Multi-specific protein molecules and uses thereof

Abstract

Disclosed herein are multi-specific binding polypeptide molecules (e.g., multi-specific antibodies) that can interact with a tumor cell and an immunosuppressive cell. In some embodiments, also described herein are methods for treating one or more diseases or conditions (e.g., cancers) using the multi-specific binding polypeptides (e.g., multi-specific antibodies).

Description

2020205100   07 Aug 2026 CROSS REFERENCE TO RELATED APPLICATIONS

[0001] This application claims the benefit under 35 U.S.C. § 119(e) of U.S. Provisional Application Serial No. 62 / 788,495, filed January 4, 2019, the contents of which are incorporated by reference herein in its entirety including all figures and tables. SUMMARY OF THE DISCLOSURE

[0002] Described herein are multi-specific binding polypeptides useful for the treatment of cancers. The multi-specific binding polypeptides comprise at least two distinct binding moieties with one of the binding moieties specific for a tumor-associated antigen, and the other specific for an antigen expressed on an immunosuppressive cell. In certain embodiments, the immunosuppressive cell is a myeloid derived suppressor cell (MDSC). In certain embodiments, the immunosuppressive cell is a tumor-associated macrophage, optionally a M2-tumor associated macrophage (M2-TAM). In certain embodiments, the multi-specific binding polypeptide further comprises a cytotoxic moiety.

[0003] In certain aspects, described herein is a multi-specific binding polypeptide comprising a tumor binding moiety that specifically binds to a tumor-associated antigen and an immune cell binding moiety that specifically binds to an antigen expressed on an immunosuppressive cell. In some embodiments, the multi-specific binding polypeptide is a multi-specific antibody. In some embodiments, the multi-specific antibody comprises a tumor binding moiety that specifically binds to a tumor-associated antigen and an immune cell binding moiety that specifically binds to an antigen expressed on an immunosuppressive cell. [0003A] Described herein is a multi-specific antibody drug conjugate comprising a first binding moiety that specifically binds to a tumor-associated antigen and a second binding moiety that specifically binds to an antigen expressed on an immunosuppressive cell, wherein: (a) the first antigen binding moiety specifically binds to TROP2 and comprises a VH domain comprising a HCDR1 as set forth in SEQ ID NO:4, a HCDR2 as set forth in SEQ ID NO:5, and a HCDR3 as set forth in SEQ ID NO:6, and a VL domain comprising a LCDR1 as set forth in SEQ ID NO:1, a LCDR2 as set forth in SEQ ID NO:2, and a LCDR3 as set forth in SEQ ID NO:3; and (b) the second antigen binding moiety specifically binds to CD33 and comprises a VH domain comprising a HCDR1 as set forth in SEQ ID NO:45, a HCDR2 as set forth in SEQ ID NO:46, and a HCDR3 as set forth in SEQ ID NO:47, and a VL domain comprising a LCDR1 as set forth in SEQ ID NO:60, a LCDR2 as set forth in SEQ ID NO:61, and a LCDR3 as set forth in SEQ ID NO:62, wherein the multi-specific antibody drug conjugate comprises at least one cytotoxic moiety.

[0004] In certain embodiments, described herein is a method of treating a disease or condition in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a multi-specific binding polypeptide (e.g., a multi-specific antibody), a pharmaceutical composition 2020205100   07 Aug 2026 comprising, consisting essentially of, or consisting of the multi-specific binding polypeptide (e.g., the multi-specific antibody), or a nucleic acid encoding the multi-specific binding polypeptide (e.g., the multi-specific antibody). In some aspects, also described herein is a method of treating an individual with a cancer, the method comprising administering to the individual with cancer the multi-specific binding polypeptide (e.g., the multi-specific antibody), the pharmaceutical composition comprising, consisting essentially of, or consisting of the multi-specific binding polypeptide (e.g., the multi-specific antibody), or a nucleic acid encoding the multi-specific binding polypeptide (e.g., the multi-specific antibody). In certain embodiments, the individual has previously been treated with an immune checkpoint inhibitor treatment.

[0005] In certain embodiments, described herein is a method of inducing tumor and immunosuppressive cell killing effect in a target cell population, the method comprises contacting the target cell population comprising at least one tumor cell and at least one immunosuppressive cell with a multi-specific binding polypeptide (e.g., a multi-specific antibody), a pharmaceutical composition comprising, consisting essentially of, or consisting of the multi-specific binding polypeptide (e.g., the multi-specific antibody), or a nucleic acid encoding the multi-specific binding polypeptide (e.g., the multi-specific antibody) for a time sufficient to induce cell kill effect, thereby killing the at least one tumor cell and the at least one immunosuppressive cell in the target cell population.

[0006] In certain embodiments, additionally described herein is a method for making a cancer treatment comprising contacting the nucleic acid encoding the multi-specific binding polypeptide to a suitable cell line to establish a transfected cell line, culturing the transfected cell line under conditions that promote secretion of the multi-specific binding polypeptide, and harvesting the multi-specific binding polypeptide from the supernatant of the transfected cell line. In certain embodiments, the method further comprises purifying the multi-specific polypeptide from the supernatant of the transfected cell line. In certain embodiments, the transfected cell line is stably transfected. In certain embodiments, described herein is a method for making a cancer treatment comprising, consisting essentially of, or consisting of admixing the multi-specific binding polypeptide and a pharmaceutically acceptable excipient, carrier, or diluent. [0006A] Described herein is a method of treating a TROP2 positive cancer or hematological malignancy in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of a multi-specific antibody drug conjugate, a pharmaceutical composition, or a nucleic acid described herein.. [0006B]   Described herein is a method of treating a cancer comprising CD33 positive immunosuppressive cells in the tumor microenvironment in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of a multi-specific antibody drug conjugate, a pharmaceutical composition, or a nucleic acid described herein. [0006C]   Described herein is a method of treating a cancer comprising at least one TROP2 positive tumor cell and at least one CD33 positive immunosuppressive cell in the tumor microenvironment in a subject in need thereof, the method comprising administering to the subject a 2020205100   07 Aug 2026 therapeutically effective amount of a multi-specific antibody drug conjugate, a pharmaceutical composition, or a nucleic acid described herein. [0006D]   Described herein is the use of a multi-specific antibody drug conjugate, or a nucleic acid described herein in the manufacture of a medicament for treating a TROP2 positive cancer or hematological malignancy, a cancer comprising CD33 positive immunosuppressive cells in the tumor microenvironment, or a cancer comprising at least one TROP2 positive tumor cell and at least one CD33 positive immunosuppressive cell in the tumor microenvironment. [0006E]       Described herein is a method of inducing tumor and immunosuppressive cell killing effect in a target cell population, comprising contacting the target cell population comprising at least one TROP2 positive tumor cell and at least one CD33 positive immunosuppressive cell with a multi-specific antibody drug conjugate, a pharmaceutical composition, or a nucleic acid described herein for a time sufficient to induce cell kill effect, thereby killing the at least one TROP2 positive tumor cell and the at least one CD33 positive immunosuppressive cell in the target cell population. [0006F]       Described herein is a pharmaceutical composition comprising the multi-specific antibody drug conjugate described herein, and a pharmaceutically acceptable excipient, carrier, or diluent, optionally wherein the pharmaceutical composition is formulated for parenteral administration, further optionally for subcutaneous, intramuscular, or intravenous administration. BRIEF DESCRIPTION OF THE DRAWINGS

[0007] Various aspects of the disclosure are set forth with particularity in the appended claims. A better understanding of the features and advantages of the present disclosure will be obtained by reference to the following detailed description that sets forth illustrative embodiments, in which the principles of the disclosure are utilized, and the accompanying drawings of which:

[0008] FIG. 1A illustrates a non-limiting embodiment of a multi-specific binding polypeptide of the current disclosure.

[0009] FIG. 1B illustrates a non-limiting embodiment of a multi-specific binding polypeptide of the current disclosure with a cytotoxic moiety.

[0010] FIG. 1C illustrates a cartoon representation of an exemplary single action bispecific antibody which binds and kills cancer cells but does not bind or kill immunosuppressor cells. The exemplary single action bispecific antibody is a non-applicant bispecific antibody.

[0011] FIG. 1D illustrates a cartoon representation of a multi-action multi-specific antibody described herein that binds and modulates killing of a cancer cell and an immunosuppressor cell.

[0012] FIG. 2 depicts an expression heat map of receptors expressed on Triple Negative Breast Cancer (TNBC) cells and immunosuppressive cells detected in the same TNBC biopsy. As shown in this figure, the TNBC cells are expressing a high level of TROP2. Select receptors are noted in the heat map chart as: TROP2 = TACSTD2; TRAIL-R2 = TNFRSF10B; PD1 = PDCD1; PDL1 = CD274.

[0013] FIG. 3 depicts an expression heat map of receptors expressed on Lung Adenocarcinoma 2020205100   07 Aug 2026 (LUAD) cells and immunosuppressive cells detected in the same LUAD biopsy. As shown in this figure, the LUAD cells are expressing high levels of TROP2.

[0014] FIG. 4 depicts an expression heat map of receptors expressed on Lung Squamous Cell Carcinoma (LUSC) cells and immunosuppressive cells detected in the same LUSC biopsy. As shown in this figure, the LUSC cells are expressing high levels of TROP2.

[0015] FIG. 5 depicts an expression heat map of receptors expressed on Prostate Adenocarcinoma (PRAD) cells and immunosuppressive cells detected in the same PRAD biopsy. As shown in this figure, the PRAD cells are expressing high levels of TROP2.

[0016] FIG. 6 depicts an expression heat map of receptors expressed on Liver Hepatocellular Carcinoma (LIHC) cells and immunosuppressive cells detected in the same LIHC biopsy. As shown in this figure, the LIHC cells are expressing high levels of GPC3.

[0017] FIG. 7 depicts an expression heat map of receptors expressed on Lung Adenocarcinoma (LU AD) cells and immunosuppressive cells detected in the same LU AD biopsy. As shown in this figure, the LU AD cells are expressing high levels of GPC3.

[0018] FIG. 8 depicts an expression heat map of receptors expressed on Lung Adenocarcinoma (LU AD) cells and immunosuppressive cells detected in the same LU AD biopsy. As shown in this figure, the LU AD cells are expressing high levels of FOLRI.

[0019] FIG. 9 depicts an expression heat map of receptors expressed on Lung Squamous Cell carcinoma (LUSC) cells and immunosuppressive cells detected in the same LUSC biopsy. As shown in this figure, the LUSC cells are expressing high levels of FOLRI.

[0020] FIG. 10 depicts an expression heat map of receptors expressed on Ovarian Cystadenocarcinoma (OV) cells and immunosuppressive cells detected in the same OV biopsy. As shown in this figure, the OV cells are expressing high levels of FOLRI.

[0021] FIG. 11 depicts an expression heat map of receptors expressed on Prostate Adenocarcinoma (PRAD) cells and immunosuppressive cells detected in the same PRAD biopsy. As shown in this figure, the PRAD cells are expressing high levels of FOLH1.

[0022] FIG. 12A-FIG. 12E illustrate expression and analytical characterization of aTROP2 x aTRAIL-R2 bispecific antibody. FIG. 12A shows transient expression titer of aTROP2 x aTRAIL-R2 bispecific antibody. The titer estimate is based on the net amount of bispecific antibody generated after Protein A purification. FIG. 12B shows solution profile analysis of Protein A purified aTROP2 x aTRAIL-R2 bispecific antibody by Size Exclusion Ultra-Performance Liquid Chromatography (SE-UPLC). The major peak with retention time = 3.596 corresponds to monomeric form of the intact bispecific antibody constructs and is -85% of all the species noted. Peaks with retention time 2.980 and 3.142 are higher molecular weight species of the bispecific antibody construct. FIG. 12C shows SDS-PAGE analysis of Protein A purified aTROP2 x aTRAIL-R2 bispecific antibody. The samples were analyzed under reducing (R) and non-reducing (NR) conditions. Two bands are observed under reducing conditions, top band corresponding to the molecular weight of the heavy chain and lower band corresponding to the molecular weight of the light chain. Under non-reducing conditions only one band was observed corresponding to the molecular weight of the intact bispecific antibody construct. FIG.12D shows intact-mass analysis of Protein A purified aTROP2 x aTRAIL-R2 bispecific antibody. The sample was reduced and de-glycosylated before analyzing the mass. Both the heavy chain (upper panel) and light chain (lower panel) exhibited a major peak corresponding to theoretical molecular mass based on the amino acid sequence. The second peak observed in the upper panel corresponds to a glycosylated heavy chain species arising from in-complete de-glycosylation. FIG. 12 E shows binding affinity measurement of aTROP2 x aTRAIL-R2 bispecific antibody toward TROP2 and TRAIL-R2 antigen. The binding affinity was measured by ELISA. The binding affinity for TROP2 antigen expressed on cancer cell was Kd = 0.0024 nM and binding affinity for TRAIL-R2 expressed on immunosuppressor cell was Kd = 0.22 nM. Rabbit anti- TROP2 and anti- TRAIL-R2 antibodies bound TROP2 and TRAIL-R2 antigens, respectively with binding affinity, Kd, 0.004 nM and 0.0009 nM. The non-binding human IgGl isotype control did not bind to TROP2 and TRAIL-R2 antigens.

[0023] FIG. 13A-FIG. 13G illustrate target mediated selective binding of aTROP2 x aTRAIL-R2 bispecific antibody on the surface of cancer cells measured by Fluorescent Activated Cell Sorting method (FACS). FIG. 13A shows selective binding of aTROP2 x aTRAIL-R2 bispecific antibody on the surface of a TROP2+ cancer cell line, SKBR3 with a binding affinity, Kd = 2.4 nM (0.463 pg / ml). The nonbinding IgGl isotype control did not bind on the surface of SKBR3. FIG. 13B shows high levels of TROP2 expressed on the surface of SKBR3 cell line. The non-binding rabbit antibody isotype control did not bind on the surface of SKBR3 cell line as detected by no fluorescent signal. FIG. 13C shows low expression of TRAIL-R2 in SKBR3 cell line. The non-binding rabbit antibody isotype control did not bind on the surface of SKBR3 cell line. FIG. 13D shows selective binding of aTROP2 x aTRAIL-R2 bispecific antibody on the surface of a TRAIL-R2+ cancer cell line, U937. There was a concentration dependent increase in fluorescence intensity by aTROP2 x aTRAIL-R2 in U937. In contrast, there was no concentration dependent increase in fluorescence intensity by Herceptin in U937 suggesting it did not bind U937. Herceptin, a therapeutic antibody which is known to bind HER2 antigen did not bind on the surface of U937, which is known not to express HER2. FIG. 13E shows no binding of aTROP2 x aTRAIL-R2 bispecific antibody on the surface of a TROP2- and TRAIL-R2- cell line, THP1. Herceptin, a therapeutic antibody known to bind HER2 did not bind on the surface of THP 1. There was no concentration dependent increase in fluorescence intensity suggesting neither aTROP2 x aTRAIL-R2 nor Herceptin bound THP1. FIG. 13F shows low levels of TRAIL-R2 and no detectable levels of expression of TROP2 in U937 cell line. Rabbit anti - TRAIL-R2 antibody bound the surface of U937 as measured by detectable level of fluorescent signal. Mouse anti- TROP2 antibody did not bind the surface of U937 as detected by no fluorescent signal. Both rabbit and mouse antibody isotype controls did not bind the surface of U937. FIG. 13G shows there is no detectable expression of TRAIL-R2 and TROP2 in U937. Rabbit anti - TRAIL-R2 antibody and mouse anti- TROP2 antibody did not bind the surface of U937 as detected by no fluorescent signal. Both rabbit and mouse antibody isotype controls did not bind the surface of U937.

[0024] FIG. 14 shows dose dependent TRAIL-R2 mediated apoptosis and cell death in U937 by aTROP2 x aTRAIL-R2 bispecific antibody with a maximum cell killing of 11% at cell passage number 7 and maximum cell killing of 15% at cell passage number 10 at IC50 = 0.65 nM (0.13 pg / ml).

[0025] FIG. 15 shows solution profile analysis of Protein A and preparative Size Exclusion WO 2020 / 142659                                                 PCT / US2020 / 012139 Chromatography (SEC) purified aTROP2 x aCD33, aTROP2 x aCSFIRand aTROP2 x aCD163, bispecific antibody by Size Exclusion Ultra-Performance Liquid Chromatography (SE-UPLC). The samples are analysed on a Waters Acquity Protein BEH SEC 125 A 1.7 pm column using PBS with 10% isopropanol as the mobile phase. A single major peak corresponding to the molecular weight of the monomer bispecific antibody assembly was observed for both aTROP2 x aCD33 and aTROP2 x aCSFIR suggesting there is no presence of low molecular and high molecular aggregates indicating stability. The major peak for aTROP2 x aCD163 corresponding to the molecular weight of the monomer bispecific antibody assembly was 81% with a preceding shoulder corresponding to possible high molecular weight forms.

[0026] FIG. 16 shows SDS-PAGE analysis of Protein A purified aTROP2 x aCD33, aTROP2 x aCSFIR and aTROP2 x aCD163 bispecific antibody. The samples were analyzed under reducing (R) and non-reducing (NR) conditions. Two bands are observed under reducing conditions, top band corresponding to the molecular weight of the heavy chain and lower band corresponding to the molecular weight of the light chain. Under non-reducing conditions only one band was observed corresponding to the molecular weight of the intact bispecific antibody construct. 2 pg of each antibody sample + / - [3-mercaptoethanol (reduced / non-reduced respectively) is loaded on aNuPage 4-12% Bis-Tris gel (ThermoFisher, Loughborough, UK) and run at 200 V for 40 minutes. Gels are stained with InstantBlue (Expedeon, Swavesey, UK). M: PAGERuler™ Plus pre-stained protein ladder (ThermoFisher, Loughborough, UK). Lanes 1,2; aTROP2 x aCSFIRbispecific antibody, reduced and non-reduced, respectively; Lanes 3, 4; aTROP2 x aCD163 bispecific antibody, reduced and non-reduced, respectively; Lanes 5,6; aTROP2 x aCD33 bispecific antibody, reduced and non-reduced, respectively; Lanes 7,8; aTROP2 monospecific antibody, reduced and non-reduced, respectively.

[0027] FIG. 17 shows Biacore multi-cycle kinetics sensorgrams of aTROP2 x aCD33 binding to TROP2 and CD33 following protein A and preparative SEC purification. A 1 to 1 model was used for fitting.

[0028] FIG. 18 shows Biacore multi-cycle kinetics sensorgrams of aTROP2 x aCD163 binding to TROP2 and CD33 following protein A and preparative SEC purification. A 1 to 1 model was used for fitting.

[0029] FIG. 19 shows Biacore multi-cycle kinetics sensorgrams of aTROP2 x aCSFIRbinding to TROP2 and CSF1R following protein A purification and preparative SEC purification. A 1 to 1 model was used for fitting.

[0030] FIG. 20A-FIG. 20D show target mediated selective binding of aTROP2 x aCD33 bispecific antibody on the surface of cancer cells measured by Fluorescent Activated Cell Sorting method (FACS). FIG. 20A shows selective binding of aTROP2 x aCD33 bispecific antibody on the surface of a TROP2+ cancer cell line, SKBR3, with a binding affinity, Kd = 2.7 nM (0.55 pg / ml). The non-binding IgGl isotype control did not bind on the surface of SKBR3. As a system control Herceptin bound on the surface of SKBR3 with a binding affinity, Kd = 4.1 nM (0.62 pg / ml) due to high expression of HER2 as manifested by high fluorescence intensity compared to the binding of aTROP2 x aCD33. FIG. 20B shows selective binding of aTROP2 x aCD33 bispecific antibody on the surface of a CD33+ cancer cell line, THP1, with abinding affinity, Kd = 6.7 nM (1.33 pg / ml). There was a concentration dependent increase in fluorescence intensity for aTROP2 x aCD33. In contrast neither the non-binding IgGl isotype control nor Herceptin exhibited any concentration dependent increase in fluorescence intensity in THP1 suggestion neither of the antibodies bound THP1. FIG. 20C shows high levels of TROP2 expressed on the surface of SKBR3 cell line. The non-binding rabbit antibody isotype control did not bind on the surface of SKBR3 cell line as detected by low fluorescent signal. FIG. 20D shows high levels of CD33 expressed on the surface of THP1 cell line. The non-binding rabbit antibody isotype control did not bind on the surface of THP1 cell line as detected by no fluorescent signal.

[0031] FIG. 21A illustrates a cartoon representation of a TRAIL-R2 mediated activation on a target cancer cell which is selectively activated by a multi-specific antibody disclosed herein. As shown in this figure, the multi-specific antibody binds to a tumor specific antigen located in the lipid raft which recruits and facilitates oligomerization (e.g., dimerization) of the TRAIL-R2 and subsequent activation of the TRAIL-R2 mediated apoptosis within the target cancer cell.

[0032] FIG. 21B illustrates a cartoon representation of an exemplary multi-specific antibody that binds to a tumor specific antigen located outside of the lipid raft of a target cancer cell. The exemplary multi-specific antibody, upon binding to the tumor specific antigen, does not activate the TRAIL-R2 mediated apoptosis within the target cancer cell.

[0033] FIG. 21C illustrates a cartoon representation of a conventional antibody such as the single action bispecific antibody illustrated in FIG. IC which generates non-tumor toxicity due to leakage of inflammatory cytokines, granzymes, and / or perforins to nearby normal cells and tissues. As shown in this figure, upon recruiting an immune effector cell to a cancer cell and activation of the immune effector cell by the conventional antibody, the activated immune effector cells not only secretes soluble TRAILs that can lead to activation of apoptosis within the cancer cell via the TRAIL-R2 pathway, but can also release cytokine, granzymes, and / or perforins which leakage to nearby normal cells leads to non-tumor toxicity.

[0034] FIG. 21D illustrates a cartoon representation of an exemplary multi-specific antibody which modulates TRAIL-R2 mediated apoptosis of the cancer cell and does not activate immune effector cells. As shown in this figure, the lack of immune effector cell activation minimizes or prevents the release of inflammatory cytokines, granzymes, and / or perforins, and as such, reduces or inhibits toxicity to nearby normal cells or tissues.

[0035] FIG. 22 illustrates no cell killing by an exemplary multi-specific antibody in a TRAILsensitive MDAMB231 cell line. DETAILED DESCRIPTION OF THE DISCLOSURE Tumor microenvironment and Immune Suppression

[0036] The tumor microenvironment (TME) plays an integral part in tumor progression, tumor cell adaptation, and resistance to anti-cancer therapy. Components of the tumor microenvironments include myeloid-derived suppressor cells (MDSCs), antigen-presenting cells (APCs), lymphocytes, neutrophils, tumor-associated macrophages (TAMs), fibroblasts, extracellular matrix composed of collagen and proteoglycans, and soluble factors (e.g., cytokines and growth factors), all of which may assist or hinder antitumor immune responses. Immunotherapy harnesses the immune system, both innate and adaptive, to attack and destroy tumor cells. However, multiple mechanisms used by tumor cells, including alteration of the antigen presentation machinery, secretion of immune suppressive factors that can induce apoptosis of lymphocytes, or activate negative regulatory pathways, resulting in evasion from anti-tumor immune response, thereby limiting the effectiveness of the immune response.

[0037] Exemplary immune suppressive factors and cells include CD4+ T regulatory (Treg) cells, MDSCs, TAMs, and inhibitory immune checkpoint molecules. In some embodiments, Treg cells are characterized with expression of FOXP3, CD4, and CD25. In some embodiments, Treg cells suppress T-effector cell responses via secretion of inhibitory cytokines (e.g., IL-10, IL-35, and TGF-J3) or via direct cell contact.

[0038] Myeloid-derived suppressor cells (MDSCs) are a heterogeneous group of immune cells originated from bone marrow stem cells. MDSCs normally differentiate into granulocytes, macrophages, or dendritic cells. However, in the tumor microenvironment, MDSCs become activated, expand rapidly, but remain undifferentiated. In some embodiments, MDSCs have been shown in clinical studies to be associated with reduced survival in several human tumors including colorectal cancer and breast cancer.

[0039] Macrophages, a plentiful myeloid derived phagocytic cell, can be classified as pro-inflammatory or anti-inflammatory, also known as classic (Ml) and alternative (M2). TAMs are a subset of macrophages that affect responses to immunotherapy and coordinate tumor-promoting angiogenesis, fibrous stroma deposition, and metastasis formation. TAMs display an alternatively activated M2 phenotype known to control tissue homeostasis and wound healing. However, in a tumor setting, this phenotype enables T cell inhibition via cytokines (e.g., IL-10), depletion of metabolites (e.g., expression of arginase, IDO), and / or by contact inhibition (e.g., via PD-L1).

[0040] Cancer therapies focus in eliminating cancer cells either directly or by engaging the body’s immune system (also see FIG. IC). For example, therapies such as chemotherapy, radiotherapy, antibody drug conjugates, CAR-Ts, or BiTEs are designed to directly eliminate cancer cells while immunotherapies such as immunecheckpoint inhibitors block certain immunecheckpoint receptors on cancer cells or T-cells to activate T-cells to eliminate cancer cells. Immuno-oncology strategies involving bispecific antibodies are also designed to eliminate cancer cells by engaging or harnessing T-cells or NK-cells. Indeed, there are 86 clinical trials evaluating bispecific antibodies and all of them are designed to engage and / or activate the immune system. As such, these cancer treatment modalities only target and kill tumor cells and do not address the immunosuppressive cells.

[0041] Disclosed herein, in certain embodiments, are multi-specific polypeptide molecules (e.g., multi-specific antibodies) that comprise a first binding moiety specific for a tumor-associated antigen or receptor expressed on a tumor cell, and a second binding moiety specific for a receptor or antigen expressed on an immunosuppressive cell. In some embodiments, the immunosuppressive cell comprises a myeloid-derived suppressor cell (MDSC), a tumor-associated macrophage, or a CD4+ T regulator (Treg) cell. In some embodiments, also described herein are multi-specific polypeptide molecules (e.g., multispecific antibodies) that comprise a first binding moiety specific for a tumor-associated antigen or receptor expressed on a tumor cell and a second binding moiety specific for a receptor or antigen expressed on a MDSC or a tumor-associated macrophage. In additional embodiments, described herein are multi-specific polypeptide molecules (e.g., multi-specific antibodies) that comprise a first binding moiety specific for a tumor-associated antigen or receptor expressed on a tumor cell and a second binding moiety specific for a receptor or antigen expressed on Treg cell.

[0042] In some embodiments, the multi-specific polypeptide molecules (e.g., multi-specific antibodies) described herein modulate an immunosuppressive cell population. In some instances, the multi-specific polypeptide molecules (e.g., multi-specific antibodies) described herein decreases an immunosuppressive cell population, induces a cell-kill effect on the immunosuppressive cell population, reduces immunosuppressive cell expansion, reduces immunosuppressive cell activation, and / or decreases immunosuppressive cell proliferation. In some cases, the multi-specific polypeptide molecules (e.g., multi-specific antibodies) described herein ameliorates, eliminates, and / or blocks an escape mechanism that the cancer cells utilize to evade an anti-tumor immune response. In some cases, the multi-specific polypeptide molecules (e.g., multi-specific antibodies) described herein does not target or specifically binds to an immune checkpoint pathway receptor.

[0043] In some embodiments, the multi-specific polypeptide molecules (e.g., multi-specific antibodies) described herein modulates a tumor-infiltrating lymphocyte (TIL) expansion by eliminating immunosuppressive factors such as, immunosuppressive cells and immunosuppressive cytokines. In some instances, the multi-specific polypeptide molecules (e.g., multi-specific antibodies) described herein enhances TIL population expansion, cytotoxic T-cell proliferation, and / or cytotoxic-T cell activation. In some cases, the multi-specific polypeptide molecules (e.g., multi-specific antibodies) described herein increases a ratio of TIL to immunosuppressive cells.

[0044] In some embodiments, the multi-specific polypeptide molecules (e.g., multi-specific antibodies) described herein modulates a population of Treg. In some instances, the multi-specific polypeptide molecules (e.g., multi-specific antibodies) described herein decreases Tregs within a target population, reduces Treg proliferation, reduces Treg expansion, and / or reduces Treg activation. In some cases, the multi-specific polypeptide molecules (e.g., multi-specific antibodies) described herein decreases a ratio of Tregs to TILs in a target population.

[0045] In some embodiments, the multi-specific polypeptide molecules (e.g., multi-specific antibodies) described herein modulates a population of MDSCs. In some instances, the multi-specific polypeptide molecules (e.g., multi-specific antibodies) described herein reduces MDSC proliferation, reduces MDSC activation, and / or reduces MDSC expansion. In some instances, the multi-specific polypeptide molecules (e.g., multi-specific antibodies) described herein reduces MDSCs in a target population. In some cases, the multi-specific polypeptide molecules (e.g., multi-specific antibodies) described herein reduces a ratio of MDSCs to TILs in a target population.

[0046] In some embodiments, the multi-specific polypeptide molecules (e.g., multi-specific antibodies) described herein modulates a Ml polarization of macrophages. In some instances, the multispecific polypeptide molecules (e.g., multi-specific antibodies) described herein increases Ml polarization. In some instances, the multi-specific polypeptide molecules (e.g., multi-specific antibodies) described herein decreases M2 polarization. In some cases, the multi-specific polypeptide molecules (e.g., multi-specific antibodies) described herein decreases TAM (e.g., M2-TAM) proliferation, expansion, and / or activation. In some cases, the multi-specific polypeptide molecules (e.g., multi-specific antibodies) described herein increases a ratio of macrophages presenting a Ml phenotype over macrophages presenting a M2 phenotype. In some cases, the multi-specific polypeptide molecules (e.g., multi-specific antibodies) described herein increases an anti-tumor macrophage proliferation and / or activation. In some cases, the multi-specific polypeptide molecules (e.g., multi-specific antibodies) described herein enhances expansion of a population of anti-tumor macrophages.

[0047] In certain embodiments, further described herein are methods of treating a subject with a multi-specific protein molecule (e.g., a multi-specific antibody) that comprise administering a multispecific polypeptide (e.g., a multi-specific antibody) comprising a first binding moiety specific for a tumor-associated antigen or receptor expressed on a tumor cell and a second binding moiety specific for a receptor or antigen expressed on an immunosuppressive cell.

[0048] In certain embodiments, additionally described herein are methods of inducing tumor and immunosuppressive cell killing in a target cell population with use of a multi-specific protein molecule (e.g., a multi-specific antibody) that comprises a first binding moiety specific for a tumor-associated antigen or receptor expressed on a tumor cell and a second binding moiety specific for a receptor or antigen expressed on an immunosuppressive cell.

[0049] In certain embodiments, additionally described herein are methods of inducing tumor and immunosuppressive cell killing in a target cell population via induce direct cell death of both cancer cells and immunosuppressive cells by activating apoptotic pathways by a multi-specific protein molecule (e.g., a multi-specific antibody) that comprises a first binding moiety specific for a tumor-associated antigen or receptor expressed on a tumor cell and a second binding moiety specific for a receptor or antigen expressed on an immunosuppressive cell. In some instances, the method comprises induction of cell death via a cytotoxic payload conjugated to the multi-specific protein molecule (e.g., the multi-specific antibody). In other instances, the method comprises utilizing antibody-dependent cellular cytotoxicity to induce cell kill effect. In additional instances, the method comprises a combination of cytotoxic payload associated cytotoxicity and antibody-dependent cellular cytotoxicity to induce cell kill effect.

[0050] In certain embodiments, further described herein are antibody-cytotoxin conjugates that specifically binds to an antigen expressed on an immunosuppressive cell. In some instances, the antibody-cytotoxin conjugates are used in a method of inducing immunosuppressive cell killing in a subject in need thereof, comprising administering the antibody-cytotoxin conjugate that specifically binds to an antigen expressed on an immunosuppressive cell, thereby killing the immunosuppressive cell in the subject. In other instances, the antibody-cytotoxin conjugates are used in a method of activating tumor cell-killing immune cells in a subject in need thereof, comprising administering an antibody-cytotoxin conjugate that specifically binds to an antigen expressed on an immunosuppressive cell, thereby killing the immunosuppressive cell and activating the tumor cell-killing immune cells in the subject. In additional instances, the antibody-cytotoxin conjugates are used in a method of reducing suppression of tumor cell-killing immune cells in a subject in need thereof, comprising administering an antibody-cytotoxin conjugate that specifically binds to an antigen expressed on an immunosuppressive cell, thereby killing the immunosuppressive cell and reducing suppression of the tumor cell-killing immune cells in the subject. In some cases, the antibody-cytotoxin conjugate further comprises a tumor-specific binding moiety. In some cases, the antibody-cytotoxin conjugate is a multi-specific binding polypeptide (e.g., a multi-specific antibody) described herein. Multi-specific binding polypeptide structures

[0051] Disclosed herein are multi-specific binding polypeptides (e.g., multi-specific antibodies) that comprise a tumor binding moiety that specifically binds to a tumor-associated antigen and an immune cell binding moiety that specifically binds to an antigen expressed on an immunosuppressive cell. In some instances, the multi-specific binding polypeptide (e.g., the multi-specific antibody) binds to and modulate killing of a cancer cell and binds to and modulate killing of an immunosuppressor cell. Also see FIG. ID. In some instances, the multi-specific binding polypeptide comprises a single polypeptide complex comprising at least two distinct non-overlapping binding domains, each of the distinct binding domains being specific for distinct non-overlapping antigens or epitopes. The antigen or epitope can be on different molecules (e.g., different proteins), or non-overlapping portions of the same molecule. A single polypeptide complex encompasses single polypeptides, polypeptides that dimerize or multimerize to form a binding unit (e.g., heavy chain and light chain of an IgG, two heavy chain and light chain pairs forming a mature IgG), polypeptides that are distinct but held together by covalent (e.g., disulfide bonds, linker molecules) or non-covalent interactions (e.g., biotin-streptavidin; affinity interactions; charge interactions, knob-in-hole).

[0052] The multi-specific binding polypeptides described herein, in certain embodiments, comprise antibodies or binding fragments of antibodies. Among the provided multi-specific binding polypeptides are monoclonal antibodies, multi-specific antibodies, for example, bispecific antibodies, and antibody fragments. The antibodies include antibody-conjugates and molecules comprising the antibodies, such as chimeric molecules. Thus, an antibody includes, but is not limited to, full-length and native antibodies, as well as fragments and portion thereof retaining the binding specificities thereof, such as any specific binding portion thereof including those having any number of, immunoglobulin classes and / or isotypes (e.g., IgGl, IgG2, IgG3, IgG4, IgM, IgA, IgD, IgE and IgM); and biologically relevant (antigen-binding) fragments or specific binding portions thereof, including but not limited to Fab, F(ab')2, Fv, and scFv (single chain or related entity). A monoclonal antibody is generally one within a composition of substantially homogeneous antibodies; thus, any individual antibodies comprised within the monoclonal antibody composition are identical except for possible naturally occurring mutations that may be present in minor amounts. A polyclonal antibody is a preparation that includes different antibodies of varying sequences that generally are directed against two or more different determinants (epitopes). The monoclonal antibody can comprise a human IgGl constant region. The monoclonal antibody can comprise a human IgG4 constant region.

[0053] In some embodiments, a multi-specific antibody described herein comprises a full-length antibody, an appended antibody, a bispecific fusion protein, or a bispecific antibody conjugate. In some cases, the multi-specific antibody comprises a nanobody, BiTE, diabody, DART, TandAb, scDiabody, scDiabody-CH3, triple body, miniantibody, minibody, TriBi minibody, scFv-CH3 KIH, Fab-scFv, scFv-CH-CL-scFv, F(ab’)2, F(ab’)2-scFv2. scFv-KIH, Fab-scFv-Fc, tetravalent HCAb, scDiabody-Fc, diabody-Fc, tandem scFv-Fc, or intrabody. In some cases, the multi-specific antibody comprises an appended antibody. In some cases, the multi-specific antibody comprises a bispecific fusion protein. In some cases, the multi-specific antibody comprises a bispecific antibody conjugate. In some cases, the multi-specific antibody comprises an appended antibody further comprising a conjugate (e.g., a payload such as a cytotoxic moiety).

[0054] The multi-specific binding polypeptides (e.g., the multi-specific antibody) of the current disclosure have certain structural attributes. FIG. 1A shows a specific non-limiting embodiment of one such multi-specific binding polypeptide (e.g., a multi-specific antibody). The polypeptide minimally comprises a first binding moiety that binds a tumor-associated antigen. This binding moiety comprises, for example, a VH / VL pair of a monoclonal antibody specific for the tumor-associated antigen. The polypeptide also comprises a second binding moiety comprising a single-chain variable region (scFv) that binds to an antigen expressed on an immunosuppressive cell. While the first binding moiety is described as being specific for a tumor-associated antigen, and the second binding moiety is described as being specific for an antigen expressed on an immunosuppressive cell, this order can be reversed with the first binding moiety binding to an antigen expressed on an immunosuppressive cell and the second binding moiety binding to a tumor-associated antigen. FIG. 1A also shows optional features for the polypeptides described herein, including an Fc region and a polypeptide linker (e.g., a flexible polypeptide linker) coupling the first binding moiety and the second binding moiety. In one embodiment, the multi-specific binding polypeptide (e.g., the multi-specific antibody) comprises a first binding moiety that comprises a monoclonal IgG antibody and a second binding moiety that comprises an scFv, wherein the scFv antibody is coupled to the C-terminus of one or both of the Fc regions of the monoclonal antibody. In certain embodiments, the IgG comprises an IgGi isotype. In certain embodiments, the IgG comprises an IgG2 isotype. In certain embodiments, the IgG comprises an IgG4 isotype. In certain embodiments, the polypeptide linker comprises a poly-Ala linker, a poly-Gly linker, or a combination of poly-Ala and poly-Gly residues. In some instances, the polypeptide linker comprises (Gly4Ser)n, wherein n is an integer from 1 to 10. In some instances, the n is an integer from 1 to 6, 1 to 4, or 1 to 2. In some cases, n is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10. In some cases, n is 1, 2, 3, 4, 5, or 6. In some cases, n is 1, 2, 3, or 4. In some cases, the polypeptide linker comprises GGGSGGGS (SEQ ID NO: 78). In some cases, the polypeptide linker comprises GGGSGGGSGGGS (SEQ ID NO: 140). In some cases, the polypeptide linker comprises GGGSGGGSGGGSGGGS (SEQ ID NO: 141). In certain embodiments, the Fc region has been modified to increase half-life in circulation. In certain embodiments, the half-life is extended to at least 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 25, 28, 30, 60, or more days. In certain embodiments, the half-life is extended to at least 7, 14 or 21 days. In certain embodiments, the Fc region has been modified to increase antibody dependent cell-cytotoxicity (ADCC). In certain embodiments, the Fc region that has been modified to reduce the affinity for human neonatal Fc receptor (FcRn). In certain embodiments, the Fc region that has been modified to reduce neutropenia. In certain embodiments, one or both hinge regions have been modified.

[0055] In addition to the configuration shown in FIG. 1A other configurations are envisioned. Exemplary multi-specific antibody configurations include, but are not limited to: (1) a Light chain c-terminal scFv format, wherein an scFv is coupled to the C-terminus of one or more light chains of an antibody, as in US 2013 / 0216543; (2) a dual-variable domain (DVD) antibody, wherein the light chain and heavy chain variable regions comprise a distinct second binding domain, as in US 8,258,268; (3) a knob-in-hole bispecific, wherein one heavy chain constant region comprises an engineered hole, and one heavy chain constant region comprises an engineered knob, as in Merchant et al. “An efficient route to human bispecific IgG.” Nat Biotechnol. 1998 Jul;16(7):677-81, and WO 2016 / 172485; (4) dual scFvs, wherein two scFvs with different binding specificities are joined by a linker; and (5) dual F(ab’)2 wherein two F(ab’)s with different binding specificities are joined by a linker or chemical crosslinking. Any bispecific antibody format can be used as a multi-specific binding polypeptide of the current disclosure. See Spiess, et al., Molecular Immunology, 67(2): 95-106 (2015). In certain embodiments, the multispecific binding polypeptide is a bispecific of a format selected from the list consisting of: IgG-scFv, nanobody, diabody, Dual-affinity Re-targeting Antibody (DART), TandAb, scDiabody, scDiabody-CH3, triple body, mini-antibody, minibody, TriBi minibody, scFv-CH3 KIH, Cross-mab Knob-in-hole (KIH), Fab-scFv, scFv-CH-CL-scFv, F(ab’)2, F(ab’)2-scFv2. scFv-KIH, Fab-scFv-Fc, tetravalent HCAb, scDiabody-Fc, diabody-Fc, tandem scFv-Fc, and intrabody.

[0056] In some embodiments, the multi-specific binding polypeptide (e.g., the multi-specific antibody) comprises a bispecific format described in Godar, et al., “Therapeutic bispecific antibody formats: a patent applications review (1994-2017),” Expert Opinion on Therapeutic Patents, 28(3): 251276 (2018).

[0057] In some embodiments, the multi-specific binding polypeptide (e.g., the multi-specific antibody) comprises a bispecific format described in Labrijn, et al., “Bispecific antibodies: a mechanistic review of the pipeline,” Nature Reviews 18: 585-608 (2019). Target Antigens

[0058] In certain embodiments, a multi-specific binding polypeptide (e.g., a multi-specific antibody) of the current disclosure possesses distinct binding moieties that bind to a tumor associated antigen and an immunosuppressive cell expressed antigen. In certain embodiments, a multi-specific binding polypeptide (e.g., a multi-specific antibody) of the current disclosure contributes to killing of both tumor / cancer cells and immunosuppressive cells.

[0059] Tumor-associated antigens (TAAs) are cell-surface antigens accessible to a multi-specific binding polypeptide (e.g., a multi-specific antibody). The multi-specific binding polypeptides (e.g., multi-specific antibodies) of the current disclosure can contribute to cell killing directly by an ADCC mechanism or by target mediated delivery of a cytotoxic payload. In certain embodiments, the tumor associated antigen is TROP2 (also referred to as TACSTD2), HER2 (also referred to as ERBB2), GPC3 (also referred to as Glypican 3), GD2, FOLRI (also referred to as Folate Receptor 1), FLT3, BCMA (also referred to as B-cell maturation antigen), MUC16, SLC4A4, STEAP1, CD19, CD20, CD22, CD25, CD33, CD38, CD30, CD47, CD123, mesothelin, MT1-MMP (also referred to as MMP14), or PSMA (also referred to as FOLH1). In certain embodiments, the tumor associated antigen is TROP2. In certain embodiments, the tumor-associated antigen is HER2. In certain embodiments, the tumor-associated antigen is CD30. In certain embodiments, the tumor-associated antigen is CD22. In certain embodiments, the tumor-associated antigen is GD2. In certain embodiments, the tumor-associated antigen is FOLRI. In certain embodiments, the tumor-associated antigen is CD33. In certain embodiments, the tumor-associated antigen is GPC3. In certain embodiments, the tumor-associated antigen is CD38. In certain embodiments, the tumor-associated antigen is FLT3. In certain embodiments, the tumor-associated antigen is CD47. In certain embodiments, the tumor-associated antigen is CD22. In certain embodiments, the tumor-associated antigen is FOLH1.

[0060] In certain embodiments, a multi-specific binding polypeptide (e.g., a multi-specific antibody) comprises a tumor binding moiety that binds to TROP2, HER2, GPC3, GD2, FOLRI, FLT3, BCMA, MUC16, SLC4A4, STEAP1, CD19, CD20, CD22, CD25, CD33, CD38, CD30, CD47, CD123, mesothelin, MT1-MMP, or PSMA. In some embodiments, the tumor binding moiety binds to TROP2, GPC3, HER2, FOLRI, CD33, CD38, FLT3, CD30, CD22, or GD2. In some embodiments, the tumor binding moiety binds to TROP2, GPC3, FOLRI, CD33, CD38, or FLT3. In some embodiments, the tumor binding moiety binds to TROP2, CD47, HER2, CD30, CD22, GD2, or FOLRI. In some cases, the tumor binding moiety binds to TROP2. In some cases, the tumor binding moiety binds to CD30. In some cases, the tumor binding moiety binds to CD22. In some cases, the tumor binding moiety binds to GD2. In some cases, the tumor binding moiety binds to FOLRI. In some cases, the tumor binding moiety binds to CD33. In some cases, the tumor binding moiety binds to GPC3. In some cases, the tumor binding moiety binds to HER2. In some cases, the tumor binding moiety binds to CD38. In some cases, the tumor binding moiety binds to FLT3. In some cases, the tumor binding moiety binds to CD47. In some cases, the tumor binding moiety binds to CD22.

[0061] Immunosuppressive cells infiltrate tumor sites and suppress the endogenous immune response to cancer cells. Killing immunosuppressive cells by a cytotoxic payload or ADCC can contribute to increased tumor killing by natural killer cells, and cytotoxic T lymphocytes. In certain embodiments, the immunosuppressive cell comprises a myeloid derived suppressor cell (MDSC), a CD4+ T regulatory cell, a CD8+ T regulatory cell, or a tumor-associated macrophage. Exemplary MDSC receptors include, but are not limited to, TNF-related apoptosis-inducing ligand receptor 2 (TRAIL-R2) and colony stimulating factor 1 receptor (CSF1R). Exemplary tumor-associated macrophage receptors WO 2020 / 142659                                                 PCT / US2020 / 012139 include, but are not limited to, Cluster of Differentiation 163 (CD 163) and macrophage receptor with collagenous structure (MARCO). Exemplary Treg receptors include, but are not limited to, tumor necrosis factor receptor 2 (TNFR2) and semaphorin 4A (SEMA4A).

