTCTGAGAAGGCCTGGCAGCCTGGCTGGACAGTCAATGTGGAGGGCAGCGGGGAG CTGTTCCGGTGGAATGTTTCGGACCTAGGTGGCCTGGGCTGTGGCCTGAAGAACAGG TCCTCAGAGGGCCCCAGCTCCCCTTCCGGGAAGCTCATGAGCCCCAAGCTGTATGTG TGGGCCAAAGACCGCCCTGAGATCTGGGAGGGAGAGCCTCCGTGTCTCCCACCGAG GGACAGCCTGAACCAGAGCCTCAGCCGGGACCTCACCGTTGCCCCTGGCTCCACACT CTGGCTGTCCTGTGGGGTACCCCCTGACTCTGTGTCCAGGGGCCCCCTCTCCTGGAC CCATGTGCACCCCAAGGGGCCTAAGTCATTGCTGAGCCTAGAGCTGAAGGACGATC GCCCGGCCAGAGATATGTGGGTAATGGGCACGAGCCTGATGTTGCCCCGGGCCACA GCTCAAGACGCTGGAAAGTGGTATTGTCACCGTGGCAACCTGACCATGTCATTCCAC CTGGAGATCACTGCTCGGCCATCTAGAGGAGGCGGAGGAAGTGGAGGAGGAGGGA GCGGGGGGGGGGGAAGTGAAGTTCAGTTGGTCGAATCTGGAGGGGGACTGGTACAG CCTGGCAACTCTCTGCGATTGTCTTGTGCTGCTAGTGGTTTTACATTTTCTTCATTTGG CATGAGTTGGGTCCGACAAGCCCCAGGAAAAGGGTTGGAATGGGTGAGCAGCATTT CTGGAAGCGGTTCTGATACACTCTACGCCGACAGCGTAAAAGGCCGGTTCACTATAT CTCGCGATAATGCAAAAACAACACTTTATCTGCAAATGAATAGTCTGCGACCTGAAG ATACAGCAGTATATTATTGTACAATAGGAGGTAGTCTCAGTAGGTCATCACAAGGA ACACTGGTAACCGTGAGTTCCGGCGGAGGCGGCTCCGGCGGCGGCGGCAGCGGAGG CGGCGGCTCTGGCGGAGGCGGATCTTTTAGCCAGCAGATCTATGGCGTGGTCTACGG CAACGTGACCTTCCACGTGCCCAGCAACGTGCCTCTGAAGGAAGTGCTCTGGAAGA AACAGAAGGACAAGGTGGCCGAGCTGGAAAACAGCGAGTTCCGGGCCTTCAGCAG CTTTAAGAACAGAGTGTACCTGGACACCGTGTCCGGCAGCCTGACAA...

Claims

Attorney Docket No. 2012106-0161CLAIMSWhat is claimed is:

1. A fusion protein comprising:(i) one or more accessory molecule(s) or a fragment thereof;(ii) one or more antigen-binding proteins or fragments that bind a tumor antigen.

2. The fusion protein of claim 1, wherein the accessory molecule or fragment thereof is or is a binding partner of an adhesion molecule, a costimulatory molecule, or a combination thereof.

3. The fusion protein of claim 2, wherein the accessory molecule is or comprises an antigen binding protein or fragment that binds an adhesion molecule, a costimulatory molecule, or a combination thereof.

4. The fusion protein of claim 3, wherein the antigen binding protein or fragment is an scFv, Fv, or VHH.

5. The fusion protein of claims 1-4, comprising two or more accessory molecule(s) or a fragment thereof.

6. The fusion protein of claims 1-5, wherein the accessory molecule is or is a binding partner of L-selectin, a4 integrin, lymphocyte function-associated antigen- 1 (LFA-1), Intercellular Adhesion Molecule 1 (1CAM-1; CD54), CD2, CD3, CD28, B7-1 (CD80), B7-2 (CD86) or LFA-3 (CD58).

7. The fusion protein of claims 1-6, wherein the fusion protein comprises at least one accessory molecule that is an antigen binding protein or fragment that binds CD3.

8. The fusion protein of claims 1-7, wherein the fusion protein comprises two LFA-3 binding domains.

9. The fusion protein of claims 1-7, wherein the fusion protein comprises B7-2 and one LFA-3 binding domain.

10. The fusion protein of claim 1, wherein the tumor antigen is a tumor specific antigen (TSA) or a tumor associated antigen (TAA).