[0062] In certain embodiments, exemplary antigens expressed by an immunosuppressive cell include, but are not limited to, TRAIL-R2 (also referred to as TNFRSF10B, Death Receptor 5, or DR5), CD33, CSF1R, SEMA4A, SEMA4D, CD163, MARCO, TNFR2 (also referred to as TNFRSF1B), TREM2, MS4A7, C5AR1, LYVE1, ABCC3, LILRB4, MRC1 (also referred to as CD206), STAB1, TMEM37, MERTK, TMEM119, SIGLEC1, SIGLEC7, SIGLEC9, IL4R, MGL1 (also referred to as CD301 or CLEC10A), MGL2 (also referred to as CD301B), CD200R, or SELPLG (also referred to as PSLG-1 or CD162). In some instances, the antigen expressed by an immunosuppressive cell comprises TRAIL-R2 (also referred to as TNFRSF10B, Death Receptor 5, or DR5), CD33, CSF1R, SEMA4A, SEMA4D, CD163, MARCO, TNFR2 (also referred to as TNFRSF1B), TREM2, MS4A7, C5AR1, LYVE1, ABCC3, LILRB4, MRC1 (also referred to as CD206), STAB1, TMEM37, MERTK, TMEM119, SIGLEC1, SIGLEC7, SIGLEC9, IL4R, MGL1 (also referred to as CD301 or CLEC10A), CD200R, or SELPLG (also referred to as PSLG-1 or CD 162). In some instances, the antigen expressed by an immunosuppressive cell comprises TRAIL-R2, CSF1R, MARCO, SELPLG, CD 163, TREM2, MS4A7, C5AR1, LYVE1, MRC1, CD200R, STAB1, MERTK, SIGLEC1, IL4R, MGL1, MGL2, CD33, ABCC3, LILRB4, TMEM37, TMEM119, SIGLEC7, or SIGLEC9. In some instances, the antigen expressed by an immunosuppressive cell comprises TRAIL-R2, CSF1R, MARCO, SELPLG, CD163, TREM2, MS4A7, C5AR1, LYVE1, MRC1, CD200R, STAB1, MERTK, SIGLEC1, IL4R, MGL1, or MGL2. In some instances, the antigen expressed by an immunosuppressive cell comprises TRAIL-R2, CSF1R, CD33, TREM2, C5AR1, LYVE1, ABCC3, LILRB4, MRC1, SIGLEC1, STAB1, TMEM37, MERTK, TMEM119, SIGLEC7, SIGLEC9, or IL4R. In some instances, the antigen expressed by an immunosuppressive cell comprises TRAIL-R2, CSF1R, TREM2, C5AR1, LYVE1, MRC1, STAB1, MERTK, SIGLEC1, or IL4R. In some instances, the antigen expressed by an immunosuppressive cell comprises MARCO, SELPLG, CD 163, MS4A7, CD200R, MGL1, or MGL2. In some instances, the antigen expressed by an immunosuppressive cell comprises CD33, ABCC3, LILRB4, TMEM37, TMEM119, SIGLEC7, or SIGLEC9. In some instances, the antigen expressed by an immunosuppressive cell comprises SEMA4A, SEMA4D, or TNFR2. In some instances, the antigen expressed by an immunosuppressive cell comprises TRAIL-R2, CD33, CD163, or CSF1R. In some instances, the antigen expressed by an immunosuppressive cell comprises CD33, CD163, or CSF1R. In some instances, the antigen expressed on the immunosuppressive cell is CSF1R. In some instances, the antigen expressed on the immunosuppressive cell is SEMA4A. In some instances, the antigen expressed on the immunosuppressive cell is CD 163. In some instances, the antigen expressed on the immunosuppressive cell is TNFR2 (or TNFRSF1B). In some instances, the antigen expressed on the immunosuppressive cell is TRAIL-R2 (or TNFRSF10B or DR5). In some instances, the antigen expressed on the immunosuppressive cell is CD33. In some instances, the antigen expressed on the immunosuppressive cell is SEMA4D. In some instances, the antigen expressed on the immunosuppressive cell is LILRB4. In some instances, the antigen expressed on the immunosuppressive cell is TREM2. In some instances, the antigen expressed on the immunosuppressive cell is C5AR1. In some instances, the antigen expressed on the immunosuppressive cell is SIGLEC1. In some instances, the antigen expressed on the immunosuppressive cell is TMEM37. In some instances, the antigen expressed on the immunosuppressive cell is TMEM119. In some instances, the antigen expressed on the immunosuppressive cell is IL4R. In some instances, the antigen expressed on the immunosuppressive cell is SIGLEC7.

[0063] In some embodiments, a multi-specific binding polypeptide (e.g., a multi-specific antibody) comprises an immune cell binding moiety that binds to TRAIL-R2 (also referred to as TNFRSF10B, Death Receptor 5, or DR5), CD33, CSF1R, SEMA4A, SEMA4D, CD163, MARCO, TNFR2 (also referred to as TNFRSF1B), TREM2, MS4A7, C5AR1, LYVE1, ABCC3, LILRB4, MRC1 (also referred to as CD206), STAB1, TMEM37, MERTK, TMEM119, SIGLEC1, SIGLEC7, SIGLEC9, IL4R, MGL1 (also referred to as CD301 or CLEC10A), MGL2 (also referred to as CD301B), CD200R, or SELPLG (also referred to as PSLG-1 or CD162). In some instances, the immune cell binding moiety binds to TRAIL-R2 (also referred to as TNFRSF10B, Death Receptor 5, or DR5), CD33, CSF1R, SEMA4A, SEMA4D, CD163, MARCO, TNFR2 (also referred to as TNFRSF1B), TREM2, MS4A7, C5AR1, LYVE1, ABCC3, LILRB4, MRC1 (also referred to as CD206), STAB1, TMEM37, MERTK, TMEM119, SIGLEC1, SIGLEC7, SIGLEC9, IL4R, MGL1 (also referred to as CD301 or CLEC10A), CD200R, or SELPLG (also referred to as PSLG-1 or CD 162). In some instances, the immune cell binding moiety binds to TRAIL-R2, CSF1R, MARCO, SELPLG, CD163, TREM2, MS4A7, C5AR1, LYVE1, MRC1, CD200R, STAB1, MERTK, SIGLEC1, IL4R, MGL1, MGL2, CD33, ABCC3, LILRB4, TMEM37, TMEM119, SIGLEC7, or SIGLEC9. In some instances, the immune cell binding moiety binds to TRAIL-R2, CSF1R, MARCO, SELPLG, CD163, TREM2, MS4A7, C5AR1, LYVE1, MRC1, CD200R, STAB1, MERTK, SIGLEC1, IL4R, MGL1, or MGL2. In some instances, the immune cell binding moiety binds to TRAIL-R2, CSF1R, CD33, TREM2, C5AR1, LYVE1, ABCC3, LILRB4, MRC1, SIGLEC1, STAB1, TMEM37, MERTK, TMEM119, SIGLEC7, SIGLEC9, or IL4R. In some instances, the immune cell binding moiety binds to TRAIL-R2, CSF1R, TREM2, C5AR1, LYVE1, MRC1, STAB1, MERTK, SIGLEC1, or IL4R. In some instances, the immune cell binding moiety binds to MARCO, SELPLG, CD163, MS4A7, CD200R, MGL1, or MGL2. In some instances, the immune cell binding moiety binds to CD33, ABCC3, LILRB4, TMEM37, TMEM119, SIGLEC7, or SIGLEC9. In some instances, the immune cell binding moiety binds to SEMA4A, SEMA4D, or TNFR2. In some instances, the immune cell binding moiety binds to TRAIL-R2, CD33, CD163, or CSF1R. In some instances, the immune cell binding moiety binds to CD33, CD163, or CSF1R. In some cases, the immune cell binding moiety binds to CSF1R. In some cases, the immune cell binding moiety binds to SEMA4A. In some cases, the immune cell binding moiety binds to CD 163. In some cases, the immune cell binding moiety binds to TNFR2 (or TNFRSF1B). In some cases, the immune cell binding moiety binds to TRAIL-R2 (or TNFRSF10B or DR5). In some cases, the immune cell binding moiety binds to CD33. In some cases, the immune cell binding moiety binds to SEMA4D. In some cases, the immune cell binding moiety binds to TREM2. In some cases, the immune cell binding moiety binds to C5AR1. In some cases, the immune cell binding moiety binds to LYVE1. In some cases, the immune cell binding moiety binds to ABCC3. In some cases, the immune cell binding moiety binds to LILRB4. In some cases, the immune cell binding moiety binds to MRC1. In some cases, the immune cell binding moiety binds to SIGLEC1. In some cases, the immune cell binding moiety binds to STAB1. In some cases, the immune cell binding moiety binds to TMEM37. In some cases, the immune cell binding moiety binds to MERTK. In some cases, the immune cell binding moiety binds to TMEM119. In some cases, the immune cell binding moiety binds to SIGLEC7. In some cases, the immune cell binding moiety binds to SIGLEC9. In some cases, the immune cell binding moiety binds to IL4R. In some cases, the immune cell binding moiety binds to MARCO. In some cases, the immune cell binding moiety binds to SELPLG (also referred to as PSLG-1 or CD162). In some cases, the immune cell binding moiety binds to CD163. In some cases, the immune cell binding moiety binds to MS4A7. In some cases, the immune cell binding moiety binds to CD200R. In some cases, the immune cell binding moiety binds to MGL1.

[0064] In certain embodiments, the multi-specific binding polypeptide is a multispecific antibody (optionally a bispecific antibody) comprising an immune cell binding moiety that binds to TRAIL-R2 (also referred to as TNFRSF10B, Death Receptor 5, or DR5), CD33, CSF1R, SEMA4A, SEMA4D, CD 163, MARCO, TNFR2 (also referred to as TNFRSF1B), TREM2, MS4A7, C5AR1, LYVE1, ABCC3, LILRB4, MRC1 (also referred to as CD206), STAB1, TMEM37, MERTK, TMEM119, SIGLEC1, SIGLEC7, SIGLEC9, IL4R, MGL1 (also referred to as CD301 or CLEC10A), MGL2 (also referred to as CD301B), CD200R, or SELPLG (also referred to as PSLG-1 or CD 162). In some instances, the immune cell binding moiety binds to TRAIL-R2 (also referred to as TNFRSF10B, Death Receptor 5, or DR5), CD33, CSF1R, SEMA4A, SEMA4D, CD163, MARCO, TNFR2 (also referred to as TNFRSF1B), TREM2, MS4A7, C5AR1, LYVE1, ABCC3, LILRB4, MRC1 (also referred to as CD206), STAB1, TMEM37, MERTK, TMEM119, SIGLEC1, SIGLEC7, SIGLEC9, IL4R, MGL1 (also referred to as CD301 or CLEC10A), CD200R, or SELPLG (also referred to as PSLG-1 or CD162). In some instances, the immune cell binding moiety binds to TRAIL-R2, CSF1R, MARCO, SELPLG, CD163, TREM2, MS4A7, C5AR1, LYVE1, MRC1, CD200R, STAB1, MERTK, SIGLEC1, IL4R, MGL1, MGL2, CD33, ABCC3, LILRB4, TMEM37, TMEM119, SIGLEC7, or SIGLEC9. In some instances, the immune cell binding moiety binds to TRAIL-R2, CSF1R, MARCO, SELPLG, CD 163, TREM2, MS4A7, C5AR1, LYVE1, MRC1, CD200R, STAB1, MERTK, SIGLEC1, IL4R, MGL1, or MGL2. In some instances, the immune cell binding moiety binds to TRAIL-R2, CSF1R, CD33, TREM2, C5AR1, LYVE1, ABCC3, LILRB4, MRC1, SIGLEC1, STAB1, TMEM37, MERTK, TMEM119, SIGLEC7, SIGLEC9, or IL4R. In some instances, the immune cell binding moiety binds to TRAIL-R2, CSF1R, TREM2, C5AR1, LYVE1, MRC1, STAB1, MERTK, SIGLEC1, or IL4R. In some instances, the immune cell binding moiety binds to MARCO, SELPLG, CD 163, MS4A7, CD200R, MGL1, or MGL2. In some instances, the immune cell binding moiety binds to CD33, ABCC3, LILRB4, TMEM37, TMEM119, SIGLEC7, or SIGLEC9. In some instances, the immune cell binding moiety binds to SEMA4A, SEMA4D, or TNFR2. In some instances, the immune cell binding moiety binds to TRAIL-R2, CD33, CD 163, or CSF1R. In some instances, the immune cell binding moiety binds to CD33, CD 163, or CSF1R. In some cases, the immune cell binding moiety binds to CSF1R. In some cases, the immune cell binding moiety binds to SEMA4A. In some cases, the immune cell binding moiety binds to CD 163. In some cases, the immune cell binding moiety binds to TNFR2 (or TNFRSF1B). In some cases, the immune cell binding moiety binds to TRAIL-R2 (or TNFRSF10B or DR5). In some cases, the immune cell binding moiety binds to CD33. In some cases, the immune cell binding moiety binds to SEMA4D. In some cases, the immune cell binding moiety binds to TREM2. In some cases, the immune cell binding moiety binds to C5AR1. In some cases, the immune cell binding moiety binds to LYVE1. In some cases, the immune cell binding moiety binds to ABCC3. In some cases, the immune cell binding moiety binds to LILRB4. In some cases, the immune cell binding moiety binds to MRC1. In some cases, the immune cell binding moiety binds to SIGLEC1. In some cases, the immune cell binding moiety binds to STAB 1. In some cases, the immune cell binding moiety binds to TMEM37. In some cases, the immune cell binding moiety binds to MERTK. In some cases, the immune cell binding moiety binds to TMEM119. In some cases, the immune cell binding moiety binds to SIGLEC7. In some cases, the immune cell binding moiety binds to SIGLEC9. In some cases, the immune cell binding moiety binds to IL4R. In some cases, the immune cell binding moiety binds to MARCO. In some cases, the immune cell binding moiety binds to SELPLG (also referred to as PSLG-1 or CD162). In some cases, the immune cell binding moiety binds to CD 163. In some cases, the immune cell binding moiety binds to MS4A7. In some cases, the immune cell binding moiety binds to CD200R. In some cases, the immune cell binding moiety binds to MGL1.

[0065] In certain embodiments, a multi-specific binding polypeptide described herein is a multispecific antibody (e.g., a bispecific antibody) comprising a tumor binding moiety that specifically binds to TROP2, HER2, GPC3, GD2, FOLRI, FLT3, BCMA, MUC16, SLC4A4, STEAP1, CD19, CD20, CD22, CD25, CD33, CD38, CD30, CD47, CD123, mesothelin, MT1-MMP, or PSMA; and an immune cell binding moiety that specifically binds to TRAIL-R2 (also referred to as TNFRSF10B, Death Receptor 5, or DR5), CD33, CSF1R, SEMA4A, SEMA4D, CD163, MARCO, TNFR2 (also referred to as TNFRSF1B), TREM2, MS4A7, C5AR1, LYVE1, ABCC3, LILRB4, MRC1 (also referred to as CD206), STAB1, TMEM37, MERTK, TMEM119, SIGLEC1, SIGLEC7, SIGLEC9, IL4R, MGL1 (also referred to as CD301 or CLEC10A), MGL2 (also referred to as CD301B), CD200R, or SELPLG (also referred to as PSLG-1 or CD162). In some embodiments, the multi-specific antibody comprises a tumor binding moiety that specifically binds to TROP2, GPC3, HER2, FOLRI, CD33, CD38, FLT3, CD30, CD22, or GD2; and an immune cell binding moiety that specifically binds to TRAIL-R2 (also referred to as TNFRSF10B, Death Receptor 5, or DR5), CD33, CSF1R, SEMA4A, SEMA4D, CD163, MARCO, TNFR2 (also referred to as TNFRSF1B), TREM2, MS4A7, C5AR1, LYVE1, ABCC3, LILRB4, MRC1 (also referred to as CD206), STAB1, TMEM37, MERTK, TMEM119, SIGLEC1, SIGLEC7, SIGLEC9, IL4R, MGL1 (also referred to as CD301 or CLEC10A), MGL2 (also referred to as CD301B), CD200R, or SELPLG (also referred to as PSLG-1 or CD 162). In some embodiments, the multi-specific antibody comprises a tumor binding moiety that specifically binds to TROP2, GPC3, FOLRI, CD33, CD38, or FLT3; and an immune cell binding moiety that specifically binds to TRAIL-R2 (also referred to as TNFRSF10B, Death Receptor 5, or DR5), CD33, CSF1R, SEMA4A, SEMA4D, CD163, MARCO, TNFR2 (also referred to as TNFRSF1B), TREM2, MS4A7, C5AR1, LYVE1, ABCC3, LILRB4, MRC1 (also referred to as CD206), STAB1, TMEM37, MERTK, TMEM119, SIGLEC1, SIGLEC7, SIGLEC9, IL4R, MGL1 (also referred to as CD301 or CLEC10A), MGL2 (also referred to as CD301B), CD200R, or SELPLG (also referred to as PSLG-1 or CD162).

[0066] In some embodiments, the multi-specific antibody comprises a tumor binding moiety that specifically binds to TROP2 and an immune cell binding moiety that specifically binds to TRAIL-R2 (also referred to as TNFRSF10B, Death Receptor 5, or DR5), CD33, CSF1R, SEMA4A, SEMA4D, CD163, MARCO, TNFR2 (also referred to as TNFRSF1B), TREM2, MS4A7, C5AR1, LYVE1, ABCC3, LILRB4, MRC1 (also referred to as CD206), STAB1, TMEM37, MERTK, TMEM119, SIGLEC1, SIGLEC7, SIGLEC9, IL4R, MGL1 (also referred to as CD301 or CLEC10A), MGL2 (also referred to as CD301B), CD200R, or SELPLG (also referred to as PSLG-1 or CD162). In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to TROP2 and an immune cell binding moiety that specifically binds to TRAIL-R2, CD33, CSF1R, SEMA4A, SEMA4D, CD163, MARCO, TNFR2, TREM2, MS4A7, C5AR1, LYVE1, ABCC3, LILRB4, MRC1, STAB1, TMEM37, MERTK, TMEM119, SIGLEC1, SIGLEC7, SIGLEC9, IL4R, MGL1, CD200R, or SELPLG. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to TROP2 and an immune cell binding moiety that specifically binds to TRAIL-R2, CSF1R, MARCO, SELPLG, CD163, TREM2, MS4A7, C5AR1, LYVE1, MRC1, CD200R, STAB1, MERTK, SIGLEC1, IL4R, MGL1, MGL2, CD33, ABCC3, LILRB4, TMEM37, TMEM119, SIGLEC7, or SIGLEC9. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to TROP2 and an immune cell binding moiety that specifically binds to TRAIL-R2, CSF1R, MARCO, SELPLG, CD163, TREM2, MS4A7, C5AR1, LYVE1, MRC1, CD200R, STAB1, MERTK, SIGLEC1, IL4R, MGL1, or MGL2. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to TROP2 and an immune cell binding moiety that specifically binds to TRAIL-R2, CSF1R, CD33, TREM2, C5AR1, LYVE1, ABCC3, LILRB4, MRC1, SIGLEC1, STAB1, TMEM37, MERTK, TMEM119, SIGLEC7, SIGLEC9, or IL4R. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to TROP2 and an immune cell binding moiety that specifically binds to TRAIL-R2, CSF1R, TREM2, C5AR1, LYVE1, MRC1, STAB1, MERTK, SIGLEC1, or IL4R. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to TROP2 and an immune cell binding moiety that specifically binds to MARCO, SELPLG, CD 163, MS4A7, CD200R, MGL1, or MGL2. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to TROP2 and an immune cell binding moiety that specifically binds to CD33, ABCC3, LILRB4, TMEM37, TMEM119, SIGLEC7, or SIGLEC9. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to TROP2 and an immune cell binding moiety that specifically binds to SEMA4A, SEMA4D, or TNFR2. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to TROP2 and an immune cell binding moiety that specifically binds to TRAIL-R2, CD33, CD163, or CSF1R. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to TROP2 and an immune cell binding moiety that specifically binds to CD33, CD163, or CSF1R. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to TROP2 and an immune cell binding moiety that specifically binds to TRAIL-R2. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to TROP2 and an immune cell binding moiety that specifically binds to CD33. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to TROP2 and an immune cell binding moiety that specifically binds to CD 163. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to TROP2 and an immune cell binding moiety that specifically binds to CSF1R. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to TROP2 and an immune cell binding moiety that specifically binds to TREM2. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to TROP2 and an immune cell binding moiety that specifically binds to C5AR1. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to TROP2 and an immune cell binding moiety that specifically binds to LYVE1. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to TROP2 and an immune cell binding moiety that specifically binds to ABCC3. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to TROP2 and an immune cell binding moiety that specifically binds to LILRB4. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to TROP2 and an immune cell binding moiety that specifically binds to MRC1. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to TROP2 and an immune cell binding moiety that specifically binds to SIGLEC1. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to TROP2 and an immune cell binding moiety that specifically binds to STAB1. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to TROP2 and an immune cell binding moiety that specifically binds to TMEM37. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to TROP2 and an immune cell binding moiety that specifically binds to MERTK. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to TROP2 and an immune cell binding moiety that specifically binds to TMEM119. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to TROP2 and an immune cell binding moiety that specifically binds to SIGLEC7. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to TROP2 and an immune cell binding moiety that specifically binds to SIGLEC9. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to TROP2 and an immune cell binding moiety that specifically binds to IL4. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to TROP2 and an immune cell binding moiety that specifically binds to MARCO. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to TROP2 and an immune cell binding moiety that specifically binds to SELPLG. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to TROP2 and an immune cell binding moiety that specifically binds to MS4A7. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to TROP2 and an immune cell binding moiety that specifically binds to CD200R. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to TROP2 and an immune cell binding moiety that specifically binds to MGL1.

[0067] In some embodiments, the multi-specific antibody comprises a tumor binding moiety that specifically binds to GPC3 and an immune cell binding moiety that specifically binds to TRAIL-R2 (also referred to as TNFRSF10B, Death Receptor 5, or DR5), CD33, CSF1R, SEMA4A, SEMA4D, CD163, MARCO, TNFR2 (also referred to as TNFRSF1B), TREM2, MS4A7, C5AR1, LYVE1, ABCC3, LILRB4, MRC1 (also referred to as CD206), STAB1, TMEM37, MERTK, TMEM119, SIGLEC1, SIGLEC7, SIGLEC9, IL4R, MGL1 (also referred to as CD301 or CLEC10A), MGL2 (also referred to as CD301B), CD200R, or SELPLG (also referred to as PSLG-1 or CD162). In some instances, the multispecific antibody comprises a tumor binding moiety that specifically binds to GPC3 and an immune cell binding moiety that specifically binds to TRAIL-R2, CD33, CSF1R, SEMA4A, SEMA4D, CD163, MARCO, TNFR2, TREM2, MS4A7, C5AR1, LYVE1, ABCC3, LILRB4, MRC1, STAB1, TMEM37, MERTK, TMEM119, SIGLEC1, SIGLEC7, SIGLEC9, IL4R, MGL1, CD200R, or SELPLG. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to GPC3 and an immune cell binding moiety that specifically binds to TRAIL-R2, CSF1R, MARCO, SELPLG, CD 163, TREM2, MS4A7, C5AR1, LYVE1, MRC1, CD200R, STAB1, MERTK, SIGLEC1, IL4R, MGL1, MGL2, CD33, ABCC3, LILRB4, TMEM37, TMEM119, SIGLEC7, or SIGLEC9. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to GPC3 and an immune cell binding moiety that specifically binds to TRAIL-R2, CSF1R, MARCO, SELPLG, CD 163, TREM2, MS4A7, C5AR1, LYVE1, MRC1, CD200R, STAB1, MERTK, SIGLEC1, IL4R, MGL1, or MGL2. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to GPC3 and an immune cell binding moiety that specifically binds to TRAIL-R2, CSF1R, CD33, TREM2, C5AR1, LYVE1, ABCC3, LILRB4, MRC1, SIGLEC1, STAB1, TMEM37, MERTK, TMEM119, SIGLEC7, SIGLEC9, or IL4R. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to GPC3 and an immune cell binding moiety that specifically binds to TRAIL-R2, CSF1R, TREM2, C5AR1, LYVE1, MRC1, STAB1, MERTK, SIGLEC1, or IL4R. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to GPC3 and an immune cell binding moiety that specifically binds to MARCO, SELPLG, CD163, MS4A7, CD200R, MGL1, or MGL2. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to GPC3 and an immune cell binding moiety that specifically binds to CD33, ABCC3, LILRB4, TMEM37, TMEM119, SIGLEC7, or SIGLEC9. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to GPC3 and an immune cell binding moiety that specifically binds to SEMA4A, SEMA4D, or TNFR2. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to GPC3 and an immune cell binding moiety that specifically binds to TRAIL-R2, CD33, CD 163, or CSF1R. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to GPC3 and an immune cell binding moiety that specifically binds to CD33, CD 163, or CSF1R. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to GPC3 and an immune cell binding moiety that specifically binds to TRAIL-R2. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to GPC3 and an immune cell binding moiety that specifically binds to CD33. In some instances, the multispecific antibody comprises a tumor binding moiety that specifically binds to GPC3 and an immune cell binding moiety that specifically binds to CD 163. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to GPC3 and an immune cell binding moiety that specifically binds to CSF1R. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to GPC3 and an immune cell binding moiety that specifically binds to TREM2. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to GPC3 and an immune cell binding moiety that specifically binds to C5AR1. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to GPC3 and an immune cell binding moiety that specifically binds to LYVE1. In some instances, the multispecific antibody comprises a tumor binding moiety that specifically binds to GPC3 and an immune cell binding moiety that specifically binds to ABCC3. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to GPC3 and an immune cell binding moiety that specifically binds to LILRB4. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to GPC3 and an immune cell binding moiety that specifically binds to MRC1. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to GPC3 and an immune cell binding moiety that specifically binds to SIGLEC1. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to GPC3 and an immune cell binding moiety that specifically binds to STAB 1. In some instances, the multispecific antibody comprises a tumor binding moiety that specifically binds to GPC3 and an immune cell binding moiety that specifically binds to TMEM37. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to GPC3 and an immune cell binding moiety that specifically binds to MERTK. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to GPC3 and an immune cell binding moiety that specifically binds to TMEM119. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to GPC3 and an immune cell binding moiety that specifically binds to SIGLEC7. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to GPC3 and an immune cell binding moiety that specifically binds to SIGLEC9. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to GPC3 and an immune cell binding moiety that specifically binds to IL4. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to GPC3 and an immune cell binding moiety that specifically binds to MARCO. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to GPC3 and an immune cell binding moiety that specifically binds to SELPLG. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to GPC3 and an immune cell binding moiety that specifically binds to MS4A7. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to GPC3 and an immune cell binding moiety that specifically binds to CD200R. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to GPC3 and an immune cell binding moiety that specifically binds to MGL1.

[0068] In some embodiments, the multi-specific antibody comprises a tumor binding moiety that specifically binds to FOLRI and an immune cell binding moiety that specifically binds to TRAIL-R2 (also referred to as TNFRSF10B, Death Receptor 5, or DR5), CD33, CSF1R, SEMA4A, SEMA4D, CD163, MARCO, TNFR2 (also referred to as TNFRSF1B), TREM2, MS4A7, C5AR1, LYVE1, ABCC3, LILRB4, MRC1 (also referred to as CD206), STAB1, TMEM37, MERTK, TMEM119, SIGLEC1, SIGLEC7, SIGLEC9, IL4R, MGL1 (also referred to as CD301 or CLEC10A), MGL2 (also referred to as CD301B), CD200R, or SELPLG (also referred to as PSLG-1 or CD162). In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to FOLRI and an immune cell binding moiety that specifically binds to TRAIL-R2, CD33, CSF1R, SEMA4A, SEMA4D, CD163, MARCO, TNFR2, TREM2, MS4A7, C5AR1, LYVE1, ABCC3, LILRB4, MRC1, STAB1, TMEM37, MERTK, TMEM119, SIGLEC1, SIGLEC7, SIGLEC9, IL4R, MGL1, CD200R, or SELPLG. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to FOLRI and an immune cell binding moiety that specifically binds to TRAIL-R2, CSF1R, MARCO, SELPLG, CD163, TREM2, MS4A7, C5AR1, LYVE1, MRC1, CD200R, STAB1, MERTK, SIGLEC1, IL4R, MGL1, MGL2, CD33, ABCC3, LILRB4, TMEM37, TMEM119, SIGLEC7, or SIGLEC9. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to FOLRI and an immune cell binding moiety that specifically binds to TRAIL-R2, CSF1R, MARCO, SELPLG, CD163, TREM2, MS4A7, C5AR1, LYVE1, MRC1, CD200R, STAB1, MERTK, SIGLEC1, IL4R, MGL1, or MGL2. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to FOLRI and an immune cell binding moiety that specifically binds to TRAIL-R2, CSF1R, CD33, TREM2, C5AR1, LYVE1, ABCC3, LILRB4, MRC1, SIGLEC1, STAB1, TMEM37, MERTK, TMEM119, SIGLEC7, SIGLEC9, or IL4R. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to FOLRI and an immune cell binding moiety that specifically binds to TRAIL-R2, CSF1R, TREM2, C5AR1, LYVE1, MRC1, STAB1, MERTK, SIGLEC1, or IL4R. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to FOLRI and an immune cell binding moiety that specifically binds to MARCO, SELPLG, CD163, MS4A7, CD200R, MGL1, or MGL2. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to FOLRI and an immune cell binding moiety that specifically binds to CD33, ABCC3, LILRB4, TMEM37, TMEM119, SIGLEC7, or SIGLEC9. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to FOLRI and an immune cell binding moiety that specifically binds to SEMA4A, SEMA4D, or TNFR2. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to FOLRI and an immune cell binding moiety that specifically binds to TRAIL-R2, CD33, CD163, or CSF1R. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to FOLRI and an immune cell binding moiety that specifically binds to CD33, CD163, or CSF1R. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to FOLRI and an immune cell binding moiety that specifically binds to TRAIL-R2. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to FOLRI and an immune cell binding moiety that specifically binds to CD33. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to FOLRI and an immune cell binding moiety that specifically binds to CD 163. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to FOLRI and an immune cell binding moiety that specifically binds to CSF1R. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to FOLRI and an immune cell binding moiety that specifically binds to TREM2. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to FOLRI and an immune cell binding moiety that specifically binds to C5AR1. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to FOLRI and an immune cell binding moiety that specifically binds to LYVE1. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to FOLRI and an immune cell binding moiety that specifically binds to ABCC3. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to FOLRI and an immune cell binding moiety that specifically binds to LILRB4. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to FOLRI and an immune cell binding moiety that specifically binds to MRC1. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to FOLRI and an immune cell binding moiety that specifically binds to SIGLEC1. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to FOLRI and an immune cell binding moiety that specifically binds to STAB1. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to FOLRI and an immune cell binding moiety that specifically binds to TMEM37. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to FOLRI and an immune cell binding moiety that specifically binds to MERTK. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to FOLRI and an immune cell binding moiety that specifically binds to TMEM119. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to FOLRI and an immune cell binding moiety that specifically binds to SIGLEC7. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to FOLRI and an immune cell binding moiety that specifically binds to SIGLEC9. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to FOLRI and an immune cell binding moiety that specifically binds to IL4. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to FOLRI and an immune cell binding moiety that specifically binds to MARCO. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to FOLRI and an immune cell binding moiety that specifically binds to SELPLG. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to FOLRI and an immune cell binding moiety that specifically binds to MS4A7. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to FOLRI and an immune cell binding moiety that specifically binds to CD200R. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to FOLRI and an immune cell binding moiety that specifically binds to MGL1.

[0069] In some embodiments, the multi-specific antibody comprises a tumor binding moiety that specifically binds to CD33 and an immune cell binding moiety that specifically binds to TRAIL-R2 (also referred to as TNFRSF10B, Death Receptor 5, or DR5), CD33, CSF1R, SEMA4A, SEMA4D, CD163, MARCO, TNFR2 (also referred to as TNFRSF1B), TREM2, MS4A7, C5AR1, LYVE1, ABCC3, LILRB4, MRC1 (also referred to as CD206), STAB1, TMEM37, MERTK, TMEM119, SIGLEC1, SIGLEC7, SIGLEC9, IL4R, MGL1 (also referred to as CD301 or CLEC10A), MGL2 (also referred to as CD301B), CD200R, or SELPLG (also referred to as PSLG-1 or CD162). In some instances, the multispecific antibody comprises a tumor binding moiety that specifically binds to CD33 and an immune cell binding moiety that specifically binds to TRAIL-R2, CD33, CSF1R, SEMA4A, SEMA4D, CD163, MARCO, TNFR2, TREM2, MS4A7, C5AR1, LYVE1, ABCC3, LILRB4, MRC1, STAB1, TMEM37, MERTK, TMEM119, SIGLEC1, SIGLEC7, SIGLEC9, IL4R, MGL1, CD200R, or SELPLG. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to CD33 and an immune cell binding moiety that specifically binds to TRAIL-R2, CSF1R, MARCO, SELPLG, CD 163, TREM2, MS4A7, C5AR1, LYVE1, MRC1, CD200R, STAB1, MERTK, SIGLEC1, IL4R, MGL1, MGL2, CD33, ABCC3, LILRB4, TMEM37, TMEM119, SIGLEC7, or SIGLEC9. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to CD33 and an immune cell binding moiety that specifically binds to TRAIL-R2, CSF1R, MARCO, SELPLG, CD 163, TREM2, MS4A7, C5AR1, LYVE1, MRC1, CD200R, STAB1, MERTK, SIGLEC1, IL4R, MGL1, or MGL2. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to CD33 and an immune cell binding moiety that specifically binds to TRAIL-R2, CSF1R, CD33, TREM2, C5AR1, LYVE1, ABCC3, LILRB4, MRC1, SIGLEC1, STAB1, TMEM37, MERTK, TMEM119, SIGLEC7, SIGLEC9, or IL4R. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to CD33 and an immune cell binding moiety that specifically binds to TRAIL-R2, CSF1R, TREM2, C5AR1, LYVE1, MRC1, STAB1, MERTK, SIGLEC1, or IL4R. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to CD33 and an immune cell binding moiety that specifically binds to MARCO, SELPLG, CD163, MS4A7, CD200R, MGL1, or MGL2. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to CD33 and an immune cell binding moiety that specifically binds to CD33, ABCC3, LILRB4, TMEM37, TMEM119, SIGLEC7, or SIGLEC9. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to CD33 and an immune cell binding moiety that specifically binds to SEMA4A, SEMA4D, or TNFR2. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to CD33 and an immune cell binding moiety that specifically binds to TRAIL-R2, CD33, CD163, or CSF1R. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to CD33 and an immune cell binding moiety that specifically binds to CD33, CD163, or CSF1R. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to CD33 and an immune cell binding moiety that specifically binds to TRAIL-R2. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to CD33 and an immune cell binding moiety that specifically binds to CD33. In some instances, the multispecific antibody comprises a tumor binding moiety that specifically binds to CD33 and an immune cell binding moiety that specifically binds to CD 163. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to CD33 and an immune cell binding moiety that specifically binds to CSF1R. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to CD33 and an immune cell binding moiety that specifically binds to TREM2. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to CD33 and an immune cell binding moiety that specifically binds to C5AR1. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to CD33 and an immune cell binding moiety that specifically binds to LYVE1. In some instances, the multispecific antibody comprises a tumor binding moiety that specifically binds to CD33 and an immune cell binding moiety that specifically binds to ABCC3. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to CD33 and an immune cell binding moiety that specifically binds to LILRB4. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to CD33 and an immune cell binding moiety that specifically binds to MRC1. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to CD33 and an immune cell binding moiety that specifically binds to SIGLEC1. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to CD33 and an immune cell binding moiety that specifically binds to STAB1. In some instances, the multispecific antibody comprises a tumor binding moiety that specifically binds to CD33 and an immune cell binding moiety that specifically binds to TMEM37. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to CD33 and an immune cell binding moiety that specifically binds to MERTK. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to CD33 and an immune cell binding moiety that specifically binds to TMEM119. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to CD33 and an immune cell binding moiety that specifically binds to SIGLEC7. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to CD33 and an immune cell binding moiety that specifically binds to SIGLEC9. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to CD33 and an immune cell binding moiety that specifically binds to IL4. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to CD33 and an immune cell binding moiety that specifically binds to MARCO. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to CD33 and an immune cell binding moiety that specifically binds to SELPLG. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to CD33 and an immune cell binding moiety that specifically binds to MS4A7. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to CD33 and an immune cell binding moiety that specifically binds to CD200R. In some instances, the multispecific antibody comprises a tumor binding moiety that specifically binds to CD33 and an immune cell binding moiety that specifically binds to MGL1.

[0070] In some embodiments, the multi-specific antibody comprises a tumor binding moiety that specifically binds to CD38 and an immune cell binding moiety that specifically binds to TRAIL-R2 (also referred to as TNFRSF10B, Death Receptor 5, or DR5), CD33, CSF1R, SEMA4A, SEMA4D, CD163, MARCO, TNFR2 (also referred to as TNFRSF1B), TREM2, MS4A7, C5AR1, LYVE1, ABCC3, LILRB4, MRC1 (also referred to as CD206), STAB1, TMEM37, MERTK, TMEM119, SIGLEC1, SIGLEC7, SIGLEC9, IL4R, MGL1 (also referred to as CD301 or CLEC10A), MGL2 (also referred to as CD301B), CD200R, or SELPLG (also referred to as PSLG-1 or CD162). In some instances, the multispecific antibody comprises a tumor binding moiety that specifically binds to CD38 and an immune cell binding moiety that specifically binds to TRAIL-R2, CD33, CSF1R, SEMA4A, SEMA4D, CD163, MARCO, TNFR2, TREM2, MS4A7, C5AR1, LYVE1, ABCC3, LILRB4, MRC1, STAB1, TMEM37, MERTK, TMEM119, SIGLEC1, SIGLEC7, SIGLEC9, IL4R, MGL1, CD200R, or SELPLG. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to CD38 and an immune cell binding moiety that specifically binds to TRAIL-R2, CSF1R, MARCO, SELPLG, CD 163, TREM2, MS4A7, C5AR1, LYVE1, MRC1, CD200R, STAB1, MERTK, SIGLEC1, IL4R, MGL1, MGL2, CD33, ABCC3, LILRB4, TMEM37, TMEM119, SIGLEC7, or SIGLEC9. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to CD38 and an immune cell binding moiety that specifically binds to TRAIL-R2, CSF1R, MARCO, SELPLG, CD 163, TREM2, MS4A7, C5AR1, LYVE1, MRC1, CD200R, STAB1, MERTK, SIGLEC1, IL4R, MGL1, or MGL2. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to CD38 and an immune cell binding moiety that specifically binds to TRAIL-R2, CSF1R, CD33, TREM2, C5AR1, LYVE1, ABCC3, LILRB4, MRC1, SIGLEC1, STAB1, TMEM37, MERTK, TMEM119, SIGLEC7, SIGLEC9, or IL4R. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to CD38 and an immune cell binding moiety that specifically binds to TRAIL-R2, CSF1R, TREM2, C5AR1, LYVE1, MRC1, STAB1, MERTK, SIGLEC1, or IL4R. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to CD38 and an immune cell binding moiety that specifically binds to MARCO, SELPLG, CD 163, MS4A7, CD200R, MGL1, or MGL2. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to CD38 and an immune cell binding moiety that specifically binds to CD33, ABCC3, LILRB4, TMEM37, TMEM119, SIGLEC7, or SIGLEC9. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to CD38 and an immune cell binding moiety that specifically binds to SEMA4A, SEMA4D, or TNFR2. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to CD38 and an immune cell binding moiety that specifically binds to TRAIL-R2, CD33, CD163, or CSF1R. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to CD38 and an immune cell binding moiety that specifically binds to CD33, CD 163, or CSF1R. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to CD38 and an immune cell binding moiety that specifically binds to TRAIL-R2. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to CD38 and an immune cell binding moiety that specifically binds to CD33. In some instances, the multi -specific antibody comprises a tumor binding moiety that specifically binds to CD3 8 and an immune cell binding moiety that specifically binds to CD 163. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to CD38 and an immune cell binding moiety that specifically binds to CSF1R. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to CD38 and an immune cell binding moiety that specifically binds to TREM2. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to CD38 and an immune cell binding moiety that specifically binds to C5AR1. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to CD38 and an immune cell binding moiety that specifically binds to LYVE1. In some instances, the multispecific antibody comprises a tumor binding moiety that specifically binds to CD38 and an immune cell binding moiety that specifically binds to ABCC3. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to CD38 and an immune cell binding moiety that specifically binds to LILRB4. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to CD38 and an immune cell binding moiety that specifically binds to MRC1. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to CD38 and an immune cell binding moiety that specifically binds to SIGLEC1. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to CD38 and an immune cell binding moiety that specifically binds to STAB1. In some instances, the multispecific antibody comprises a tumor binding moiety that specifically binds to CD38 and an immune cell binding moiety that specifically binds to TMEM37. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to CD38 and an immune cell binding moiety that specifically binds to MERTK. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to CD38 and an immune cell binding moiety that specifically binds to TMEM119. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to CD38 and an immune cell binding moiety that specifically binds to SIGLEC7. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to CD38 and an immune cell binding moiety that specifically binds to SIGLEC9. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to CD38 and an immune cell binding moiety that specifically binds to IL4. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to CD38 and an immune cell binding moiety that specifically binds to MARCO. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to CD38 and an immune cell binding moiety that specifically binds to SELPLG. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to CD38 and an immune cell binding moiety that specifically binds to MS4A7. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to CD38 and an immune cell binding moiety that specifically binds to CD200R. In some instances, the multispecific antibody comprises a tumor binding moiety that specifically binds to CD38 and an immune cell binding moiety that specifically binds to MGL1.

[0071] In some embodiments, the multi-specific antibody comprises a tumor binding moiety that specifically binds to FLT3 and an immune cell binding moiety that specifically binds to TRAIL-R2 (also referred to as TNFRSF10B, Death Receptor 5, or DR5), CD33, CSF1R, SEMA4A, SEMA4D, CD163, MARCO, TNFR2 (also referred to as TNFRSF1B), TREM2, MS4A7, C5AR1, LYVE1, ABCC3, LILRB4, MRC1 (also referred to as CD206), STAB1, TMEM37, MERTK, TMEM119, SIGLEC1, SIGLEC7, SIGLEC9, IL4R, MGL1 (also referred to as CD301 or CLEC10A), MGL2 (also referred to as CD301B), CD200R, or SELPLG (also referred to as PSLG-1 or CD162). In some instances, the multispecific antibody comprises a tumor binding moiety that specifically binds to FLT3 and an immune cell binding moiety that specifically binds to TRAIL-R2, CD33, CSF1R, SEMA4A, SEMA4D, CD163, MARCO, TNFR2, TREM2, MS4A7, C5AR1, LYVE1, ABCC3, LILRB4, MRC1, STAB1, TMEM37, MERTK, TMEM119, SIGLEC1, SIGLEC7, SIGLEC9, IL4R, MGL1, CD200R, or SELPLG. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to FLT3 and an immune cell binding moiety that specifically binds to TRAIL-R2, CSF1R, MARCO, SELPLG, CD 163, TREM2, MS4A7, C5AR1, LYVE1, MRC1, CD200R, STAB1, MERTK, SIGLEC1, IL4R, MGL1, MGL2, CD33, ABCC3, LILRB4, TMEM37, TMEM119, SIGLEC7, or SIGLEC9. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to FLT3 and an immune cell binding moiety that specifically binds to TRAIL-R2, CSF1R, MARCO, SELPLG, CD 163, TREM2, MS4A7, C5AR1, LYVE1, MRC1, CD200R, STAB1, MERTK, SIGLEC1, IL4R, MGL1, or MGL2. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to FLT3 and an immune cell binding moiety that specifically binds to TRAIL-R2, CSF1R, CD33, TREM2, C5AR1, LYVE1, ABCC3, LILRB4, MRC1, SIGLEC1, STAB1, TMEM37, MERTK, TMEM119, SIGLEC7, SIGLEC9, or IL4R. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to FLT3 and an immune cell binding moiety that specifically binds to TRAIL-R2, CSF1R, TREM2, C5AR1, LYVE1, MRC1, STAB1, MERTK, SIGLEC1, or IL4R. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to FLT3 and an immune cell binding moiety that specifically binds to MARCO, SELPLG, CD163, MS4A7, CD200R, MGL1, or MGL2. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to FLT3 and an immune cell binding moiety that specifically binds to CD33, ABCC3, LILRB4, TMEM37, TMEM119, SIGLEC7, or SIGLEC9. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to FLT3 and an immune cell binding moiety that specifically binds to SEMA4A, SEMA4D, or TNFR2. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to FLT3 and an immune cell binding moiety that specifically binds to TRAIL-R2, CD33, CD163, or CSF1R. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to FLT3 and an immune cell binding moiety that specifically binds to CD33, CD 163, or CSF1R. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to FLT3 and an immune cell binding moiety that specifically binds to TRAIL-R2. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to FLT3 and an immune cell binding moiety that specifically binds to CD33. In some instances, the multispecific antibody comprises a tumor binding moiety that specifically binds to FLT3 and an immune cell binding moiety that specifically binds to CD 163. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to FLT3 and an immune cell binding moiety that specifically binds to CSF1R. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to FLT3 and an immune cell binding moiety that specifically binds to TREM2. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to FLT3 and an immune cell binding moiety that specifically binds to C5 ARI. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to FLT3 and an immune cell binding moiety that specifically binds to LYVE1. In some instances, the multispecific antibody comprises a tumor binding moiety that specifically binds to FLT3 and an immune cell binding moiety that specifically binds to ABCC3. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to FLT3 and an immune cell binding moiety that specifically binds to LILRB4. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to FLT3 and an immune cell binding moiety that specifically binds to MRC1. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to FLT3 and an immune cell binding moiety that specifically binds to SIGLEC1. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to FLT3 and an immune cell binding moiety that specifically binds to STAB1. In some instances, the multispecific antibody comprises a tumor binding moiety that specifically binds to FLT3 and an immune cell binding moiety that specifically binds to TMEM37. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to FLT3 and an immune cell binding moiety that specifically binds to MERTK. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to FLT3 and an immune cell binding moiety that specifically binds to TMEM119. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to FLT3 and an immune cell binding moiety that specifically binds to SIGLEC7. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to FLT3 and an immune cell binding moiety that specifically binds to SIGLEC9. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to FLT3 and an immune cell binding moiety that specifically binds to IL4. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to FLT3 and an immune cell binding moiety that specifically binds to MARCO. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to FLT3 and an immune cell binding moiety that specifically binds to SELPLG. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to FLT3 and an immune cell binding moiety that specifically binds to MS4A7. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to FLT3 and an immune cell binding moiety that specifically binds to CD200R. In some instances, the multispecific antibody comprises a tumor binding moiety that specifically binds to FLT3 and an immune cell binding moiety that specifically binds to MGL1.

[0072] In some embodiments, the multi-specific antibody comprises a tumor binding moiety that specifically binds to HER2 and an immune cell binding moiety that specifically binds to TRAIL-R2 (also referred to as TNFRSF10B, Death Receptor 5, or DR5), CD33, CSF1R, SEMA4A, SEMA4D, CD163, MARCO, TNFR2 (also referred to as TNFRSF1B), TREM2, MS4A7, C5AR1, LYVE1, ABCC3, LILRB4, MRC1 (also referred to as CD206), STAB1, TMEM37, MERTK, TMEM119, SIGLEC1, SIGLEC7, SIGLEC9, IL4R, MGL1 (also referred to as CD301 or CLEC10A), MGL2 (also referred to as CD301B), CD200R, or SELPLG (also referred to as PSLG-1 or CD162). In some instances, the multispecific antibody comprises a tumor binding moiety that specifically binds to HER2 and an immune cell binding moiety that specifically binds to TRAIL-R2, CD33, CSF1R, SEMA4A, SEMA4D, CD163, MARCO, TNFR2, TREM2, MS4A7, C5AR1, LYVE1, ABCC3, LILRB4, MRC1, STAB1, TMEM37, MERTK, TMEM119, SIGLEC1, SIGLEC7, SIGLEC9, IL4R, MGL1, CD200R, or SELPLG. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to HER2 and an immune cell binding moiety that specifically binds to TRAIL-R2, CSF1R, MARCO, SELPLG, CD 163, TREM2, MS4A7, C5AR1, LYVE1, MRC1, CD200R, STAB1, MERTK, SIGLEC1, IL4R, MGL1, MGL2, CD33, ABCC3, LILRB4, TMEM37, TMEM119, SIGLEC7, or SIGLEC9. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to HER2 and an immune cell binding moiety that specifically binds to TRAIL-R2, CSF1R, MARCO, SELPLG, CD 163, TREM2, MS4A7, C5AR1, LYVE1, MRC1, CD200R, STAB1, MERTK, SIGLEC1, IL4R, MGL1, or MGL2. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to HER2 and an immune cell binding moiety that specifically binds to TRAIL-R2, CSF1R, CD33, TREM2, C5AR1, LYVE1, ABCC3, LILRB4, MRC1, SIGLEC1, STAB1, TMEM37, MERTK, TMEM119, SIGLEC7, SIGLEC9, or IL4R. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to HER2 and an immune cell binding moiety that specifically binds to TRAIL-R2, CSF1R, TREM2, C5AR1, LYVE1, MRC1, STAB1, MERTK, SIGLEC1, or IL4R. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to HER2 and an immune cell binding moiety that specifically binds to MARCO, SELPLG, CD163, MS4A7, CD200R, MGL1, or MGL2. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to HER2 and an immune cell binding moiety that specifically binds to CD33, ABCC3, LILRB4, TMEM37, TMEM119, SIGLEC7, or SIGLEC9. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to HER2 and an immune cell binding moiety that specifically binds to SEMA4A, SEMA4D, or TNFR2. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to HER2 and an immune cell binding moiety that specifically binds to TRAIL-R2, CD33, CD163, or CSF1R. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to HER2 and an immune cell binding moiety that specifically binds to CD33, CD 163, or CSF1R. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to HER2 and an immune cell binding moiety that specifically binds to TRAIL-R2. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to HER2 and an immune cell binding moiety that specifically binds to CD33. In some instances, the multispecific antibody comprises a tumor binding moiety that specifically binds to HER2 and an immune cell binding moiety that specifically binds to CD 163. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to HER2 and an immune cell binding moiety that specifically binds to CSF1R. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to HER2 and an immune cell binding moiety that specifically binds to TREM2. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to HER2 and an immune cell binding moiety that specifically binds to C5 ARI. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to HER2 and an immune cell binding moiety that specifically binds to LYVE1. In some instances, the multispecific antibody comprises a tumor binding moiety that specifically binds to HER2 and an immune cell binding moiety that specifically binds to ABCC3. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to HER2 and an immune cell binding moiety that specifically binds to LILRB4. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to HER2 and an immune cell binding moiety that specifically binds to MRC1. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to HER2 and an immune cell binding moiety that specifically binds to SIGLEC1. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to HER2 and an immune cell binding moiety that specifically binds to STAB 1. In some instances, the multispecific antibody comprises a tumor binding moiety that specifically binds to HER2 and an immune cell binding moiety that specifically binds to TMEM37. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to HER2 and an immune cell binding moiety that specifically binds to MERTK. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to HER2 and an immune cell binding moiety that specifically binds to TMEM119. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to HER2 and an immune cell binding moiety that specifically binds to SIGLEC7. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to HER2 and an immune cell binding moiety that specifically binds to SIGLEC9. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to HER2 and an immune cell binding moiety that specifically binds to IL4. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to HER2 and an immune cell binding moiety that specifically binds to MARCO. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to HER2 and an immune cell binding moiety that specifically binds to SELPLG. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to HER2 and an immune cell binding moiety that specifically binds to MS4A7. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to HER2 and an immune cell binding moiety that specifically binds to CD200R. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to HER2 and an immune cell binding moiety that specifically binds to MGL1.

[0073] In some embodiments, the multi-specific antibody comprises a tumor binding moiety that specifically binds to CD30 and an immune cell binding moiety that specifically binds to TRAIL-R2 (also referred to as TNFRSF10B, Death Receptor 5, or DR5), CD33, CSF1R, SEMA4A, SEMA4D, CD163, MARCO, TNFR2 (also referred to as TNFRSF1B), TREM2, MS4A7, C5AR1, LYVE1, ABCC3, LILRB4, MRC1 (also referred to as CD206), STAB1, TMEM37, MERTK, TMEM119, SIGLEC1, SIGLEC7, SIGLEC9, IL4R, MGL1 (also referred to as CD301 or CLEC10A), MGL2 (also referred to as CD301B), CD200R, or SELPLG (also referred to as PSLG-1 or CD162). In some instances, the multispecific antibody comprises a tumor binding moiety that specifically binds to CD30 and an immune cell binding moiety that specifically binds to TRAIL-R2, CD33, CSF1R, SEMA4A, SEMA4D, CD163, MARCO, TNFR2, TREM2, MS4A7, C5AR1, LYVE1, ABCC3, LILRB4, MRC1, STAB1, TMEM37, MERTK, TMEM119, SIGLEC1, SIGLEC7, SIGLEC9, IL4R, MGL1, CD200R, or SELPLG. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to CD30 and an immune cell binding moiety that specifically binds to TRAIL-R2, CSF1R, MARCO, SELPLG, CD 163, TREM2, MS4A7, C5AR1, LYVE1, MRC1, CD200R, STAB1, MERTK, SIGLEC1, IL4R, MGL1, MGL2, CD33, ABCC3, LILRB4, TMEM37, TMEM119, SIGLEC7, or SIGLEC9. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to CD30 and an immune cell binding moiety that specifically binds to TRAIL-R2, CSF1R, MARCO, SELPLG, CD 163, TREM2, MS4A7, C5AR1, LYVE1, MRC1, CD200R, STAB1, MERTK, SIGLEC1, IL4R, MGL1, or MGL2. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to CD30 and an immune cell binding moiety that specifically binds to TRAIL-R2, CSF1R, CD33, TREM2, C5AR1, LYVE1, ABCC3, LILRB4, MRC1, SIGLEC1, STAB1, TMEM37, MERTK, TMEM119, SIGLEC7, SIGLEC9, or IL4R. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to CD30 and an immune cell binding moiety that specifically binds to TRAIL-R2, CSF1R, TREM2, C5AR1, LYVE1, MRC1, STAB1, MERTK, SIGLEC1, or IL4R. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to CD30 and an immune cell binding moiety that specifically binds to MARCO, SELPLG, CD163, MS4A7, CD200R, MGL1, or MGL2. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to CD30 and an immune cell binding moiety that specifically binds to CD33, ABCC3, LILRB4, TMEM37, TMEM119, SIGLEC7, or SIGLEC9. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to CD30 and an immune cell binding moiety that specifically binds to SEMA4A, SEMA4D, or TNFR2. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to CD30 and an immune cell binding moiety that specifically binds to TRAIL-R2, CD33, CD163, or CSF1R. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to CD30 and an immune cell binding moiety that specifically binds to CD33, CD 163, or CSF1R. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to CD30 and an immune cell binding moiety that specifically binds to TRAIL-R2. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to CD30 and an immune cell binding moiety that specifically binds to CD33. In some instances, the multispecific antibody comprises a tumor binding moiety that specifically binds to CD30 and an immune cell binding moiety that specifically binds to CD 163. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to CD30 and an immune cell binding moiety that specifically binds to CSF1R. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to CD30 and an immune cell binding moiety that specifically binds to TREM2. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to CD30 and an immune cell binding moiety that specifically binds to C5AR1. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to CD30 and an immune cell binding moiety that specifically binds to LYVE1. In some instances, the multispecific antibody comprises a tumor binding moiety that specifically binds to CD30 and an immune cell binding moiety that specifically binds to ABCC3. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to CD30 and an immune cell binding moiety that specifically binds to LILRB4. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to CD30 and an immune cell binding moiety that specifically binds to MRC1. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to CD30 and an immune cell binding moiety that specifically binds to SIGLEC1. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to CD30 and an immune cell binding moiety that specifically binds to STAB1. In some instances, the multispecific antibody comprises a tumor binding moiety that specifically binds to CD30 and an immune cell binding moiety that specifically binds to TMEM37. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to CD30 and an immune cell binding moiety that specifically binds to MERTK. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to CD30 and an immune cell binding moiety that specifically binds to TMEM119. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to CD30 and an immune cell binding moiety that specifically binds to SIGLEC7. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to CD30 and an immune cell binding moiety that specifically binds to SIGLEC9. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to CD30 and an immune cell binding moiety that specifically binds to IL4. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to CD30 and an immune cell binding moiety that specifically binds to MARCO. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to CD30 and an immune cell binding moiety that specifically binds to SELPLG. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to CD30 and an immune cell binding moiety that specifically binds to MS4A7. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to CD30 and an immune cell binding moiety that specifically binds to CD200R. In some instances, the multispecific antibody comprises a tumor binding moiety that specifically binds to CD30 and an immune cell binding moiety that specifically binds to MGL1.

[0074] In some embodiments, the multi-specific antibody comprises a tumor binding moiety that specifically binds to CD22 and an immune cell binding moiety that specifically binds to TRAIL-R2 (also referred to as TNFRSF10B, Death Receptor 5, or DR5), CD33, CSF1R, SEMA4A, SEMA4D, CD163, MARCO, TNFR2 (also referred to as TNFRSF1B), TREM2, MS4A7, C5AR1, LYVE1, ABCC3, LILRB4, MRC1 (also referred to as CD206), STAB1, TMEM37, MERTK, TMEM119, SIGLEC1, SIGLEC7, SIGLEC9, IL4R, MGL1 (also referred to as CD301 or CLEC10A), MGL2 (also referred to as CD301B), CD200R, or SELPLG (also referred to as PSLG-1 or CD162). In some instances, the multispecific antibody comprises a tumor binding moiety that specifically binds to CD22 and an immune cell binding moiety that specifically binds to TRAIL-R2, CD33, CSF1R, SEMA4A, SEMA4D, CD163, MARCO, TNFR2, TREM2, MS4A7, C5AR1, LYVE1, ABCC3, LILRB4, MRC1, STAB1, TMEM37, MERTK, TMEM119, SIGLEC1, SIGLEC7, SIGLEC9, IL4R, MGL1, CD200R, or SELPLG. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to CD22 and an immune cell binding moiety that specifically binds to TRAIL-R2, CSF1R, MARCO, SELPLG, CD 163, TREM2, MS4A7, C5AR1, LYVE1, MRC1, CD200R, STAB1, MERTK, SIGLEC1, IL4R, MGL1, MGL2, CD33, ABCC3, LILRB4, TMEM37, TMEM119, SIGLEC7, or SIGLEC9. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to CD22 and an immune cell binding moiety that specifically binds to TRAIL-R2, CSF1R, MARCO, SELPLG, CD 163, TREM2, MS4A7, C5AR1, LYVE1, MRC1, CD200R, STAB1, MERTK, SIGLEC1, IL4R, MGL1, or MGL2. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to CD22 and an immune cell binding moiety that specifically binds to TRAIL-R2, CSF1R, CD33, TREM2, C5AR1, LYVE1, ABCC3, LILRB4, MRC1, SIGLEC1, STAB1, TMEM37, MERTK, TMEM119, SIGLEC7, SIGLEC9, or IL4R. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to CD22 and an immune cell binding moiety that specifically binds to TRAIL-R2, CSF1R, TREM2, C5AR1, LYVE1, MRC1, STAB1, MERTK, SIGLEC1, or IL4R. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to CD22 and an immune cell binding moiety that specifically binds to MARCO, SELPLG, CD 163, MS4A7, CD200R, MGL1, or MGL2. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to CD22 and an immune cell binding moiety that specifically binds to CD33, ABCC3, LILRB4, TMEM37, TMEM119, SIGLEC7, or SIGLEC9. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to CD22 and an immune cell binding moiety that specifically binds to SEMA4A, SEMA4D, or TNFR2. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to CD22 and an immune cell binding moiety that specifically binds to TRAIL-R2, CD33, CD163, or CSF1R. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to CD22 and an immune cell binding moiety that specifically binds to CD33, CD 163, or CSF1R. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to CD22 and an immune cell binding moiety that specifically binds to TRAIL-R2. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to CD22 and an immune cell binding moiety that specifically binds to CD33. In some instances, the multispecific antibody comprises a tumor binding moiety that specifically binds to CD22 and an immune cell binding moiety that specifically binds to CD 163. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to CD22 and an immune cell binding moiety that specifically binds to CSF1R. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to CD22 and an immune cell binding moiety that specifically binds to TREM2. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to CD22 and an immune cell binding moiety that specifically binds to C5 ARI. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to CD22 and an immune cell binding moiety that specifically binds to LYVE1. In some instances, the multispecific antibody comprises a tumor binding moiety that specifically binds to CD22 and an immune cell binding moiety that specifically binds to ABCC3. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to CD22 and an immune cell binding moiety that specifically binds to LILRB4. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to CD22 and an immune cell binding moiety that specifically binds to MRC1. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to CD22 and an immune cell binding moiety that specifically binds to SIGLEC1. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to CD22 and an immune cell binding moiety that specifically binds to STAB1. In some instances, the multispecific antibody comprises a tumor binding moiety that specifically binds to CD22 and an immune cell binding moiety that specifically binds to TMEM37. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to CD22 and an immune cell binding moiety that specifically binds to MERTK. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to CD22 and an immune cell binding moiety that specifically binds to TMEM119. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to CD22 and an immune cell binding moiety that specifically binds to SIGLEC7. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to CD22 and an immune cell binding moiety that specifically binds to SIGLEC9. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to CD22 and an immune cell binding moiety that specifically binds to IL4. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to CD22 and an immune cell binding moiety that specifically binds to MARCO. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to CD22 and an immune cell binding moiety that specifically binds to SELPLG. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to CD22 and an immune cell binding moiety that specifically binds to MS4A7. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to CD22 and an immune cell binding moiety that specifically binds to CD200R. In some instances, the multispecific antibody comprises a tumor binding moiety that specifically binds to CD22 and an immune cell binding moiety that specifically binds to MGL1.

[0075] In some embodiments, the multi-specific antibody comprises a tumor binding moiety that specifically binds to GD2 and an immune cell binding moiety that specifically binds to TRAIL-R2 (also referred to as TNFRSF10B, Death Receptor 5, or DR5), CD33, CSF1R, SEMA4A, SEMA4D, CD163, MARCO, TNFR2 (also referred to as TNFRSF1B), TREM2, MS4A7, C5AR1, LYVE1, ABCC3, LILRB4, MRC1 (also referred to as CD206), STAB1, TMEM37, MERTK, TMEM119, SIGLEC1, SIGLEC7, SIGLEC9, IL4R, MGL1 (also referred to as CD301 or CLEC10A), MGL2 (also referred to as CD301B), CD200R, or SELPLG (also referred to as PSLG-1 or CD162). In some instances, the multispecific antibody comprises a tumor binding moiety that specifically binds to GD2 and an immune cell binding moiety that specifically binds to TRAIL-R2, CD33, CSF1R, SEMA4A, SEMA4D, CD163, MARCO, TNFR2, TREM2, MS4A7, C5AR1, LYVE1, ABCC3, LILRB4, MRC1, STAB1, TMEM37, MERTK, TMEM119, SIGLEC1, SIGLEC7, SIGLEC9, IL4R, MGL1, CD200R, or SELPLG. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to GD2 and an immune cell binding moiety that specifically binds to TRAIL-R2, CSF1R, MARCO, SELPLG, CD 163, TREM2, MS4A7, C5AR1, LYVE1, MRC1, CD200R, STAB1, MERTK, SIGLEC1, IL4R, MGL1, MGL2, CD33, ABCC3, LILRB4, TMEM37, TMEM119, SIGLEC7, or SIGLEC9. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to GD2 and an immune cell binding moiety that specifically binds to TRAIL-R2, CSF1R, MARCO, SELPLG, CD 163, TREM2, MS4A7, C5AR1, LYVE1, MRC1, CD200R, STAB1, MERTK, SIGLEC1, IL4R, MGL1, or MGL2. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to GD2 and an immune cell binding moiety that specifically binds to TRAIL-R2, CSF1R, CD33, TREM2, C5AR1, LYVE1, ABCC3, LILRB4, MRC1, SIGLEC1, STAB1, TMEM37, MERTK, TMEM119, SIGLEC7, SIGLEC9, or IL4R. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to GD2 and an immune cell binding moiety that specifically binds to TRAIL-R2, CSF1R, TREM2, C5AR1, LYVE1, MRC1, STAB1, MERTK, SIGLEC1, or IL4R. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to GD2 and an immune cell binding moiety that specifically binds to MARCO, SELPLG, CD163, MS4A7, CD200R, MGL1, or MGL2. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to GD2 and an immune cell binding moiety that specifically binds to CD33, ABCC3, LILRB4, TMEM37, TMEM119, SIGLEC7, or SIGLEC9. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to GD2 and an immune cell binding moiety that specifically binds to SEMA4A, SEMA4D, or TNFR2. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to GD2 and an immune cell binding moiety that specifically binds to TRAIL-R2, CD33, CD163, or CSF1R. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to GD2 and an immune cell binding moiety that specifically binds to CD33, CD163, or CSF1R. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to GD2 and an immune cell binding moiety that specifically binds to TRAIL-R2. In some instances, the multispecific antibody comprises a tumor binding moiety that specifically binds to GD2 and an immune cell binding moiety that specifically binds to CD33. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to GD2 and an immune cell binding moiety that specifically binds to CD 163. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to GD2 and an immune cell binding moiety that specifically binds to CSF1R. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to GD2 and an immune cell binding moiety that specifically binds to TREM2. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to GD2 and an immune cell binding moiety that specifically binds to C5AR1. In some instances, the multispecific antibody comprises a tumor binding moiety that specifically binds to GD2 and an immune cell binding moiety that specifically binds to LYVE1. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to GD2 and an immune cell binding moiety that specifically binds to ABCC3. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to GD2 and an immune cell binding moiety that specifically binds to LILRB4. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to GD2 and an immune cell binding moiety that specifically binds to MRC1. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to GD2 and an immune cell binding moiety that specifically binds to SIGLEC1. In some instances, the multispecific antibody comprises a tumor binding moiety that specifically binds to GD2 and an immune cell binding moiety that specifically binds to STAB1. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to GD2 and an immune cell binding moiety that specifically binds to TMEM37. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to GD2 and an immune cell binding moiety that specifically binds to MERTK. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to GD2 and an immune cell binding moiety that specifically binds to TMEM119. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to GD2 and an immune cell binding moiety that specifically binds to SIGLEC7. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to GD2 and an immune cell binding moiety that specifically binds to SIGLEC9. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to GD2 and an immune cell binding moiety that specifically binds to IL4. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to GD2 and an immune cell binding moiety that specifically binds to MARCO. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to GD2 and an immune cell binding moiety that specifically binds to SELPLG. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to GD2 and an immune cell binding moiety that specifically binds to MS4A7. In some instances, the multispecific antibody comprises a tumor binding moiety that specifically binds to GD2 and an immune cell binding moiety that specifically binds to CD200R. In some instances, the multi-specific antibody comprises a tumor binding moiety that specifically binds to GD2 and an immune cell binding moiety that specifically binds to MGL1.

[0076] In certain embodiments, the multi-specific binding polypeptide (e.g., the multi-specific antibody) comprises a tumor binding moiety that specifically binds to TROP2 / TACSTD2. In some embodiments, the multi-specific binding polypeptide (e.g., the multi-specific antibody) comprises an immune cell binding moiety that specifically binds to TRAIL-R2, CSF1R, SEMA4A, TNFR2, TREM2, ABCC3, STAB1, or IL4R; TRAIL-R2, CSF1R, SEMA4A, CD163, TNFR2, TREM2, MS4A7, ABCC3, LILRB4, STAB1, TMEM119, or IL4R; TRAIL-R2, CSF1R, SEMA4A, TNFR2, CD163, TREM2, LILRB4, STAB1, TMEM119, or IL4R; TRAIL-R2, CSF1R, SEMA4A, TNFR2, TREM2, ABCC3, LILRB4, STAB1, TMEM37, or IL4R; TRAIL-R2, CSF1R, TNFR2, TREM2, MS4A7, CSAR1, ABCC3, LILRB4, STAB1, TMEM37, or IL4R; TRAIL-R2, TNFR2, ABCC3, STAB1, or IL4R; TRAIL-R2, CSF1R, SEMA4A, CD163, TNFR2, TREM2, MS4A7, C5AR1, ABCC3, LILRB4, STAB1, TMEM37, or IL4R; TRAIL-R2, CSF1R, SEMA4A, CD163, MARCO, TNFR2, TREM2, MS4A7, C5AR1, ABCC3, LILRB4, MRC1, STAB1, TMEM37, TMEM119, CLEC10A, or IL4R; TRAIL-R2, CSF1R, SEMA4A, TNFR2, TREM2, C5AR1, ABCC3, LILRB4, MRC1, STAB1, MERTK, TMEM37, TMEM119, or IL4R; TRAIL-R2, CSF1R, SEMA4A, CD163, MARCO, TNFR2, TREM2, C5AR1, ABCC3, LILRB4, MRC1, STAB1, MERTK, TMEM37, or IL4R; TRAIL-R2, CSF1R, SEMA4A, CD 163, TNFR2, TREM2, MS4A7, C5AR1, ABCC3, LILRB4, MRC1, STAB1, MERTK, TMEM37, TMEM119, or IL4R; or TRAIL-R2, CSF1R, TNFR2, ABCC3, STAB1, TMEM37, TMEM119, or IL4R. In certain embodiments, the multi-specific binding polypeptide (e.g., the multi-specific antibody) comprises a tumor binding moiety that specifically binds to TROP2 / TACSTD2 and an immune cell binding moiety that specifically binds to TRAIL-R2, CSF1R, SEMA4A, TNFR2, TREM2, ABCC3, STAB1, or IL4R. In certain embodiments, the multi-specific binding polypeptide (e.g., the multi-specific antibody) comprises a tumor binding moiety that specifically binds to TROP2 / TACSTD2 and an immune cell binding moiety that specifically binds to TRAIL-R2, CSF1R, SEMA4A, CD 163, TNFR2, TREM2, MS4A7, ABCC3, LILRB4, STAB1, TMEM119, or IL4R. In certain embodiments, the multi-specific binding polypeptide (e.g., the multi-specific antibody) comprises a tumor binding moiety that specifically binds to TROP2 / TACSTD2 and an immune cell binding moiety that specifically binds to TRAIL-R2, CSF1R, SEMA4A, TNFR2, CD 163, TREM2, LILRB4, STAB1, TMEM119, or IL4R. In certain embodiments, the multi-specific binding polypeptide (e.g., the multi-specific antibody) comprises a tumor binding moiety that specifically binds to TROP2 / TACSTD2 and an immune cell binding moiety that specifically binds to TRAIL-R2, CSF1R, SEMA4A, TNFR2, TREM2, ABCC3, LILRB4, STAB1, TMEM37, or IL4R. In certain embodiments, the multi-specific binding polypeptide (e.g., the multi-specific antibody) comprises a tumor binding moiety that specifically binds to TROP2 / TACSTD2 and an immune cell binding moiety that specifically binds to TRAIL-R2, CSF1R, TNFR2, TREM2, MS4A7, CSAR1, ABCC3, LILRB4, STAB1, TMEM37, or IL4R. In certain embodiments, the multi-specific binding polypeptide (e.g., the multi-specific antibody) comprises a tumor binding moiety that specifically binds to TROP2 / TACSTD2 and an immune cell binding moiety that specifically binds to TRAIL-R2, TNFR2, ABCC3, STAB1, or IL4R. In certain embodiments, the multi-specific binding polypeptide (e.g., the multi-specific antibody) comprises a tumor binding moiety that specifically binds to TROP2 / TACSTD2 and an immune cell binding moiety that specifically binds to TRAIL-R2, CSF1R, SEMA4A, CD163, TNFR2, TREM2, MS4A7, C5AR1, ABCC3, LILRB4, STAB1, TMEM37, or IL4R. In certain embodiments, the multi-specific binding polypeptide (e.g., the multi-specific antibody) comprises a tumor binding moiety that specifically binds to TROP2 / TACSTD2 and an immune cell binding moiety that specifically binds to TRAIL-R2, CSF1R, SEMA4A, CD 163, MARCO, TNFR2, TREM2, MS4A7, C5AR1, ABCC3, LILRB4, MRC1, STAB1, TMEM37, TMEM119, CLEC10A, or IL4R. In certain embodiments, the multi-specific binding polypeptide (e.g., the multi-specific antibody) comprises a tumor binding moiety that specifically binds to TROP2 / TACSTD2 and an immune cell binding moiety that specifically binds to TRAIL-R2, CSF1R, SEMA4A, TNFR2, TREM2, C5AR1, ABCC3, LILRB4, MRC1, STAB1, MERTK, TMEM37, TMEM119, or IL4R. In certain embodiments, the multi-specific binding polypeptide (e.g., the multi-specific antibody) comprises a tumor binding moiety that specifically binds to TROP2 / TACSTD2 and an immune cell binding moiety that specifically binds to TRAIL-R2, CSF1R, SEMA4A, CD163, MARCO, TNFR2, TREM2, C5AR1, ABCC3, LILRB4, MRC1, STAB1, MERTK, TMEM37, or IL4R. In certain embodiments, the multi-specific binding polypeptide (e.g., the multi-specific antibody) comprises a tumor binding moiety that specifically binds to TROP2 / TACSTD2 and an immune cell binding moiety that specifically binds to TRAIL-R2, CSF1R, SEMA4A, CD163, TNFR2, TREM2, MS4A7, C5AR1, ABCC3, LILRB4, MRC1, STAB1, MERTK, TMEM37, TMEM119, or IL4R. In certain embodiments, the multi-specific binding polypeptide (e.g., the multi-specific antibody) comprises a tumor binding moiety that specifically binds to TROP2 / TACSTD2 and an immune cell binding moiety that specifically binds to TRAIL-R2, CSF1R, TNFR2, ABCC3, STAB1, TMEM37, TMEM119, or IL4R. In some embodiments, the multi-specific binding polypeptide (e.g., the multi specific antibody) is a bispecific antibody.

[0077] In certain embodiments, the multi-specific binding polypeptide (e.g., the multi-specific antibody) comprises a tumor binding moiety that specifically binds to TROP2 / TACSTD2 or FOLRI. In some embodiments, the multi-specific binding polypeptide (e.g., the multi-specific antibody) comprises an immune cell binding moiety that specifically binds to TRAIL-R2, CD33, CSF1R, CD 163, MARCO, TNFR2, TREM2, MS4A7, C5AR1, ABCC3, LILRB4, STAB1, TMEM37, or IL4R; TNFRSF10B, TNFRSF1B, CSF1R, SEMA4A, CD163, MARCO, TNFRSF1B, TREM2, MS4A7, C5AR1, ABCC3, LILRB4, MRC1, STAB1, TMEM37, MERTK, TMEM119, SIGLEC1, or CLEC10A; or TRAIL-R2, CSF1R, SEMA4A, CD163, TNFR2, TREM2, STAB1, LILRB4, TMEM37, MERTK, TMEM119, or IL4R. In certain embodiments, the multi-specific binding polypeptide (e.g., the multi-specific antibody) comprises a tumor binding moiety that specifically binds to TROP2 / TACSTD2 or FOLRI, and an immune cell binding moiety that specifically binds to TRAIL-R2, CSF1R, CD 163, MARCO, TNFR2, TREM2, MS4A7, C5AR1, ABCC3, LILRB4, STAB1, TMEM37, or IL4R. In certain embodiments, the multi-specific binding polypeptide (e.g., the multi-specific antibody) comprises a tumor binding moiety that specifically binds to TROP2 / TACSTD2 or FOLRI, and an immune cell binding moiety that specifically binds to TNFRSF10B, TNFRSF1B, CSF1R, SEMA4A, CD163, MARCO, TNFRSF1B, TREM2, MS4A7, C5AR1, ABCC3, LILRB4, MRC1, STAB1, TMEM37, MERTK, TMEM119, SIGLEC1, or CLEC10A. In certain embodiments, the multi-specific binding polypeptide (e.g., the multispecific antibody) comprises a tumor binding moiety that specifically binds to TROP2 / TACSTD2 or FOLRI, and an immune cell binding moiety that specifically binds to TRAIL-R2, CSF1R, SEMA4A, CD 163, TNFR2, TREM2, STAB1, LILRB4, TMEM37, MERTK, TMEM119, or IL4R. In some embodiments, the multi-specific binding polypeptide (e.g., the multi-specific antibody) is a bispecific antibody.

[0078] In certain embodiments, the multi-specific binding polypeptide (e.g., the multi-specific antibody) comprises a tumor binding moiety that specifically binds to TROP2 / TACSTD2 or FOLH1 / PSMA. In some embodiments, the multi-specific binding polypeptide (e.g., the multi-specific antibody) comprises an immune cell binding moiety that specifically binds to TRAIL-R2, CSF1R, SEMA4A, TNFR2, TREM2, MS4A7, C5AR1, ABCC3, STAB1, LILRB4, TMEM37, MERTK, TMEM119, or IL4R. In some embodiments, the multi-specific binding polypeptide (e.g., the multispecific antibody) is a bispecific antibody.

[0079] In certain embodiments, the multi-specific binding polypeptide (e.g., the multi-specific antibody) comprises a tumor binding moiety that specifically binds to FOLRI. In some embodiments, the multi-specific binding polypeptide (e.g., the multi-specific antibody) comprises an immune cell binding moiety that specifically binds to TRAIL-R2, CSF1R, SEMA4A, CD163, TNFR2, TREM2, MS4A7, C5AR1, LYVE1, ABCC3, LILRB4, MRC1, STAB1, MERTK, TMEM37, TMEM119, IL4R, or SIGLEC1. In some embodiments, the multi-specific binding polypeptide (e.g., the multi-specific antibody) is a bispecific antibody.

[0080] In certain embodiments, the multi-specific binding polypeptide (e.g., the multi-specific antibody) comprises a tumor binding moiety that specifically binds to CD33 or CD 123. In some embodiments, the multi-specific binding polypeptide (e.g., the multi-specific antibody) comprises an immune cell binding moiety that specifically binds to TNFRSF10B / TRAIL-R2, TNFRSF1B / TNFR2, CSF1R, SEMA4A, MS4A7, C5AR1, LILRB4, STAB1, MERTK, SIGLEC7, SIGLEC9, CLEC10A, or CD200R1. In some embodiments, the multi-specific binding polypeptide (e.g., the multi-specific antibody) is a bispecific antibody.

[0081] In certain embodiments, the multi-specific binding polypeptide (e.g., the multi-specific antibody) comprises a tumor binding moiety that specifically binds to CD 123, CD30, CD22, or CD 19. In some embodiments, the multi-specific binding polypeptide (e.g., the multi-specific antibody) comprises an immune cell binding moiety that specifically binds to TNFRSF10B, CSF1R, SEMA4A, CD163, MARCO, TNFRSF1B, C5AR1, ABCC3, LILRB4, STAB1, TMEM119, SIGLEC1, SIGLEC7, SIGLEC9, CLEC10A, or IL4R. In some embodiments, the multi-specific binding polypeptide (e.g., the multi-specific antibody) is a bispecific antibody. Multi-specific Antibodies

[0082] In certain embodiments, a multi-specific binding polypeptide (e.g., a multi-specific antibody) described herein comprises a tumor binding moiety comprising sequences at least about 80%, 85%, 90%, 95%, 97%, 98%, 99%, or 100% identical to or consist of sequences (e.g., CDRs or VH / VL sequences) selected from Tables 1 and / or 2. Table 1 NAME* VH SEQUENCE* HCDR1 HCDR2 HCDR3 VH aTROP2 QVQLQQSGSELKKPGASVKVSCKASGY TFTNYGMNWVKOAPGOGLKWMGWIN TYTGEPTYTDDFKGRFAFSLDTSVSTAY LQIS SLKADDTAVYFCARGGFGS SYWY FDVWGOGSLVTVSS SEQID NO: 4 SEQID NO: 5 SEQ ID NO: 6 SEQID NO: 9 «GPC3 QVQLVQSGAEVKKPGASVKVSCKASG YTFTDYEMHWVRQAPGQGLEWMGAL DPKTGDTAYSQKFKGRVTLTADKSTST AYMELSSLTSEDTAVYYCTRFYSYTYW GQGTLVTVSS SEQID NO: 13 SEQID NO: 14 SEQ ID NO: 15 SEQID NO: 16 aFOLRl QVQLVQSGAEWKPGASVKISCKASGY TFTGYFMNWVKQSPGQSLEWIGRIHPY DGDTFYNQKFQGKATLTVDKS SNTAH MELLSLTSEDFAVYYCTRYDGSRAMDY WGQGTTVTVSS SEQID NO: 17 SEQID NO: 18 SEQ ID NO: 19 SEQID NO: 20 aCD38 EVQLLESGGGLVQPGGSLRLSCAVSGFT FNSFAMSWVROAPGKGLEWVSAISGSG GGTYYADSVKGRFTISRDNSKNTLYLQ MNSLRAEDTAVYFCAKDKILWFGEPVF DYWGQGTLVTVSS SEQID NO: 21 SEQID NO: 22 SEQ ID NO: 23 SEQID NO: 24 aFLT3 EVQLVQSGAEVKKPGASVKVSCKASG YTFTSYYMHWVRQAPGQGLEWMGIIN SEQID NO: 25 SEQID NO: 26 SEQ ID NO: 27 SEQID NO: 28 PSGGSTSYAQKFQGRVTMTRDTSTSTV YMELS SLRSEDTAVYYCARGVGAHDA FDIWGQGTTVTVSS *The notation “a” in front of an exemplary tumor-associated antigen denotes an antibody that specifically binds to the tumor-associated antigen. For example, aTROP2 refers to an antibody that specifically binds to TROP2; aGPC3 refers to an antibody that specifically binds to GPC3; aFOLRl refers to an antibody that specifically binds to FOLRI; aCD38 refers to an antibody that specifically binds to CD38; and aFLT3 refers to an antibody that specifically binds to FLT3. #The underlined sequences denote the respective HCDR1, HCDR2, and HCDR3. For example, the first underlined sequence in aTROP2 denotes HCDR1; the second underlined sequence in aTROP2 denotes HCDR2; and the third underlined sequence in aTROP2 denotes HCDR3. Table 2 NAME" VL SEQUENCE* LCDR1 LCDR2 LCDR3 VL aTROP2 DIQLTQSPSSLSASVGDRVSITCKASQD VSIAVAWYQQKPGKAPKLLIYSASYRY TGVPDRFSGSGSGTDFTLTISSLQPEDFA VYYCQQHYITPLTFGAGTKVEIK SEQ ID NO: 1 SEQ ID NO: 2 SEQ ID NO: 3 SEQ ID NO: 10 aGPC3 DVVMTOSPLSLPVTPGEPASISCRSSQSL VHSNRNTYLHWYLOKPGOSPOLLIYKV SNRFSGVPDRFSGSGSGTDFTLKISRVE AEDVGVYYCSQNTHVPPTFGQGTKLEI K SEQ ID NO: 29 SEQ ID NO: 30 SEQ ID NO: 31 SEQ ID NO: 32 aFOLRl DIVLTQSPLSLAVSLGQPAIISCKASOSV SFAGTSLMHWYHOKPGOQPRLLIYRAS NLEAGVPDRFSGSGSKTDFTLTISPVEA EDAATYYCQQSREYPYTFGGGTKLEIK SEQ ID NO: 33 SEQ ID NO: 34 SEQ ID NO: 35 SEQ ID NO: 36 aCD38 EIVLTOSPATLSLSPGERATLSCRASQSV SSYLAWYOQKPGOAPRLLIYDASNRAT GIPARFSGSGSGTDFTLTISSLEPEDFAV YYCQQRSNWPPTFGOGTKVEIK SEQ ID NO: 37 SEQ ID NO: 38 SEQ ID NO: 39 SEQ ID NO: 40 aFLT3 DVVMTQSPLSLPVTPGEPASISCRSSQSL LHSNGNNYLDWYLQKPGQSPQLLIYLG SNRASGVPDRFSGSGSDTDFTLOISRVE AEDVGVYYCMQGTHPAISFGOGTRLEI K SEQ ID NO: 41 SEQ ID NO: 42 SEQ ID NO: 43 SEQ ID NO: 44 *The notation “a” in front of an exemplary tumor-associated antigen denotes an antibody that specifically binds to the tumor-associated antigen. #The underlined sequences denote the respective LCDR1, LCDR2, and LCDR3. For example, the first underlined sequence in aTROP2 denotes LCDR1; the second underlined sequence in aTROP2 denotes LCDR2; and the third underlined sequence in aTROP2 denotes LCDR3.

[0083] In some embodiments, a multi-specific binding polypeptide (e.g., a multi-specific antibody) described herein comprises a tumor binding moiety comprising an immunoglobulin heavy chain variable region comprising an amino acid sequence at least about 80%, 85%, 90%, 95%, 97%, 98%, 99%, or 100% identical to or consist of the amino acid sequence set forth in SEQ ID NOs: 9, 16, 20, 24, or 28; and an immunoglobulin light chain variable region at least about 80%, 85%, 90%, 95%, 97%, 98%, 99%, or 100% identical to or consist of the amino acid sequence set forth in SEQ ID NOs: 10, 32, 36, 40, or 44. In some instances, the amino acid differences contributing to the at least about 80%, 85%, 90%, 95%, 97%, 98%, or 99% sequence identity resides in a framework region of the immunoglobulin heavy chain variable region and the CDRs remain unchanged relative to the CDRs set forth in SEQ ID NOs: 9, 16, 20, 24, or 28. In some instances, the amino acid differences contributing to the at least about 80%, 85%, 90%, 95%, 97%, 98%, or 99% sequence identity resides in a framework region of the immunoglobulin light chain variable region and the CDRs remain unchanged relative to the CDRs set forth in SEQ ID NOs: 10, 32, 36, 40, or 44. In some cases, the amino acid differences contributing to the at least about 80%, 85%, 90%, 95%, 97%, 98%, or 99% sequence identity comprises conservative substitutions in the framework region, CDRs region, or a combination thereof, of the immunoglobulin heavy chain variable region but the multi-specific binding polypeptide (e.g., the multi-specific antibody) retains binding to a target tumor antigen. In some cases, the amino acid differences contributing to the at least about 80%, 85%, 90%, 95%, 97%, 98%, or 99% sequence identity comprises conservative substitutions in the framework region, CDRs region, or a combination thereof, of the immunoglobulin light chain variable region but the multi-specific binding polypeptide (e.g., the multi-specific antibody) retains binding to a target tumor antigen.

[0084] In some embodiments, a multi-specific binding polypeptide (e.g., a multi-specific antibody) described herein comprises a tumor binding moiety comprising an immunoglobulin heavy chain variable region comprising an amino acid sequence at least about 80%, 85%, 90%, 95%, 97%, 98%, 99%, or 100% identical to or consist of the amino acid sequence set forth in SEQ ID NO: 9; and an immunoglobulin light chain variable region at least about 80%, 85%, 90%, 95%, 97%, 98%, 99%, or 100% identical to or consist of the amino acid sequence set forth in SEQ ID NO: 10. In some instances, the amino acid differences contributing to the at least about 80%, 85%, 90%, 95%, 97%, 98%, or 99% sequence identity resides in a framework region of the immunoglobulin heavy chain variable region and the CDRs remain unchanged relative to the CDRs set forth in SEQ ID NO: 9. In some instances, the amino acid differences contributing to the at least about 80%, 85%, 90%, 95%, 97%, 98%, or 99% sequence identity resides in a framework region of the immunoglobulin light chain variable region and the CDRs remain unchanged relative to the CDRs set forth in SEQ ID NO: 10. In some cases, the amino acid differences contributing to the at least about 80%, 85%, 90%, 95%, 97%, 98%, or 99% sequence identity comprises conservative substitutions in the framework region, CDRs region, or a combination thereof, of the immunoglobulin heavy chain variable region but the multi-specific binding polypeptide (e.g., the multi-specific antibody) retains binding to a target tumor antigen. In some cases, the amino acid differences contributing to the at least about 80%, 85%, 90%, 95%, 97%, 98%, or 99% sequence identity comprises conservative substitutions in the framework region, CDRs region, or a combination thereof, of the immunoglobulin light chain variable region but the multi-specific binding polypeptide (e.g., the multi-specific antibody) retains binding to a target tumor antigen.

[0085] In some embodiments, a multi-specific binding polypeptide (e.g., a multi-specific antibody) described herein comprises a tumor binding moiety comprising an immunoglobulin heavy chain variable region comprising an amino acid sequence at least about 80%, 85%, 90%, 95%, 97%, 98%, 99%, or 100% identical to or consist of the amino acid sequence set forth in SEQ ID NO: 16; and an immunoglobulin light chain variable region at least about 80%, 85%, 90%, 95%, 97%, 98%, 99%, or 100% identical to or consist of the amino acid sequence set forth in SEQ ID NO: 32. In some instances, the amino acid differences contributing to the at least about 80%, 85%, 90%, 95%, 97%, 98%, or 99% sequence identity resides in a framework region of the immunoglobulin heavy chain variable region and the CDRs remain unchanged relative to the CDRs set forth in SEQ ID NO: 16. In some instances, the amino acid differences contributing to the at least about 80%, 85%, 90%, 95%, 97%, 98%, or 99% sequence identity resides in a framework region of the immunoglobulin light chain variable region and the CDRs remain unchanged relative to the CDRs set forth in SEQ ID NO: 32. In some cases, the amino acid differences contributing to the at least about 80%, 85%, 90%, 95%, 97%, 98%, or 99% sequence identity comprises conservative substitutions in the framework region, CDRs region, or a combination thereof, of the immunoglobulin heavy chain variable region but the multi-specific binding polypeptide (e.g., the multi-specific antibody) retains binding to a target tumor antigen. In some cases, the amino acid differences contributing to the at least about 80%, 85%, 90%, 95%, 97%, 98%, or 99% sequence identity comprises conservative substitutions in the framework region, CDRs region, or a combination thereof, of the immunoglobulin light chain variable region but the multi-specific binding polypeptide (e.g., the multi-specific antibody) retains binding to a target tumor antigen.

[0086] In some embodiments, a multi-specific binding polypeptide (e.g., a multi-specific antibody) described herein comprises a tumor binding moiety comprising an immunoglobulin heavy chain variable region comprising an amino acid sequence at least about 80%, 85%, 90%, 95%, 97%, 98%, 99%, or 100% identical to or consist of the amino acid sequence set forth in SEQ ID NO: 20; and an immunoglobulin light chain variable region at least about 80%, 85%, 90%, 95%, 97%, 98%, 99%, or 100% identical to or consist of the amino acid sequence set forth in SEQ ID NO: 36. In some instances, the amino acid differences contributing to the at least about 80%, 85%, 90%, 95%, 97%, 98%, or 99% sequence identity resides in a framework region of the immunoglobulin heavy chain variable region and the CDRs remain unchanged relative to the CDRs set forth in SEQ ID NO: 20. In some instances, the amino acid differences contributing to the at least about 80%, 85%, 90%, 95%, 97%, 98%, or 99% sequence identity resides in a framework region of the immunoglobulin light chain variable region and the CDRs remain unchanged relative to the CDRs set forth in SEQ ID NO: 36. In some cases, the amino acid differences contributing to the at least about 80%, 85%, 90%, 95%, 97%, 98%, or 99% sequence identity comprises conservative substitutions in the framework region, CDRs region, or a combination thereof, of the immunoglobulin heavy chain variable region but the multi-specific binding polypeptide (e.g., the multi-specific antibody) retains binding to a target tumor antigen. In some cases, the amino acid differences contributing to the at least about 80%, 85%, 90%, 95%, 97%, 98%, or 99% sequence identity comprises conservative substitutions in the framework region, CDRs region, or a combination thereof, of the immunoglobulin light chain variable region but the multi-specific binding polypeptide (e.g., the multi-specific antibody) retains binding to a target tumor antigen.