11. The fusion protein of claim 10, wherein the tumor antigen is MART- 1 / Mel anA (MART-I), gplOO (Pmel 17), tyrosinase, TRP-1, TRP-2, MAGE-1, MAGE-3, BAGE, GAGE-1, GAGE-2, pl 5, CEA, p53, Ras, HER-2 / neu, BCR-ABL, E2A-PRL, H4-RET, IGH-IGK, MYL-RAR, Epstein Barr virus antigens EBVA, human papillomavirus (HPV) antigen E6 or E7, TSP-180, MAGE-4, MAGE-5, MAGE-6, RAGE, NY-ESO, erbB, pl85erbB2, pl80erbB-3, c-met, nm-23H1, PSA, TAG-72, CA 19-9, CA 72-4, CAM 17.1, NuMa, K-ras, beta-Catenin, CDK4, Mum- - 223 - 13321680vlAttorney Docket No. 2012106-01611, p 15, p 16, 43-9F, 5T4, 791Tgp72, alpha-fetoprotein, beta-HCG, BCA225, BTAA, CA 125, CA 15-3\CA 27.29\BCAA, CA 195, CA 242, CA-50, CAM43, CD68VP1, CO-029, FGF-5, G250, Ga733\EpCAM, HTgp-175, M344, MA-50, MG7-Ag, M0V18, NB / 70K, NY-CO-1, RCAS1, SDCCAG16, TA-90\Mac-2 binding protein\cyclophilin C-associated protein, TAAL6, TAG72, TLP, MUC16, IL13Ra2, FRa, VEGFR2, Lewis Y, FAP, EphA2, CEACAM5, EGFR, CA6, CA9, GPNMB, EGP1, F0LR1, endothelial receptor, STEAP1, SLC44A4, Nectin-4, AGS-16, guanalyl cyclase C, MUC-1, CFC1B, integrin alpha 3 chain (of a3bl, a laminin receptor chain), TPS, CD19, CD20, CD22, CD30, CD72, CD180, CD171 (L1CAM), CD123, CD133, CD138, CD37, CD70, CD79a, CD79b, CD56, CD74, CD166, CD71, CLL-1 / CLEC12A, R0R1, Glypican 3 (GPC3), Mesothelin, CD33 / IL3Ra, c-Met, PSCA, PSMA, Glycolipid F77, EGFRvIII, BCMA, GD-2, MY-ESO-1, B7-H3 (CD276) or MAGE A3.

12. The fusion protein of claim 11, wherein the tumor antigen is CD19, CD20, B7-H3, HER-2 or IL-13Ra2.

13. The fusion protein of claim 1, wherein the antigen-binding protein or fragment that binds a tumor antigen is an scFv, Fv, or VHH.

14. The fusion protein of claim 1, wherein the fusion protein comprises two or more antigenbinding proteins or fragments that bind a tumor antigen.

15. The fusion protein of claims 1-14, wherein the fusion protein comprises an anti-B7-H3 VHH and an anti-IL-13Ra2 VHH.

16. The fusion protein of claims 1-14, wherein the fusion protein comprises an anti-B7-H3 VHH and an anti-HER2-scFv.

17. The fusion protein of claims 1-16, wherein the fusion protein further comprises a target polypeptide for a cellular therapeutic.