[0087] In some embodiments, a multi-specific binding polypeptide (e.g., a multi-specific antibody) described herein comprises a tumor binding moiety comprising an immunoglobulin heavy chain variable region comprising an amino acid sequence at least about 80%, 85%, 90%, 95%, 97%, 98%, 99%, or 100% identical to or consist of the amino acid sequence set forth in SEQ ID NO: 24; and an immunoglobulin light chain variable region at least about 80%, 85%, 90%, 95%, 97%, 98%, 99%, or 100% identical to or consist of the amino acid sequence set forth in SEQ ID NO: 40. In some instances, the amino acid differences contributing to the at least about 80%, 85%, 90%, 95%, 97%, 98%, or 99% sequence identity resides in a framework region of the immunoglobulin heavy chain variable region and the CDRs remain unchanged relative to the CDRs set forth in SEQ ID NO: 24. In some instances, the amino acid differences contributing to the at least about 80%, 85%, 90%, 95%, 97%, 98%, or 99% sequence identity resides in a framework region of the immunoglobulin light chain variable region and the CDRs remain unchanged relative to the CDRs set forth in SEQ ID NO: 40. In some cases, the amino acid differences contributing to the at least about 80%, 85%, 90%, 95%, 97%, 98%, or 99% sequence identity comprises conservative substitutions in the framework region, CDRs region, or a combination thereof, of the immunoglobulin heavy chain variable region but the multi-specific binding polypeptide (e.g., the multi-specific antibody) retains binding to a target tumor antigen. In some cases, the amino acid differences contributing to the at least about 80%, 85%, 90%, 95%, 97%, 98%, or 99% sequence identity comprises conservative substitutions in the framework region, CDRs region, or a combination thereof, of the immunoglobulin light chain variable region but the multi-specific binding polypeptide (e.g., the multi-specific antibody) retains binding to a target tumor antigen.

[0088] In some embodiments, a multi-specific binding polypeptide (e.g., a multi-specific antibody) described herein comprises a tumor binding moiety comprising an immunoglobulin heavy chain variable region comprising an amino acid sequence at least about 80%, 85%, 90%, 95%, 97%, 98%, 99%, or 100% identical to or consist of the amino acid sequence set forth in SEQ ID NO: 28; and an immunoglobulin light chain variable region at least about 80%, 85%, 90%, 95%, 97%, 98%, 99%, or 100% identical to or consist of the amino acid sequence set forth in SEQ ID NO: 44. In some instances, the amino acid differences contributing to the at least about 80%, 85%, 90%, 95%, 97%, 98%, or 99% sequence identity resides in a framework region of the immunoglobulin heavy chain variable region and the CDRs remain unchanged relative to the CDRs set forth in SEQ ID NO: 28. In some instances, the amino acid differences contributing to the at least about 80%, 85%, 90%, 95%, 97%, 98%, or 99% sequence identity resides in a framework region of the immunoglobulin light chain variable region and the CDRs remain unchanged relative to the CDRs set forth in SEQ ID NO: 44. In some cases, the amino acid differences contributing to the at least about 80%, 85%, 90%, 95%, 97%, 98%, or 99% sequence identity comprises conservative substitutions in the framework region, CDRs region, or a combination thereof, of the immunoglobulin heavy chain variable region but the multi-specific binding polypeptide (e.g., the multi-specific antibody) retains binding to a target tumor antigen. In some cases, the amino acid differences contributing to the at least about 80%, 85%, 90%, 95%, 97%, 98%, or 99% sequence identity comprises conservative substitutions in the framework region, CDRs region, or a combination thereof, of WO 2020 / 142659                                                 PCT / US2020 / 012139 the immunoglobulin light chain variable region but the multi-specific binding polypeptide (e.g., the multi-specific antibody) retains binding to a target tumor antigen.

[0089] In certain embodiments, a multi-specific binding polypeptide (e.g., a multi-specific antibody) described herein comprises a tumor binding moiety comprising sequences (e.g., CDRs or VH / VL sequences) derived from a known antibody such as RS7, MAAP-9001a, 7E6, 4F6 (TROP2), Codrituzumab / GC33 / RO5137382, HN3, YP7, HS20, 4G5, MDX-1414 (GPC3) Trastuzumab (HER2), brentuximab, ramucirumab, iratumumab, olinvacimab, vesencumab (CD30), dinutuximab, Hu3F8, MAb-3F8, MORAb-028 / MT228, KM666 (GD2), mirvetuximab / huFR107, farletuzumab, MORAb-003, SP8166, 26B3.F2, HuRA15 (FOLRI), daratumumab, isatuximab, mezagitamab (CD38), or IMC-EB10 (FLT3).

[0090] In certain embodiments, a multi-specific binding polypeptide (e.g., a multi-specific antibody) described herein comprises an immune cell binding moiety comprising sequences at least about 80%, 85%, 90%, 95%, 97%, 98%, 99%, or 100% identical to or consist of sequences (e.g., CDRs or VH / VL sequences) selected from Tables 3 and / or 4. Table 3 NAME* VH SEQUENCE* HCDR1 HCDR2 HCDR3 VH aCD33 EVQLVQSGAEVKKPGSSVKVSCKASGY TITDSNIHWVRQAPGQSLEWIGYIYPYN GGTDYNQKFKNRATLTVDNPTNTAYM ELS SLRSEDTAFYYCVNGNPWLAYWG QGTLVTVSS SEQID NO: 45 SEQID NO: 46 SEQ ID NO: 47 SEQID NO: 48 aCD163 QVQLQESGPGLVKPSETLSLTCTVSGYS ITSDYAWNWIRQFPGNKLEWMGYITYS GSTYYNPSLKSRVTISVDTSKNQFSLKL SSVTAADTATYYCVSGTYYFDYWGOG TTLTVSS SEQID NO: 49 SEQID NO: 50 SEQ ID NO: 51 SEQID NO: 52 aTRAIL-R2 (aDR5) QVQLQESGPGLVKPSQTLSLTCTVSGGS IS SGDYFWSWIROLPGKGLEWIGHIHNS GTTYYNPSLKSRVTISVDTSKKQFSLRL S SVTAADTAVYYCARDRGGDYYYGMD VWGQGTTVTVSS SEQID NO: 53 SEQID NO: 54 SEQ ID NO: 55 SEQID NO: 11 aCSFIR QVQLVQSGAEVKKPGSSVKVSCKASGY TFTDNYMIWVRQAPGQGLEWMGDINP YNGGTTFNQKFKGRVTITADKSTSTAY MELSSLRSEDTAVYYCARESPYFSNLY VMDYWGQGTLVTVS S SEQID NO: 56 SEQID NO: 57 SEQ ID NO: 58 SEQID NO: 59 *The notation “a” in front of an exemplary antigen expressed on an immunosuppressive cell denotes an antibody that specifically binds to the antigen expressed on an immunosuppressive cell. #The underlined sequences denote the respective HCDR1, HCDR2, and HCDR3. Table 4 NAME* VL SEQUENCE* LCDR1 LCDR2 LCDR3 VL aCD33 DIOLTOSPSTLSASVGDRVTITCRASES LDNYGIRFLTWFOQKPGKAPKLLMYA ASNQGSGVPSRFSGSGSGTEFTLTISSL SEQID NO: 60 SEQID NO: 61 SEQ ID NO: 62 SEQID NO: 63 QPDDFATYYCQQTKEVPWSFGOGTKV EVK aCD163 DIVMTQSPSSLSASVGDRVTITCRASOS VSSDVAWFOQKPGKSPKPLIYYASNR YSGVPSRFSGSGSGTDFTLTISSLQAED FAVYFCGQDYTSPRTFGGGTKLEIK SEQ ID NO: 64 SEQ ID NO: 65 SEQ ID NO: 66 SEQ ID NO: 67 aTRAIL-R2 (aDR5) EIVLTOSPGTLSLSPGERATLSCRASQGI SRSYLAWYQQKPGQAPSLLIYGASSRA TGIPDRFSGSGSGTDFTLTISRLEPEDFA VYYCQQFGSSPWTFGQGTKVEI SEQ ID NO: 68 SEQ ID NO: 69 SEQ ID NO: 70 SEQ ID NO: 12 aCSFIR EIVLTQSPATLSLSPGERATLSCKASOS VDYDGDNYMNWYOQKPGOAPRLLIY AASNLESGIPARFSGSGSGTDFTLTIS SL EPEDFAVYYCHLSNEDLSTFGGGTKVE IK SEQ ID NO: 71 SEQ ID NO: 72 SEQ ID NO: 73 SEQ ID NO: 74 *The notation “a” in front of an exemplary antigen expressed on an immunosuppressive cell denotes an antibody that specifically binds to the antigen expressed on an immunosuppressive cell. #The underlined sequences denote the respective LCDR1, LCDR2, and LCDR3.

[0091] In some embodiments, a multi-specific binding polypeptide (e.g., a multi-specific antibody) described herein comprises an immune cell binding moiety comprising an immunoglobulin heavy chain variable region comprising an amino acid sequence at least about 80%, 85%, 90%, 95%, 97%, 98%, 99%, or 100% identical to the amino acid sequence set forth in SEQ ID NOs: 11, 48, 52, or 59; and an immunoglobulin light chain variable region at least about 80%, 85%, 90%, 95%, 97%, 98%, 99%, or 100% identical to the amino acid sequence set forth in SEQ ID NOs: 12, 63, 67, or 74. In some instances, the amino acid differences contributing to the at least about 80%, 85%, 90%, 95%, 97%, 98%, or 99% sequence identity resides in a framework region of the immunoglobulin heavy chain variable region and the CDRs remain unchanged relative to the CDRs set forth in SEQ ID NOs: 11, 48, 52, or 59. In some instances, the amino acid differences contributing to the at least about 80%, 85%, 90%, 95%, 97%, 98%, or 99% sequence identity resides in a framework region of the immunoglobulin light chain variable region and the CDRs remain unchanged relative to the CDRs set forth in SEQ ID NOs: 12, 63, 67, or 74. In some cases, the amino acid differences contributing to the at least about 80%, 85%, 90%, 95%, 97%, 98%, or 99% sequence identity comprises conservative substitutions in the framework region, CDRs region, or a combination thereof, of the immunoglobulin heavy chain variable region but the multispecific binding polypeptide (e.g., the multi-specific antibody) retains binding to a target antigen expressed on an immunosuppressive cell. In some cases, the amino acid differences contributing to the at least about 80%, 85%, 90%, 95%, 97%, 98%, or 99% sequence identity comprises conservative substitutions in the framework region, CDRs region, or a combination thereof, of the immunoglobulin light chain variable region but the multi-specific binding polypeptide (e.g., the multi-specific antibody) retains binding to a target antigen expressed on an immunosuppressive cell.

[0092] In some embodiments, a multi-specific binding polypeptide (e.g., a multi-specific antibody) described herein comprises an immune cell binding moiety comprising an immunoglobulin heavy chain variable region comprising an amino acid sequence at least about 80%, 85%, 90%, 95%, 97%, 98%, 99%, or 100% identical to the amino acid sequence set forth in SEQ ID NO: 11; and an immunoglobulin light chain variable region at least about 80%, 85%, 90%, 95%, 97%, 98%, 99%, or 100% identical to the amino acid sequence set forth in SEQ ID NO: 12. In some instances, the amino acid differences contributing to the at least about 80%, 85%, 90%, 95%, 97%, 98%, or 99% sequence identity resides in a framework region of the immunoglobulin heavy chain variable region and the CDRs remain unchanged relative to the CDRs set forth in SEQ ID NO: 11. In some instances, the amino acid differences contributing to the at least about 80%, 85%, 90%, 95%, 97%, 98%, or 99% sequence identity resides in a framework region of the immunoglobulin light chain variable region and the CDRs remain unchanged relative to the CDRs set forth in SEQ ID NO: 12. In some cases, the amino acid differences contributing to the at least about 80%, 85%, 90%, 95%, 97%, 98%, or 99% sequence identity comprises conservative substitutions in the framework region, CDRs region, or a combination thereof, of the immunoglobulin heavy chain variable region but the multi-specific binding polypeptide (e.g., the multi-specific antibody) retains binding to a target antigen expressed on an immunosuppressive cell. In some cases, the amino acid differences contributing to the at least about 80%, 85%, 90%, 95%, 97%, 98%, or 99% sequence identity comprises conservative substitutions in the framework region, CDRs region, or a combination thereof, of the immunoglobulin light chain variable region but the multi-specific binding polypeptide (e.g., the multi-specific antibody) retains binding to a target antigen expressed on an immunosuppressive cell.

[0093] In some embodiments, a multi-specific binding polypeptide (e.g., a multi-specific antibody) described herein comprises an immune cell binding moiety comprising an immunoglobulin heavy chain variable region comprising an amino acid sequence at least about 80%, 85%, 90%, 95%, 97%, 98%, 99%, or 100% identical to the amino acid sequence set forth in SEQ ID NO: 48; and an immunoglobulin light chain variable region at least about 80%, 85%, 90%, 95%, 97%, 98%, 99%, or 100% identical to the amino acid sequence set forth in SEQ ID NO: 63. In some instances, the amino acid differences contributing to the at least about 80%, 85%, 90%, 95%, 97%, 98%, or 99% sequence identity resides in a framework region of the immunoglobulin heavy chain variable region and the CDRs remain unchanged relative to the CDRs set forth in SEQ ID NO: 48. In some instances, the amino acid differences contributing to the at least about 80%, 85%, 90%, 95%, 97%, 98%, or 99% sequence identity resides in a framework region of the immunoglobulin light chain variable region and the CDRs remain unchanged relative to the CDRs set forth in SEQ ID NO: 63. In some cases, the amino acid differences contributing to the at least about 80%, 85%, 90%, 95%, 97%, 98%, or 99% sequence identity comprises conservative substitutions in the framework region, CDRs region, or a combination thereof, of the immunoglobulin heavy chain variable region but the multi-specific binding polypeptide (e.g., the multi-specific antibody) retains binding to a target antigen expressed on an immunosuppressive cell. In some cases, the amino acid differences contributing to the at least about 80%, 85%, 90%, 95%, 97%, 98%, or 99% sequence identity comprises conservative substitutions in the framework region, CDRs region, or a combination thereof, of the immunoglobulin light chain variable region but the multi-specific binding polypeptide (e.g., the multi-specific antibody) retains binding to a target antigen expressed on an immunosuppressive cell.

[0094] In some embodiments, a multi-specific binding polypeptide (e.g., a multi-specific antibody) described herein comprises an immune cell binding moiety comprising an immunoglobulin heavy chain variable region comprising an amino acid sequence at least about 80%, 85%, 90%, 95%, 97%, 98%, 99%, or 100% identical to the amino acid sequence set forth in SEQ ID NO: 52; and an immunoglobulin light chain variable region at least about 80%, 85%, 90%, 95%, 97%, 98%, 99%, or 100% identical to the amino acid sequence set forth in SEQ ID NO: 67. In some instances, the amino acid differences contributing to the at least about 80%, 85%, 90%, 95%, 97%, 98%, or 99% sequence identity resides in a framework region of the immunoglobulin heavy chain variable region and the CDRs remain unchanged relative to the CDRs set forth in SEQ ID NO: 52. In some instances, the amino acid differences contributing to the at least about 80%, 85%, 90%, 95%, 97%, 98%, or 99% sequence identity resides in a framework region of the immunoglobulin light chain variable region and the CDRs remain unchanged relative to the CDRs set forth in SEQ ID NO: 67. In some cases, the amino acid differences contributing to the at least about 80%, 85%, 90%, 95%, 97%, 98%, or 99% sequence identity comprises conservative substitutions in the framework region, CDRs region, or a combination thereof, of the immunoglobulin heavy chain variable region but the multi-specific binding polypeptide (e.g., the multi-specific antibody) retains binding to a target antigen expressed on an immunosuppressive cell. In some cases, the amino acid differences contributing to the at least about 80%, 85%, 90%, 95%, 97%, 98%, or 99% sequence identity comprises conservative substitutions in the framework region, CDRs region, or a combination thereof, of the immunoglobulin light chain variable region but the multi-specific binding polypeptide (e.g., the multi-specific antibody) retains binding to a target antigen expressed on an immunosuppressive cell.

[0095] In some embodiments, a multi-specific binding polypeptide (e.g., a multi-specific antibody) described herein comprises an immune cell binding moiety comprising an immunoglobulin heavy chain variable region comprising an amino acid sequence at least about 80%, 85%, 90%, 95%, 97%, 98%, 99%, or 100% identical to the amino acid sequence set forth in SEQ ID NO: 59; and an immunoglobulin light chain variable region at least about 80%, 85%, 90%, 95%, 97%, 98%, 99%, or 100% identical to the amino acid sequence set forth in SEQ ID NO: 74. In some instances, the amino acid differences contributing to the at least about 80%, 85%, 90%, 95%, 97%, 98%, or 99% sequence identity resides in a framework region of the immunoglobulin heavy chain variable region and the CDRs remain unchanged relative to the CDRs set forth in SEQ ID NO: 59. In some instances, the amino acid differences contributing to the at least about 80%, 85%, 90%, 95%, 97%, 98%, or 99% sequence identity resides in a framework region of the immunoglobulin light chain variable region and the CDRs remain unchanged relative to the CDRs set forth in SEQ ID NO: 74. In some cases, the amino acid differences contributing to the at least about 80%, 85%, 90%, 95%, 97%, 98%, or 99% sequence identity comprises conservative substitutions in the framework region, CDRs region, or a combination thereof, of the immunoglobulin heavy chain variable region but the multi-specific binding polypeptide (e.g., the multi-specific antibody) retains binding to a target antigen expressed on an immunosuppressive cell. In some cases, the amino acid differences contributing to the at least about 80%, 85%, 90%, 95%, 97%, 98%, or 99% sequence identity comprises conservative substitutions in the framework region, CDRs region, or a combination thereof, of the immunoglobulin light chain variable region but the multi-specific binding polypeptide (e.g., the multi-specific antibody) retains binding to a target antigen expressed on an immunosuppressive cell.

[0096] In certain embodiments, the immune cell binding moiety of the multi-specific binding polypeptide (e.g., multi-specific antibody) comprises sequences (e.g., CDRs or VH / VL sequences) derived from a known antibody such as Conatumumab, Tigatuzumab, drozitumab, Lexatumumab, Benufutamab, Zaptuzumab (TRAIL-R2), Cabiralizumab, Emactuzumab, LY3022855, AMG820 (CSF1R), Gemtuzumab, Vadastuximab, Lintuzumab (CD33), or TBI 304H (CD163).

[0097] In certain embodiments, a multi-specific binding polypeptide (e.g., a multi-specific antibody) described herein comprises an immunoglobulin heavy chain comprising an amino acid sequence at least about 80%, 85%, 90%, 95%, 97%, 98%, 99%, or 100% identical to or consist of the amino acid sequence set forth under the heavy chain (HC) sequences of Table 5; and an immunoglobulin light chain comprising an amino acid sequence at least about 80%, 85%, 90%, 95%, 97%, 98%, 99%, or 100% identical to or consist of the amino acid sequence set forth under the light chain (LC) sequences of Table 5. In some instances, the amino acid differences contributing to the at least about 80%, 85%, 90%, 95%, 97%, 98%, or 99% sequence identity resides in a framework region of the immunoglobulin heavy chain and the CDRs remain unchanged relative to the CDRs set forth in the respective HC sequence in Table 5. In some instances, the amino acid differences contributing to the at least about 80%, 85%, 90%, 95%, 97%, 98%, or 99% sequence identity resides in a framework region of the immunoglobulin light chain and the CDRs remain unchanged relative to the CDRs set forth in the respective LC sequence in Table 5. In some cases, the amino acid differences contributing to the at least about 80%, 85%, 90%, 95%, 97%, 98%, or 99% sequence identity comprises conservative substitutions in the framework region, CDRs region, or a combination thereof, of the immunoglobulin heavy chain but the multi-specific binding polypeptide (e.g., the multi-specific antibody) retains binding to a target tumor antigen and / or a target antigen expressed on an immunosuppressive cell. In some cases, the amino acid differences contributing to the at least about 80%, 85%, 90%, 95%, 97%, 98%, or 99% sequence identity comprises conservative substitutions in the framework region, CDRs region, or a combination thereof, of the immunoglobulin light chain but the multi-specific binding polypeptide (e.g., the multi-specific antibody) retains binding to a target tumor antigen and / or a target antigen expressed on an immunosuppressive cell.

[0098] As shown below in Table 5, the underlined portion in the HC sequences indicate the scFv portion of the multi-specific antibody. The respective linker sequences in the heavy chain are shown in lower cases. Table 5 NAME HC SEQ ID NO: LC SEQ ID NO: DABA 1 (aTROP2 x aTRAIL-R2 in IgGl backbone) QVQLQQSGSELKKPGASVKVSCKASGYT FTNYGMNWVKQAPGQGLKWMGWINTY TGEPTYTDDFKGRFAFSLDTSVSTAYLQIS SLKADDTAVYFCARGGFGS SYWYFDVW GQGSLVTVSSASTKGPSVFPLAPSSKSTSG GTAALGCLVKDYFPEPVTVSWNSGALTS GVHTFPAVLQSSGLYSLSSVVTVPSSSLGT QTYICNVNHKPSNTKVDKKVEPKSCDKT HTCPPCPAPELLGGPSVFLFPPKPKDTLMI SRTPEVTCVVVDVEHEDPEVKFNWYVDG VEVHNAKTKPREEQYNSTYRVVSVLTVL HQDWLNGKEYKCKVSNKALPAPIEKTIS KAKGQPREPQVYTLPPSREEMTKNQVSL TCLVKGFYPSDIAVEWESNGQPENNYKT TPPVLDSDGSFFLYSKLTVDKSRWQQGN VFSCSVMHEALHNHYTQKSLSLSLggggsg ^sEIVLTOSPGTLSLSPGERATLSCRASOGI SRSYLA WYOQKPGOAPSLLIYGASSRATGIP DRFSGSGSGTDFTLTISRLEPEDFA VYYCQO FGSSPWTFGQGTKVEIKggggsggggsggg^^ ^OVOLOESGPGLVKPSQTLSLTCTVSGGSI SSGDYFWSWIRQLPGKGLEWIGHIHNSGTT YYNPSLKSRVTISVDTSKKOFSLRLSSVTAAD TA VYYCARDRGGDYYYGMDVWGQGTTVTV SS 7 DIQLTQSPSSLS ASVGDRVSITC KASQDVSIAVA WYQQKPGKAP KLLIYSASYRYT GVPDRFSGSGS GTDFTLTISSLQ PEDFAVYYCQQ HYITPLTFGAGT KVEIKRTVAAP SVFIFPPSDEQL KSGTASVVCLL NNFYPREAKVQ WKVDNALQSG NSQESVTEQDS KDSTYSLSSTLT LSKADYEKHKV YACEVTHQGLS SPVTKSFNRGE C 8 DABA 2 (aTROP2 x aCD33 in IgGl Backbone) QVQLQQSGSELKKPGASVKVSCKASGYT FTNYGMNWVKQAPGQGLKWMGWINTY TGEPTYTDDFKGRFAFSLDTSVSTAYLQIS SLKADDTAVYFCARGGFGS SYWYFDVW GQGSLVTVSSASTKGPSVFPLAPSSKSTSG GTAALGCLVKDYFPEPVTVSWNSGALTS GVHTFPAVLQSSGLYSLSSVVTVPSSSLGT QTYICNVNHKPSNTKVDKKVEPKSCDKT HTCPPCPAPELLGGPSVFLFPPKPKDTLMI SRTPEVTCVVVDVSHEDPEVKFNWYVDG VEVHNAKTKPREEQYNSTYRVVSVLTVL HQDWLNGKEYKCKVSNKALPAPIEKTIS KAKGQPREPQVYTLPPSREEMTKNQVSL TCLVKGFYPSDIAVEWESNGQPENNYKT TPPVLDSDGSFFLYSKLTVDKSRWQQGN VFSCSVMHEALHNHYTQKSLSLSLggggsg ^sEVOLVQSGAEVKKPGSSVKVSCKASGY TITDSNIHWVRQAPGQSLEWIGYIYPYNGGT DYNQKFKNRATLTVDNPTNTAYMELSSLRSE DTAFYYCVNGNP WLAYWGQGTLVTVSS^^ s^s^s^sDIQLTQSPSTLSASVGDRV TITCRASESLDNYGIRFLTWFOQKPGKAPKL LMYAASNQGSGVPSRFSGSGSGTEFTLTISSL OPDDFATYYCOQTKEVPWSFGOGTKVEVK 75 DIQLTQSPSSLS ASVGDRVSITC KASQDVSIAVA WYQQKPGKAP KLLIYSASYRYT GVPDRFSGSGS GTDFTLTISSLQ PEDFAVYYCQQ HYITPLTFGAGT KVEIKRTVAAP SVFIFPPSDEQL KSGTASVVCLL NNFYPREAKVQ WKVDNALQSG NSQESVTEQDS KDSTYSLSSTLT LSKADYEKHKV YACEVTHQGLS SPVTKSFNRGE C 8 DABA 3 (aTROP2 x aCSFIR in IgGl Backbone) QVQLQQSGSELKKPGASVKVSCKASGYT FTNYGMNWVKQAPGQGLKWMGWINTY TGEPTYTDDFKGRFAFSLDTSVSTAYLQIS SLKADDTAVYFCARGGFGS SYWYFDVW GQGSLVTVSSASTKGPSVFPLAPSSKSTSG GTAALGCLVKDYFPEPVTVSWNSGALTS 76 DIQLTQSPSSLS ASVGDRVSITC KASQDVSIAVA WYQQKPGKAP KLLIYSASYRYT GVPDRFSGSGS 8 GVHTFPAVLQSSGLYSLSSVVTVPSSSLGT QTYICNVNHKPSNTKVDKRVEPKSCDKT HTCPPCPAPELLGGPSVFLFPPKPKDTLMI SRTPEVTCVVVDVSHEDPEVKFNWYVDG VEVHNAKTKPREEQYNSTYRVVSVLTVL HQDWLNGKEYKCKVSNKALPAPIEKTIS KAKGQPREPQVYTLPPSREEMTKNQVSL TCLVKGFYPSDIAVEWESNGQPENNYKT TPPVLDSDGSFFLYSKLTVDKSRWQQGN VFSCSVMHEALHNHYTQKSLSLSLggggsg ^sOVOLVQSGAEVKKPGSSVKVSCKASGY TFTDNYMIWVRQAPGQGLEWMGDINPYNG GTTFNQKFKGRVTITADKSTSTAYMELSSLR SEPTA VYYCARESPYFSNLYVMDYWGQGTL VTVSSpgggsggggsggggsggggsEIVLTQSPATLS LSPGERATLSCKASOSVDYDGDNYMNWYO OKPGOAPRLLIYAASNLESGIPARFSGSGSGT DFTLTISSLEPEDFA VYYCHLSNEDLSTFGG GTKVEIK GTDFTLTISSLQ PEDFAVYYCQQ HYITPLTFGAGT KVEIKRTVAAP SVFIFPPSDEQL KSGTASVVCLL NNFYPREAKVQ WKVDNALQSG NSQESVTEQDS KDSTYSLSSTLT LSKADYEKHKV YACEVTHQGLS SPVTKSFNRGE C DABA 4 (aTROP2 x aCD163 in IgGl Backbone) QVQLQQSGSELKKPGASVKVSCKASGYT FTNYGMNWVKQAPGQGLKWMGWINTY TGEPTYTDDFKGRFAFSLDTSVSTAYLQIS SLKADDTAVYFCARGGFGS SYWYFDVW GQGSLVTVSSASTKGPSVFPLAPSSKSTSG GTAALGCLVKDYFPEPVTVSWNSGALTS GVHTFPAVLQSSGLYSLSSVVTVPSSSLGT QTYICNVNHKPSNTKVDKRVEPKSCDKT HTCPPCPAPELLGGPSVFLFPPKPKDTLMI SRTPEVTCVVVDVSHEDPEVKFNWYVDG VEVHNAKTKPREEQYNSTYRVVSVLTVL HQDWLNGKEYKCKVSNKALPAPIEKTIS KAKGQPREPQVYTLPPSREEMTKNQVSL TCLVKGFYPSDIAVEWESNGQPENNYKT TPPVLDSDGSFFLYSKLTVDKSRWQQGN VFSCSVMHEALHNHYTQKSLSLSLggggsg ^sQVOLQESGPGLVKPSETLSLTCTVSGYS ITSDYA WNWIRQFPGNKLEWMGYITYSGSTY YNPSLKSRVTISVDTSKNQFSLKLSSVTAADT ATYYCVSGTYYFDYWGQGTTLTVSS^^^ ^^^^DIVMTQSPSSLSASVGDRVTIT CRASQSVSSDVA WFQQKPGKSPKPLIYYASN RYSGVPSRFSGSGSGTDFTLTISSLOAEDFA V YFCGQDYTSPRTFGGGTKLEIK 77 DIQLTQSPSSLS ASVGDRVSITC KASQDVSIAVA WYQQKPGKAP KLLIYSASYRYT GVPDRFSGSGS GTDFTLTISSLQ PEDFAVYYCQQ HYITPLTFGAGT KVEIKRTVAAP SVFIFPPSDEQL KSGTASVVCLL NNFYPREAKVQ WKVDNALQSG NSQESVTEQDS KDSTYSLSSTLT LSKADYEKHKV YACEVTHQGLS SPVTKSFNRGE C 8 DABA 5 («GPC3 x aCD33 in IgGl Backbone) QVQLVQSGAEVKKPGASVKVSCKASGYT FTDYEMHWVRQAPGQGLEWMGALDPKT GDTAYSQKFKGRVTLTADKSTSTAYMEL S SLTSEDTAVYYCTRFYSYTYWGQGTLV TVSSASTKGPSVFPLAPSSKSTSGGTAALG CLVKDYFPEPVTVSWNSGALTSGVHTFP AVLQSSGLYSLSSVVTVPSSSLGTQTYICN VNHKPSNTKVDKRVEPKSCDKTHTCPPC PAPELLGGPSVFLFPPKPKDTLMISRTPEV TCVVVDVSHEDPEVKFNWYVDGVEVHN AKTKPREEQYNSTYRVVSVLTVLHQDWL NGKEYKCKVSNKALPAPIEKTISKAKGQP 79 DVVMTQSPLSL PVTPGEPASISC RSSQSLVHSNR NTYLHWYLQK PGQSPQLLIYKV SNRFSGVPDRFS GSGSGTDFTLKI SRVEAEDVGVY YCSQNTHVPPT FGQGTKLEIKR TVAAPSVFIFPP SDEQLKSGTAS 80 REPQVYTLPPSREEMTKNQVSLTCLVKGF YPSDIAVEWESNGQPENNYKTTPPVLDSD GSFFLYSKLTVDKSRWQQGNVFSCSVMH EALHNHYTQKSLSLSLggggsggggsEH2£H2 SGAEVKKPGSSVKVSCKASGYTITDSNIHWV ROAPGOSLEWIGYIYPYNGGTDYNOKFKNR ATLTVDNPTNTAYMELSSLRSEDTAFYYCVN GNPWLAYWGQGTLVTVSSggRgwgsggRgs ggggsDIQLTQSPSTLSASVGDR VTITCRASES LDNYGIRFLTWFOOKPGKAPKLLMYAASNQ GSGVPSRFSGSGSGTEFTLTISSLQPDDFATY YCOQTKEVP WSFGQGTKVEVK VVCLLNNFYPR EAKVQWKVDN ALQSGNSQESV TEQDSKDSTYS LSSTLTLSKAD YEKHKVYACE VTHQGLSSPVT KSFNRGEC DABA 6 («GPC3 x aCSFIR in IgGl Backbone) QVQLVQSGAEVKKPGASVKVSCKASGYT FTDYEMHWVRQAPGQGLEWMGALDPKT GDTAYSQKFKGRVTLTADKSTSTAYMEL S SLTSEDTAVYYCTRFYSYTYWGQGTLV TVSSASTKGPSVFPLAPSSKSTSGGTAALG CLVKDYFPEPVTVSWNSGALTSGVHTFP AVLQSSGLYSLSSVVTVPSSSLGTQTYICN VNHKPSNTKVDKRVEPKSCDKTHTCPPC PAPELLGGPSVFLFPPKPKDTLMISRTPEV TCVVVDVSHEDPEVKFNWYVDGVEVHN AKTKPREEQYNSTYRVVSVLTVLHQDWL NGKEYKCKVSNKALPAPIEKTISKAKGQP REPQVYTLPPSREEMTKNQVSLTCLVKGF YPSDIAVEWESNGQPENNYKTTPPVLDSD GSFFLYSKLTVDKSRWQQGNVFSCSVMH EALHNHYTOKSLSLSLggggsggggsCWLE QSGAEVKKPGSSVKVSCKASGYTFTDNYMI WVROAPGOGLEWMGDINPYNGGTTFNQK FKGR VTITADKSTSTA YMELSSLRSEDTA VYY CARESPYFSNLYVMDYWGOGTL VTVSSggggs ggggsggggsggggsEIVLTQSPATESLSPGERAT LSCKASOSVDYDGDNYMNWYOOKPGQAPR LLIYAASNLESGIPARFSGSGSGTDFTLTISSL EPEDFA VYYCHLSNEDLSTFGGGTKVEIK 81 DVVMTQSPLSL PVTPGEPASISC RSSQSLVHSNR NTYLHWYLQK PGQSPQLLIYKV SNRFSGVPDRFS GSGSGTDFTLKI SRVEAEDVGVY YCSQNTHVPPT FGQGTKLEIKR TVAAPSVFIFPP SDEQLKSGTAS VVCLLNNFYPR EAKVQWKVDN ALQSGNSQESV TEQDSKDSTYS LSSTLTLSKAD YEKHKVYACE VTHQGLSSPVT KSFNRGEC 80 DABA 7 («GPC3 x aCD163 in IgGl Backbone) QVQLVQSGAEVKKPGASVKVSCKASGYT FTDYEMHWVRQAPGQGLEWMGALDPKT GDTAYSQKFKGRVTLTADKSTSTAYMEL S SLTSEDTAVYYCTRFYSYTYWGQGTLV TVSSASTKGPSVFPLAPSSKSTSGGTAALG CLVKDYFPEPVTVSWNSGALTSGVHTFP AVLQSSGLYSLSSVVTVPSSSLGTQTYICN VNHKPSNTKVDKRVEPKSCDKTHTCPPC PAPELLGGPSVFLFPPKPKDTLMISRTPEV TCVVVDVSHEDPEVKFNWYVDGVEVHN AKTKPREEQYNSTYRVVSVLTVLHQDWL NGKEYKCKVSNKALPAPIEKTISKAKGQP REPQVYTLPPSREEMTKNQVSLTCLVKGF YPSDIAVEWESNGQPENNYKTTPPVLDSD GSFFLYSKLTVDKSRWQQGNVFSCSVMH EALHNHYTOKSLSLSLggggsggggsOEpLp ESGPGL VKPSETLSLTCTVSGYSITSDYA WN WIRQFPGNKLEWMGYITYSGSTYYNPSLKSR VTISVDTSKNQFSLKLSSVTAADTATYYCVSG 82 DVVMTQSPLSL PVTPGEPASISC RSSQSLVHSNR NTYLHWYLQK PGQSPQLLIYKV SNRFSGVPDRFS GSGSGTDFTLKI SRVEAEDVGVY YCSQNTHVPPT FGQGTKLEIKR TVAAPSVFIFPP SDEQLKSGTAS VVCLLNNFYPR EAKVQWKVDN ALQSGNSQESV TEQDSKDSTYS LSSTLTLSKAD YEKHKVYACE 80 / ymmRzfiGZZIIKSSggggsggggsggggsggg gsDIVMTQSPSSLSASVGDRVTITCRASQSVSS DVA WFOQKPGKSPKPUYYASNRYSGVPSRF SGSGSGTDFTLTISSLQAEDFA VYFCGQDYT SPRTFGGGTKLEIK VTHQGLSSPVT KSFNRGEC DABA 8 («GPC3 x aTRAIL-R2 in IgGl Backbone) QVQLVQSGAEVKKPGASVKVSCKASGYT FTDYEMHWVRQAPGQGLEWMGALDPKT GDTAYSQKFKGRVTLTADKSTSTAYMEL S SLTSEDTAVYYCTRFYSYTYWGQGTLV TVSSASTKGPSVFPLAPSSKSTSGGTAALG CLVKDYFPEPVTVSWNSGALTSGVHTFP AVLQSSGLYSLSSVVTVPSSSLGTQTYICN VNHKPSNTKVDKRVEPKSCDKTHTCPPC PAPELLGGPSVFLFPPKPKDTLMISRTPEV TCVVVDVSHEDPEVKFNWYVDGVEVHN AKTKPREEQYNSTYRVVSVLTVLHQDWL NGKEYKCKVSNKALPAPIEKTISKAKGQP REPQVYTLPPSREEMTKNQVSLTCLVKGF YPSDIAVEWESNGQPENNYKTTPPVLDSD GSFFLYSKLTVDKSRWQQGNVFSCSVMH EALHNHYTQKSLSLSLggggsggggsEnZZe^ PGTLSLSPGERATLSCRASQGISRSYLA WYQ QKPGQAPSLLIYGASSRATGIPDRFSGSGSG TDFTLTISRLEPEDFA VYYCQQFGSSP WTFG QGTKVEIKggggsggggsggggsggggsQVQLQES GPGL VKPSQTLSLTCTVSGGSISSGDYFWSW IRQLPGKGLEWIGHIHNSGTTYYNPSLKSRV TISVDTSKKQFSLRLSSVTAADTA VYYCARDR GGDYYYGMDVWGOGTTVTVSS 83 DVVMTQSPLSL PVTPGEPASISC RSSQSLVHSNR NTYLHWYLQK PGQSPQLLIYKV SNRFSGVPDRFS GSGSGTDFTLKI SRVEAEDVGVY YCSQNTHVPPT FGQGTKLEIKR TVAAPSVFIFPP SDEQLKSGTAS VVCLLNNFYPR EAKVQWKVDN ALQSGNSQESV TEQDSKDSTYS LSSTLTLSKAD YEKHKVYACE VTHQGLSSPVT KSFNRGEC 80 DABA 9 (aFOLRl x aCD33 in IgGl Backbone) QVQLVQSGAEWKPGASVKISCKASGYT FTGYFMNWVKQSPGQSLEWIGRIHPYDG DTFYNQKFQGKATLTVDKS SNTAHMELL SLTSEDFAVYYCTRYDGSRAMDYWGQG TTVTVSSASTKGPSVFPLAPSSKSTSGGTA ALGCLVKDYFPEPVTVSWNSGALTSGVH TFPAVLQSSGLYSLSSVVTVPSSSLGTQTY ICNVNHKPSNTKVDKRVEPKSCDKTHTCP PCPAPELLGGPSVFLFPPKPKDTLMISRTP EVTCVVVDVSHEDPEVKFNWYVDGVEV HNAKTKPREEQYNSTYRVVSVLTVLHQD WLNGKEYKCKVSNKALPAPIEKTISKAK GQPREPQVYTLPPSREEMTKNQVSLTCLV KGFYPSDIAVEWESNGQPENNYKTTPPVL DSDGSFFLYSKLTVDKSRWQQGNVFSCS VMHEALHNHYTQKSLSLSLggggsggggsEy OL VOSGAEVKKPGSSVKVSCKASGYTITDSN IHWVROAPGOSLEWIGYIYPYNGGTDYNQK FKNRATLTVDNPTNTAYMELSSLRSEDTAFY YCVNGNP WLA YWGQGTL VTVSSggggsggggs ggggsggggsDIOLTQSPSTLSASVGDRVTITCR ASESLDNYGIRFLTWFOQKPGKAPKLEMYA ASNQGSGVPSRFSGSGSGTEFTLTISSLQPD DFATYYCOQTKEVP WSFGQGTKVEVK 84 DIVLTQSPLSLA VSLGQPAIISCK ASQSVSFAGTS LMHWYHQKPG QQPRLLIYRAS NLEAGVPDRFS GSGSKTDFTLTI SPVEAEDAATY YCQQSREYPYT FGGGTKLEIKR TVAAPSVFIFPP SDEQLKSGTAS VVCLLNNFYPR EAKVQWKVDN ALQSGNSQESV TEQDSKDSTYS LSSTLTLSKAD YEKHKVYACE VTHQGLSSPVT KSFNRGEC 85 DABA 10 (aFOLRl x aCSFIR in IgGl Backbone) QVQLVQSGAEWKPGASVKISCKASGYT FTGYFMNWVKQSPGQSLEWIGRIHPYDG DTFYNQKFQGKATLTVDKS SNTAHMELL SLTSEDFAVYYCTRYDGSRAMDYWGQG TTVTVSSASTKGPSVFPLAPSSKSTSGGTA ALGCLVKDYFPEPVTVSWNSGALTSGVH TFPAVLQSSGLYSLSSVVTVPSSSLGTQTY ICNVNHKPSNTKVDKRVEPKSCDKTHTCP PCPAPELLGGPSVFLFPPKPKDTLMISRTP EVTCVVVDVSHEDPEVKFNWYVDGVEV HNAKTKPREEQYNSTYRVVSVLTVLHQD WLNGKEYKCKVSNKALPAPIEKTISKAK GQPREPQVYTLPPSREEMTKNQVSLTCLV KGFYPSDIAVEWESNGQPENNYKTTPPVL DSDGSFFLYSKLTVDKSRWQQGNVFSCS VMHEALHNHYTQKSLSLSLggggsggggs^E OL VQSGAEVKKPGSSVKVSCKASGYTFTDN YMIWVRQAPGQGLEWMGDINPYNGGTTFN QKFKGRVTITADKSTSTAYMELSSLRSEDTA VYYCARESPYFSNLYVMDYWGQGTL FTVSSg RATLSCKASQSVDYDGDNYMNWYOOKPGQ APRLLIYAASNLESGIPARFSGSGSGTDFTLTI SSLEPEDFA VYYCHLSNEDLSTFGGGTKVEI K 86 DIVLTQSPLSLA VSLGQPAIISCK ASQSVSFAGTS LMHWYHQKPG QQPRLLIYRAS NLEAGVPDRFS GSGSKTDFTLTI SPVEAEDAATY YCQQSREYPYT FGGGTKLEIKR TVAAPSVFIFPP SDEQLKSGTAS VVCLLNNFYPR EAKVQWKVDN ALQSGNSQESV TEQDSKDSTYS LSSTLTLSKAD YEKHKVYACE VTHQGLSSPVT KSFNRGEC 85 DABA 11 (aFOLRl x aCD163 in IgGl Backbone) QVQLVQSGAEWKPGASVKISCKASGYT FTGYFMNWVKQSPGQSLEWIGRIHPYDG DTFYNQKFQGKATLTVDKS SNTAHMELL SLTSEDFAVYYCTRYDGSRAMDYWGQG TTVTVSSASTKGPSVFPLAPSSKSTSGGTA ALGCLVKDYFPEPVTVSWNSGALTSGVH TFPAVLQSSGLYSLSSVVTVPSSSLGTQTY ICNVNHKPSNTKVDKRVEPKSCDKTHTCP PCPAPELLGGPSVFLFPPKPKDTLMISRTP EVTCVVVDVSHEDPEVKFNWYVDGVEV HNAKTKPREEQYNSTYRVVSVLTVLHQD WLNGKEYKCKVSNKALPAPIEKTISKAK GQPREPQWTLPPSREEMTKNQVSLTCLV KGFYPSDIAVEWESNGQPENNYKTTPPVL DSDGSFFLYSKLTVDKSRWQQGNVFSCS VMHEALHNHYTQKSLSLSLggggsggggs^E QLQESGPGLVKPSETLSLTCTVSGYSITSDYA WNWIRQFPGNKLEWMGYITYSGSTYYNPSL KSRVTISVDTSKNQFSLKLSSVTAADTATYYC VSGTYYFDYWGQGTTLTVSS^^^^ swwsDIVMTQSPSSIAASVGDRVTITCRASO SVSSDVA WFQQKPGKSPKPLIYYASNRYSGV PSRFSGSGSGTDFTLTISSLQAEDFA VYFCG ODYTSPRTFGGGTKLEIK 87 DIVLTQSPLSLA VSLGQPAIISCK ASQSVSFAGTS LMHWYHQKPG QQPRLLIYRAS NLEAGVPDRFS GSGSKTDFTLTI SPVEAEDAATY YCQQSREYPYT FGGGTKLEIKR TVAAPSVFIFPP SDEQLKSGTAS VVCLLNNFYPR EAKVQWKVDN ALQSGNSQESV TEQDSKDSTYS LSSTLTLSKAD YEKHKVYACE VTHQGLSSPVT KSFNRGEC 85 DABA 12 (aFOLRl x aTRAIL-R2 in IgGl Backbone) QVQLVQSGAEWKPGASVKISCKASGYT FTGYFMNWVKQSPGQSLEWIGRIHPYDG DTFYNQKFQGKATLTVDKS SNTAHMELL SLTSEDFAVYYCTRYDGSRAMDYWGQG TTVTVSSASTKGPSVFPLAPSSKSTSGGTA ALGCLVKDYFPEPVTVSWNSGALTSGVH 88 DIVLTQSPLSLA VSLGQPAIISCK ASQSVSFAGTS LMHWYHQKPG QQPRLLIYRAS NLEAGVPDRFS 85 TFPAVLQSSGLYSLSSVVTVPSSSLGTQTY ICNVNHKPSNTKVDKRVEPKSCDKTHTCP PCPAPELLGGPSVFLFPPKPKDTLMISRTP EVTCVVVDVSHEDPEVKFNWYVDGVEV HNAKTKPREEQYNSTYRVVSVLTVLHQD WLNGKEYKCKVSNKALPAPIEKTISKAK GQPREPQVYTLPPSREEMTKNQVSLTCLV KGFYPSDIAVEWESNGQPENNYKTTPPVL DSDGSFFLYSKLTVDKSRWQQGNVFSCS VMHEALHNHYTQKSLSLSLggggsggggsE / E LTQSPGTLSLSPGERATLSCRASQGISRSYLA WYQQKPGQAPSLLIYGASSRA TGIPDRFSGS GSGTDFTLTISRLEPEDFA VYYCQQFGSSP W TFGQGTKVEIK%%%%S£%%^^^^ QESGPGL VKPSQTLSLTCTVSGGSISSGDYF WSWIRQLPGKGLEWIGHIHNSGTTYYNPSLK SRVTISVDTSKKQFSLRLSSVTAADTA VYYCA RDRGGDYYYGMDVWGQGTTVTVSS GSGSKTDFTLTI SPVEAEDAATY YCQQSREYPYT FGGGTKLEIKR TVAAPSVFIFPP SDEQLKSGTAS VVCLLNNFYPR EAKVQWKVDN ALQSGNSQESV TEQDSKDSTYS LSSTLTLSKAD YEKHKVYACE VTHQGLSSPVT KSFNRGEC DABA13 («CD33 x aTRAIL-R2 in IgGl Backbone) EVQLVQSGAEVKKPGSSVKVSCKASGYTI TDSNIHWVRQAPGQSLEWIGYIYPYNGGT DYNQKFKNRATLTVDNPTNTAYMELSSL RSEDTAFYYCVNGNPWLAYWGQGTLVT VSSASTKGPSVFPLAPSSKSTSGGTAALGC LVKDYFPEPVTVSWNSGALTSGVHTFPA VLQSSGLYSLSSVVTVPSSSLGTQTYICNV NHKPSNTKVDKKVEPKSCDKTHTCPPCP APELLGGPSVFLFPPKPKDTLMISRTPEVT CVVVDVEHEDPEVKFNWYVDGVEVHNA KTKPREEQYNSTYRVVSVLTVLHQDWLN GKEYKCKVSNKALPAPIEKTISKAKGQPR EPQVYTLPPSREEMTKNQVSLTCLVKGFY PSDIAVEWESNGQPENNYKTTPPVLDSDG SFFLYSKLTVDKSRWQQGNVFSCSVMHE ALHNHYTQKSLSLSLggggsggggsEnZZe^ GTLSLSPGERATLSCRASQGISRSYLA WYQQ KPGQAPSLLIYGASSRATGIPDRFSGSGSGT DFTLTISRLEPEDFA VYYCQQFGSSP WTFGQ PGL VKPSQTLSLTCTVSGGSISSGDYFWSWIR QLPGKGLEWIGHIHNSGTTYYNPSLKSRVTIS VDTSKKQFSLRLSSVTAADTA VYYCARDRGG DYYYGMDVWGQGTTVTVSS 89 DIQLTQSPSTLS ASVGDRVTITC RASESLDNYGI RFLTWFQQKPG KAPKLLMYAAS NQGSGVPSRFS GSGSGTEFTLTI SSLQPDDFATY YCQQTKEVPWS FGQGTKVEVKR TVAAPSVFIFPP SDEQLKSGTAS VVCLLNNFYPR EAKVQWKVDN ALQSGNSQESV TEQDSKDSTYS LSSTLTLSKAD YEKHKVYACE VTHQGLSSPVT KSFNRGEC 90 DABA14 («CD38 x aTRAIL-R2 in IgGl Backbone) EVQLLESGGGLVQPGGSLRLSCAVSGFTF NSFAMSWVRQAPGKGLEWVSAISGSGGG TYYADSVKGRFTISRDNSKNTLYLQMNS LRAEDTAVYFCAKDKILWFGEPVFDYWG QGTLVTVSSASTKGPSVFPLAPSSKSTSGG TAALGCLVKDYFPEPVTVSWNSGALTSG VHTFPAVLQSSGLYSLSSVVTVPSSSLGTQ TYICNVNHKPSNTKVDKRVEPKSCDKTH TCPPCPAPELLGGPSVFLFPPKPKDTLMIS RTPEVTCVVVDVSHEDPEVKFNWYVDG VEVHNAKTKPREEQYNSTYRVVSVLTVL HQDWLNGKEYKCKVSNKALPAPIEKTIS KAKGQPREPQVYTLPPSREEMTKNQVSL 91 EIVLTQSPATLS LSPGERATLSCR ASQSVSSYLAW YQQKPGQAPRL LIYDASNRATGI PARFSGSGSGT DFTLTISSLEPE DFAVYYCQQRS NWPPTFGQGTK VEIKRTVAAPS VFIFPPSDEQLK SGTASVVCLLN NFYPREAKVQ 92 TCLVKGFYPSDIAVEWESNGQPENNYKT TPPVLDSDGSFFLYSKLTVDKSRWQQGN VFSCSVMHEALHNHYTQKSLSLSLggggsg ^sEIVLTQSPGTLSLSPGERATLSCRASQGI SRSYLA WYOQKPGOAPSLLIYGASSRATGIP DRFSGSGSGTDFTLTISRLEPEDFA VYYCQQ FGSSPWTFGQGTKVEIK^^^^^^^ ^OVOLOESGPGLVKPSQTLSLTCTVSGGSI SSGDYFWSWIROLPGKGLEWIGHIHNSGTT YYNPSLKSRVTISVDTSKKQFSLRLSSVTAAD TA VYYCARDRGGDYYYGMDVWGQGTTVTV SS WKVDNALQSG NSQESVTEQDS KDSTYSLSSTLT LSKADYEKHKV YACEVTHQGLS SPVTKSFNRGE C DABA15 (aFLT3 x aTRAIL-R2 in IgGl Backbone) EVQLVQSGAEVKKPGASVKVSCKASGYT FTSYYMHWVRQAPGQGLEWMGIINPSGG STSYAQKFQGRVTMTRDTSTSTVYMELS SLRSEDTAVYYCARGVGAHDAFDIWGQG TTVTVSSASTKGPSVFPLAPSSKSTSGGTA ALGCLVKDYFPEPVTVSWNSGALTSGVH TFPAVLQSSGLYSLSSVVTVPSSSLGTQTY ICNVNHKPSNTKVDKRVEPKSCDKTHTCP PCPAPELLGGPSVFLFPPKPKDTLMISRTP EVTCVVVDVSHEDPEVKFNWYVDGVEV HNAKTKPREEQYNSTYRVVSVLTVLHQD WLNGKEYKCKVSNKALPAPIEKTISKAK GQPREPQVYTLPPSREEMTKNQVSLTCLV KGFYPSDIAVEWESNGQPENNYKTTPPVL DSDGSFFLYSKLTVDKSRWQQGNVFSCS VMHEALHNHYTQKSLSLSLggggsggggsL / E LTQSPGTLSLSPGERATLSCRASQGISRSYLA WYQOKPGQAPSLUYGASSRATGIPDRFSGS GSGTDFTLTISRLEPEDFA VYYCQQFGSSP W TYGSGTXEEZKggggsggggsggggsggggsfilSE QESGPGL VKPSQTLSLTCTVSGGSISSGDYF WSWIROLPGKGLEWIGHIHNSGTTYYNPSLK SRVTISVDTSKKQFSLRLSSVTAADTA VYYCA RDRGGDYYYGMDVWGOGTTVTVSS 93 DVVMTQSPLSL PVTPGEPASISC RSSQSLLHSNG NNYLDWYLQK PGQSPQLLIYLG SNRASGVPDRF SGSGSDTDFTL QISRVEAEDVG VYYCMQGTHP AISFGQGTRLEI KRTVAAPSVFIF PPSDEQLKSGT ASVVCLLNNFY PREAKVQWKV DNALQSGNSQE SVTEQDSKDST YSLSSTLTLSKA DYEKHKVYAC EVTHQGLSSPV TKSFNRGEC 94 DABA16 (aTROP2 x aCD33 in IgGl Backbone with Impaired Fc) QVQLQQSGSELKKPGASVKVSCKASGYT FTNYGMNWVKQAPGQGLKWMGWINTY TGEPTYTDDFKGRFAFSLDTSVSTAYLQIS SLKADDTAVYFCARGGFGS SYWYFDVW GQGSLVTVSSASTKGPSVFPLAPCSRSTSE STAALGCLVKDYFPEPVTVSWNSGALTS GVHTFPAVLQSSGLYSLSSVVTVPSSNFG TQTYICNVNHKPSNTKVDKRVEPKSCDK THTCPPCPAPEAAGGS S VFLFPPKPKDTL MISRTPEVTCVVVDVSHEDPEVKFNWYV DGVEVHNAKTKPREEQYNSTYRVVSVLT VLHQDWLNGKEYKCKVSNKALPASIEKT ISKAKGQPREPQVYTLPPSREEMTKNQVS LTCLVKGFYPSDIAVEWESNGQPENNYK TTPPVLDSDGSFFLYSKLTVDKSRWQQG NVFSCSVMHEALHNHYTQKSLSLSLggggs ^^EVOL VQSGAEVKKPGSSVKVSCKASGY TITDSNIHWVROAPGQSLEWIGYIYPYNGGT DYNQKFKNRATLTVDNPTNTAYMELSSLRSE 95 DIQLTQSPSSLS ASVGDRVSITC KASQDVSIAVA WYQQKPGKAP KLLIYSASYRYT GVPDRFSGSGS GTDFTLTISSLQ PEDFAVYYCQQ HYITPLTFGAGT KVEIKRTVAAP SVFIFPPSDEQL KSGTASVVCLL NNFYPREAKVQ WKVDNALQSG NSQESVTEQDS KDSTYSLSSTLT LSKADYEKHKV YACEVTHQGLS 8 DTAFYYCVNGNP WLAYWGQGTLVTVSSgggg sggggsggggsggggsDIQLTQSPSTLSASVGDRV TITCRASESLDNYGIRFLTWFOOKPGKAPKL LMYAASNQGSGVPSRFSGSGSGTEFTLTISSL OPDDFATYYCOQTKEVPWSFGOGTKVEVK SPVTKSFNRGE C DAB A 17 (aTROP2 x aCSFIR in IgGl Backbone with impaired Fc) QVQLQQSGSELKKPGASVKVSCKASGYT FTNYGMNWVKQAPGQGLKWMGWINTY TGEPTYTDDFKGRFAFSLDTSVSTAYLQIS SLKADDTAVYFCARGGFGS SYWYFDVW GQGSLVTVSSASTKGPSVFPLAPCSRSTSE STAALGCLVKDYFPEPVTVSWNSGALTS GVHTFPAVLQSSGLYSLSSVVTVPSSNFG TQTYICNVNHKPSNTKVDKRVEPKSCDK THTCPPCPAPEAAGGSSVFLFPPKPKDTL MISRTPEVTCVVVDVSHEDPEVKFNWYV DGVEVHNAKTKPREEQYNSTYRVVSVLT VLHQDWLNGKEYKCKVSNKALPASIEKT ISKAKGQPREPQVYTLPPSREEMTKNQVS LTCLVKGFYPSDIAVEWESNGQPENNYK TTPPVLDSDGSFFLYSKLTVDKSRWQQG NVFSCSVMHEALHNHYTQKSLSLSLggggs ggggsQVOLVQSGAEVKKPGSSVKVSCKASG YTFTDNYMIWVROAPGOGLEWMGDINPYN GGTTFNQKFKGRVTITADKSTSTAYMELSSL RSEDTA VYYCARESPYFSNLYVMDYWGQGT LVTVSSggggsggggsggggsggggsEIVLTQSPAT LSLSPGERATLSCKASQSVDYDGDNYMNWY OOKPGQAPRLLIYAASNLESGIPARFSGSGS GTDFTLTISSLEPEDFA VYYCHLSNEDLSTFG GGTKVEIK 96 DIQLTQSPSSLS ASVGDRVSITC KASQDVSIAVA WYQQKPGKAP KLLIYSASYRYT GVPDRFSGSGS GTDFTLTISSLQ PEDFAVYYCQQ HYITPLTFGAGT KVEIKRTVAAP SVFIFPPSDEQL KSGTASVVCLL NNFYPREAKVQ WKVDNALQSG NSQESVTEQDS KDSTYSLSSTLT LSKADYEKHKV YACEVTHQGLS SPVTKSFNRGE C 8 DABA18 (aTROP2 x aCD163 in IgGl Backbone with impaired Fc) QVQLQQSGSELKKPGASVKVSCKASGYT FTNYGMNWVKQAPGQGLKWMGWINTY TGEPTYTDDFKGRFAFSLDTSVSTAYLQIS SLKADDTAVYFCARGGFGS SYWYFDVW GQGSLVTVSSASTKGPSVFPLAPCSRSTSE STAALGCLVKDYFPEPVTVSWNSGALTS GVHTFPAVLQSSGLYSLSSVVTVPSSNFG TQTYICNVNHKPSNTKVDKRVEPKSCDK THTCPPCPAPEAAGGS S VFLFPPKPKDTL MISRTPEVTCVVVDVSHEDPEVKFNWYV DGVEVHNAKTKPREEQYNSTYRVVSVLT VLHQDWLNGKEYKCKVSNKALPASIEKT ISKAKGQPREPQVYTLPPSREEMTKNQVS LTCLVKGFYPSDIAVEWESNGQPENNYK TTPPVLDSDGSFFLYSKLTVDKSRWQQG NVFSCSVMHEALHNHYTQKSLSLSLggggs ggggsQVOLQESGPGLVKPSETLSLTCTVSGY SITSDYA WNWIRQFPGNKLEWMGYITYSGST YYNPSLKSRVTISVDTSKNQFSLKLSSVTAAD TATYYCVSGTYYFDYWGQGTTLTVSSgggg^ gggsggggsggggsDIVMTQSPSSLSASVGDRVTI TCRASQSVSSDVA WFQQKPGKSPKPLIYYAS NRYSGVPSRFSGSGSGTDFTLTISSLQAEDFA VYFCGODYTSPRTFGGGTKLEIK 97 DIQLTQSPSSLS ASVGDRVSITC KASQDVSIAVA WYQQKPGKAP KLLIYSASYRYT GVPDRFSGSGS GTDFTLTISSLQ PEDFAVYYCQQ HYITPLTFGAGT KVEIKRTVAAP SVFIFPPSDEQL KSGTASVVCLL NNFYPREAKVQ WKVDNALQSG NSQESVTEQDS KDSTYSLSSTLT LSKADYEKHKV YACEVTHQGLS SPVTKSFNRGE C 8 DABA 19 (aTROP2 x aDR5 in IgGl Backbone with impaired Fc) QVQLQQSGSELKKPGASVKVSCKASGYT FTNYGMNWVKQAPGQGLKWMGWINTY TGEPTYTDDFKGRFAFSLDTSVSTAYLQIS SLKADDTAVYFCARGGFGS SYWYFDVW GQGSLVTVSSASTKGPSVFPLAPCSRSTSE STAALGCLVKDYFPEPVTVSWNSGALTS GVHTFPAVLQSSGLYSLSSVVTVPSSNFG TQTYICNVNHKPSNTKVDKRVEPKSCDK THTCPPCPAPEAAGGS S VFLFPPKPKDTL MISRTPEVTCVVVDVSHEDPEVKFNWYV DGVEVHNAKTKPREEQYNSTYRVVSVLT VLHQDWLNGKEYKCKVSNKALPASIEKT ISKAKGQPREPQVYTLPPSREEMTKNQVS LTCLVKGFYPSDIAVEWESNGQPENNYK TTPPVLDSDGSFFLYSKLTVDKSRWQQG NVFSCSVMHEALHNHYTQKSLSLSLggggs Z^sEIVLTQSPGTLSLSPGERATLSCRASQG ISRSYLA WYOQKPGOAPSLLIYGASSRATGIP DRFSGSGSGTDFTLTISRLEPEDFA VYYCQO FGSSPWTFGQGTKVEIKggggsggggsggg^^ ^OVOLOESGPGLVKPSQTLSLTCTVSGGSI SSGDYFWSWIRQLPGKGLEWIGHIHNSGTT YYNPSLKSRVTISVDTSKKOFSLRLSSVTAAD TA VYYCARDRGGDYYYGMDVWGQGTTVTV SS 98 DIQLTQSPSSLS ASVGDRVSITC KASQDVSIAVA WYQQKPGKAP KLLIYSASYRYT GVPDRFSGSGS GTDFTLTISSLQ PEDFAVYYCQQ HYITPLTFGAGT KVEIKRTVAAP SVFIFPPSDEQL KSGTASVVCLL NNFYPREAKVQ WKVDNALQSG NSQESVTEQDS KDSTYSLSSTLT LSKADYEKHKV YACEVTHQGLS SPVTKSFNRGE C 8 DABA 20 («GPC3 x aCD33 in IgGl Backbone with impaired Fc) QVQLVQSGAEVKKPGASVKVSCKASGYT FTDYEMHWVRQAPGQGLEWMGALDPKT GDTAYSQKFKGRVTLTADKSTSTAYMEL S SLTSEDTAVYYCTRFYSYTYWGQGTLV TVSSASTKGPSVFPLAPSSKSTSGGTAALG CLVKDYFPEPVTVSWNSGALTSGVHTFP AVLQSSGLYSLSSVVTVPSSSLGTQTYICN VNHKPSNTKVDKRVEPKSCDKTHTCPPC PAPEAAGGSSVFLFPPKPKDTLMISRTPEV TCVVVDVSHEDPEVKFNWYVDGVEVHN AKTKPREEQYNSTYRVVSVLTVLHQDWL NGKEYKCKVSNKALPASIEKTISKAKGQP REPQVYTLPPSREEMTKNQVSLTCLVKGF YPSDIAVEWESNGQPENNYKTTPPVLDSD GSFFLYSKLTVDKSRWQQGNVFSCSVMH EALHNHYTOKSLSLSLggggsggggsEEpZFp SGAEVKKPGSSVKVSCKASGYTITDSNIHWV ROAPGQSLEWIGYIYPYNGGTDYNQKFKNR ATLTVDNPTNTAYMELSSLRSEDTAFYYCVN GNPWIAYWGQGTLVWSS^s*^^ ^^DIQLTQSPSTLSASVGDR VTITCRASES LDNYGIRFLTWFOQKPGKAPKLLMYAASNO GSGVPSRFSGSGSGTEFTLTISSLQPDDFATY YCQQTKEVP WSFGQGTKVEVK 99 DVVMTQSPLSL PVTPGEPASISC RSSQSLVHSNR NTYLHWYLQK PGQSPQLLIYKV SNRFSGVPDRFS GSGSGTDFTLKI SRVEAEDVGVY YCSQNTHVPPT FGQGTKLEIKR TVAAPSVFIFPP SDEQLKSGTAS VVCLLNNFYPR EAKVQWKVDN ALQSGNSQESV TEQDSKDSTYS LSSTLTLSKAD YEKHKVYACE VTHQGLSSPVT KSFNRGEC 80 DABA 21 («GPC3 x aCSFIR in IgGl Backbone QVQLVQSGAEVKKPGASVKVSCKASGYT FTDYEMHWVRQAPGQGLEWMGALDPKT GDTAYSQKFKGRVTLTADKSTSTAYMEL S SLTSEDTAVYYCTRFYSYTYWGQGTLV TVSSASTKGPSVFPLAPSSKSTSGGTAALG CLVKDYFPEPVTVSWNSGALTSGVHTFP 100 DVVMTQSPLSL PVTPGEPASISC RSSQSLVHSNR NTYLHWYLQK PGQSPQLLIYKV SNRFSGVPDRFS 80 with impaired Fc) AVLQSSGLYSLSSVVTVPSSSLGTQTYICN VNHKPSNTKVDKRVEPKSCDKTHTCPPC PAPEAAGGS SVFLFPPKPKDTLMISRTPEV TCVVVDVSHEDPEVKFNWYVDGVEVHN AKTKPREEQYNSTYRVVSVLTVLHQDWL NGKEYKCKVSNKALPASIEKTISKAKGQP REPQVYTLPPSREEMTKNQVSLTCLVKGF YPSDIAVEWESNGQPENNYKTTPPVLDSD GSFFLYSKLTVDKSRWQQGNVFSCSVMH EALHNHYTQKSLSLSLggggsggggs(WZF QSGAEVKKPGSSVKVSCKASGYTFTDNYMI WVRQAPGQGLEWMGDINPYNGGTTFNQK FKGR VTITADKSTSTA YMELSSLRSEDTA VYY CARESPYFSNLYVMDYWGQGTLVTVSS^^ ^s^s^sEIVLTOSPATLSLSPGERAT LSCKASOSVDYDGDNYMNWYOOKPGOAPR LLIYAASNLESGIPARFSGSGSGTDFTLTISSL EPEDFA VYYCHLSNEDLSTFGGGTKVEIK GSGSGTDFTLKI SRVEAEDVGVY YCSQNTHVPPT FGQGTKLEIKR TVAAPSVFIFPP SDEQLKSGTAS VVCLLNNFYPR EAKVQWKVDN ALQSGNSQESV TEQDSKDSTYS LSSTLTLSKAD YEKHKVYACE VTHQGLSSPVT KSFNRGEC DABA 22 («GPC3 x aCD163 in IgGl Backbone with impaired Fc) QVQLVQSGAEVKKPGASVKVSCKASGYT FTDYEMHWVRQAPGQGLEWMGALDPKT GDTAYSQKFKGRVTLTADKSTSTAYMEL S SLTSEDTAVYYCTRFYSYTYWGQGTLV TVSSASTKGPSVFPLAPSSKSTSGGTAALG CLVKDYFPEPVTVSWNSGALTSGVHTFP AVLQSSGLYSLSSVVTVPSSSLGTQTYICN VNHKPSNTKVDKRVEPKSCDKTHTCPPC PAPEAAGGS SVFLFPPKPKDTLMISRTPEV TCVVVDVSHEDPEVKFNWYVDGVEVHN AKTKPREEQYNSTYRVVSVLTVLHQDWL NGKEYKCKVSNKALPASIEKTISKAKGQP REPQVYTLPPSREEMTKNQVSLTCLVKGF YPSDIAVEWESNGQPENNYKTTPPVLDSD GSFFLYSKLTVDKSRWQQGNVFSCSVMH EALHNHYTOKSLSLSLggggsggggsOEpLp ESGPGL VKPSETLSLTCTVSGYSITSDYA WN WIRQFPGNKLEWMGYITYSGSTYYNPSLKSR VTISVDTSKNQFSLKLSSVTAADTATYYCVSG TYYFDYWGQGTTLTVSS^^^^^^ gsDIVMTOSPSSLSASVGDRVTITCRASOSVSS DVA WFQQKPGKSPKPLIYYASNRYSGVPSRF SGSGSGTDFTLTISSLQAEDFA VYFCGQDYT SPRTFGGGTKLEIK 101 DVVMTQSPLSL PVTPGEPASISC RSSQSLVHSNR NTYLHWYLQK PGQSPQLLIYKV SNRFSGVPDRFS GSGSGTDFTLKI SRVEAEDVGVY YCSQNTHVPPT FGQGTKLEIKR TVAAPSVFIFPP SDEQLKSGTAS VVCLLNNFYPR EAKVQWKVDN ALQSGNSQESV TEQDSKDSTYS LSSTLTLSKAD YEKHKVYACE VTHQGLSSPVT KSFNRGEC 80 DABA 23 («GPC3 x aDR5 in IgGl Backbone with impaired Fc) QVQLVQSGAEVKKPGASVKVSCKASGYT FTDYEMHWVRQAPGQGLEWMGALDPKT GDTAYSQKFKGRVTLTADKSTSTAYMEL S SLTSEDTAVYYCTRFYSYTYWGQGTLV TVSSASTKGPSVFPLAPSSKSTSGGTAALG CLVKDYFPEPVTVSWNSGALTSGVHTFP AVLQSSGLYSLSSVVTVPSSSLGTQTYICN VNHKPSNTKVDKRVEPKSCDKTHTCPPC PAPEAAGGS SVFLFPPKPKDTLMISRTPEV TCVVVDVSHEDPEVKFNWYVDGVEVHN AKTKPREEQYNSTYRVVSVLTVLHQDWL NGKEYKCKVSNKALPASIEKTISKAKGQP REPQVYTLPPSREEMTKNQVSLTCLVKGF 102 DVVMTQSPLSL PVTPGEPASISC RSSQSLVHSNR NTYLHWYLQK PGQSPQLLIYKV SNRFSGVPDRFS GSGSGTDFTLKI SRVEAEDVGVY YCSQNTHVPPT FGQGTKLEIKR TVAAPSVFIFPP SDEQLKSGTAS VVCLLNNFYPR 80 YPSDIAVEWESNGQPENNYKTTPPVLDSD GSFFLYSKLTVDKSRWQQGNVFSCSVMH EALHNHYTQKSLSLSLggggsggggsEnYTgS PGTLSLSPGERATLSCRASQGISRSYLA WYQ OKPGQAPSLLIYGASSRATGIPDRFSGSGSG TDFTLTISRLEPEDFA VYYCQQFGSSP WTFG QGTKVEIKggggszgggsggg^^^ GPGL VKPSQTLSLTCTVSGGSISSGDYFWSW IROLPGKGLEWIGHIHNSGTTYYNPSLKSRV TISVDTSKKQFSLRLSSVTAADTA VYYCARDR GGDYYYGMDVWGQGTTVTVSS EAKVQWKVDN ALQSGNSQESV TEQDSKDSTYS LSSTLTLSKAD YEKHKVYACE VTHQGLSSPVT KSFNRGEC DABA 24 (aFOLRl x aCD33 in IgGl Backbone with impaired Fc) QVQLVQSGAEWKPGASVKISCKASGYT FTGYFMNWVKQSPGQSLEWIGRIHPYDG DTFYNQKFQGKATLTVDKS SNTAHMELL SLTSEDFAVYYCTRYDGSRAMDYWGQG TTVTVSSASTKGPSVFPLAPSSKSTSGGTA ALGCLVKDYFPEPVTVSWNSGALTSGVH TFPAVLQSSGLYSLSSVVTVPSSSLGTQTY ICNVNHKPSNTKVDKRVEPKSCDKTHTCP PCPAPEAAGGS SVFLFPPKPKDTLMISRTP EVTCVVVDVSHEDPEVKFNWYVDGVEV HNAKTKPREEQYNSTYRVVSVLTVLHQD WLNGKEYKCKVSNKALPASIEKTISKAK GQPREPQVYTLPPSREEMTKNQVSLTCLV KGFYPSDIAVEWESNGQPENNYKTTPPVL DSDGSFFLYSKLTVDKSRWQQGNVFSCS VMHEALHNHYTQKSLSLSLggggsggggsEy QL VQSGAEVKKPGSSVKVSCKASGYTITDSN IHWVROAPGOSLEWIGYIYPYNGGTDYNOK FKNRATLTVDNPTNTAYMELSSLRSEDTAFY YCVNGNP WLA YWGQGTL VTVSS^^^^ ^^^sDIOLTOSPSTLSASVGDRVTITCR ASESLDNYGIRFLTWFOOKPGKAPKLLMYA ASNQGSGVPSRFSGSGSGTEFTLTISSLQPD DFATYYCQQTKEVP WSFGQGTKVEVK 103 DIVLTQSPLSLA VSLGQPAIISCK ASQSVSFAGTS LMHWYHQKPG QQPRLLIYRAS NLEAGVPDRFS GSGSKTDFTLTI SPVEAEDAATY YCQQSREYPYT FGGGTKLEIKR TVAAPSVFIFPP SDEQLKSGTAS VVCLLNNFYPR EAKVQWKVDN ALQSGNSQESV TEQDSKDSTYS LSSTLTLSKAD YEKHKVYACE VTHQGLSSPVT KSFNRGEC 85 DABA 25 (aFOLRl x aCSFIR in IgGl Backbone with impaired Fc) QVQLVQSGAEWKPGASVKISCKASGYT FTGYFMNWVKQSPGQSLEWIGRIHPYDG DTFYNQKFQGKATLTVDKS SNTAHMELL SLTSEDFAVYYCTRYDGSRAMDYWGQG TTVTVSSASTKGPSVFPLAPSSKSTSGGTA ALGCLVKDYFPEPVTVSWNSGALTSGVH TFPAVLQSSGLYSLSSVVTVPSSSLGTQTY ICNVNHKPSNTKVDKRVEPKSCDKTHTCP PCPAPEAAGGS SVFLFPPKPKDTLMISRTP EVTCVVVDVSHEDPEVKFNWYVDGVEV HNAKTKPREEQYNSTYRVVSVLTVLHQD WLNGKEYKCKVSNKALPASIEKTISKAK GQPREPQWTLPPSREEMTKNQVSLTCLV KGFYPSDIAVEWESNGQPENNYKTTPPVL DSDGSFFLYSKLTVDKSRWQQGNVFSCS VMHEALHNHYTQKSLSLSLggggsggggs^E QL VQSGAEVKKPGSSVKVSCKASGYTFTDN YMIWVRQAPGQGLEWMGDINPYNGGTTFN OKFKGRVTITADKSTSTAYMELSSLRSEDTA VYYCARESPYFSNLYVMDYWGQGTL VTVSSg 104 DIVLTQSPLSLA VSLGQPAIISCK ASQSVSFAGTS LMHWYHQKPG QQPRLLIYRAS NLEAGVPDRFS GSGSKTDFTLTI SPVEAEDAATY YCQQSREYPYT FGGGTKLEIKR TVAAPSVFIFPP SDEQLKSGTAS VVCLLNNFYPR EAKVQWKVDN ALQSGNSQESV TEQDSKDSTYS LSSTLTLSKAD YEKHKVYACE VTHQGLSSPVT KSFNRGEC 85 gggsggggsggggsggggsE / mW^ZLSZAPGE RATLSCKASQSVDYDGDNYMNWYOOKPGQ APRLLIYAASNLESGIPARFSGSGSGTDFTLTI SSLEPEDFA VYYCHLSNEDLSTFGGGTKVEI K DABA 26 (aFOLRl x aCD163 in IgGl Backbone with impaired Fc) QVQLVQSGAEWKPGASVKISCKASGYT FTGYFMNWVKQSPGQSLEWIGRIHPYDG DTFYNQKFQGKATLTVDKS SNTAHMELL SLTSEDFAVYYCTRYDGSRAMDYWGQG TTVTVSSASTKGPSVFPLAPSSKSTSGGTA ALGCLVKDYFPEPVTVSWNSGALTSGVH TFPAVLQSSGLYSLSSVVTVPSSSLGTQTY ICNVNHKPSNTKVDKRVEPKSCDKTHTCP PCPAPEAAGGS SVFLFPPKPKDTLMISRTP EVTCVVVDVSHEDPEVKFNWYVDGVEV HNAKTKPREEQYNSTYRVVSVLTVLHQD WLNGKEYKCKVSNKALPASIEKTISKAK GQPREPQVYTLPPSREEMTKNQVSLTCLV KGFYPSDIAVEWESNGQPENNYKTTPPVL DSDGSFFLYSKLTVDKSRWQQGNVFSCS VMHEALHNHYTQKSLSLSLggggsggggs^E QLQESGPGLVKPSETLSLTCTVSGYSITSDYA WNWIRQFPGNKLEWMGYITYSGSTYYNPSL KSRVTISVDTSKNQFSLKLSSVTAADTATYYC VSGTYYFDYWGQGTTLTVSS^w^m swwsDIVMTQSPSSIAASVGDRVTITCRASO SVSSDVA WFQQKPGKSPKPLIYYASNRYSGV PSRFSGSGSGTDFTLTISSLQAEDFA VYFCG ODYTSPRTFGGGTKLEIK 105 DIVLTQSPLSLA VSLGQPAIISCK ASQSVSFAGTS LMHWYHQKPG QQPRLLIYRAS NLEAGVPDRFS GSGSKTDFTLTI SPVEAEDAATY YCQQSREYPYT FGGGTKLEIKR TVAAPSVFIFPP SDEQLKSGTAS VVCLLNNFYPR EAKVQWKVDN ALQSGNSQESV TEQDSKDSTYS LSSTLTLSKAD YEKHKVYACE VTHQGLSSPVT KSFNRGEC 85 DABA 27 (aFOLRl x aDR5 in IgGl Backbone with impaired Fc) QVQLVQSGAEWKPGASVKISCKASGYT FTGYFMNWVKQSPGQSLEWIGRIHPYDG DTFYNQKFQGKATLTVDKS SNTAHMELL SLTSEDFAVYYCTRYDGSRAMDYWGQG TTVTVSSASTKGPSVFPLAPSSKSTSGGTA ALGCLVKDYFPEPVTVSWNSGALTSGVH TFPAVLQSSGLYSLSSVVTVPSSSLGTQTY ICNVNHKPSNTKVDKRVEPKSCDKTHTCP PCPAPEAAGGS SVFLFPPKPKDTLMISRTP EVTCVVVDVSHEDPEVKFNWYVDGVEV HNAKTKPREEQYNSTYRVVSVLTVLHQD WLNGKEYKCKVSNKALPASIEKTISKAK GQPREPQWTLPPSREEMTKNQVSLTCLV KGFYPSDIAVEWESNGQPENNYKTTPPVL DSDGSFFLYSKLTVDKSRWQQGNVFSCS VMHEALHNHYTQKSLSLSLggggsggggsE / E LTOSPGTLSLSPGERATLSCRASQGISRSYLA WYOQKPGOAPSLLIYGASSRATGIPDRFSGS GSGTDFTLTISRLEPEDFA VYYCQQFGSSP W TFGOGTKVEIK££££S££^^ QESGPGL VKPSQTLSLTCTVSGGSISSGDYF WSWIROLPGKGLEWIGHIHNSGTTYYNPSLK SRVTISVDTSKKQFSLRLSSVTAADTA VYYCA RDRGGDYYYGMDVWGOGTTVTVSS 106 DIVLTQSPLSLA VSLGQPAIISCK ASQSVSFAGTS LMHWYHQKPG QQPRLLIYRAS NLEAGVPDRFS GSGSKTDFTLTI SPVEAEDAATY YCQQSREYPYT FGGGTKLEIKR TVAAPSVFIFPP SDEQLKSGTAS VVCLLNNFYPR EAKVQWKVDN ALQSGNSQESV TEQDSKDSTYS LSSTLTLSKAD YEKHKVYACE VTHQGLSSPVT KSFNRGEC 85 DABA 28 («CD33 x aDR5 in IgGl Backbone with impaired Fc) EVQLVQSGAEVKKPGSSVKVSCKASGYTI TDSNIHWVRQAPGQSLEWIGYIYPYNGGT DYNQKFKNRATLTVDNPTNTAYMELSSL RSEDTAFYYCVNGNPWLAYWGQGTLVT VSSASTKGPSVFPLAPSSKSTSGGTAALGC LVKDYFPEPVTVSWNSGALTSGVHTFPA VLQSSGLYSLSSVVTVPSSSLGTQTYICNV NHKPSNTKVDKRVEPKSCDKTHTCPPCP APEAAGGS SVFLFPPKPKDTLMISRTPEVT CVVVDVSHEDPEVKFNWYVDGVEVHNA KTKPREEQYNSTYRVVSVLTVLHQDWLN GKEYKCKVSNKALPASIEKTISKAKGQPR EPQVYTLPPSREEMTKNQVSLTCLVKGFY PSDIAVEWESNGQPENNYKTTPPVLDSDG SFFLYSKLTVDKSRWQQGNVFSCSVMHE ALHNHYTQKSLSLSLggggsggggsE / mSSP GTLSLSPGERATLSCRASQGISRSYLA WYQQ KPGQAPSLLIYGASSRATGIPDRFSGSGSGT DFTLTISRLEPEDFA VYYCQQFGSSP WTFGQ GZ^EE^ggggsggggsggggsggggsfilSEQ^G PGL VKPSOTLSLTCTVSGGSISSGDYFWSWIR OLPGKGLEWIGHIHNSGTTYYNPSLKSRVTIS VDTSKKOFSLRLSSVTAADTA VYYCARDRGG DYYYGMDVWGQGTTVTVSS 107 DIQLTQSPSTLS ASVGDRVTITC RASESLDNYGI RFLTWFQQKPG KAPKLLMYAAS NQGSGVPSRFS GSGSGTEFTLTI SSLQPDDFATY YCQQTKEVPWS FGQGTKVEVKR TVAAPSVFIFPP SDEQLKSGTAS VVCLLNNFYPR EAKVQWKVDN ALQSGNSQESV TEQDSKDSTYS LSSTLTLSKAD YEKHKVYACE VTHQGLSSPVT KSFNRGEC 90 DABA 29 (aCD38 x aDR5 in IgGl Backbone with impaired Fc) EVQLLESGGGLVQPGGSLRLSCAVSGFTF NSFAMSWVRQAPGKGLEWVSAISGSGGG TYYADSVKGRFTISRDNSKNTLYLQMNS LRAEDTAVYFCAKDKILWFGEPVFDYWG QGTLVTVSSASTKGPSVFPLAPSSKSTSGG TAALGCLVKDYFPEPVTVSWNSGALTSG VHTFPAVLQSSGLYSLSSVVTVPSSSLGTQ TYICNVNHKPSNTKVDKRVEPKSCDKTH TCPPCPAPEAAGGS SVFLFPPKPKDTLMIS RTPEVTCVVVDVSHEDPEVKFNWYVDG VEVHNAKTKPREEQYNSTYRVVSVLTVL HQDWLNGKEYKCKVSNKALPASIEKTIS KAKGQPREPQVYTLPPSREEMTKNQVSL TCLVKGFYPSDIAVEWESNGQPENNYKT TPPVLDSDGSFFLYSKLTVDKSRWQQGN VFSCSVMHEALHNHYTQKSLSLSLggggsg ^sEIVLTQSPGTLSLSPGERATLSCRASQGI SRSYLA WYQQKPGQAPSLLIYGASSRATGIP DRFSGSGSGTDFTLTISRLEPEDFA VYYCQQ FGSSPWTFGQGTKVEIKzgggsgg^ ^sOVQLQESGPGLVKPSQTLSLTCTVSGGSl SSGDYFWSWIROLPGKGLEWIGHIHNSGTT YYNPSLKSRVTISVDTSKKQFSLRLSSVTAAD TA VYYCARDRGGDYYYGMDVWGQGTTVTV ss 108 EIVLTQSPATLS LSPGERATLSCR ASQSVSSYLAW YQQKPGQAPRL LIYDASNRATGI PARFSGSGSGT DFTLTISSLEPE DFAVYYCQQRS NWPPTFGQGTK VEIKRTVAAPS VFIFPPSDEQLK SGTASVVCLLN NFYPREAKVQ WKVDNALQSG NSQESVTEQDS KDSTYSLSSTLT LSKADYEKHKV YACEVTHQGLS SPVTKSFNRGE C 92 DABA 30 (aFLT3 x oDR5 in IgGl Backbone EVQLVQSGAEVKKPGASVKVSCKASGYT FTSYYMHWVRQAPGQGLEWMGIINPSGG STSYAQKFQGRVTMTRDTSTSTVYMELS SLRSEDTAVYYCARGVGAHDAFDIWGQG TTVTVSSASTKGPSVFPLAPSSKSTSGGTA ALGCLVKDYFPEPVTVSWNSGALTSGVH 109 DVVMTQSPLSL PVTPGEPASISC RSSQSLLHSNG NNYLDWYLQK PGQSPQLLIYLG SNRASGVPDRF 94 with impaired Fc) TFPAVLQSSGLYSLSSVVTVPSSSLGTQTY ICNVNHKPSNTKVDKRVEPKSCDKTHTCP PCPAPEAAGGS SVFLFPPKPKDTLMISRTP EVTCVVVDVSHEDPEVKFNWYVDGVEV HNAKTKPREEQYNSTYRVVSVLTVLHQD WLNGKEYKCKVSNKALPASIEKTISKAK GQPREPQVYTLPPSREEMTKNQVSLTCLV KGFYPSDIAVEWESNGQPENNYKTTPPVL DSDGSFFLYSKLTVDKSRWQQGNVFSCS VMHEALHNHYTQKSLSLSLggggsggggsE / E LTQSPGTLSLSPGERATLSCRASQGISRSYLA WYOQKPGOAPSLLIYGASSRATGIPDRFSGS GSGTDFTLTISRLEPEDFA VYYCQQFGSSP W TFGfiGZKEE^ggggsggggsggggsggggs^ISE QESGPGL VKPSQTLSLTCTVSGGSISSGDYF WSWIRQLPGKGLEWIGHIHNSGTTYYNPSLK SRVTISVDTSKKQFSLRLSSVTAADTA VYYCA RDRGGDYYYGMDVWGOGTTVTVSS SGSGSDTDFTL QISRVEAEDVG VYYCMQGTHP AISFGQGTRLEI KRTVAAPSVFIF PPSDEQLKSGT ASVVCLLNNFY PREAKVQWKV DNALQSGNSQE SVTEQDSKDST YSLSSTLTLSKA DYEKHKVYAC EVTHQGLSSPV TKSFNRGEC DABA31 (aTROP2 x aCD33 in IgGl Backbone with ADCC Enhanced) QVQLQQSGSELKKPGASVKVSCKASGYT FTNYGMNWVKQAPGQGLKWMGWINTY TGEPTYTDDFKGRFAFSLDTSVSTAYLQIS SLKADDTAVYFCARGGFGS SYWYFDVW GQGSLVTVSSASTKGPSVFPLAPCSRSTSE STAALGCLVKDYFPEPVTVSWNSGALTS GVHTFPAVLQSSGLYSLSSVVTVPSSNFG TQTYICNVNHKPSNTKVDKRVEPKSCDK THTCPPCPAPELLGGPDVFLFPPKPKDTL MISRTPEVTCVVVDVSHEDPEVKFNWYV DGVEVHNAKTKPREEQYNSTYRVVSVLT VLHQDWLNGKEYKCKVSNKALPLPEEKT ISKAKGQPREPQVYTLPPSREEMTKNQVS LTCLVKGFYPSDIAVEWESNGQPENNYK TTPPVLDSDGSFFLYSKLTVDKSRWQQG NVFSCSVMHEALHNHYTQKSLSLSLggggg sggggsEEQL VQSGAEVKKPGSSVKVSCKASG YTITDSNIHWVROAPGOSLEWIGYIYPYNGG TDYNQKFKNRATLTVDNPTNTA YMELSSLRS EDTAFYYCVNGNP WLAYWGQGTLVTVSS^ ^s^^s^^^sDIOLTOSPSTLSASVGD RVTITCRASESLDNYGIRFLTWFQQKPGKAP KLLMYAASNQGSGVPSRFSGSGSGTEFTLTI SSLOPDDFATYYCOOTKEVPWSFGOGTKVE VK 110 DIQLTQSPSSLS ASVGDRVSITC KASQDVSIAVA WYQQKPGKAP KLLIYSASYRYT GVPDRFSGSGS GTDFTLTISSLQ PEDFAVYYCQQ HYITPLTFGAGT KVEIKRTVAAP SVFIFPPSDEQL KSGTASVVCLL NNFYPREAKVQ WKVDNALQSG NSQESVTEQDS KDSTYSLSSTLT LSKADYEKHKV YACEVTHQGLS SPVTKSFNRGE C 8 DABA 32 (aTROP2 x aCSFIR in IgGl Backbone with ADCC Enhanced) QVQLQQSGSELKKPGASVKVSCKASGYT FTNYGMNWVKQAPGQGLKWMGWINTY TGEPTYTDDFKGRFAFSLDTSVSTAYLQIS SLKADDTAVYFCARGGFGS SYWYFDVW GQGSLVTVSSASTKGPSVFPLAPCSRSTSE STAALGCLVKDYFPEPVTVSWNSGALTS GVHTFPAVLQSSGLYSLSSVVTVPSSNFG TQTYICNVNHKPSNTKVDKRVEPKSCDK THTCPPCPAPELLGGPDVFLFPPKPKDTL MISRTPEVTCVVVDVSHEDPEVKFNWYV DGVEVHNAKTKPREEQYNSTYRVVSVLT VLHQDWLNGKEYKCKVSNKALPLPEEKT 111 DIQLTQSPSSLS ASVGDRVSITC KASQDVSIAVA WYQQKPGKAP KLLIYSASYRYT GVPDRFSGSGS GTDFTLTISSLQ PEDFAVYYCQQ HYITPLTFGAGT KVEIKRTVAAP SVFIFPPSDEQL KSGTASVVCLL 8 ISKAKGQPREPQVYTLPPSREEMTKNQVS LTCLVKGFYPSDIAVEWESNGQPENNYK TTPPVLDSDGSFFLYSKLTVDKSRWQQG NVFSCSVMHEALHNHYTQKSLSLSLggggs ^^sOVOLVQSGAEVKKPGSSVKVSCKASG YTFTDNYMIWVROAPGOGLEWMGDINPYN GGTTFNQKFKGRVTITADKSTSTAYMELSSL RSEDTA VYYCARESPYFSNLYVMDYWGQGT TJYTOSggggsggggsggggsggggsEnYTgSMI LSLSPGERATLSCKASQSVDYDGDNYMNWY OOKPGQAPRLLIYAASNLESGIPARFSGSGS GTDFTLTISSLEPEDFA VYYCHLSNEDLSTFG GGTKVEIK NNFYPREAKVQ WKVDNALQSG NSQESVTEQDS KDSTYSLSSTLT LSKADYEKHKV YACEVTHQGLS SPVTKSFNRGE C DABA33 (aTROP2 x aCD163 in IgGl Backbone with ADCC Enhanced) QVQLQQSGSELKKPGASVKVSCKASGYT FTNYGMNWVKQAPGQGLKWMGWINTY TGEPTYTDDFKGRFAFSLDTSVSTAYLQIS SLKADDTAVYFCARGGFGS SYWYFDVW GQGSLVTVSSASTKGPSVFPLAPCSRSTSE STAALGCLVKDYFPEPVTVSWNSGALTS GVHTFPAVLQSSGLYSLSSVVTVPSSNFG TQTYICNVNHKPSNTKVDKRVEPKSCDK THTCPPCPAPELLGGPDVFLFPPKPKDTL MISRTPEVTCVVVDVSHEDPEVKFNWYV DGVEVHNAKTKPREEQYNSTYRVVSVLT VLHQDWLNGKEYKCKVSNKALPLPEEKT ISKAKGQPREPQVYTLPPSREEMTKNQVS LTCLVKGFYPSDIAVEWESNGQPENNYK TTPPVLDSDGSFFLYSKLTVDKSRWQQG NVFSCSVMHEALHNHYTQKSLSLSLggggs ^^OVOLQESGPGLVKPSETLSLTCTVSGY SITSDYA WNWIRQFPGNKLEWMGYITYSGST YYNPSLKSRVTISVDTSKNOFSLKLSSVTAAD TATYYCJ^GTYYFDYWGQGTPI^^ ^^^^^DIVMTQSPSSLSASVGDRVTI TCRASQSVSSDVA WFQQKPGKSPKPLIYYAS NRYSGVPSRFSGSGSGTDFTLTISSLQAEDFA VYFCGQDYTSPRTFGGGTKLEIK 112 DIQLTQSPSSLS ASVGDRVSITC KASQDVSIAVA WYQQKPGKAP KLLIYSASYRYT GVPDRFSGSGS GTDFTLTISSLQ PEDFAVYYCQQ HYITPLTFGAGT KVEIKRTVAAP SVFIFPPSDEQL KSGTASVVCLL NNFYPREAKVQ WKVDNALQSG NSQESVTEQDS KDSTYSLSSTLT LSKADYEKHKV YACEVTHQGLS SPVTKSFNRGE C 8 DABA 34 (aTROP2 x aDR5 in IgGl Backbone with ADCC Enhanced) QVQLQQSGSELKKPGASVKVSCKASGYT FTNYGMNWVKQAPGQGLKWMGWINTY TGEPTYTDDFKGRFAFSLDTSVSTAYLQIS SLKADDTAVYFCARGGFGS SYWYFDVW GQGSLVTVSSASTKGPSVFPLAPCSRSTSE STAALGCLVKDYFPEPVTVSWNSGALTS GVHTFPAVLQSSGLYSLSSVVTVPSSNFG TQTYICNVNHKPSNTKVDKRVEPKSCDK THTCPPCPAPELLGGPDVFLFPPKPKDTL MISRTPEVTCVVVDVSHEDPEVKFNWYV DGVEVHNAKTKPREEQYNSTYRVVSVLT VLHQDWLNGKEYKCKVSNKALPLPEEKT ISKAKGQPREPQVYTLPPSREEMTKNQVS LTCLVKGFYPSDIAVEWESNGQPENNYK TTPPVLDSDGSFFLYSKLTVDKSRWQQG NVFSCSVMHEALHNHYTQKSLSLSLggggs WWsEIVLTOSPGTLSLSPGERATLSCRASQG ISRSYLA WYOOKPGOAPSLLIYGASSRATGIP 113 DIQLTQSPSSLS ASVGDRVSITC KASQDVSIAVA WYQQKPGKAP KLLIYSASYRYT GVPDRFSGSGS GTDFTLTISSLQ PEDFAVYYCQQ HYITPLTFGAGT KVEIKRTVAAP SVFIFPPSDEQL KSGTASVVCLL NNFYPREAKVQ WKVDNALQSG NSQESVTEQDS KDSTYSLSSTLT LSKADYEKHKV YACEVTHQGLS 8 DRFSGSGSGTDFTLTISRLEPEDFA VYYCQQ FGSSPWTFGQGTKVEIKzgggsgg^ ^sOVOLOESGPGLVKPSOTLSLTCTVSGGSI SSGDYFWSWIRQLPGKGLEWIGHIHNSGTT YYNPSLKSRVTISVDTSKKQFSLRLSSVTAAD TA VYYCARDRGGDYYYGMDVWGOGTTVTV ss SPVTKSFNRGE C DABA 35 («GPC3 x aCD33 in IgGl Backbone with ADCC Enhanced) QVQLVQSGAEVKKPGASVKVSCKASGYT FTDYEMHWVRQAPGQGLEWMGALDPKT GDTAYSQKFKGRVTLTADKSTSTAYMEL S SLTSEDTAVYYCTRFYSYTYWGQGTLV TVSSASTKGPSVFPLAPSSKSTSGGTAALG CLVKDYFPEPVTVSWNSGALTSGVHTFP AVLQSSGLYSLSSVVTVPSSSLGTQTYICN VNHKPSNTKVDKRVEPKSCDKTHTCPPC PAPELLGGPDVFLFPPKPKDTLMISRTPEV TCVVVDVSHEDPEVKFNWYVDGVEVHN AKTKPREEQYNSTYRVVSVLTVLHQDWL NGKEYKCKVSNKALPLPEEKTISKAKGQP REPQVYTLPPSREEMTKNQVSLTCLVKGF YPSDIAVEWESNGQPENNYKTTPPVLDSD GSFFLYSKLTVDKSRWQQGNVFSCSVMH EALHNHYTQKSLSLSLggggsggggsEOJ^ SGAEVKKPGSSVKVSCKASGYTITDSNIHWV ROAPGOSLEWIGYIYPYNGGTDYNQKFKNR ATLTVDNPTNTAYMELSSLRSEDTAFYYCVN GNPWLAYWGQGTLVTVSS^w^^ ^^DIOLTQSPSTLSASVGDR VTITCRASES LDNYGIRFLTWFOQKPGKAPKLLMYAASNQ GSGVPSRFSGSGSGTEFTLTISSLQPDDFATY YCOQTKEVP WSFGQGTKVEVK 114 DVVMTQSPLSL PVTPGEPASISC RSSQSLVHSNR NTYLHWYLQK PGQSPQLLIYKV SNRFSGVPDRFS GSGSGTDFTLKI SRVEAEDVGVY YCSQNTHVPPT FGQGTKLEIKR TVAAPSVFIFPP SDEQLKSGTAS VVCLLNNFYPR EAKVQWKVDN ALQSGNSQESV TEQDSKDSTYS LSSTLTLSKAD YEKHKVYACE VTHQGLSSPVT KSFNRGEC 80 DABA 36 («GPC3 x aCSFIR in IgGl Backbone with ADCC Enhanced) QVQLVQSGAEVKKPGASVKVSCKASGYT FTDYEMHWVRQAPGQGLEWMGALDPKT GDTAYSQKFKGRVTLTADKSTSTAYMEL S SLTSEDTAVYYCTRFYSYTYWGQGTLV TVSSASTKGPSVFPLAPSSKSTSGGTAALG CLVKDYFPEPVTVSWNSGALTSGVHTFP AVLQSSGLYSLSSVVTVPSSSLGTQTYICN VNHKPSNTKVDKRVEPKSCDKTHTCPPC PAPELLGGPDVFLFPPKPKDTLMISRTPEV TCVVVDVSHEDPEVKFNWYVDGVEVHN AKTKPREEQYNSTYRVVSVLTVLHQDWL NGKEYKCKVSNKALPLPEEKTISKAKGQP REPQVYTLPPSREEMTKNQVSLTCLVKGF YPSDIAVEWESNGQPENNYKTTPPVLDSD GSFFLYSKLTVDKSRWQQGNVFSCSVMH EALHNHYTOKSLSLSLggggsggggsCWLE QSGAEVKKPGSSVKVSCKASGYTFTDNYMI WVROAPGOGLEWMGDINPYNGGTTFNQK FKGR VTITADKSTSTA YMELSSLRSEDTA VYY CARESPYFSNLYVMDYWGQGTLVTVSS^^ ^s^s^sEIVLTOSPATLSLSPGERAT LSCKASOSVDYDGDNYMNWYOOKPGQAPR LLIYAASNLESGIPARFSGSGSGTDFTLTISSL EPEDFA VYYCHLSNEDLSTFGGGTKVEIK 115 DVVMTQSPLSL PVTPGEPASISC RSSQSLVHSNR NTYLHWYLQK PGQSPQLLIYKV SNRFSGVPDRFS GSGSGTDFTLKI SRVEAEDVGVY YCSQNTHVPPT FGQGTKLEIKR TVAAPSVFIFPP SDEQLKSGTAS VVCLLNNFYPR EAKVQWKVDN ALQSGNSQESV TEQDSKDSTYS LSSTLTLSKAD YEKHKVYACE VTHQGLSSPVT KSFNRGEC 80 DABA37 («GPC3 x aCD163 in IgGl Backbone with ADCC Enhanced) QVQLVQSGAEVKKPGASVKVSCKASGYT FTDYEMHWVRQAPGQGLEWMGALDPKT GDTAYSQKFKGRVTLTADKSTSTAYMEL S SLTSEDTAVYYCTRFYSYTYWGQGTLV TVSSASTKGPSVFPLAPSSKSTSGGTAALG CLVKDYFPEPVTVSWNSGALTSGVHTFP AVLQSSGLYSLSSVVTVPSSSLGTQTYICN VNHKPSNTKVDKRVEPKSCDKTHTCPPC PAPELLGGPDVFLFPPKPKDTLMISRTPEV TCVVVDVSHEDPEVKFNWYVDGVEVHN AKTKPREEQYNSTYRVVSVLTVLHQDWL NGKEYKCKVSNKALPLPEEKTISKAKGQP REPQVYTLPPSREEMTKNQVSLTCLVKGF YPSDIAVEWESNGQPENNYKTTPPVLDSD GSFFLYSKLTVDKSRWQQGNVFSCSVMH EALHNHYTQKSLSLSLggggsggggs(WZC> ESGPGL VKPSETLSLTCTVSGYSITSDYA WN WIRQFPGNKLEWMGYITYSGSTYYNPSLKSR VTISVDTSKNOFSLKLSSVTAADTATYYCVSG 7THY)lMz2G77Z72<SSggggsggggsggggsggg ^DIVMTQSPSSLSASVGDRVTITCRASQSVSS DVA WFOOKPGKSPKPUYYASNRYSGVPSRF SGSGSGTDFTLTISSLQAEDFA VYFCGQDYT SPRTFGGGTKLEIK 116 DVVMTQSPLSL PVTPGEPASISC RSSQSLVHSNR NTYLHWYLQK PGQSPQLLIYKV SNRFSGVPDRFS GSGSGTDFTLKI SRVEAEDVGVY YCSQNTHVPPT FGQGTKLEIKR TVAAPSVFIFPP SDEQLKSGTAS VVCLLNNFYPR EAKVQWKVDN ALQSGNSQESV TEQDSKDSTYS LSSTLTLSKAD YEKHKVYACE VTHQGLSSPVT KSFNRGEC 80 DABA 38 («GPC3 x aDR5 in IgGl Backbone with ADCC Enhanced) QVQLVQSGAEVKKPGASVKVSCKASGYT FTDYEMHWVRQAPGQGLEWMGALDPKT GDTAYSQKFKGRVTLTADKSTSTAYMEL S SLTSEDTAVYYCTRFYSYTYWGQGTLV TVSSASTKGPSVFPLAPSSKSTSGGTAALG CLVKDYFPEPVTVSWNSGALTSGVHTFP AVLQSSGLYSLSSVVTVPSSSLGTQTYICN VNHKPSNTKVDKRVEPKSCDKTHTCPPC PAPELLGGPDVFLFPPKPKDTLMISRTPEV TCVVVDVSHEDPEVKFNWYVDGVEVHN AKTKPREEQYNSTYRVVSVLTVLHQDWL NGKEYKCKVSNKALPLPEEKTISKAKGQP REPQVYTLPPSREEMTKNQVSLTCLVKGF YPSDIAVEWESNGQPENNYKTTPPVLDSD GSFFLYSKLTVDKSRWQQGNVFSCSVMH EALHNHYTOKSLSLSLggggsggggsE’HYTO PGTLSLSPGERATLSCRASQGISRSYLA WYQ OKPGOAPSLLIYGASSRATGIPDRFSGSGSG TDFTLTISRLEPEDFA VYYCQQFGSSP WTFG QGTJGGE[^gggsggggsggggsggggsQVQ^ GPGL VKPSQTLSLTCTVSGGSISSGDYFWSW IRQLPGKGLEWIGHIHNSGTTYYNPSLKSRV TISVDTSKKQFSLRLSSVTAADTA VYYCARDR GGDYYYGMDVWGOGTTVTVSS 117 DVVMTQSPLSL PVTPGEPASISC RSSQSLVHSNR NTYLHWYLQK PGQSPQLLIYKV SNRFSGVPDRFS GSGSGTDFTLKI SRVEAEDVGVY YCSQNTHVPPT FGQGTKLEIKR TVAAPSVFIFPP SDEQLKSGTAS VVCLLNNFYPR EAKVQWKVDN ALQSGNSQESV TEQDSKDSTYS LSSTLTLSKAD YEKHKVYACE VTHQGLSSPVT KSFNRGEC 80 DABA 39 (aFOLRl x aCD33 in IgGl Backbone QVQLVQSGAEWKPGASVKISCKASGYT FTGYFMNWVKQSPGQSLEWIGRIHPYDG DTFYNQKFQGKATLTVDKS SNTAHMELL SLTSEDFAVYYCTRYDGSRAMDYWGQG TTVTVSSASTKGPSVFPLAPSSKSTSGGTA ALGCLVKDYFPEPVTVSWNSGALTSGVH 118 DIVLTQSPLSLA VSLGQPAIISCK ASQSVSFAGTS LMHWYHQKPG QQPRLLIYRAS NLEAGVPDRFS 85 with ADCC Enhanced) TFPAVLQSSGLYSLSSVVTVPSSSLGTQTY ICNVNHKPSNTKVDKRVEPKSCDKTHTCP PCPAPELLGGPDVFLFPPKPKDTLMISRTP EVTCVVVDVSHEDPEVKFNWYVDGVEV HNAKTKPREEQYNSTYRVVSVLTVLHQD WLNGKEYKCKVSNKALPLPEEKTISKAK GQPREPQVYTLPPSREEMTKNQVSLTCLV KGFYPSDIAVEWESNGQPENNYKTTPPVL DSDGSFFLYSKLTVDKSRWQQGNVFSCS VMHEALHNHYTQKSLSLSLggggsggggsEy QL VQSGAEVKKPGSSVKVSCKASGYTITDSN IHWVRQAPGQSLEWIGYIYPYNGGTDYNQK FKNRATLTVDNPTNTAYMELSSLRSEDTAFY YCVNGNP WLA YWGQGTL VTVSS^^^^ ^^^^DIQLTQSPSTLSASVGDRVTITCR ASESLDNYGIRFLTWFQQKPGKAPKLLMYA ASNQGSGVPSRFSGSGSGTEFTLTISSLQPD DFATYYCQQTKEVP WSFGQGTKVEVK GSGSKTDFTLTI SPVEAEDAATY YCQQSREYPYT FGGGTKLEIKR TVAAPSVFIFPP SDEQLKSGTAS VVCLLNNFYPR EAKVQWKVDN ALQSGNSQESV TEQDSKDSTYS LSSTLTLSKAD YEKHKVYACE VTHQGLSSPVT KSFNRGEC DABA 40 (aFOLRl x aCSFIR in IgGl Backbone with ADCC Enhanced) QVQLVQSGAEWKPGASVKISCKASGYT FTGYFMNWVKQSPGQSLEWIGRIHPYDG DTFYNQKFQGKATLTVDKS SNTAHMELL SLTSEDFAVYYCTRYDGSRAMDYWGQG TTVTVSSASTKGPSVFPLAPSSKSTSGGTA ALGCLVKDYFPEPVTVSWNSGALTSGVH TFPAVLQSSGLYSLSSVVTVPSSSLGTQTY ICNVNHKPSNTKVDKRVEPKSCDKTHTCP PCPAPELLGGPDVFLFPPKPKDTLMISRTP EVTCVVVDVSHEDPEVKFNWYVDGVEV HNAKTKPREEQYNSTYRVVSVLTVLHQD WLNGKEYKCKVSNKALPLPEEKTISKAK GQPREPQVYTLPPSREEMTKNQVSLTCLV KGFYPSDIAVEWESNGQPENNYKTTPPVL DSDGSFFLYSKLTVDKSRWQQGNVFSCS VMHEALHNHYTQKSLSLSLggggsggggs^E QL VQSGAEVKKPGSSVKVSCKASGYTFTDN YMIWVRQAPGQGLEWMGDINPYNGGTTFN QKFKGRVTITADKSTSTAYMELSSLRSEDTA VYYCARESPYFSNLYVMDYWGQGTL VTVSSz gggsggggsggggsggggsEIVLTQSPATLSLS RATLSCKASQSVDYDGDNYMNWYQQKPGQ APRLLIYAASNLESGIPARFSGSGSGTDFTLTI SSLEPEDFA VYYCHLSNEDLSTFGGGTKVEI K 119 DIVLTQSPLSLA VSLGQPAIISCK ASQSVSFAGTS LMHWYHQKPG QQPRLLIYRAS NLEAGVPDRFS GSGSKTDFTLTI SPVEAEDAATY YCQQSREYPYT FGGGTKLEIKR TVAAPSVFIFPP SDEQLKSGTAS VVCLLNNFYPR EAKVQWKVDN ALQSGNSQESV TEQDSKDSTYS LSSTLTLSKAD YEKHKVYACE VTHQGLSSPVT KSFNRGEC 85 DABA 41 (aFOLRl x aCD163 in IgGl Backbone with ADCC Enhanced) QVQLVQSGAEWKPGASVKISCKASGYT FTGYFMNWVKQSPGQSLEWIGRIHPYDG DTFYNQKFQGKATLTVDKS SNTAHMELL SLTSEDFAVYYCTRYDGSRAMDYWGQG TTVTVSSASTKGPSVFPLAPSSKSTSGGTA ALGCLVKDYFPEPVTVSWNSGALTSGVH TFPAVLQSSGLYSLSSVVTVPSSSLGTQTY ICNVNHKPSNTKVDKRVEPKSCDKTHTCP PCPAPELLGGPDVFLFPPKPKDTLMISRTP EVTCVVVDVSHEDPEVKFNWYVDGVEV HNAKTKPREEQYNSTYRVVSVLTVLHQD WLNGKEYKCKVSNKALPLPEEKTISKAK 120 DIVLTQSPLSLA VSLGQPAIISCK ASQSVSFAGTS LMHWYHQKPG QQPRLLIYRAS NLEAGVPDRFS GSGSKTDFTLTI SPVEAEDAATY YCQQSREYPYT FGGGTKLEIKR TVAAPSVFIFPP SDEQLKSGTAS 85 GQPREPQVYTLPPSREEMTKNQVSLTCLV KGFYPSDIAVEWESNGQPENNYKTTPPVL DSDGSFFLYSKLTVDKSRWQQGNVFSCS VMHEALHNHYTQKSLSLSLggggsggggseE QLQESGPGLVKPSETLSLTCTVSGYSITSDYA WNWIRQFPGNKLEWMGYITYSGSTYYNPSL KSRVTISVDTSKNQFSLKLSSVTAADTATYYC VSGTYYFDYWGQGTTLTVSS^w^m swwsDIVMTQSPSSIAASVGDRVTITCRASQ SVSSDVA WFQQKPGKSPKPLIYYASNRYSGV PSRFSGSGSGTDFTLTISSLQAEDFA VYFCG QDYTSPRTFGGGTKLEIK VVCLLNNFYPR EAKVQWKVDN ALQSGNSQESV TEQDSKDSTYS LSSTLTLSKAD YEKHKVYACE VTHQGLSSPVT KSFNRGEC DABA 42 (aFOLRl x aDR5 in IgGl Backbone with ADCC Enhanced) QVQLVQSGAEWKPGASVKISCKASGYT FTGYFMNWVKQSPGQSLEWIGRIHPYDG DTFYNQKFQGKATLTVDKS SNTAHMELL SLTSEDFAVYYCTRYDGSRAMDYWGQG TTVTVSSASTKGPSVFPLAPSSKSTSGGTA ALGCLVKDYFPEPVTVSWNSGALTSGVH TFPAVLQSSGLYSLSSVVTVPSSSLGTQTY ICNVNHKPSNTKVDKRVEPKSCDKTHTCP PCPAPELLGGPDVFLFPPKPKDTLMISRTP EVTCVVVDVSHEDPEVKFNWYVDGVEV HNAKTKPREEQYNSTYRVVSVLTVLHQD WLNGKEYKCKVSNKALPLPEEKTISKAK GQPREPQVYTLPPSREEMTKNQVSLTCLV KGFYPSDIAVEWESNGQPENNYKTTPPVL DSDGSFFLYSKLTVDKSRWQQGNVFSCS VMHEALHNHYTQKSLSLSLggggsggggsL / E LTQSPGTLSLSPGERATLSCRASQGISRSYLA WYQQKPGQAPSLUYGASSRATGIPDRFSGS GSGTDFTLTISRLEPEDFA VYYCQQFGSSP W TFGQGTKVEIK^^^^^^^OVQL QESGPGL VKPSQTLSLTCTVSGGSISSGDYF WSWIROLPGKGLEWIGHIHNSGTTYYNPSLK SRVTISVDTSKKQFSLRLSSVTAADTA VYYCA RDRGGDYYYGMDVWGOGTTVTVSS 121 DIVLTQSPLSLA VSLGQPAIISCK ASQSVSFAGTS LMHWYHQKPG QQPRLLIYRAS NLEAGVPDRFS GSGSKTDFTLTI SPVEAEDAATY YCQQSREYPYT FGGGTKLEIKR TVAAPSVFIFPP SDEQLKSGTAS VVCLLNNFYPR EAKVQWKVDN ALQSGNSQESV TEQDSKDSTYS LSSTLTLSKAD YEKHKVYACE VTHQGLSSPVT KSFNRGEC 85 DABA 43 (aCD33 x aDR5 in IgGl Backbone with ADCC Enhanced) EVQLVQSGAEVKKPGSSVKVSCKASGYTI TDSNIHWVRQAPGQSLEWIGYIYPYNGGT DYNQKFKNRATLTVDNPTNTAYMELSSL RSEDTAFYYCVNGNPWLAYWGQGTLVT VSSASTKGPSVFPLAPCSRSTSESTAALGC LVKDYFPEPVTVSWNSGALTSGVHTFPA VLQSSGLYSLSSVVTVPSSNFGTQTYICNV NHKPSNTKVDKRVEPKSCDKTHTCPPCP APELLGGPDVFLFPPKPKDTLMISRTPEVT CVVVDVSHEDPEVKFNWYVDGVEVHNA KTKPREEQYNSTYRVVSVLTVLHQDWLN GKEYKCKVSNKALPLPEEKTISKAKGQPR EPQVYTLPPSREEMTKNQVSLTCLVKGFY PSDIAVEWESNGQPENNYKTTPPVLDSDG SFFLYSKLTVDKSRWQQGNVFSCSVMHE ALHNHYTQKSLSLSLggggsggggsEnZTe^ GTLSLSPGERATLSCRASQGISRSYLA WYQQ KPGOAPSLLIYGASSRATGIPDRFSGSGSGT DFTLTISRLEPEDFA VYYCQQFGSSP WTFGQ 122 DIQLTQSPSTLS ASVGDRVTITC RASESLDNYGI RFLTWFQQKPG KAPKLLMYAAS NQGSGVPSRFS GSGSGTEFTLTI SSLQPDDFATY YCQQTKEVPWS FGQGTKVEVKR TVAAPSVFIFPP SDEQLKSGTAS VVCLLNNFYPR EAKVQWKVDN ALQSGNSQESV TEQDSKDSTYS LSSTLTLSKAD YEKHKVYACE 90 GTKVEIK^^^s^^^sQVQLQESG PGL VKPSOTLSLTCTVSGGSISSGDYFWSWIR QLPGKGLEWIGHIHNSGTTYYNPSLKSRVTIS VDTSKKQFSLRLSSVTAADTA VYYCARDRGG DYYYGMDVWGQGTTVTVSS VTHQGLSSPVT KSFNRGEC DABA44 («CD38 x aDR5 in IgGl Backbone with ADCC Enhanced) EVQLLESGGGLVQPGGSLRLSCAVSGFTF NSFAMSWVRQAPGKGLEWVSAISGSGGG TYYADSVKGRFTISRDNSKNTLYLQMNS LRAEDTAVYFCAKDKILWFGEPVFDYWG QGTLVTVSSASTKGPSVFPLAPSSKSTSGG TAALGCLVKDYFPEPVTVSWNSGALTSG VHTFPAVLQSSGLYSLSSVVTVPSSSLGTQ TYICNVNHKPSNTKVDKRVEPKSCDKTH TCPPCPAPELLGGPDVFLFPPKPKDTLMIS RTPEVTCVVVDVSHEDPEVKFNWYVDG VEVHNAKTKPREEQYNSTYRVVSVLTVL HQDWLNGKEYKCKVSNKALPLPEEKTIS KAKGQPREPQVYTLPPSREEMTKNQVSL TCLVKGFYPSDIAVEWESNGQPENNYKT TPPVLDSDGSFFLYSKLTVDKSRWQQGN VFSCSVMHEALHNHYTQKSLSLSLggggsg ^sEIVLTQSPGTLSLSPGERATLSCRASQGI SRSYLA WYQQKPGQAPSLLIYGASSRATGIP DRFSGSGSGTDFTLTISRLEPEDFA VYYCQQ FGSSPWTFGQGTKVEIKzgggsgg^ ^sOVOLOESGPGLVKPSOTLSLTCTVSGGSI SSGDYFWSWIRQLPGKGLEWIGHIHNSGTT YYNPSLKSRVTISVDTSKKQFSLRLSSVTAAD TA VYYCARDRGGDYYYGMDVWGOGTTVTV ss 123 EIVLTQSPATLS LSPGERATLSCR ASQSVSSYLAW YQQKPGQAPRL LIYDASNRATGI PARFSGSGSGT DFTLTISSLEPE DFAVYYCQQRS NWPPTFGQGTK VEIKRTVAAPS VFIFPPSDEQLK SGTASVVCLLN NFYPREAKVQ WKVDNALQSG NSQESVTEQDS KDSTYSLSSTLT LSKADYEKHKV YACEVTHQGLS SPVTKSFNRGE C 92 DABA45 (aFLT3 x aDR5 in IgGl Backbone with ADCC Enhanced) EVQLVQSGAEVKKPGASVKVSCKASGYT FTSYYMHWVRQAPGQGLEWMGIINPSGG STSYAQKFQGRVTMTRDTSTSTVYMELS SLRSEDTAVYYCARGVGAHDAFDIWGQG TTVTVSSASTKGPSVFPLAPSSKSTSGGTA ALGCLVKDYFPEPVTVSWNSGALTSGVH TFPAVLQSSGLYSLSSVVTVPSSSLGTQTY ICNVNHKPSNTKVDKRVEPKSCDKTHTCP PCPAPELLGGPDVFLFPPKPKDTLMISRTP EVTCVVVDVSHEDPEVKFNWYVDGVEV HNAKTKPREEQYNSTYRVVSVLTVLHQD WLNGKEYKCKVSNKALPLPEEKTISKAK GQPREPQVYTLPPSREEMTKNQVSLTCLV KGFYPSDIAVEWESNGQPENNYKTTPPVL DSDGSFFLYSKLTVDKSRWQQGNVFSCS VMHEALHNHYTQKSLSLSLggggsggggsE£E LTQSPGTLSLSPGERATLSCRASQGISRSYLA WYOOKPGOAPSLUYGASSRATGIPDRFSGS GSGTDFTLTISRLEPEDFA VYYCQQFGSSP W TFGOGTKVEIK^^^^^^^OVQL QESGPGL VKPSQTLSLTCTVSGGSISSGDYF WSWIROLPGKGLEWIGHIHNSGTTYYNPSLK SRVTISVDTSKKQFSLRLSSVTAADTA VYYCA RDRGGDYYYGMDVWGOGTTVTVSS 124 DVVMTQSPLSL PVTPGEPASISC RSSQSLLHSNG NNYLDWYLQK PGQSPQLLIYLG SNRASGVPDRF SGSGSDTDFTL QISRVEAEDVG VYYCMQGTHP AISFGQGTRLEI KRTVAAPSVFIF PPSDEQLKSGT ASVVCLLNNFY PREAKVQWKV DNALQSGNSQE SVTEQDSKDST YSLSSTLTLSKA DYEKHKVYAC EVTHQGLSSPV TKSFNRGEC 94 DABA 46 (aTROP2 x aCD33 in IgG4 Backbone with S228P Mutation) QVQLQQSGSELKKPGASVKVSCKASGYT FTNYGMNWVKQAPGQGLKWMGWINTY TGEPTYTDDFKGRFAFSLDTSVSTAYLQIS SLKADDTAVYFCARGGFGS SYWYFDVW GQGSLVTVSSASTKGPSVFPLAPCSRSTSE STAALGCLVKDYFPEPVTVSWNSGALTS GVHTFPAVLQSSGLYSLSSVVTVPSSSLGT KTYTCNVDHKPSNTKVDKRVESKYGPPC PPCPAPEFLGGPSVFLFPPKPKDTLMISRTP EVTCVVVDVSQEDPEVQFNWYVDGVEV HNAKTKPREEQFNSTYRVVSVLTVLHQD WLNGKEYKCKVSNKGLPSSIEKTISKAKG QPREPQVYTLPPSQEEMTKNQVSLTCLVK GFYPSDIAVEWESNGQPENNYKTTPPVLD SDGSFFLYSRLTVDKSRWQEGNVFSCSV MHEALHNHYTQKSLSLSLggggsggggsEP^ L VOSGAEVKKPGSSVKVSCKASGYTITDSNI HWVROAPGQSLEWIGYIYPYNGGTDYNQKF KNRATLTVDNPTNTAYMELSSLRSEDTAFYY CVNGNPWTAYWGQGTU^rV^ ^^^DIQLTQSPSTLSASVGDRVTITCRA SESLDNYGIRFLTWFOQKPGKAPKLLMYAA SNQGSGVPSRFSGSGSGTEFTLTISSLOPDD FATYYCOQTKEVP WSFGQGTKVEVK 125 DIQLTQSPSSLS ASVGDRVSITC KASQDVSIAVA WYQQKPGKAP KLLIYSASYRYT GVPDRFSGSGS GTDFTLTISSLQ PEDFAVYYCQQ HYITPLTFGAGT KVEIKRTVAAP SVFIFPPSDEQL KSGTASVVCLL NNFYPREAKVQ WKVDNALQSG NSQESVTEQDS KDSTYSLSSTLT LSKADYEKHKV YACEVTHQGLS SPVTKSFNRGE C 8 DABA 47 (aTROP2 x aCSFIR in IgG4 Backbone with S228P Mutation) QVQLQQSGSELKKPGASVKVSCKASGYT FTNYGMNWVKQAPGQGLKWMGWINTY TGEPTYTDDFKGRFAFSLDTSVSTAYLQIS SLKADDTAVYFCARGGFGS SYWYFDVW GQGSLVTVSSASTKGPSVFPLAPCSRSTSE STAALGCLVKDYFPEPVTVSWNSGALTS GVHTFPAVLQSSGLYSLSSVVTVPSSSLGT KTYTCNVDHKPSNTKVDKRVESKYGPPC PPCPAPEFLGGPSVFLFPPKPKDTLMISRTP EVTCVVVDVSQEDPEVQFNWYVDGVEV HNAKTKPREEQFNSTYRVVSVLTVLHQD WLNGKEYKCKVSNKGLPSSIEKTISKAKG QPREPQVYTLPPSQEEMTKNQVSLTCLVK GFYPSDIAVEWESNGQPENNYKTTPPVLD SDGSFFLYSRLTVDKSRWQEGNVFSCSV MHEALHNHYTQKSLSLSLggggsggggsOEQ L VQSGAEVKKPGSSVKVSCKASGYTFTDNY MIWVROAPGOGLEWMGDINPYNGGTTFNO KFKGR VTITADKSTSTA YMELSSLRSEDTA VY YCARESPYFSNLYVMDYWGQGT^^ gsggggsggggsggggsEIVLTQSPATLSLSPGERA TLSCKASOSVDYDGDNYMNWYOOKPGOAP RLLIYAASNLESGIPARFSGSGSGTDFTLTISS LEPEDFA VYYCHLSNEDLSTFGGGTKVEIK 126 DIQLTQSPSSLS ASVGDRVSITC KASQDVSIAVA WYQQKPGKAP KLLIYSASYRYT GVPDRFSGSGS GTDFTLTISSLQ PEDFAVYYCQQ HYITPLTFGAGT KVEIKRTVAAP SVFIFPPSDEQL KSGTASVVCLL NNFYPREAKVQ WKVDNALQSG NSQESVTEQDS KDSTYSLSSTLT LSKADYEKHKV YACEVTHQGLS SPVTKSFNRGE C 8 DABA 48 (aTROP2 x aCD163 in IgG4 Backbone QVQLQQSGSELKKPGASVKVSCKASGYT FTNYGMNWVKQAPGQGLKWMGWINTY TGEPTYTDDFKGRFAFSLDTSVSTAYLQIS SLKADDTAVYFCARGGFGS SYWYFDVW GQGSLVTVSSASTKGPSVFPLAPCSRSTSE STAALGCLVKDYFPEPVTVSWNSGALTS GVHTFPAVLQSSGLYSLSSVVTVPSSSLGT 127 DIQLTQSPSSLS ASVGDRVSITC KASQDVSIAVA WYQQKPGKAP KLLIYSASYRYT GVPDRFSGSGS GTDFTLTISSLQ 8 with S228P Mutation) KTYTCNVDHKPSNTKVDKRVESKYGPPC PPCPAPEFLGGPSVFLFPPKPKDTLMISRTP EVTCVVVDVSQEDPEVQFNWYVDGVEV HNAKTKPREEQFNSTYRVVSVLTVLHQD WLNGKEYKCKVSNKGLPSSIEKTISKAKG QPREPQVYTLPPSQEEMTKNQVSLTCLVK GFYPSDIAVEWESNGQPENNYKTTPPVLD SDGSFFLYSRLTVDKSRWQEGNVFSCSV MHEALHNHYTQKSLSLSLggggsggggs(W LOESGPGLVKPSETLSLTCTVSGYSITSDYA W NWIRQFPGNKLEWMGYITYSGSTYYNPSLKS RVTISVDTSKNQFSLKLSSVTAADTATYYCVS GTYYFDYWGQGTTLTVSS^w^^ ^sDIVMTQSPSSLSASVGDRVTITCRASQSV SSDVA WFOQKPGKSPKPLIYYASNRYSGVPS RFSGSGSGTDFTLTISSLOAEDFA VYFCGQD YTSPRTFGGGTKLEIK PEDFAVYYCQQ HYITPLTFGAGT KVEIKRTVAAP SVFIFPPSDEQL KSGTASVVCLL NNFYPREAKVQ WKVDNALQSG NSQESVTEQDS KDSTYSLSSTLT LSKADYEKHKV YACEVTHQGLS SPVTKSFNRGE C DABA 49 (aTROP2 x aDR5 in IgG4 Backbone with S228P Mutation) QVQLQQSGSELKKPGASVKVSCKASGYT FTNYGMNWVKQAPGQGLKWMGWINTY TGEPTYTDDFKGRFAFSLDTSVSTAYLQIS SLKADDTAVYFCARGGFGS SYWYFDVW GQGSLVTVSSASTKGPSVFPLAPCSRSTSE STAALGCLVKDYFPEPVTVSWNSGALTS GVHTFPAVLQSSGLYSLSSVVTVPSSSLGT KTYTCNVDHKPSNTKVDKRVESKYGPPC PPCPAPEFLGGPSVFLFPPKPKDTLMISRTP EVTCVVVDVSQEDPEVQFNWYVDGVEV HNAKTKPREEQFNSTYRVVSVLTVLHQD WLNGKEYKCKVSNKGLPSSIEKTISKAKG QPREPQVYTLPPSQEEMTKNQVSLTCLVK GFYPSDIAVEWESNGQPENNYKTTPPVLD SDGSFFLYSRLTVDKSRWQEGNVFSCSV MHEALHNHYTQKSLSLSLggggsggggsE / IY TQSPGTLSLSPGERATLSCRASQGISRSYLA W YOQKPGOAPSLLIYGASSRATGIPDRFSGSG SGTDFTLTISRLEPEDFA VYYCQQFGSSP WT FGSGTKEE^ggggsggggsggggsggggs^ISE QESGPGL VKPSQTLSLTCTVSGGSISSGDYF WSWIRQLPGKGLEWIGHIHNSGTTYYNPSLK SRVTISVDTSKKQFSLRLSSVTAADTA VYYCA RDRGGDYYYGMDVWGOGTTVTVSS 128 DIQLTQSPSSLS ASVGDRVSITC KASQDVSIAVA WYQQKPGKAP KLLIYSASYRYT GVPDRFSGSGS GTDFTLTISSLQ PEDFAVYYCQQ HYITPLTFGAGT KVEIKRTVAAP SVFIFPPSDEQL KSGTASVVCLL NNFYPREAKVQ WKVDNALQSG NSQESVTEQDS KDSTYSLSSTLT LSKADYEKHKV YACEVTHQGLS SPVTKSFNRGE C 8 DABA 50 («GPC3 x aCD33 in IgG4 Backbone with S228P Mutation) QVQLVQSGAEVKKPGASVKVSCKASGYT FTDYEMHWVRQAPGQGLEWMGALDPKT GDTAYSQKFKGRVTLTADKSTSTAYMEL S SLTSEDTAVYYCTRFYSYTYWGQGTLV TVSSASTKGPSVFPLAPCSRSTSESTAALG CLVKDYFPEPVTVSWNSGALTSGVHTFP AVLQSSGLYSLSSVVTVPSSSLGTKTYTC NVDHKPSNTKVDKRVESKYGPPCPPCPAP EFLGGPSVFLFPPKPKDTLMISRTPEVTCV VVDVSQEDPEVQFNWYVDGVEVHNAKT KPREEQFNSTYRVVSVLTVLHQDWLNGK EYKCKVSNKGLPS SIEKTISKAKGQPREPQ VYTLPPSQEEMTKNQVSLTCLVKGFYPSD IAVEWESNGQPENNYKTTPPVLDSDGSFF 129 DVVMTQSPLSL PVTPGEPASISC RSSQSLVHSNR NTYLHWYLQK PGQSPQLLIYKV SNRFSGVPDRFS GSGSGTDFTLKI SRVEAEDVGVY YCSQNTHVPPT FGQGTKLEIKR TVAAPSVFIFPP SDEQLKSGTAS VVCLLNNFYPR EAKVQWKVDN 80 LYSRLTVDKSRWQEGNVFSCSVMHEALH NHYTQKSLSLSLggggsggggsEP^EI^SGAE VKKPGSSVKVSCKASGYTITDSNIHWVRQAP GOSLEWIGYIYPYNGGTDYNOKFKNRATLTV DNPTNTA YMELSSLRSEDTAFYYCVNGNP W LAYWGQGTLVTVSS^^s^^s^^s^^sD IQLTQSPSTLSASVGDR VTITCRASESLDNYGI RFLTWFOOKPGKAPKLLMYAASNOGSGVPS RFSGSGSGTEFTLTISSLOPDDFATYYCOOT KEVP WSFGQGTKVEVK ALQSGNSQESV TEQDSKDSTYS LSSTLTLSKAD YEKHKVYACE VTHQGLSSPVT KSFNRGEC DABA 51 («GPC3 x aCSFIR in IgG4 Backbone with S228P Mutation) QVQLVQSGAEVKKPGASVKVSCKASGYT FTDYEMHWVRQAPGQGLEWMGALDPKT GDTAYSQKFKGRVTLTADKSTSTAYMEL S SLTSEDTAVYYCTRFYSYTYWGQGTLV TVSSASTKGPSVFPLAPCSRSTSESTAALG CLVKDYFPEPVTVSWNSGALTSGVHTFP AVLQSSGLYSLSSVVTVPSSSLGTKTYTC NVDHKPSNTKVDKRVESKYGPPCPPCPAP EFLGGPSVFLFPPKPKDTLMISRTPEVTCV VVDVSQEDPEVQFNWYVDGVEVHNAKT KPREEQFNSTYRVVSVLTVLHQDWLNGK EYKCKVSNKGLPS SIEKTISKAKGQPREPQ VYTLPPSQEEMTKNQVSLTCLVKGFYPSD IAVEWESNGQPENNYKTTPPVLDSDGSFF LYSRLTVDKSRWQEGNVFSCSVMHEALH NHYTQKSLSLSLggggsggggsOEOLEQSGAE VKKPGSSVKVSCKASGYTFTDNYMIWVRQA PGOGLEWMGDINPYNGGTTFNQKFKGRVT ITADKSTSTAYMELSSLRSEDTA VYYCARESP YFSN]J[VMDYWGQ^ ^^^EIVLTQSPATLSLSPGERATLSCKAS OSVDYDGDNYMNWYOOKPGOAPRLLIYAA SNLESGIPARFSGSGSGTDFTLTISSLEPEDF A VYYCHLSNEDLSTFGGGTKVEIK 130 DVVMTQSPLSL PVTPGEPASISC RSSQSLVHSNR NTYLHWYLQK PGQSPQLLIYKV SNRFSGVPDRFS GSGSGTDFTLKI SRVEAEDVGVY YCSQNTHVPPT FGQGTKLEIKR TVAAPSVFIFPP SDEQLKSGTAS VVCLLNNFYPR EAKVQWKVDN ALQSGNSQESV TEQDSKDSTYS LSSTLTLSKAD YEKHKVYACE VTHQGLSSPVT KSFNRGEC 80 DABA 52 («GPC3 x aCD163 in IgG4 Backbone with S228P Mutation) QVQLVQSGAEVKKPGASVKVSCKASGYT FTDYEMHWVRQAPGQGLEWMGALDPKT GDTAYSQKFKGRVTLTADKSTSTAYMEL S SLTSEDTAVYYCTRFYSYTYWGQGTLV TVSSASTKGPSVFPLAPCSRSTSESTAALG CLVKDYFPEPVTVSWNSGALTSGVHTFP AVLQSSGLYSLSSVVTVPSSSLGTKTYTC NVDHKPSNTKVDKRVESKYGPPCPPCPAP EFLGGPSVFLFPPKPKDTLMISRTPEVTCV VVDVSQEDPEVQFNWYVDGVEVHNAKT KPREEQFNSTYRVVSVLTVLHQDWLNGK EYKCKVSNKGLPS SIEKTISKAKGQPREPQ VYTLPPSQEEMTKNQVSLTCLVKGFYPSD IAVEWESNGQPENNYKTTPPVLDSDGSFF LYSRLTVDKSRWQEGNVFSCSVMHEALH NHYTOKSLSLSLggggsggggsOEpLpESGPG L VKPSETLSLTCTVSGYSITSDYA WNWIRQFP GNKLEWMGYITYSGSTYYNPSLKSRVTISVDT SKNQFSLKLSSVTAADTATYYCVSGTYYFDY WGQGTTLIVSSr^s^zs^^^ TQSPSSLSASVGDRVTITCRASQSVSSDVA WF 131 DVVMTQSPLSL PVTPGEPASISC RSSQSLVHSNR NTYLHWYLQK PGQSPQLLIYKV SNRFSGVPDRFS GSGSGTDFTLKI SRVEAEDVGVY YCSQNTHVPPT FGQGTKLEIKR TVAAPSVFIFPP SDEQLKSGTAS VVCLLNNFYPR EAKVQWKVDN ALQSGNSQESV TEQDSKDSTYS LSSTLTLSKAD YEKHKVYACE VTHQGLSSPVT KSFNRGEC 80 OQKPGKSPKPLIYYASNRYSGVPSRFSGSGS GTDFTLTISSLQAEDFA VYFCGQDYTSPRTF GGGTKLEIK DABA53 («GPC3 x aDR5 in IgG4 Backbone with S228P Mutation) QVQLVQSGAEVKKPGASVKVSCKASGYT FTDYEMHWVRQAPGQGLEWMGALDPKT GDTAYSQKFKGRVTLTADKSTSTAYMEL S SLTSEDTAVYYCTRFYSYTYWGQGTLV TVSSASTKGPSVFPLAPCSRSTSESTAALG CLVKDYFPEPVTVSWNSGALTSGVHTFP AVLQSSGLYSLSSVVTVPSSSLGTKTYTC NVDHKPSNTKVDKRVESKYGPPCPPCPAP EFLGGPSVFLFPPKPKDTLMISRTPEVTCV VVDVSQEDPEVQFNWYVDGVEVHNAKT KPREEQFNSTYRVVSVLTVLHQDWLNGK EYKCKVSNKGLPSSIEKTISKAKGQPREPQ VYTLPPSQEEMTKNQVSLTCLVKGFYPSD IAVEWESNGQPENNYKTTPPVLDSDGSFF LYSRLTVDKSRWQEGNVFSCSVMHEALH NHYTQKSLSLSLggggsggggsffZELWPGTZ SLSPGERATLSCRASQGISRSYLA WYQQKPG QAPSLLIYGASSRATGIPDRFSGSGSGTDFTL TISRLEPEDFA VYYCQQFGSSP WTFGQGTK VEIKggggsggggsggggs^ VKPSOTLSLTCTVSGGSISSGDYFWSWIROLP GKGLEWIGHIHNSGTTYYNPSLKSRVTISVDT SKKQFSLRLSSVTAADTA VYYCARDRGGDYY YGMDVWGQGTTVTVSS 132 DVVMTQSPLSL PVTPGEPASISC RSSQSLVHSNR NTYLHWYLQK PGQSPQLLIYKV SNRFSGVPDRFS GSGSGTDFTLKI SRVEAEDVGVY YCSQNTHVPPT FGQGTKLEIKR TVAAPSVFIFPP SDEQLKSGTAS VVCLLNNFYPR EAKVQWKVDN ALQSGNSQESV TEQDSKDSTYS LSSTLTLSKAD YEKHKVYACE VTHQGLSSPVT KSFNRGEC 80 DABA54 (aFOLRl x aCD33 in IgG4 Backbone with S228P Mutation) QVQLVQSGAEWKPGASVKISCKASGYT FTGYFMNWVKQSPGQSLEWIGRIHPYDG DTFYNQKFQGKATLTVDKS SNTAHMELL SLTSEDFAVYYCTRYDGSRAMDYWGQG TTVTVSSASTKGPSVFPLAPCSRSTSESTA ALGCLVKDYFPEPVTVSWNSGALTSGVH TFPAVLQSSGLYSLSSVVTVPSSSLGTKTY TCNVDHKPSNTKVDKRVESKYGPPCPPCP APEFLGGPSVFLFPPKPKDTLMISRTPEVT CVVVDVSQEDPEVQFNWYVDGVEVHNA KTKPREEQFNSTYRVVSVLTVLHQDWLN GKEYKCKVSNKGLPSSIEKTISKAKGQPR EPQWTLPPSQEEMTKNQVSLTCLVKGFY PSDIAVEWESNGQPENNYKTTPPVLDSDG SFFLYSRLTVDKSRWQEGNVFSCSVMHE ALHNHYTOKSLSLSLggggsggggsEECYmS GAEVKKPGSSVKVSCKASGYTITDSNIHWVR OAPGOSLEWIGYIYPYNGGTDYNOKFKNRA TLTVDNPTNTA YMELSSLRSEDTAFYYCVNG NP WLA YWGQGTLVTVSS^^^^^^ ^sDIOLTOSPSTLSASVGDRVTITCRASESLD NYGIRFLTWFOQKPGKAPKLLMYAASNQGS GVPSRFSGSGSGTEFTLTISSLQPDDFATYYC OOTKEVP WSFGQGTKVEVK 133 DIVLTQSPLSLA VSLGQPAIISCK ASQSVSFAGTS LMHWYHQKPG QQPRLLIYRAS NLEAGVPDRFS GSGSKTDFTLTI SPVEAEDAATY YCQQSREYPYT FGGGTKLEIKR TVAAPSVFIFPP SDEQLKSGTAS VVCLLNNFYPR EAKVQWKVDN ALQSGNSQESV TEQDSKDSTYS LSSTLTLSKAD YEKHKVYACE VTHQGLSSPVT KSFNRGEC 85 DABA55 QVQLVQSGAEWKPGASVKISCKASGYT FTGYFMNWVKQSPGQSLEWIGRIHPYDG DTFYNQKFQGKATLTVDKS SNTAHMELL 134 DIVLTQSPLSLA VSLGQPAIISCK ASQSVSFAGTS 85 (aFOLRl x aCSFIR in IgG4 Backbone with S228P Mutation) SLTSEDFAVYYCTRYDGSRAMDYWGQG TTVTVSSASTKGPSVFPLAPCSRSTSESTA ALGCLVKDYFPEPVTVSWNSGALTSGVH TFPAVLQSSGLYSLSSVVTVPSSSLGTKTY TCNVDHKPSNTKVDKRVESKYGPPCPPCP APEFLGGPSVFLFPPKPKDTLMISRTPEVT CVVVDVSQEDPEVQFNWYVDGVEVHNA KTKPREEQFNSTYRVVSVLTVLHQDWLN GKEYKCKVSNKGLPSSIEKTISKAKGQPR EPQVYTLPPSQEEMTKNQVSLTCLVKGFY PSDIAVEWESNGQPENNYKTTPPVLDSDG SFFLYSRLTVDKSRWQEGNVFSCSVMHE ALHNHYTQKSLSLSLggggsggggsfilW^ GAEVKKPGSSVKVSCKASGYTFTDNYMIWV ROAPGOGLEWMGDINPYNGGTTFNOKFKG R VTITADKSTSTA YMELSSLRSEDTA VYYCAR ESPYFSNEYV^^ ^^^^EIVLTQSPATLSLSPGERATLSC KASOSVDYDGDNYMNWYOQKPGQAPRLLI YAASNLESGIPARFSGSGSGTDFTLTISSLEPE DFA VYYCHLSNEDLSTFGGGTKVEIK LMHWYHQKPG QQPRLLIYRAS NLEAGVPDRFS GSGSKTDFTLTI SPVEAEDAATY YCQQSREYPYT FGGGTKLEIKR TVAAPSVFIFPP SDEQLKSGTAS VVCLLNNFYPR EAKVQWKVDN ALQSGNSQESV TEQDSKDSTYS LSSTLTLSKAD YEKHKVYACE VTHQGLSSPVT KSFNRGEC DABA 56 (aFOLRl x aCD163 in IgG4 Backbone with S228P Mutation) QVQLVQSGAEWKPGASVKISCKASGYT FTGYFMNWVKQSPGQSLEWIGRIHPYDG DTFYNQKFQGKATLTVDKS SNTAHMELL SLTSEDFAVYYCTRYDGSRAMDYWGQG TTVTVSSASTKGPSVFPLAPCSRSTSESTA ALGCLVKDYFPEPVTVSWNSGALTSGVH TFPAVLQSSGLYSLSSVVTVPSSSLGTKTY TCNVDHKPSNTKVDKRVESKYGPPCPPCP APEFLGGPSVFLFPPKPKDTLMISRTPEVT CVVVDVSQEDPEVQFNWYVDGVEVHNA KTKPREEQFNSTYRVVSVLTVLHQDWLN GKEYKCKVSNKGLPSSIEKTISKAKGQPR EPQVYTLPPSQEEMTKNQVSLTCLVKGFY PSDIAVEWESNGQPENNYKTTPPVLDSDG SFFLYSRLTVDKSRWQEGNVFSCSVMHE ALHNHYTOKSLSLSLggggsggggsCWZpES GPGLVKPSETLSLTCTVSGYSITSDYA WNWIR QFPGNKLEWMGYITYSGSTYYNPSLKSRVTIS VDTSKNOFSLKLSSVTAADTATYYCVSGTYYF DYWGQGTTLTVSS^^^^^^^DI VMTOSPSSLSASVGDRVTITCRASOSVSSDVA WFQQKPGKSPKPLIYYASNRYSGVPSRFSGS GSGTDFTLTISSLQAEDFA VYFCGQDYTSPR TFGGGTKLEIK 135 DIVLTQSPLSLA VSLGQPAIISCK ASQSVSFAGTS LMHWYHQKPG QQPRLLIYRAS NLEAGVPDRFS GSGSKTDFTLTI SPVEAEDAATY YCQQSREYPYT FGGGTKLEIKR TVAAPSVFIFPP SDEQLKSGTAS VVCLLNNFYPR EAKVQWKVDN ALQSGNSQESV TEQDSKDSTYS LSSTLTLSKAD YEKHKVYACE VTHQGLSSPVT KSFNRGEC 85 DABA 57 (aFOLRl x aDR5 in IgG4 Backbone with S228P Mutation) QVQLVQSGAEWKPGASVKISCKASGYT FTGYFMNWVKQSPGQSLEWIGRIHPYDG DTFYNQKFQGKATLTVDKS SNTAHMELL SLTSEDFAVYYCTRYDGSRAMDYWGQG TTVTVSSASTKGPSVFPLAPCSRSTSESTA ALGCLVKDYFPEPVTVSWNSGALTSGVH TFPAVLQSSGLYSLSSVVTVPSSSLGTKTY TCNVDHKPSNTKVDKRVESKYGPPCPPCP APEFLGGPSVFLFPPKPKDTLMISRTPEVT CVVVDVSQEDPEVQFNWYVDGVEVHNA 136 DIVLTQSPLSLA VSLGQPAIISCK ASQSVSFAGTS LMHWYHQKPG QQPRLLIYRAS NLEAGVPDRFS GSGSKTDFTLTI SPVEAEDAATY YCQQSREYPYT FGGGTKLEIKR 85 KTKPREEQFNSTYRVVSVLTVLHQDWLN GKEYKCKVSNKGLPSSIEKTISKAKGQPR EPQVYTLPPSQEEMTKNQVSLTCLVKGFY PSDIAVEWESNGQPENNYKTTPPVLDSDG SFFLYSRLTVDKSRWQEGNVFSCSVMHE ALHNHYTQKSLSLSLggggsggggsEnYTgSP GTLSLSPGERATLSCRASQGISRSYLA WYQQ KPGQAPSLLIYGASSRATGIPDRFSGSGSGT DFTLTISRLEPEDFA VYYCQQFGSSP WTFGQ GTKVEIKggggsggggsggg^^ PGL VKPSOTLSLTCTVSGGSISSGDYFWSWIR OLPGKGLEWIGHIHNSGTTYYNPSLKSRVTIS VDTSKKQFSLRLSSVTAADTA VYYCARDRGG DYYYGMDVWGQGTTVTVSS TVAAPSVFIFPP SDEQLKSGTAS VVCLLNNFYPR EAKVQWKVDN ALQSGNSQESV TEQDSKDSTYS LSSTLTLSKAD YEKHKVYACE VTHQGLSSPVT KSFNRGEC DABA 58 («CD33 x aDR5 in IgG4 Backbone with S228P Mutation) EVQLVQSGAEVKKPGSSVKVSCKASGYTI TDSNIHWVRQAPGQSLEWIGYIYPYNGGT DYNQKFKNRATLTVDNPTNTAYMELSSL RSEDTAFYYCVNGNPWLAYWGQGTLVT VSSASTKGPSVFPLAPCSRSTSESTAALGC LVKDYFPEPVTVSWNSGALTSGVHTFPA VLQSSGLYSLSSVVTVPSSSLGTKTYTCN VDHKPSNTKVDKRVESKYGPPCPPCPAPE FLGGPSVFLFPPKPKDTLMISRTPEVTCVV VDVSQEDPEVQFNWYVDGVEVHNAKTK PREEQFNSTYRVVSVLTVLHQDWLNGKE YKCKVSNKGLPSSIEKTISKAKGQPREPQ VYTLPPSQEEMTKNQVSLTCLVKGFYPSD IAVEWESNGQPENNYKTTPPVLDSDGSFF LYSRLTVDKSRWQEGNVFSCSVMHEALH NHYTQKSLSLSLggggsggggsffZELWPGTZ SLSPGERATLSCRASQGISRSYLA WYQQKPG OAPSLUYGASSRATGIPDRFSGSGSGTDFTL TISRLEPEDFA VYYCQQFGSSP WTFGQGTK VEIK^^^^^^^QVQLQESGPGL VKPSOTLSLTCTVSGGSISSGDYFWSWIROLP GKGLEWIGHIHNSGTTYYNPSLKSRVTISVDT SKKQFSLRLSSVTAADTA VYYCARDRGGDYY YGMDVWGQGTTVTVSS 137 DIQLTQSPSTLS ASVGDRVTITC RASESLDNYGI RFLTWFQQKPG KAPKLLMYAAS NQGSGVPSRFS GSGSGTEFTLTI SSLQPDDFATY YCQQTKEVPWS FGQGTKVEVKR TVAAPSVFIFPP SDEQLKSGTAS VVCLLNNFYPR EAKVQWKVDN ALQSGNSQESV TEQDSKDSTYS LSSTLTLSKAD YEKHKVYACE VTHQGLSSPVT KSFNRGEC 90 DABA_59 (aCD38 x aDR5 in IgG4 Backbone with S228P Mutation) EVQLLESGGGLVQPGGSLRLSCAVSGFTF NSFAMSWVRQAPGKGLEWVSAISGSGGG TYYADSVKGRFTISRDNSKNTLYLQMNS LRAEDTAVYFCAKDKILWFGEPVFDYWG QGTLVTVSSASTKGPSVFPLAPCSRSTSES TAALGCLVKDYFPEPVTVSWNSGALTSG VHTFPAVLQSSGLYSLSSVVTVPSSSLGTK TYTCNVDHKPSNTKVDKRVESKYGPPCP PCPAPEFLGGPSVFLFPPKPKDTLMISRTP EVTCVVVDVSQEDPEVQFNWYVDGVEV HNAKTKPREEQFNSTYRVVSVLTVLHQD WLNGKEYKCKVSNKGLPSSIEKTISKAKG QPREPQVYTLPPSQEEMTKNQVSLTCLVK GFYPSDIAVEWESNGQPENNYKTTPPVLD SDGSFFLYSRLTVDKSRWQEGNVFSCSV MHEALHNHYTQKSLSLSLggggsggggsEZFL TQSPGTLSLSPGERATLSCRASQGISRSYLA W 138 EIVLTQSPATLS LSPGERATLSCR ASQSVSSYLAW YQQKPGQAPRL LIYDASNRATGI PARFSGSGSGT DFTLTISSLEPE DFAVYYCQQRS NWPPTFGQGTK VEIKRTVAAPS VFIFPPSDEQLK SGTASVVCLLN NFYPREAKVQ WKVDNALQSG NSQESVTEQDS KDSTYSLSSTLT LSKADYEKHKV 92 YQQKPGQAPSLLIYGASSRATGIPDRFSGSG SGTDFTLTISRLEPEDFA VYYCQQFGSSP WT FGQGTKVEIKggggsggggsggggsggggsQVQL QESGPGL VKPSQTLSLTCTVSGGSISSGDYF WSWIROLPGKGLEWIGHIHNSGTTYYNPSLK SRVTISVDTSKKQFSLRLSSVTAADTA VYYCA RDRGGDYYYGMDVWGQGTTVTVSS YACEVTHQGLS SPVTKSFNRGE C DABA 60 (aFLT3 x aDR5 in IgG4 Backbone with S228P Mutation) EVQLVQSGAEVKKPGASVKVSCKASGYT FTSYYMHWVRQAPGQGLEWMGIINPSGG STSYAQKFQGRVTMTRDTSTSTVYMELS SLRSEDTAVYYCARGVGAHDAFDIWGQG TTVTVSSASTKGPSVFPLAPCSRSTSESTA ALGCLVKDYFPEPVTVSWNSGALTSGVH TFPAVLQSSGLYSLSSVVTVPSSSLGTKTY TCNVDHKPSNTKVDKRVESKYGPPCPPCP APEFLGGPSVFLFPPKPKDTLMISRTPEVT CVVVDVSQEDPEVQFNWYVDGVEVHNA KTKPREEQFNSTYRVVSVLTVLHQDWLN GKEYKCKVSNKGLPSSIEKTISKAKGQPR EPQVYTLPPSQEEMTKNQVSLTCLVKGFY PSDIAVEWESNGQPENNYKTTPPVLDSDG SFFLYSRLTVDKSRWQEGNVFSCSVMHE ALHNHYTQKSLSLSLggggsggggsEZmaiSP GTLSLSPGERATLSCRASQGISRSYLA WYQQ KPGQAPSLLIYGASSRATGIPDRFSGSGSGT DFTLTISRLEPEDFA VYYCQQFGSSP WTFGQ PGL VKPSOTLSLTCTVSGGSISSGDYFWSWIR QLPGKGLEWIGHIHNSGTTYYNPSLKSRVTIS VDTSKKQFSLRLSSVTAADTA VYYCARDRGG DYYYGMDVWGQGTTVTVSS 139 DVVMTQSPLSL PVTPGEPASISC RSSQSLLHSNG NNYLDWYLQK PGQSPQLLIYLG SNRASGVPDRF SGSGSDTDFTL QISRVEAEDVG VYYCMQGTHP AISFGQGTRLEI KRTVAAPSVFIF PPSDEQLKSGT ASVVCLLNNFY PREAKVQWKV DNALQSGNSQE SVTEQDSKDST YSLSSTLTLSKA DYEKHKVYAC EVTHQGLSSPV TKSFNRGEC 94