18. The fusion protein of claim 17, wherein the target polypeptide is a tumor antigen or fragment thereof.

19. The fusion protein of claim 18, wherein the tumor antigen is or is a fragment of MART-1 / MelanA (MART-I), gplOO (Pmel 17), tyrosinase, TRP-1, TRP-2, MAGE-1, MAGE-3, BAGE, GAGE-1, GAGE-2, pl5, CEA, p53, Ras, HER-2 / neu, BCR-ABL, E2A-PRL, H4-RET, IGH-IGK, MYL-RAR, Epstein Barr virus antigens EBVA, human papillomavirus (HPV) antigen E6 orE7, TSP-180, MAGE-4, MAGE-5, MAGE-6, RAGE, NY-ESO, erbB, pl85erbB2, pl80erbB-3, c-met, nm-23Hl, PSA, TAG-72, CA 19-9, CA 72-4, CAM 17.1, NuMa, K-ras, beta-Catenin, CDK4, Mum-1, p 15, p 16, 43-9F, 5T4, 791Tgp72, alpha-fetoprotein, beta-HCG, BCA225, BTAA, CA 125, CA 15-3\CA 27.29\BCAA, CA 195, CA 242, CA-50, CAM43, CD68\P1, CO-029, FGF-5, G250, Ga733\EpCAM, HTgp-175, M344, MA-50, MG7-Ag, M0V18, NB / 70K, NY-CO-1, RCAS1, SDCCAG16, TA-90\Mac-2 binding protein\cyclophilin C-associated protein, TAAL6, TAG72, TLP, MUC16, IL13Ra2, FRa, VEGFR2, Lewis Y, FAP, EphA2, CEACAM5, EGFR, CA6, CA9, GPNMB, EGP1, FOLR1, endothelial receptor, STEAP1, SLC44A4, Nectin-4, AGS-16, guanalyl cyclase C, MUC-1, CFC1B, integrin alpha 3 chain (of - 224 - 13321680vlAttorney Docket No. 2012106-0161a3bl, a laminin receptor chain), TPS, CD 19, CD20, CD22, CD30, CD72, CD 180, CD171 (L1CAM), CD123, CD133, CD138, CD37, CD70, CD79a, CD79b, CD56, CD74, CD166, CD71, CLL-1 / CLEC12A, R0R1, Glypican 3 (GPC3), Mesothelin, CD33 / IL3Ra, c-Met, PSCA, PSMA, Glycolipid F77, EGFRvIII, BCMA, GD-2, MY-ESO-1, B7-H3 (CD276) or MAGE A3.

20. The fusion protein of claim 19, wherein the tumor antigen is or is a fragment of CD20, CD 19, or BCMA.

21. The fusion protein of claim 17, wherein the cellular therapeutic is a CAR-T cell, CAR-NK cell, TCR-T cell, TIL cell, allogenic NK cell, or autologous NK cell.

22. The fusion protein of any of claims 1-21, further comprising a half-life extension polypeptide.

23. The fusion protein of claim 22, wherein the half-life extension polypeptide is any one of a hyaluronan binding motif, PAS polypeptide, proline / alanine random coil polypeptide, and an anti-albumin antibody or antigen binding fragment thereof.

24. The fusion protein of claim 23, wherein the half-life extension polypeptide is an anti-albumin antibody or antigen binding fragment thereof.

25. The fusion protein of claim 24, wherein the anti-albumin antibody or antigen binding fragment thereof comprises an anti -albumin VHH.

26. The fusion protein of any of claims 1-25, further comprising an immune suppression blockade domain.

27. The fusion protein of claim 26, wherein the immune suppression blockade domain is a receptor for an immune suppression molecule.

28. The fusion protein of claim 27, wherein the immune suppression blockade domain is TGF0 receptor 2 ECD (TGFβR2 ECD).

29. A nucleic acid encoding the fusion protein of any of the preceding claims.

30. A vector comprising the nucleic acid of claim 29.

31. An immune cell comprising an expression construct encoding a fusion protein of any of the preceding claims.

32. The immune cell of claim 31, wherein the fusion protein is secreted from the immune cell.

33. A method of treating a subject having a tumor, comprising administering to the subject an effective amount of the fusion protein of any one of claims 1-28, thereby treating the subject.

34. The method of claim 33, wherein the tumor expresses the tumor antigen.- 225 - 13321680vlAttorney Docket No. 2012106-016135. The method of claim 29 or 30, wherein upon administration, the fusion protein binds to the tumor antigen.

36. The method of any one of claim 33-35, further comprising administering to the subject an immune cell therapeutic (e.g., CAR-T cell) that specifically recognizes the target polypeptide.

37. The method of claim 36, wherein upon administration to the subject, the immune cell therapeutic (e.g., CAR-T cell) binds to the fusion protein.

38. The method of claim 33, wherein binding of the immune cell therapeutic (e.g., CAR-T cell) to the fusion protein induces killing of the tumor.

39. A method of treating a subject having a tumor, comprising administering to the subject an effective amount of the immune cell of claim 31.

40. The method of claim 39, wherein the tumor expresses the tumor antigen.

41. The method of claim 35, where a T-cell binds the one or more accessory molecule(s) or a fragment thereof.

42. The method of claim 41, wherein binding of the T-cell to the fusion protein induces killing of the tumor.- 226 - 13321680vl

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