[0099] In certain embodiments, a multi-specific binding polypeptide (e.g., a multi-specific antibody) described herein comprises an immunoglobulin heavy chain comprising an amino acid sequence at least about 80%, 85%, 90%, 95%, 97%, 98%, 99%, or 100% identical to or consist of the amino acid sequence set forth in SEQ ID NOs: 7 or 75-77; and an immunoglobulin light chain at least about 80%, 85%, 90%, 95%, 97%, 98%, 99%, or 100% identical to or consist of the amino acid sequence set forth in SEQ ID NO: 8. In some instances, the amino acid differences contributing to the at least about 80%, 85%, 90%, 95%, 97%, 98%, or 99% sequence identity resides in a framework region of the immunoglobulin heavy chain and the CDRs remain unchanged relative to the CDRs set forth in SEQ ID NOs: 7 or 75-77. In some instances, the amino acid differences contributing to the at least about 80%, 85%, 90%, 95%, 97%, 98%, or 99% sequence identity resides in a framework region of the immunoglobulin light chain and the CDRs remain unchanged relative to the CDRs set forth in SEQ ID NO: 8. In some cases, the amino acid differences contributing to the at least about 80%, 85%, 90%, 95%, 97%, 98%, or 99% sequence identity comprises conservative substitutions in the framework region, CDRs region, or a combination thereof, of the immunoglobulin heavy chain but the multi-specific binding polypeptide (e.g., the multi-specific antibody) retains binding to a target tumor antigen and / or a target antigen expressed on an immunosuppressive cell. In some cases, the amino acid differences contributing to the at least about 80%, 85%, 90%, 95%, 97%, 98%, or 99% sequence identity comprises conservative substitutions in the framework region, CDRs region, or a combination thereof, of the immunoglobulin light chain but the multi-specific binding polypeptide (e.g., the multi-specific antibody) retains binding to a target tumor antigen and / or a target antigen expressed on an immunosuppressive cell.

[0100] In certain embodiments, a multi-specific binding polypeptide (e.g., a multi-specific antibody) described herein comprises an immunoglobulin heavy chain comprising an amino acid sequence at least about 80%, 85%, 90%, 95%, 97%, 98%, 99%, or 100% identical to or consist of the amino acid sequence set forth in SEQ ID NOs: 79-83; and an immunoglobulin light chain at least about 80%, 85%, 90%, 95%, 97%, 98%, 99%, or 100% identical to or consist of the amino acid sequence set forth in SEQ ID NO: 80. In some instances, the amino acid differences contributing to the at least about 80%, 85%, 90%, 95%, 97%, 98%, or 99% sequence identity resides in a framework region of the immunoglobulin heavy chain and the CDRs remain unchanged relative to the CDRs set forth in SEQ ID NOs: 79-83. In some instances, the amino acid differences contributing to the at least about 80%, 85%, 90%, 95%, 97%, 98%, or 99% sequence identity resides in a framework region of the immunoglobulin light chain and the CDRs remain unchanged relative to the CDRs set forth in SEQ ID NO: 80. In some cases, the amino acid differences contributing to the at least about 80%, 85%, 90%, 95%, 97%, 98%, or 99% sequence identity comprises conservative substitutions in the framework region, CDRs region, or a combination thereof, of the immunoglobulin heavy chain but the multi-specific binding polypeptide (e.g., the multi-specific antibody) retains binding to a target tumor antigen and / or a target antigen expressed on an immunosuppressive cell. In some cases, the amino acid differences contributing to the at least about 80%, 85%, 90%, 95%, 97%, 98%, or 99% sequence identity comprises conservative substitutions in the framework region, CDRs region, or a combination thereof, of the immunoglobulin light chain but the multi-specific binding polypeptide (e.g., the multi-specific antibody) retains binding to a target tumor antigen and / or a target antigen expressed on an immunosuppressive cell.

[0101] In certain embodiments, a multi-specific binding polypeptide (e.g., a multi-specific antibody) described herein comprises an immunoglobulin heavy chain comprising an amino acid sequence at least about 80%, 85%, 90%, 95%, 97%, 98%, 99%, or 100% identical to or consist of the amino acid sequence set forth in SEQ ID NOs: 84-88; and an immunoglobulin light chain at least about 80%, 85%, 90%, 95%, 97%, 98%, 99%, or 100% identical to or consist of the amino acid sequence set forth in SEQ ID NO: 85. In some instances, the amino acid differences contributing to the at least about 80%, 85%, 90%, 95%, 97%, 98%, or 99% sequence identity resides in a framework region of the immunoglobulin heavy chain and the CDRs remain unchanged relative to the CDRs set forth in SEQ ID NOs: 84-88. In some instances, the amino acid differences contributing to the at least about 80%, 85%, 90%, 95%, 97%, 98%, or 99% sequence identity resides in a framework region of the immunoglobulin light chain and the CDRs remain unchanged relative to the CDRs set forth in SEQ ID NO: 85. In some cases, the amino acid differences contributing to the at least about 80%, 85%, 90%, 95%, 97%, 98%, or 99% sequence identity comprises conservative substitutions in the framework region, CDRs region, or a combination thereof, of the immunoglobulin heavy chain but the multi-specific binding polypeptide (e.g., the multi-specific antibody) retains binding to a target tumor antigen and / or a target antigen expressed on an immunosuppressive cell. In some cases, the amino acid differences contributing to the at least about 80%, 85%, 90%, 95%, 97%, 98%, or 99% sequence identity comprises conservative substitutions in the framework region, CDRs region, or a combination thereof, of the immunoglobulin light chain but the multi-specific binding polypeptide (e.g., the multi-specific antibody) retains binding to a target tumor antigen and / or a target antigen expressed on an immunosuppressive cell.

[0102] In certain embodiments, a multi-specific binding polypeptide (e.g., a multi-specific antibody) described herein comprises an immunoglobulin heavy chain comprising an amino acid sequence at least about 80%, 85%, 90%, 95%, 97%, 98%, 99%, or 100% identical to or consist of the amino acid sequence set forth in SEQ ID NOs: 89, 91, or 93; and an immunoglobulin light chain at least about 80%, 85%, 90%, 95%, 97%, 98%, 99%, or 100% identical to or consist of the amino acid sequence set forth in SEQ ID NO: 90, 92, or 94. In some instances, the amino acid differences contributing to the at least about 80%, 85%, 90%, 95%, 97%, 98%, or 99% sequence identity resides in a framework region of the immunoglobulin heavy chain and the CDRs remain unchanged relative to the CDRs set forth in SEQ ID NO: 89, 91, or 93. In some instances, the amino acid differences contributing to the at least about 80%, 85%, 90%, 95%, 97%, 98%, or 99% sequence identity resides in a framework region of the immunoglobulin light chain and the CDRs remain unchanged relative to the CDRs set forth in SEQ ID NO: 90, 92, or 94. In some cases, the amino acid differences contributing to the at least about 80%, 85%, 90%, 95%, 97%, 98%, or 99% sequence identity comprises conservative substitutions in the framework region, CDRs region, or a combination thereof, of the immunoglobulin heavy chain but the multi-specific binding polypeptide (e.g., the multi-specific antibody) retains binding to a target tumor antigen and / or a target antigen expressed on an immunosuppressive cell. In some cases, the amino acid differences contributing to the at least about 80%, 85%, 90%, 95%, 97%, 98%, or 99% sequence identity comprises conservative substitutions in the framework region, CDRs region, or a combination thereof, of the immunoglobulin light chain but the multi-specific binding polypeptide (e.g., the multi-specific antibody) retains binding to a target tumor antigen and / or a target antigen expressed on an immunosuppressive cell.

[0103] In certain embodiments, the multi-specific binding polypeptide (e.g., multi-specific antibody) comprises a tumor binding moiety that specifically binds to TROP2 / TACSTD2 and an immune cell binding moiety that specifically binds to TRAIL-R2. In certain embodiments the moiety that binds to TROP2 / TACSTD2 comprises a heavy chain variable region that comprises an amino acid sequence at least about 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to that set forth in SEQ ID NO: 9; and a light chain variable region that comprises an amino acid sequence at least about 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to that set forth in SEQ ID NO: 10. In certain embodiments the moiety that binds to TRAIL-R2 comprises a heavy chain variable region that comprises an amino acid sequence at least about 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to that set forth in SEQ ID NO: 11; and a light chain variable region that comprises an amino acid sequence at least about 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to that set forth in SEQ ID NO: 12. In certain embodiments, the heavy chain of the multi-specific binding polypeptide comprises an amino acid sequence at least about 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to that set forth in SEQ ID NO: 7; and the light chain comprises an amino acid sequence at least about 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to that set forth in SEQ ID NO: 8. SEQ ID NO: 7 and 8 form a bispecific molecule depicted as in FIG. 1A that can be conjugated to a cytotoxic moiety as in FIG. IB. In some instances, the multi-specific binding polypeptide (e.g., the multi-specific antibody) further comprises one or more Fc modifications (e.g., substitutions) to reduce the affinity for human neonatal Fc receptor (FcRn), to reduce ADCC functionality (e.g., a modification at L234, L235, P238, or P331, or a combination thereof), to reduce neutropenia (e.g., a modification at L234, S239, S442, or a combination thereof), to enhance ADCC (e.g., a modification at S239, A330,1332, or a combination thereof), and / or to modulate hinge region rigidity (e.g., a modification at S228).

[0104] In certain embodiments, the multi-specific binding polypeptide comprises an IgG that specifically binds TROP2 comprising a light chain variable region and a heavy chain variable region, wherein the light chain variable region comprises a CDR1 with the amino acid sequence set forth as KASQDVSIAVA (SEQ ID NO: 1), a CDR2 with the amino acid sequence set forth as SASYRYT (SEQ ID NO: 2), and a CDR3 with the amino acid sequence set forth as QQHYITPLT (SEQ ID NO: 3); and the heavy chain variable region comprises a CDRlwith the amino acid sequence set forth as NYGMN (SEQ ID NO: 4), a CDR2 with the amino acid sequence set forth as WINTYTGEPTYTDDFKG (SEQ ID NO: 5), and a CDR3 with the amino acid sequence set forth as GGFGSSYWYFDV (SEQ ID NO: 6). In certain embodiments, the multi-specific binding polypeptide comprises an scFv that specifically binds TROP2 comprising a light chain variable region and a heavy chain variable region, wherein the light chain variable region comprises a CDR1 with the amino acid sequence set forth as KASQDVSIAVA (SEQ ID NO: 1), a CDR2 with the amino acid sequence set forth as SASYRYT (SEQ ID NO: 2), and a CDR3 with the amino acid sequence set forth as QQHYITPLT (SEQ ID NO: 3); and the heavy chain variable region comprises a CDRlwith the amino acid sequence set forth as NYGMN (SEQ ID NO: 4), a CDR2 with the amino acid sequence set forth as WINTYTGEPTYTDDFKG (SEQ ID NO: 5), and a CDR3 with the amino acid sequence set forth as GGFGSSYWYFDV (SEQ ID NO: 6).

[0105] In some embodiments, a multi-specific binding polypeptide (e.g., a multi-specific antibody) described herein comprises a tumor binding moiety that specifically binds to TROP2 and an immune cell binding moiety that specifically binds to CD33. In some instances, the tumor biding moiety that specifically binds to TROP2 comprises an immunoglobulin heavy chain variable region comprising CDR1 (HCDR1), CDR2 (HCDR2), and CDR3 (HCDR3), in which HCDR1 comprises SEQ ID NO: 4, HCDR2 comprises SEQ ID NO: 5, and HCDR3 comprises SEQ ID NO: 6. In some instances, the tumor biding moiety that specifically binds to TROP2 comprises an immunoglobulin light chain variable region comprising CDR1 (LCDR1), CDR2 (LCDR2), and CDR3 (LCDR3), in which LCDR1 comprises SEQ ID NO: 1, LCDR2 comprises SEQ ID NO: 2, and LCDR3 comprises SEQ ID NO: 3. In some instances, the tumor biding moiety comprises an immunoglobulin heavy chain variable region comprising an amino acid sequence at least about 80%, 85%, 90%, 95%, 97%, 98%, 99%, or 100% identical to or consist of the amino acid sequence set forth in SEQ ID NO: 9; and an immunoglobulin light chain variable region at least about 80%, 85%, 90%, 95%, 97%, 98%, 99%, or 100% identical to or consist of the amino acid sequence set forth in SEQ ID NO: 10. In some instances, the immune cell binding moiety that specifically binds to CD33 comprises an immunoglobulin heavy chain variable region comprising CDR1 (HCDR1), CDR2 (HCDR2), and CDR3 (HCDR3), in which HCDR1 comprises SEQ ID NO: 45, HCDR2 comprises SEQ ID NO: 46, and HCDR3 comprises SEQ ID NO: 47. In some instances, the immune cell binding moiety that specifically binds to CD33 comprises an immunoglobulin light chain variable region comprising CDR1 (LCDR1), CDR2 (LCDR2), and CDR3 (LCDR3), in which LCDR1 comprises SEQ ID NO: 60, LCDR2 comprises SEQ ID NO: 61, and LCDR3 comprises SEQ ID NO: 62. In some instances, the immune cell binding moiety comprises an immunoglobulin heavy chain variable region comprising an amino acid sequence at least about 80%, 85%, 90%, 95%, 97%, 98%, 99%, or 100% identical to or consist of the amino acid sequence set forth in SEQ ID NO: 48; and an immunoglobulin light chain variable region at least about 80%, 85%, 90%, 95%, 97%, 98%, 99%, or 100% identical to or consist of the amino acid sequence set forth in SEQ ID NO: 63. In some cases, the multi-specific binding polypeptide (e.g., the multi-specific antibody) comprises an immunoglobulin heavy chain comprising an amino acid sequence at least about 80%, 85%, 90%, 95%, 97%, 98%, 99%, or 100% identical to or consist of the amino acid sequence set forth in SEQ ID NO: 75; and an immunoglobulin light chain at least about 80%, 85%, 90%, 95%, 97%, 98%, 99%, or 100% identical to or consist of the amino acid sequence set forth in SEQ ID NO: 8. In some cases, the multi-specific binding polypeptide (e.g., the multi-specific antibody) further comprises a payload (e.g., a cytotoxic moiety). In some instances, the multi-specific binding polypeptide (e.g., the multi-specific antibody) further comprises one or more Fc modifications (e.g., substitutions) to reduce the affinity for human neonatal Fc receptor (FcRn), to reduce ADCC functionality (e.g., a modification at L234, L235, P238, or P331, or a combination thereof), to reduce neutropenia (e.g., a modification at L234, S239, S442, or a combination thereof), to enhance ADCC (e.g., a modification at S239, A330,1332, or a combination thereof), and / or to modulate hinge region rigidity (e.g., a modification at S228).

[0106] In some embodiments, a multi-specific binding polypeptide (e.g., a multi-specific antibody) described herein comprises a tumor binding moiety that specifically binds to TROP2 and an immune cell binding moiety that specifically binds to CD 163. In some instances, the tumor biding moiety that specifically binds to TROP2 comprises an immunoglobulin heavy chain variable region comprising CDR1 (HCDR1), CDR2 (HCDR2), and CDR3 (HCDR3), in which HCDR1 comprises SEQ ID NO: 4, HCDR2 comprises SEQ ID NO: 5, and HCDR3 comprises SEQ ID NO: 6. In some instances, the tumor biding moiety that specifically binds to TROP2 comprises an immunoglobulin light chain variable region comprising CDR1 (LCDR1), CDR2 (LCDR2), and CDR3 (LCDR3), in which LCDR1 comprises SEQ ID NO: 1, LCDR2 comprises SEQ ID NO: 2, and LCDR3 comprises SEQ ID NO: 3. In some instances, the tumor biding moiety comprises an immunoglobulin heavy chain variable region comprising an amino acid sequence at least about 80%, 85%, 90%, 95%, 97%, 98%, 99%, or 100% identical to or consist of the amino acid sequence set forth in SEQ ID NO: 9; and an immunoglobulin light chain variable region at least about 80%, 85%, 90%, 95%, 97%, 98%, 99%, or 100% identical to or consist of the amino acid sequence set forth in SEQ ID NO: 10. In some instances, the immune cell binding moiety that specifically binds to CD 163 comprises an immunoglobulin heavy chain variable region comprising CDR1 (HCDR1), CDR2 (HCDR2), and CDR3 (HCDR3), in which HCDR1 comprises SEQ ID NO: 49, HCDR2 comprises SEQ ID NO: 50, and HCDR3 comprises SEQ ID NO: 51. In some instances, the immune cell binding moiety that specifically binds to CD 163 comprises an immunoglobulin light chain variable region comprising CDR1 (LCDR1), CDR2 (LCDR2), and CDR3 (LCDR3), in which LCDR1 comprises SEQ ID NO: 64, LCDR2 comprises SEQ ID NO: 65, and LCDR3 comprises SEQ ID NO: 66. In some instances, the immune cell binding moiety comprises an immunoglobulin heavy chain variable region comprising an amino acid sequence at least about 80%, 85%, 90%, 95%, 97%, 98%, 99%, or 100% identical to or consist of the amino acid sequence set forth in SEQ ID NO: 52; and an immunoglobulin light chain variable region at least about 80%, 85%, 90%, 95%, 97%, 98%, 99%, or 100% identical to or consist of the amino acid sequence set forth in SEQ ID NO: 67. In some cases, the multi-specific binding polypeptide (e.g., the multi-specific antibody) comprises an immunoglobulin heavy chain comprising an amino acid sequence at least about 80%, 85%, 90%, 95%, 97%, 98%, 99%, or 100% identical to or consist of the amino acid sequence set forth in SEQ ID NO: 77; and an immunoglobulin light chain at least about 80%, 85%, 90%, 95%, 97%, 98%, 99%, or 100% identical to or consist of the amino acid sequence set forth in SEQ ID NO: 8. In some cases, the multi-specific binding polypeptide (e.g., the multi-specific antibody) further comprises a payload (e.g., a cytotoxic moiety). In some instances, the multi-specific binding polypeptide (e.g., the multi-specific antibody) further comprises one or more Fc modifications (e.g., substitutions) to reduce the affinity for human neonatal Fc receptor (FcRn), to reduce ADCC functionality (e.g., a modification at L234, L235, P238, or P331, or a combination thereof), to reduce neutropenia (e.g., a modification at L234, S239, S442, or a combination thereof), to enhance ADCC (e.g., a modification at S239, A330,1332, or a combination thereof), and / or to modulate hinge region rigidity (e.g., a modification at S228).

[0107] In some embodiments, a multi-specific binding polypeptide (e.g., a multi-specific antibody) described herein comprises a tumor binding moiety that specifically binds to TROP2 and an immune cell binding moiety that specifically binds to CSF1R. In some instances, the tumor biding moiety that specifically binds to TROP2 comprises an immunoglobulin heavy chain variable region comprising CDR1 (HCDR1), CDR2 (HCDR2), and CDR3 (HCDR3), in which HCDR1 comprises SEQ ID NO: 4, HCDR2 comprises SEQ ID NO: 5, and HCDR3 comprises SEQ ID NO: 6. In some instances, the tumor biding moiety that specifically binds to TROP2 comprises an immunoglobulin light chain variable region comprising CDR1 (LCDR1), CDR2 (LCDR2), and CDR3 (LCDR3), in which LCDR1 comprises SEQ ID NO: 1, LCDR2 comprises SEQ ID NO: 2, and LCDR3 comprises SEQ ID NO: 3. In some instances, the tumor biding moiety comprises an immunoglobulin heavy chain variable region comprising an amino acid sequence at least about 80%, 85%, 90%, 95%, 97%, 98%, 99%, or 100% identical to or consist of the amino acid sequence set forth in SEQ ID NO: 9; and an immunoglobulin light chain variable region at least about 80%, 85%, 90%, 95%, 97%, 98%, 99%, or 100% identical to or consist of the amino acid sequence set forth in SEQ ID NO: 10. In some instances, the immune cell binding moiety that specifically binds to CSF1R comprises an immunoglobulin heavy chain variable region comprising CDR1 (HCDR1), CDR2 (HCDR2), and CDR3 (HCDR3), in which HCDR1 comprises SEQ ID NO: 56, HCDR2 comprises SEQ ID NO: 57, and HCDR3 comprises SEQ ID NO: 58. In some instances, the immune cell binding moiety that specifically binds to CSF1R comprises an immunoglobulin light chain variable region comprising CDR1 (LCDR1), CDR2 (LCDR2), and CDR3 (LCDR3), in which LCDR1 comprises SEQ ID NO: 71, LCDR2 comprises SEQ ID NO: 72, and LCDR3 comprises SEQ ID NO: 73. In some instances, the immune cell binding moiety comprises an immunoglobulin heavy chain variable region comprising an amino acid sequence at least about 80%, 85%, 90%, 95%, 97%, 98%, 99%, or 100% identical to or consist of the amino acid sequence set forth in SEQ ID NO: 59; and an immunoglobulin light chain variable region at least about 80%, 85%, 90%, 95%, 97%, 98%, 99%, or 100% identical to or consist of the amino acid sequence set forth in SEQ ID NO: 74. In some cases, the multi-specific binding polypeptide (e.g., the multi-specific antibody) comprises an immunoglobulin heavy chain comprising an amino acid sequence at least about 80%, 85%, 90%, 95%, 97%, 98%, 99%, or 100% identical to or consist of the amino acid sequence set forth in SEQ ID NO: 76; and an immunoglobulin light chain at least about 80%, 85%, 90%, 95%, 97%, 98%, 99%, or 100% identical to or consist of the amino acid sequence set forth in SEQ ID NO: 8. In some cases, the multi-specific binding polypeptide (e.g., the multi-specific antibody) further comprises a payload (e.g., a cytotoxic moiety). In some instances, the multi-specific binding polypeptide (e.g., the multi-specific antibody) further comprises one or more Fc modifications (e.g., substitutions) to reduce the affinity for human neonatal Fc receptor (FcRn), to reduce ADCC functionality (e.g., a modification at L234, L235, P238, or P331, or a combination thereof), to reduce neutropenia (e.g., a modification at L234, S239, S442, or a combination thereof), to enhance ADCC (e.g., a modification at S239, A330,1332, or a combination thereof), and / or to modulate hinge region rigidity (e.g., a modification at S228).

[0108] In some embodiments, a multi-specific binding polypeptide (e.g., a multi-specific antibody) described herein comprises a tumor binding moiety that specifically binds to TROP2 and an immune cell binding moiety that specifically binds to TRAIL-R2. In some instances, the tumor biding moiety that specifically binds to TROP2 comprises an immunoglobulin heavy chain variable region comprising CDR1 (HCDR1), CDR2 (HCDR2), and CDR3 (HCDR3), in which HCDR1 comprises SEQ ID NO: 4, HCDR2 comprises SEQ ID NO: 5, and HCDR3 comprises SEQ ID NO: 6. In some instances, the tumor biding moiety that specifically binds to TROP2 comprises an immunoglobulin light chain variable region comprising CDR1 (LCDR1), CDR2 (LCDR2), and CDR3 (LCDR3), in which LCDR1 comprises SEQ ID NO: 1, LCDR2 comprises SEQ ID NO: 2, and LCDR3 comprises SEQ ID NO: 3. In some instances, the tumor biding moiety comprises an immunoglobulin heavy chain variable region comprising an amino acid sequence at least about 80%, 85%, 90%, 95%, 97%, 98%, 99%, or 100% identical to or consist of the amino acid sequence set forth in SEQ ID NO: 9; and an immunoglobulin light chain variable region at least about 80%, 85%, 90%, 95%, 97%, 98%, 99%, or 100% identical to or consist of the amino acid sequence set forth in SEQ ID NO: 10. In some instances, the immune cell binding moiety that specifically binds to TRAIL-R2 comprises an immunoglobulin heavy chain variable region comprising CDR1 (HCDR1), CDR2 (HCDR2), and CDR3 (HCDR3), in which HCDR1 comprises SEQ ID NO: 53, HCDR2 comprises SEQ ID NO: 54, and HCDR3 comprises SEQ ID NO: 55. In some instances, the immune cell binding moiety that specifically binds to TRAIL-R2 comprises an immunoglobulin light chain variable region comprising CDR1 (LCDR1), CDR2 (LCDR2), and CDR3 (LCDR3), in which LCDR1 comprises SEQ ID NO: 68, LCDR2 comprises SEQ ID NO: 69, and LCDR3 comprises SEQ ID NO: 70. In some instances, the immune cell binding moiety comprises an immunoglobulin heavy chain variable region comprising an amino acid sequence at least about 80%, 85%, 90%, 95%, 97%, 98%, 99%, or 100% identical to or consist of the amino acid sequence set forth in SEQ ID NO: 11; and an immunoglobulin light chain variable region at least about 80%, 85%, 90%, 95%, 97%, 98%, 99%, or 100% identical to or consist of the amino acid sequence set forth in SEQ ID NO: 12. In some cases, the multi-specific binding polypeptide (e.g., the multi-specific antibody) comprises an immunoglobulin heavy chain comprising an amino acid sequence at least about 80%, 85%, 90%, 95%, 97%, 98%, 99%, or 100% identical to or consist of the amino acid sequence set forth in SEQ ID NO: 78; and an immunoglobulin light chain at least about 80%, 85%, 90%, 95%, 97%, 98%, 99%, or 100% identical to or consist of the amino acid sequence set forth in SEQ ID NO: 8. In some cases, the multi-specific binding polypeptide (e.g., the multi-specific antibody) further comprises a payload (e.g., a cytotoxic moiety). In some instances, the multi-specific binding polypeptide (e.g., the multi-specific antibody) further comprises one or more Fc modifications (e.g., substitutions) to reduce the affinity for human neonatal Fc receptor (FcRn), to reduce ADCC functionality (e.g., a modification at L234, L235, P238, or P331, or a combination thereof), to reduce neutropenia (e.g., a modification at L234, S239, S442, or a combination thereof), to enhance ADCC (e.g., a modification at S239, A330,1332, or a combination thereof), and / or to modulate hinge region rigidity (e.g., a modification at S228).

[0109] In some embodiments, a multi-specific binding polypeptide (e.g., a multi-specific antibody) described herein comprises a tumor binding moiety that specifically binds to GPC3 and an immune cell binding moiety that specifically binds to CD33. In some instances, the tumor biding moiety that specifically binds to GPC3 comprises an immunoglobulin heavy chain variable region comprising CDR1 (HCDR1), CDR2 (HCDR2), and CDR3 (HCDR3), in which HCDR1 comprises SEQ ID NO: 13, HCDR2 comprises SEQ ID NO: 14, and HCDR3 comprises SEQ ID NO: 15. In some instances, the tumor biding moiety that specifically binds to GPC3 comprises an immunoglobulin light chain variable region comprising CDR1 (LCDR1), CDR2 (LCDR2), and CDR3 (LCDR3), in which LCDR1 comprises SEQ ID NO: 29, LCDR2 comprises SEQ ID NO: 30, and LCDR3 comprises SEQ ID NO: 31. In some instances, the tumor biding moiety comprises an immunoglobulin heavy chain variable region comprising an amino acid sequence at least about 80%, 85%, 90%, 95%, 97%, 98%, 99%, or 100% identical to or consist of the amino acid sequence set forth in SEQ ID NO: 16; and an immunoglobulin light chain variable region at least about 80%, 85%, 90%, 95%, 97%, 98%, 99%, or 100% identical to or consist of the amino acid sequence set forth in SEQ ID NO: 32. In some instances, the immune cell binding moiety that specifically binds to CD33 comprises an immunoglobulin heavy chain variable region comprising CDR1 (HCDR1), CDR2 (HCDR2), and CDR3 (HCDR3), in which HCDR1 comprises SEQ ID NO: 45, HCDR2 comprises SEQ ID NO: 46, and HCDR3 comprises SEQ ID NO: 47. In some instances, the immune cell binding moiety that specifically binds to CD33 comprises an immunoglobulin light chain variable region comprising CDR1 (LCDR1), CDR2 (LCDR2), and CDR3 (LCDR3), in which LCDR1 comprises SEQ ID NO: 60, LCDR2 comprises SEQ ID NO: 61, and LCDR3 comprises SEQ ID NO: 62. In some instances, the immune cell binding moiety comprises an immunoglobulin heavy chain variable region comprising an amino acid sequence at least about 80%, 85%, 90%, 95%, 97%, 98%, 99%, or 100% identical to or consist of the amino acid sequence set forth in SEQ ID NO: 48; and an immunoglobulin light chain variable region at least about 80%, 85%, 90%, 95%, 97%, 98%, 99%, or 100% identical to or consist of the amino acid sequence set forth in SEQ ID NO: 63. In some cases, the multi-specific binding polypeptide (e.g., the multi-specific antibody) comprises an immunoglobulin heavy chain comprising an amino acid sequence at least about 80%, 85%, 90%, 95%, 97%, 98%, 99%, or 100% identical to or consist of the amino acid sequence set forth in SEQ ID NO: 79; and an immunoglobulin light chain at least about 80%, 85%, 90%, 95%, 97%, 98%, 99%, or 100% identical to or consist of the amino acid sequence set forth in SEQ ID NO: 80. In some cases, the multi-specific binding polypeptide (e.g., the multi-specific antibody) further comprises a payload (e.g., a cytotoxic moiety). In some instances, the multi-specific binding polypeptide (e.g., the multi-specific antibody) further comprises one or more Fc modifications (e.g., substitutions) to reduce the affinity for human neonatal Fc receptor (FcRn), to reduce ADCC functionality (e.g., a modification at L234, L235, P238, or P331, or a combination thereof), to reduce neutropenia (e.g., a modification at L234, S239, S442, or a combination thereof), to enhance ADCC (e.g., a modification at S239, A330,1332, or a combination thereof), and / or to modulate hinge region rigidity (e.g., a modification at S228).

[0110] In some embodiments, a multi-specific binding polypeptide (e.g., a multi-specific antibody) described herein comprises a tumor binding moiety that specifically binds to GPC3 and an immune cell binding moiety that specifically binds to CD 163. In some instances, the tumor biding moiety that specifically binds to GPC3 comprises an immunoglobulin heavy chain variable region comprising CDR1 (HCDR1), CDR2 (HCDR2), and CDR3 (HCDR3), in which HCDR1 comprises SEQ ID NO: 13, HCDR2 comprises SEQ ID NO: 14, and HCDR3 comprises SEQ ID NO: 15. In some instances, the tumor biding moiety that specifically binds to GPC3 comprises an immunoglobulin light chain variable region comprising CDR1 (LCDR1), CDR2 (LCDR2), and CDR3 (LCDR3), in which LCDR1 comprises SEQ ID NO: 29, LCDR2 comprises SEQ ID NO: 30, and LCDR3 comprises SEQ ID NO: 31. In some instances, the tumor biding moiety comprises an immunoglobulin heavy chain variable region comprising an amino acid sequence at least about 80%, 85%, 90%, 95%, 97%, 98%, 99%, or 100% identical to or consist of the amino acid sequence set forth in SEQ ID NO: 16; and an immunoglobulin light chain variable region at least about 80%, 85%, 90%, 95%, 97%, 98%, 99%, or 100% identical to or consist of the amino acid sequence set forth in SEQ ID NO: 32. In some instances, the immune cell binding moiety that specifically binds to CD 163 comprises an immunoglobulin heavy chain variable region comprising CDR1 (HCDR1), CDR2 (HCDR2), and CDR3 (HCDR3), in which HCDR1 comprises SEQ ID NO: 49, HCDR2 comprises SEQ ID NO: 50, and HCDR3 comprises SEQ ID NO: 51. In some instances, the immune cell binding moiety that specifically binds to CD 163 comprises an immunoglobulin light chain variable region comprising CDR1 (LCDR1), CDR2 (LCDR2), and CDR3 (LCDR3), in which LCDR1 comprises SEQ ID NO: 64, LCDR2 comprises SEQ ID NO: 65, and LCDR3 comprises SEQ ID NO: 66. In some instances, the immune cell binding moiety comprises an immunoglobulin heavy chain variable region comprising an amino acid sequence at least about 80%, 85%, 90%, 95%, 97%, 98%, 99%, or 100% identical to or consist of the amino acid sequence set forth in SEQ ID NO: 52; and an immunoglobulin light chain variable region at least about 80%, 85%, 90%, 95%, 97%, 98%, 99%, or 100% identical to or consist of the amino acid sequence set forth in SEQ ID NO: 67. In some cases, the multi-specific binding polypeptide (e.g., the multi-specific antibody) comprises an immunoglobulin heavy chain comprising an amino acid sequence at least about 80%, 85%, 90%, 95%, 97%, 98%, 99%, or 100% identical to or consist of the amino acid sequence set forth in SEQ ID NO: 82; and an immunoglobulin light chain at least about 80%, 85%, 90%, 95%, 97%, 98%, 99%, or 100% identical to or consist of the amino acid sequence set forth in SEQ ID NO: 80. In some cases, the multispecific binding polypeptide (e.g., the multi-specific antibody) further comprises a payload (e.g., a cytotoxic moiety). In some instances, the multi-specific binding polypeptide (e.g., the multi-specific antibody) further comprises one or more Fc modifications (e.g., substitutions) to reduce the affinity for human neonatal Fc receptor (FcRn), to reduce ADCC functionality (e.g., a modification at L234, L235, P238, or P331, or a combination thereof), to reduce neutropenia (e.g., a modification at L234, S239, S442, or a combination thereof), to enhance ADCC (e.g., a modification at S239, A330,1332, or a combination thereof), and / or to modulate hinge region rigidity (e.g., a modification at S228).

[0111] In some embodiments, a multi-specific binding polypeptide (e.g., a multi-specific antibody) described herein comprises a tumor binding moiety that specifically binds to GPC3 and an immune cell binding moiety that specifically binds to CSF1R. In some instances, the tumor biding moiety that specifically binds to GPC3 comprises an immunoglobulin heavy chain variable region comprising CDR1 (HCDR1), CDR2 (HCDR2), and CDR3 (HCDR3), in which HCDR1 comprises SEQ ID NO: 13, HCDR2 comprises SEQ ID NO: 14, and HCDR3 comprises SEQ ID NO: 15. In some instances, the tumor biding moiety that specifically binds to GPC3 comprises an immunoglobulin light chain variable region comprising CDR1 (LCDR1), CDR2 (LCDR2), and CDR3 (LCDR3), in which LCDR1 comprises SEQ ID NO: 29, LCDR2 comprises SEQ ID NO: 30, and LCDR3 comprises SEQ ID NO: 31. In some instances, the tumor biding moiety comprises an immunoglobulin heavy chain variable region comprising an amino acid sequence at least about 80%, 85%, 90%, 95%, 97%, 98%, 99%, or 100% identical to or consist of the amino acid sequence set forth in SEQ ID NO: 16; and an immunoglobulin light chain variable region at least about 80%, 85%, 90%, 95%, 97%, 98%, 99%, or 100% identical to or consist of the amino acid sequence set forth in SEQ ID NO: 32. In some instances, the immune cell binding moiety that specifically binds to CSF1R comprises an immunoglobulin heavy chain variable region comprising CDR1 (HCDR1), CDR2 (HCDR2), and CDR3 (HCDR3), in which HCDR1 comprises SEQ ID NO: 56, HCDR2 comprises SEQ ID NO: 57, and HCDR3 comprises SEQ ID NO: 58. In some instances, the immune cell binding moiety that specifically binds to CSF1R comprises an immunoglobulin light chain variable region comprising CDR1 (LCDR1), CDR2 (LCDR2), and CDR3 (LCDR3), in which LCDR1 comprises SEQ ID NO: 71, LCDR2 comprises SEQ ID NO: 72, and LCDR3 comprises SEQ ID NO: 73. In some instances, the immune cell binding moiety comprises an immunoglobulin heavy chain variable region comprising an amino acid sequence at least about 80%, 85%, 90%, 95%, 97%, 98%, 99%, or 100% identical to or consist of the amino acid sequence set forth in SEQ ID NO: 59; and an immunoglobulin light chain variable region at least about 80%, 85%, 90%, 95%, 97%, 98%, 99%, or 100% identical to or consist of the amino acid sequence set forth in SEQ ID NO: 74. In some cases, the multi-specific binding polypeptide (e.g., the multi-specific antibody) comprises an immunoglobulin heavy chain comprising an amino acid sequence at least about 80%, 85%, 90%, 95%, 97%, 98%, 99%, or 100% identical to or consist of the amino acid sequence set forth in SEQ ID NO: 81; and an immunoglobulin light chain at least about 80%, 85%, 90%, 95%, 97%, 98%, 99%, or 100% identical to or consist of the amino acid sequence set forth in SEQ ID NO: 80. In some cases, the multispecific binding polypeptide (e.g., the multi-specific antibody) further comprises a payload (e.g., a cytotoxic moiety). In some instances, the multi-specific binding polypeptide (e.g., the multi-specific antibody) further comprises one or more Fc modifications (e.g., substitutions) to reduce the affinity for human neonatal Fc receptor (FcRn), to reduce ADCC functionality (e.g., a modification at L234, L235, P238, or P331, or a combination thereof), to reduce neutropenia (e.g., a modification at L234, S239, S442, or a combination thereof), to enhance ADCC (e.g., a modification at S239, A330,1332, or a combination thereof), and / or to modulate hinge region rigidity (e.g., a modification at S228).

[0112] In some embodiments, a multi-specific binding polypeptide (e.g., a multi-specific antibody) described herein comprises a tumor binding moiety that specifically binds to FOLRI and an immune cell binding moiety that specifically binds to CD33. In some instances, the tumor biding moiety that specifically binds to FOLRI comprises an immunoglobulin heavy chain variable region comprising CDR1 (HCDR1), CDR2 (HCDR2), and CDR3 (HCDR3), in which HCDR1 comprises SEQ ID NO: 17, HCDR2 comprises SEQ ID NO: 18, and HCDR3 comprises SEQ ID NO: 19. In some instances, the tumor biding moiety that specifically binds to FOLRI comprises an immunoglobulin light chain variable region comprising CDR1 (LCDR1), CDR2 (LCDR2), and CDR3 (LCDR3), in which LCDR1 comprises SEQ ID NO: 33, LCDR2 comprises SEQ ID NO: 34, and LCDR3 comprises SEQ ID NO: 35. In some instances, the tumor biding moiety comprises an immunoglobulin heavy chain variable region comprising an amino acid sequence at least about 80%, 85%, 90%, 95%, 97%, 98%, 99%, or 100% identical to or consist of the amino acid sequence set forth in SEQ ID NO: 20; and an immunoglobulin light chain variable region at least about 80%, 85%, 90%, 95%, 97%, 98%, 99%, or 100% identical to or consist of the amino acid sequence set forth in SEQ ID NO: 36. In some instances, the immune cell binding moiety that specifically binds to CD33 comprises an immunoglobulin heavy chain variable region comprising CDR1 (HCDR1), CDR2 (HCDR2), and CDR3 (HCDR3), in which HCDR1 comprises SEQ ID NO: 45, HCDR2 comprises SEQ ID NO: 46, and HCDR3 comprises SEQ ID NO: 47. In some instances, the immune cell binding moiety that specifically binds to CD33 comprises an immunoglobulin light chain variable region comprising CDR1 (LCDR1), CDR2 (LCDR2), and CDR3 (LCDR3), in which LCDR1 comprises SEQ ID NO: 60, LCDR2 comprises SEQ ID NO: 61, and LCDR3 comprises SEQ ID NO: 62. In some instances, the immune cell binding moiety comprises an immunoglobulin heavy chain variable region comprising an amino acid sequence at least about 80%, 85%, 90%, 95%, 97%, 98%, 99%, or 100% identical to or consist of the amino acid sequence set forth in SEQ ID NO: 48; and an immunoglobulin light chain variable region at least about 80%, 85%, 90%, 95%, 97%, 98%, 99%, or 100% identical to or consist of the amino acid sequence set forth in SEQ ID NO: 63. In some cases, the multi-specific binding polypeptide (e.g., the multi-specific antibody) comprises an immunoglobulin heavy chain comprising an amino acid sequence at least about 80%, 85%, 90%, 95%, 97%, 98%, 99%, or 100% identical to or consist of the amino acid sequence set forth in SEQ ID NO: 84; and an immunoglobulin light chain at least about 80%, 85%, 90%, 95%, 97%, 98%, 99%, or 100% identical to or consist of the amino acid sequence set forth in SEQ ID NO: 85. In some cases, the multi-specific binding polypeptide (e.g., the multi-specific antibody) further comprises a payload (e.g., a cytotoxic moiety). In some instances, the multi-specific binding polypeptide (e.g., the multi-specific antibody) further comprises one or more Fc modifications (e.g., substitutions) to reduce the affinity for human neonatal Fc receptor (FcRn), to reduce ADCC functionality (e.g., a modification at L234, L235, P238, or P331, or a combination thereof), to reduce neutropenia (e.g., a modification at L234, S239, S442, or a combination thereof), to enhance ADCC (e.g., a modification at S239, A330,1332, or a combination thereof), and / or to modulate hinge region rigidity (e.g., a modification at S228).

[0113] In some embodiments, a multi-specific binding polypeptide (e.g., a multi-specific antibody) described herein comprises a tumor binding moiety that specifically binds to FOLRI and an immune cell binding moiety that specifically binds to CD 163. In some instances, the tumor biding moiety that specifically binds to FOLRI comprises an immunoglobulin heavy chain variable region comprising CDR1 (HCDR1), CDR2 (HCDR2), and CDR3 (HCDR3), in which HCDR1 comprises SEQ ID NO: 17, HCDR2 comprises SEQ ID NO: 18, and HCDR3 comprises SEQ ID NO: 19. In some instances, the tumor biding moiety that specifically binds to FOLRI comprises an immunoglobulin light chain variable region comprising CDR1 (LCDR1), CDR2 (LCDR2), and CDR3 (LCDR3), in which LCDR1 comprises SEQ ID NO: 33, LCDR2 comprises SEQ ID NO: 34, and LCDR3 comprises SEQ ID NO: 35. In some instances, the tumor biding moiety comprises an immunoglobulin heavy chain variable region comprising an amino acid sequence at least about 80%, 85%, 90%, 95%, 97%, 98%, 99%, or 100% identical to or consist of the amino acid sequence set forth in SEQ ID NO: 20; and an immunoglobulin light chain variable region at least about 80%, 85%, 90%, 95%, 97%, 98%, 99%, or 100% identical to or consist of the amino acid sequence set forth in SEQ ID NO: 36. In some instances, the immune cell binding moiety that specifically binds to CD 163 comprises an immunoglobulin heavy chain variable region comprising CDR1 (HCDR1), CDR2 (HCDR2), and CDR3 (HCDR3), in which HCDR1 comprises SEQ ID NO: 49, HCDR2 comprises SEQ ID NO: 50, and HCDR3 comprises SEQ ID NO: 51. In some instances, the immune cell binding moiety that specifically binds to CD 163 comprises an immunoglobulin light chain variable region comprising CDR1 (LCDR1), CDR2 (LCDR2), and CDR3 (LCDR3), in which LCDR1 comprises SEQ ID NO: 64, LCDR2 comprises SEQ ID NO: 65, and LCDR3 comprises SEQ ID NO: 66. In some instances, the immune cell binding moiety comprises an immunoglobulin heavy chain variable region comprising an amino acid sequence at least about 80%, 85%, 90%, 95%, 97%, 98%, 99%, or 100% identical to or consist of the amino acid sequence set forth in SEQ ID NO: 52; and an immunoglobulin light chain variable region at least about 80%, 85%, 90%, 95%, 97%, 98%, 99%, or 100% identical to or consist of the amino acid sequence set forth in SEQ ID NO: 67. In some cases, the multi-specific binding polypeptide (e.g., the multi-specific antibody) comprises an immunoglobulin heavy chain comprising an amino acid sequence at least about 80%, 85%, 90%, 95%, 97%, 98%, 99%, or 100% identical to or consist of the amino acid sequence set forth in SEQ ID NO: 87; and an immunoglobulin light chain at least about 80%, 85%, 90%, 95%, 97%, 98%, 99%, or 100% identical to or consist of the amino acid sequence set forth in SEQ ID NO: 85. In some cases, the multispecific binding polypeptide (e.g., the multi-specific antibody) further comprises a payload (e.g., a cytotoxic moiety). In some instances, the multi-specific binding polypeptide (e.g., the multi-specific antibody) further comprises one or more Fc modifications (e.g., substitutions) to reduce the affinity for human neonatal Fc receptor (FcRn), to reduce ADCC functionality (e.g., a modification at L234, L235, P238, or P331, or a combination thereof), to reduce neutropenia (e.g., a modification at L234, S239, S442, or a combination thereof), to enhance ADCC (e.g., a modification at S239, A330,1332, or a combination thereof), and / or to modulate hinge region rigidity (e.g., a modification at S228).

[0114] In some embodiments, a multi-specific binding polypeptide (e.g., a multi-specific antibody) described herein comprises a tumor binding moiety that specifically binds to FOLRI and an immune cell binding moiety that specifically binds to CSF1R. In some instances, the tumor biding moiety that specifically binds to FOLRI comprises an immunoglobulin heavy chain variable region comprising CDR1 (HCDR1), CDR2 (HCDR2), and CDR3 (HCDR3), in which HCDR1 comprises SEQ ID NO: 17, HCDR2 comprises SEQ ID NO: 18, and HCDR3 comprises SEQ ID NO: 19. In some instances, the tumor biding moiety that specifically binds to FOLRI comprises an immunoglobulin light chain variable region comprising CDR1 (LCDR1), CDR2 (LCDR2), and CDR3 (LCDR3), in which LCDR1 comprises SEQ ID NO: 33, LCDR2 comprises SEQ ID NO: 34, and LCDR3 comprises SEQ ID NO: 35. In some instances, the tumor biding moiety comprises an immunoglobulin heavy chain variable region comprising an amino acid sequence at least about 80%, 85%, 90%, 95%, 97%, 98%, 99%, or 100% identical to or consist of the amino acid sequence set forth in SEQ ID NO: 20; and an immunoglobulin light chain variable region at least about 80%, 85%, 90%, 95%, 97%, 98%, 99%, or 100% identical to or consist of the amino acid sequence set forth in SEQ ID NO: 36. In some instances, the immune cell binding moiety that specifically binds to CSF1R comprises an immunoglobulin heavy chain variable region comprising CDR1 (HCDR1), CDR2 (HCDR2), and CDR3 (HCDR3), in which HCDR1 comprises SEQ ID NO: 56, HCDR2 comprises SEQ ID NO: 57, and HCDR3 comprises SEQ ID NO: 58. In some instances, the immune cell binding moiety that specifically binds to CSF1R comprises an immunoglobulin light chain variable region comprising CDR1 (LCDR1), CDR2 (LCDR2), and CDR3 (LCDR3), in which LCDR1 comprises SEQ ID NO: 71, LCDR2 comprises SEQ ID NO: 72, and LCDR3 comprises SEQ ID NO: 73. In some instances, the immune cell binding moiety comprises an immunoglobulin heavy chain variable region comprising an amino acid sequence at least about 80%, 85%, 90%, 95%, 97%, 98%, 99%, or 100% identical to or consist of the amino acid sequence set forth in SEQ ID NO: 59; and an immunoglobulin light chain variable region at least about 80%, 85%, 90%, 95%, 97%, 98%, 99%, or 100% identical to or consist of the amino acid sequence set forth in SEQ ID NO: 74. In some cases, the multi-specific binding polypeptide (e.g., the multi-specific antibody) comprises an immunoglobulin heavy chain comprising an amino acid sequence at least about 80%, 85%, 90%, 95%, 97%, 98%, 99%, or 100% identical to or consist of the amino acid sequence set forth in SEQ ID NO: 86; and an immunoglobulin light chain at least about 80%, 85%, 90%, 95%, 97%, 98%, 99%, or 100% identical to or consist of the amino acid sequence set forth in SEQ ID NO: 85. In some cases, the multispecific binding polypeptide (e.g., the multi-specific antibody) further comprises a payload (e.g., a cytotoxic moiety). In some instances, the multi-specific binding polypeptide (e.g., the multi-specific antibody) further comprises one or more Fc modifications (e.g., substitutions) to reduce the affinity for human neonatal Fc receptor (FcRn), to reduce ADCC functionality (e.g., a modification at L234, L235, P238, or P331, or a combination thereof), to reduce neutropenia (e.g., a modification at L234, S239, S442, or a combination thereof), to enhance ADCC (e.g., a modification at S239, A330,1332, or a combination thereof), and / or to modulate hinge region rigidity (e.g., a modification at S228).

[0115] In some embodiments, a multi-specific binding polypeptide (e.g., a multi-specific antibody) described herein comprises a tumor binding moiety that specifically binds to GPC3 and an immune cell binding moiety that specifically binds to TRAIL-R2. In some instances, the tumor biding moiety that specifically binds to GPC3 comprises an immunoglobulin heavy chain variable region comprising CDR1 (HCDR1), CDR2 (HCDR2), and CDR3 (HCDR3), in which HCDR1 comprises SEQ ID NO: 13, HCDR2 comprises SEQ ID NO: 14, and HCDR3 comprises SEQ ID NO: 15. In some instances, the tumor biding moiety that specifically binds to GPC3 comprises an immunoglobulin light chain variable region comprising CDR1 (LCDR1), CDR2 (LCDR2), and CDR3 (LCDR3), in which LCDR1 comprises SEQ ID NO: 29, LCDR2 comprises SEQ ID NO: 30, and LCDR3 comprises SEQ ID NO: 31. In some instances, the tumor biding moiety comprises an immunoglobulin heavy chain variable region comprising an amino acid sequence at least about 80%, 85%, 90%, 95%, 97%, 98%, 99%, or 100% identical to or consist of the amino acid sequence set forth in SEQ ID NO: 16; and an immunoglobulin light chain variable region at least about 80%, 85%, 90%, 95%, 97%, 98%, 99%, or 100% identical to or consist of the amino acid sequence set forth in SEQ ID NO: 32. In some instances, the immune cell binding moiety that specifically binds to TRAIL-R2 comprises an immunoglobulin heavy chain variable region comprising CDR1 (HCDR1), CDR2 (HCDR2), and CDR3 (HCDR3), in which HCDR1 comprises SEQ ID NO: 53, HCDR2 comprises SEQ ID NO: 54, and HCDR3 comprises SEQ ID NO: 55. In some instances, the immune cell binding moiety that specifically binds to TRAIL-R2 comprises an immunoglobulin light chain variable region comprising CDR1 (LCDR1), CDR2 (LCDR2), and CDR3 (LCDR3), in which LCDR1 comprises SEQ ID NO: 68, LCDR2 comprises SEQ ID NO: 69, and LCDR3 comprises SEQ ID NO: 70. In some instances, the immune cell binding moiety comprises an immunoglobulin heavy chain variable region comprising an amino acid sequence at least about 80%, 85%, 90%, 95%, 97%, 98%, 99%, or 100% identical to or consist of the amino acid sequence set forth in SEQ ID NO: 11; and an immunoglobulin light chain variable region at least about 80%, 85%, 90%, 95%, 97%, 98%, 99%, or 100% identical to or consist of the amino acid sequence set forth in SEQ ID NO: 12. In some cases, the multi-specific binding polypeptide (e.g., the multi-specific antibody) comprises an immunoglobulin heavy chain comprising an amino acid sequence at least about 80%, 85%, 90%, 95%, 97%, 98%, 99%, or 100% identical to or consist of the amino acid sequence set forth in SEQ ID NO: 83; and an immunoglobulin light chain at least about 80%, 85%, 90%, 95%, 97%, 98%, 99%, or 100% identical to or consist of the amino acid sequence set forth in SEQ ID NO: 80. In some cases, the multispecific binding polypeptide (e.g., the multi-specific antibody) further comprises a payload (e.g., a cytotoxic moiety). In some instances, the multi-specific binding polypeptide (e.g., the multi-specific antibody) further comprises one or more Fc modifications (e.g., substitutions) to reduce the affinity for human neonatal Fc receptor (FcRn), to reduce ADCC functionality (e.g., a modification at L234, L235, P238, or P331, or a combination thereof), to reduce neutropenia (e.g., a modification at L234, S239, S442, or a combination thereof), to enhance ADCC (e.g., a modification at S239, A330,1332, or a combination thereof), and / or to modulate hinge region rigidity (e.g., a modification at S228).

[0116] In some embodiments, a multi-specific binding polypeptide (e.g., a multi-specific antibody) described herein comprises a tumor binding moiety that specifically binds to FOLRI and an immune cell binding moiety that specifically binds to TRAIL-R2. In some instances, the tumor biding moiety that specifically binds to FOLRI comprises an immunoglobulin heavy chain variable region comprising CDR1 (HCDR1), CDR2 (HCDR2), and CDR3 (HCDR3), in which HCDR1 comprises SEQ ID NO: 17, HCDR2 comprises SEQ ID NO: 18, and HCDR3 comprises SEQ ID NO: 19. In some instances, the tumor biding moiety that specifically binds to FOLRI comprises an immunoglobulin light chain variable region comprising CDR1 (LCDR1), CDR2 (LCDR2), and CDR3 (LCDR3), in which LCDR1 comprises SEQ ID NO: 33, LCDR2 comprises SEQ ID NO: 34, and LCDR3 comprises SEQ ID NO: 35. In some instances, the tumor biding moiety comprises an immunoglobulin heavy chain variable region comprising an amino acid sequence at least about 80%, 85%, 90%, 95%, 97%, 98%, 99%, or 100% identical to or consist of the amino acid sequence set forth in SEQ ID NO: 20; and an immunoglobulin light chain variable region at least about 80%, 85%, 90%, 95%, 97%, 98%, 99%, or 100% identical to or consist of the amino acid sequence set forth in SEQ ID NO: 36. In some instances, the immune cell binding moiety that specifically binds to TRAIL-R2 comprises an immunoglobulin heavy chain variable region comprising CDR1 (HCDR1), CDR2 (HCDR2), and CDR3 (HCDR3), in which HCDR1 comprises SEQ ID NO: 53, HCDR2 comprises SEQ ID NO: 54, and HCDR3 comprises SEQ ID NO: 55. In some instances, the immune cell binding moiety that specifically binds to TRAIL-R2 comprises an immunoglobulin light chain variable region comprising CDR1 (LCDR1), CDR2 (LCDR2), and CDR3 (LCDR3), in which LCDR1 comprises SEQ ID NO: 68, LCDR2 comprises SEQ ID NO: 69, and LCDR3 comprises SEQ ID NO: 70. In some instances, the immune cell binding moiety comprises an immunoglobulin heavy chain variable region comprising an amino acid sequence at least about 80%, 85%, 90%, 95%, 97%, 98%, 99%, or 100% identical to or consist of the amino acid sequence set forth in SEQ ID NO: 11; and an immunoglobulin light chain variable region at least about 80%, 85%, 90%, 95%, 97%, 98%, 99%, or 100% identical to or consist of the amino acid sequence set forth in SEQ ID NO: 12. In some cases, the multi-specific binding polypeptide (e.g., the multi-specific antibody) comprises an immunoglobulin heavy chain comprising an amino acid sequence at least about 80%, 85%, 90%, 95%, 97%, 98%, 99%, or 100% identical to or consist of the amino acid sequence set forth in SEQ ID NO: 88; and an immunoglobulin light chain at least about 80%, 85%, 90%, 95%, 97%, 98%, 99%, or 100% identical to or consist of the amino acid sequence set forth in SEQ ID NO: 85. In some cases, the multispecific binding polypeptide (e.g., the multi-specific antibody) further comprises a payload (e.g., a cytotoxic moiety). In some instances, the multi-specific binding polypeptide (e.g., the multi-specific antibody) further comprises one or more Fc modifications (e.g., substitutions) to reduce the affinity for human neonatal Fc receptor (FcRn), to reduce ADCC functionality (e.g., a modification at L234, L235, P238, or P331, or a combination thereof), to reduce neutropenia (e.g., a modification at L234, S239, S442, or a combination thereof), to enhance ADCC (e.g., a modification at S239, A330,1332, or a combination thereof), and / or to modulate hinge region rigidity (e.g., a modification at S228).

[0117] In some embodiments, a multi-specific binding polypeptide (e.g., a multi-specific antibody) described herein comprises a tumor binding moiety that specifically binds to CD33 and an immune cell binding moiety that specifically binds to TRAIL-R2. In some instances, the tumor biding moiety that specifically binds to CD33 comprises an immunoglobulin heavy chain variable region comprising CDR1 (HCDR1), CDR2 (HCDR2), and CDR3 (HCDR3), in which HCDR1 comprises SEQ ID NO: 45, HCDR2 comprises SEQ ID NO: 46, and HCDR3 comprises SEQ ID NO: 47. In some instances, the tumor biding moiety that specifically binds to CD33 comprises an immunoglobulin light chain variable region comprising CDR1 (LCDR1), CDR2 (LCDR2), and CDR3 (LCDR3), in which LCDR1 comprises SEQ ID NO: 60, LCDR2 comprises SEQ ID NO: 61, and LCDR3 comprises SEQ ID NO: 62. In some instances, the tumor biding moiety comprises an immunoglobulin heavy chain variable region comprising an amino acid sequence at least about 80%, 85%, 90%, 95%, 97%, 98%, 99%, or 100% identical to or consist of the amino acid sequence set forth in SEQ ID NO: 48; and an immunoglobulin light chain variable region at least about 80%, 85%, 90%, 95%, 97%, 98%, 99%, or 100% identical to or consist of the amino acid sequence set forth in SEQ ID NO: 63. In some instances, the immune cell binding moiety that specifically binds to TRAIL-R2 comprises an immunoglobulin heavy chain variable region comprising CDR1 (HCDR1), CDR2 (HCDR2), and CDR3 (HCDR3), in which HCDR1 comprises SEQ ID NO: 53, HCDR2 comprises SEQ ID NO: 54, and HCDR3 comprises SEQ ID NO: 55. In some instances, the immune cell binding moiety that specifically binds to TRAIL-R2 comprises an immunoglobulin light chain variable region comprising CDR1 (LCDR1), CDR2 (LCDR2), and CDR3 (LCDR3), in which LCDR1 comprises SEQ ID NO: 68, LCDR2 comprises SEQ ID NO: 69, and LCDR3 comprises SEQ ID NO: 70. In some instances, the immune cell binding moiety comprises an immunoglobulin heavy chain variable region comprising an amino acid sequence at least about 80%, 85%, 90%, 95%, 97%, 98%, 99%, or 100% identical to or consist of the amino acid sequence set forth in SEQ ID NO: 11; and an immunoglobulin light chain variable region at least about 80%, 85%, 90%, 95%, 97%, 98%, 99%, or 100% identical to or consist of the amino acid sequence set forth in SEQ ID NO: 12. In some cases, the multi-specific binding polypeptide (e.g., the multi-specific antibody) comprises an immunoglobulin heavy chain comprising an amino acid sequence at least about 80%, 85%, 90%, 95%, 97%, 98%, 99%, or 100% identical to or consist of the amino acid sequence set forth in SEQ ID NO: 89; and an immunoglobulin light chain at least about 80%, 85%, 90%, 95%, 97%, 98%, 99%, or 100% identical to or consist of the amino acid sequence set forth in SEQ ID NO: 90. In some cases, the multispecific binding polypeptide (e.g., the multi-specific antibody) further comprises a payload (e.g., a cytotoxic moiety). In some instances, the multi-specific binding polypeptide (e.g., the multi-specific antibody) further comprises one or more Fc modifications (e.g., substitutions) to reduce the affinity for human neonatal Fc receptor (FcRn), to reduce ADCC functionality (e.g., a modification at L234, L235, P238, or P331, or a combination thereof), to reduce neutropenia (e.g., a modification at L234, S239, S442, or a combination thereof), to enhance ADCC (e.g., a modification at S239, A330,1332, or a combination thereof), and / or to modulate hinge region rigidity (e.g., a modification at S228).

[0118] In some embodiments, a multi-specific binding polypeptide (e.g., a multi-specific antibody) described herein comprises a tumor binding moiety that specifically binds to CD38 and an immune cell binding moiety that specifically binds to TRAIL-R2. In some instances, the tumor biding moiety that specifically binds to CD38 comprises an immunoglobulin heavy chain variable region comprising CDR1 (HCDR1), CDR2 (HCDR2), and CDR3 (HCDR3), in which HCDR1 comprises SEQ ID NO: 21, HCDR2 comprises SEQ ID NO: 22, and HCDR3 comprises SEQ ID NO: 23. In some instances, the tumor biding moiety that specifically binds to CD38 comprises an immunoglobulin light chain variable region comprising CDR1 (LCDR1), CDR2 (LCDR2), and CDR3 (LCDR3), in which LCDR1 comprises SEQ ID NO: 37, LCDR2 comprises SEQ ID NO: 38, and LCDR3 comprises SEQ ID NO: 39. In some instances, the tumor biding moiety comprises an immunoglobulin heavy chain variable region comprising an amino acid sequence at least about 80%, 85%, 90%, 95%, 97%, 98%, 99%, or 100% identical to or consist of the amino acid sequence set forth in SEQ ID NO: 24; and an immunoglobulin light chain variable region at least about 80%, 85%, 90%, 95%, 97%, 98%, 99%, or 100% identical to or consist of the amino acid sequence set forth in SEQ ID NO: 40. In some instances, the immune cell binding moiety that specifically binds to TRAIL-R2 comprises an immunoglobulin heavy chain variable region comprising CDR1 (HCDR1), CDR2 (HCDR2), and CDR3 (HCDR3), in which HCDR1 comprises SEQ ID NO: 53, HCDR2 comprises SEQ ID NO: 54, and HCDR3 comprises SEQ ID NO: 55. In some instances, the immune cell binding moiety that specifically binds to TRAIL-R2 comprises an immunoglobulin light chain variable region comprising CDR1 (LCDR1), CDR2 (LCDR2), and CDR3 (LCDR3), in which LCDR1 comprises SEQ ID NO: 68, LCDR2 comprises SEQ ID NO: 69, and LCDR3 comprises SEQ ID NO: 70. In some instances, the immune cell binding moiety comprises an immunoglobulin heavy chain variable region comprising an amino acid sequence at least about 80%, 85%, 90%, 95%, 97%, 98%, 99%, or 100% identical to or consist of the amino acid sequence set forth in SEQ ID NO: 11; and an immunoglobulin light chain variable region at least about 80%, 85%, 90%, 95%, 97%, 98%, 99%, or 100% identical to or consist of the amino acid sequence set forth in SEQ ID NO: 12. In some cas...

Claims

1. A multi-specific antibody drug conjugate comprising a first binding moiety that specificallybinds to a tumor-associated antigen and a second binding moiety that specifically binds to an antigen expressed on an immunosuppressive cell, wherein:(a) the first antigen binding moiety specifically binds to TROP2 and comprises a VH domain comprising a HCDR1 as set forth in SEQ ID NO:4, a HCDR2 as set forth in SEQ ID NO:5, and a HCDR3 as set forth in SEQ ID NO:6, and a VL domain comprising a LCDR1 as set forth in SEQ ID NO:1, a LCDR2 as set forth in SEQ ID NO:2, and a LCDR3 as set forth in SEQ ID NO:3; and(b) the second antigen binding moiety specifically binds to CD33 and comprises a VH domain comprising a HCDR1 as set forth in SEQ ID NO:45, a HCDR2 as set forth in SEQ ID NO:46, and a HCDR3 as set forth in SEQ ID NO:47, and a VL domain comprising a LCDR1 as set forth in SEQ ID NO:60, a LCDR2 as set forth in SEQ ID NO:61, and a LCDR3 as set forth in SEQ ID NO:62,wherein the multi-specific antibody drug conjugate comprises at least one cytotoxic moiety.

2. The multi-specific antibody drug conjugate of claim 1, wherein the immunosuppressive cellis a Myeloid-derived suppressor cell (MDSC) or a tumor-associated macrophage (TAM), optionally a M2 polarized TAM (M2-TAM).

3. The multi-specific antibody drug conjugate of claim 1 or claim 2, wherein the first bindingmoiety comprises a full-length antibody.

4. The multi-specific antibody drug conjugate of any one of claims 1-3, wherein the firstbinding moiety comprises an immunoglobulin heavy chain variable region comprising an amino acid sequence at least about 90%, 95%, 97%, 98%, 99%, or 100% identical to or consisting of the amino acid sequence set forth in SEQ ID NO: 9; and an immunoglobulin light chain variable region at least about 90%, 95%, 97%, 98%, 99%, or 100% identical to or consisting of the amino acid sequence set forth in SEQ ID NO: 10.2020205100   07 Aug 20265.     The multi-specific antibody drug conjugate of any one of claims 1-4, wherein the secondbinding moiety comprises a Fab, F(ab)2, single-domain antibody, or a single chain variable fragment (scFv).

6. The multi-specific antibody drug conjugate of any one of claims 1-5, wherein the secondbinding moiety comprises an immunoglobulin heavy chain variable region comprising an amino acid sequence at least about 90%, 95%, 97%, 98%, 99%, or 100% identical to the amino acid sequence set forth in SEQ ID NO: 48; and an immunoglobulin light chain variable region at least about 90%, 95%, 97%, 98%, 99%, or 100% identical to the amino acid sequence set forth in SEQ ID NO: 63.

7. The multi-specific antibody drug conjugate of any one of claims 1-6, wherein:the first binding moiety comprises an IgG antibody framework, optionally a full-length IgG antibody framework, the second binding moiety is an scFv, and the second binding moiety is coupled to the C-terminus of the first binding moiety, optionally recombinantly fused to the C-terminus of the first binding moiety; orthe first binding moiety comprises an IgG antibody framework, optionally a full-length IgG antibody framework, the second binding moiety is an scFv, and the second binding moiety is coupled to the N-terminus of the first binding moiety, optionally recombinantly fused to the N-terminus of the first binding moiety optionally wherein the second binding moiety is coupled to the first binding moiety by a polypeptide linker, optionally wherein the polypeptide linker comprises (Gly4Ser)n, wherein n is an integer from 1 to 10, further optionally from 1 to 6, 1 to 4, or 1 to 2, further optionally 1, 2, 3, or 4.

8. The multi-specific antibody drug conjugate of any one of claims 1-7, wherein the firstbinding moiety has a KD for the tumor associated antigen that is lower than the KD of the second binding moiety for the antigen expressed on the immunosuppressive cell.

9. The multi-specific antibody drug conjugate of any one of claims 1-8, wherein the multispecific antibody drug conjugate comprises an Fc region that has been modified to reduce the affinity for human neonatal Fc receptor (FcRn).2020205100   07 Aug 202610. The multi-specific antibody drug conjugate of any one of claims 1-9, wherein the multispecific antibody drug conjugate comprises an Fc region comprising a modification to reduce antibody-dependent cellular cytotoxicity (ADCC), wherein the modification optionally comprises L234, L235, P238, or P331, or a combination thereof, wherein L234, L235, P238, and P331 correspond to positions 234, 235, 238, and 331 of a wild-type IgG1, according to the EU numbering convention.

11. The multi-specific antibody drug conjugate of any one of claims 1-10, wherein the multispecific antibody drug conjugate comprises an Fc region that has been modified to reduce neutropenia, wherein the Fc region optionally comprises a modification at L234, S239, S442, or a combination thereof, wherein L234, S239, and S442 correspond to positions 234, 239, 442 of a wildtype IgG1, according to the EU numbering convention.

12. The multi-specific antibody drug conjugate of any one of claims 1-11, wherein the multispecific antibody drug conjugate comprises an Fc region that has been modified to enhance antibody-dependent cellular cytotoxicity (ADCC), optionally wherein the Fc region comprises a modification at S239, A330, I332, or a combination thereof, wherein S239, A330, and I332 correspond to positions 239, 330, and 332 of a wild-type IgG1, according to the EU numbering convention.

13. The multi-specific antibody drug conjugate of any one of claims 1-12, wherein the multispecific antibody drug conjugate comprises a modification to a hinge region, optionally wherein the hinge region comprises a modification at S228, wherein S228 corresponds to position 228 of a wildtype IgG4, according to the EU numbering convention.

14. The multi-specific antibody drug conjugate of claim 1, wherein the antibody drug conjugate comprises:an immunoglobulin heavy chain comprising an amino acid sequence at least about 90%, 95%, 97%, 98%, 99%, or 100% identical to or consisting of the amino acid sequence set forth in SEQ ID NOs: 7 or 75-77; and an immunoglobulin light chain at least about 90%, 95%, 97%, 98%, 99%, or 100% identical to or consist of the amino acid sequence set forth in SEQ ID NO: 8.2020205100   07 Aug 202615. The multi-specific antibody drug conjugate of any one of claims 1-14, wherein the cytotoxic moiety comprises an auristatin, an auristatin derivative, maytansine, a maytansinoid, a taxane, a calicheamicin, cemadotin, a duocarmycin, a pyrrolobenzodiazepine (PBD), tubulysin, dexamethasone, or dasatinib, further optionally wherein:the auristatin derivative is monomethyl auristatin E (MMAE) or monomethyl auristatin F (MMAF);the maytansinoid is DM1, DM2, or DM4; orthe pyrrolobenzodiazepine is a pyrrolobenzodiazepine dimer.

16. The multi-specific antibody drug conjugate of claims 1-15, wherein the at least one cytotoxic moiety is attached to the multi-specific antibody drug conjugate via a linker, optionally a cleavable linker or a non-cleavable linker.

17. A pharmaceutical composition comprising the multi-specific antibody drug conjugate of any one of claims 1 to 16, and a pharmaceutically acceptable excipient, carrier, or diluent, optionally wherein the pharmaceutical composition is formulated for parenteral administration, further optionally for subcutaneous, intramuscular, or intravenous administration.

18. A nucleic acid encoding a multi-specific antibody comprising an immunoglobulin heavy chain comprising an amino acid sequence at least about 90%, 95%, 97%, 98%, 99%, or 100% identical to or consisting of the amino acid sequence set forth in SEQ ID NOs: 7 or 75-77; and an immunoglobulin light chain at least about 90%, 95%, 97%, 98%, 99%, or 100% identical to or consisting of the amino acid sequence set forth in SEQ ID NO: 8.

19. A method of treating a TROP2 positive cancer or hematological malignancy in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of a multi-specific antibody drug conjugate of any one of claims 1-16, a pharmaceutical composition of claim 17, or a nucleic acid of claim 18.

20. A method of treating a cancer comprising CD33 positive immunosuppressive cells in the tumor microenvironment in a subject in need thereof, the method comprising administering to the2020205100   07 Aug 2026subject a therapeutically effective amount of a multi-specific antibody drug conjugate of any one of claims 1-16, a pharmaceutical composition of claim 17, or a nucleic acid of claim 18.

21. A method of treating a cancer comprising at least one TROP2 positive tumor cell and at least one CD33 positive immunosuppressive cell in the tumor microenvironment in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of a multi-specific antibody drug conjugate of any one of claims 1-16, a pharmaceutical composition of claim 17, or a nucleic acid of claim 18.

22. Use of a multi-specific antibody drug conjugate of any one of claims 1-16, or a nucleic acid of claim 18 in the manufacture of a medicament for treating a TROP2 positive cancer or hematological malignancy, a cancer comprising CD33 positive immunosuppressive cells in the tumor microenvironment, or a cancer comprising at least one TROP2 positive tumor cell and at least one CD33 positive immunosuppressive cell in the tumor microenvironment.

23. The method of any one of claims 19-21 or the use of claim 22, wherein the cancer is bladder cancer, bone cancer, brain cancer, breast cancer, cervical cancer, cholangiocarcinoma, colorectal cancer, endometrial cancer, esophageal cancer, eye cancer, head and neck cancer, kidney cancer, liver cancer, lung cancer, melanoma, ovarian cancer, pancreatic cancer, prostate cancer, sarcoma, stomach cancer, testicular cancer, or thyroid cancer, optionally wherein the breast cancer is luminal A breast cancer, luminal B breast cancer, triple-negative breast cancer, HER2-enriched breast cancer, or normal-like breast cancer, further optionally wherein the breast cancer is triple-negative breast cancer.

24. The method of claim 19 or the use of claim 22, wherein the hematological malignancy comprises chronic lymphocytic leukemia (CLL), acute myeloid leukemia (AML), diffuse large B-cell lymphoma (DLBCL), chronic myeloid leukemia, or acute myeloid leukemia.

25. The method or the use of any one of claims 19-24, wherein the subject has previously been treated with an immune checkpoint inhibitor treatment, optionally wherein the subject is insensitive to treatment with an immune checkpoint inhibitor, has failed to respond to treatment with an immune2020205100   07 Aug 2026checkpoint inhibitor, or who expresses low level of or does not express an immune checkpoint protein.

26. A method of inducing tumor and immunosuppressive cell killing effect in a target cell population, comprising contacting the target cell population comprising at least one TROP2 positive tumor cell and at least one CD33 positive immunosuppressive cell with a multi-specific antibody drug conjugate of claims 1-16, a pharmaceutical composition of claim 17, or a nucleic acid of claim 18 for a time sufficient to induce cell kill effect, thereby killing the at least one TROP2 positive tumor cell and the at least one CD33 positive immunosuppressive cell in the target cell population.

27. The method of claim 26, wherein the tumor cell is a cell from a solid tumor or a hematological malignancy, optionally wherein the solid tumor is a bladder cancer, bone cancer, brain cancer, breast cancer, cervical cancer, cholangiocarcinoma, colorectal cancer, endometrial cancer, esophageal cancer, eye cancer, head and neck cancer, kidney cancer, liver cancer, lung cancer, melanoma, ovarian cancer, pancreatic cancer, prostate cancer, sarcoma, stomach cancer, testicular cancer, or thyroid cancer.

28. The method of claim 26 or claim 27, wherein the immunosuppressive cell is MDSC, or a tumor-associated macrophage.

29. The method of any one of the claims 26-28, wherein the multi-specific antibody drug conjugate of claims 1-16, the pharmaceutical composition of claim 17, or the nucleic acid of claim 18 decreases tumor cells, optionally tumor cell proliferation, in the target cell population by about 1fold, 2-fold, 3-fold, 4-fold, 5-fold, 6-fold, 7-fold, 8-fold, 9-fold, 10-fold, 20-fold, 30-fold, 40-fold, 50-fold, or more.

30. The method of any one of the claims 26-29, wherein the multi-specific antibody drug conjugate of claims 1-16, the pharmaceutical composition of claim 17, or the nucleic acid of claim 18 decreases immunosuppressive cells, optionally immunosuppressives cell proliferation in the target cell population by about 1-fold, 2-fold, 3-fold, 4-fold, 5-fold, 6-fold, 7-fold, 8-fold, 9-fold, 10fold, 20-fold, 30-fold, 40-fold, 50-fold, or more.2020205100   07 Aug 202631. The method of any one of the claims 26-30, wherein the multi-specific antibody drug conjugate of claims 1-16, the pharmaceutical composition of claim 17, or the nucleic acid of claim 18 enhances T-cell proliferation, optionally tumor-infiltrating lymphocyte (TIL) proliferation.

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