WO2023055376A1

WO2023055376A1 - Multispecific antibodies for targeting cd47 and icam1 and methods of use thereof

Info

Publication number: WO2023055376A1
Application number: PCT/US2021/052887
Authority: WO
Inventors: Xinhua Wang; Xiaocheng Chen; Oi Kwan WONG; Leonard Post
Original assignee: Virtuoso Binco, Inc.
Priority date: 2021-09-30
Filing date: 2021-09-30
Publication date: 2023-04-06
Also published as: WO2023056378A1

Abstract

Provided herein are compositions and methods that comprise a multispecific antibody that targets CD47 and ICAM1. Also provided are methods of using the multispecific antibody for treatment of diseases and conditions.

Description

MULTISPECIFIC ANTIBODIES FOR TARGETING CD47 AND ICAM1 AND METHODS OF USE THEREOF

SUMMARY

[0001] Disclosed herein, in some embodiments, are multispecific antibodies comprising a CD47 binding domain and an Intercellular Adhesion Molecule 1 (ICAM1) binding domain. In some embodiments, the multispecific antibody is bispecific, trispecific, or tetraspecific. In some embodiments, the multispecific antibody is bispecific. In some embodiments, the multispecific antibody is bivalent, trivalent, or tetravalent. In some embodiments, the multispecific antibody is bivalent. In some embodiments, the CD47 binding domain comprises an antibody, or functional fragment or functional variant thereof, that binds specifically to CD47. In some embodiments, the antibody, or functional fragment or functional variant thereof that binds specifically to CD47 comprises an anti-CD47 heavy chain and an anti-CD47 light chain. In some embodiments, the anti-CD47 heavy chain comprises an anti-CD47 heavy chain variable domain. In some embodiments, the anti-CD47 heavy chain variable domain comprises a variable domain of an IgGl, IgG2, IgG3, or IgG4 heavy chain. In some embodiments, the anti-CD47 light chain comprises an anti-CD47 light chain variable domain. In some embodiments, the anti-CD47 light chain variable domain comprises a variable domain of a Kappa or Lambda light chain. In some embodiments, the anti-CD47 heavy chain variable domain comprises the variable domain of an IgGl heavy chain and the anti-CD47 light chain variable domain comprises the variable domain of a Kappa or Lambda light chain. In some embodiments, the anti-CD47 heavy chain variable domain comprises the variable domain of an IgGl heavy chain and the anti-CD47 light chain variable domain comprises the variable domain of a Kappa light chain. In some embodiments, the anti-CD47 heavy chain variable domain comprises the variable domain of an IgGl heavy chain and the anti-CD47 light chain variable domain comprises the variable domain of a Lambda light chain. In some embodiments, the antibody, or functional fragment or functional variant thereof, that binds specifically to CD47 comprises a single -chain variable fragment (scFv) or an antigen-binding fragment (Fab). In some embodiments, the anti-CD47 heavy chain variable domain comprises complementarity determining regions (CDRs): HC-CDR1, HC-CDR2, and HC-CDR3, and wherein the HC-CDR1, the HC-CDR2, and the HC-CDR3 of the anti-CD47 heavy chain variable domain comprise amino acid sequences according to HC-CDR1: SEQ ID NO: 1; HC-CDR2: SEQ ID NO: 2; HC-CDR3: SEQ ID NO: 3; and wherein the CDRs comprise from 0-2 amino acid modification(s) (e.g., 0 or 1 amino acid modification(s)) in at least one of the HC-CDR1, HC-CDR2, or HC-CDR3. In some embodiments, the anti-CD47 light chain variable domain comprises complementarity determining regions (CDRs): LC-CDR1, LC-CDR2, and LC-CDR3, and wherein the LC-CDR1, the LC-CDR2, and the LC-CDR3 of the anti-CD47 light chain variable domain comprises amino acid sequences according to LC-CDR1: SEQ ID NO: 4; LC-CDR2: SEQ ID NO: 5; LC-CDR3: SEQ ID NO: 6; and wherein the CDRs comprise from 0-2 amino acid modification(s) (e.g., 0 or 1 amino acid modification(s)) in at least one of the LC-CDR1, LC-CDR2, or LC-CDR3. In some embodiments, the HC-CDR1, the HC-CDR2, and the HC-CDR3 of the anti-CD47 heavy chain variable domain comprises amino acid sequences according to HC-CDR1: SEQ ID NO: 1; HC-CDR2: SEQ ID NO: 2; HC-CDR3: SEQ ID NO: 3 and the LC-CDR1, the LC-CDR2, and the LC-CDR3 of the anti-CD47 light chain variable domain comprises amino acid sequences according to LC-CDR1: SEQ ID NO: 4; LC-CDR2: SEQ ID NO: 5; LC-CDR3: SEQ ID NO: 6. In some embodiments, the anti-CD47 heavy chain comprises an amino acid sequence with at least 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to of SEQ ID NO: 7. In some embodiments, the anti-CD47 heavy chain comprises an amino acid sequence with at least 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to of SEQ ID NO: A. In some embodiments, the anti-CD47 light chain comprises an amino acid sequence with at least 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NO: 9. In some embodiments, the anti-CD47 light chain comprises an amino acid sequence with at least 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NOs: B. In some embodiments, the anti-CD47 heavy chain comprises an amino acid sequence with at least 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NO: 7; and the anti-CD47 light chain comprises an amino acid sequence with at least 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NO: 9. In some embodiments, the anti-CD47 heavy chain comprises an amino acid sequence with at least 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NO: A; and the anti-CD47 light chain comprises an amino acid sequence with at least 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NO: B. In some embodiments, the ICAM1 binding domain comprises an antibody, or functional fragment or functional variant thereof, that binds specifically to ICAM1. In some embodiments, the antibody, or functional fragment or functional variant thereof that binds specifically to ICAM1 comprises an anti-ICAMl heavy chain and an anti-ICAMl light chain. In some embodiments, the anti-ICAMl heavy chain comprises an anti-ICAMl heavy chain variable domain. In some embodiments, the anti-ICAMl heavy chain variable domain comprises a variable domain of an IgGl, IgG2, IgG3, or IgG4 heavy chain. In some embodiments, the anti-ICAMl light chain comprises an anti-ICAMl light chain variable domain. In some embodiments, the anti-ICAMl light chain variable domain comprises a variable domain of a Kappa or Lambda light chain. In some embodiments, the anti- ICAMl heavy chain variable domain comprises the variable domain of an IgGl heavy chain and the anti- ICAMl light chain variable domain comprises the variable domain of a Kappa or Lambda light chain. In some embodiments, the anti-ICAMl heavy chain variable domain comprises the variable domain of an IgGl heavy chain and the anti-ICAMl light chain variable domain comprises the variable domain of a Kappa light chain. In some embodiments, the anti-ICAMl heavy chain variable domain comprises the variable domain of an IgGl heavy chain and the anti-ICAMl light chain variable domain comprises the variable domain of a Lambda light chain. In some embodiments, the antibody, or functional fragment or functional variant thereof, that binds specifically to ICAM1 comprises a single -chain variable fragment (scFv) or an antigen-binding fragment (Fab). In some embodiments, the anti-ICAMl heavy chain variable domain comprises complementarity determining regions (CDRs): HC-CDR1, HC-CDR2, and HC-CDR3, and wherein the HC-CDR1, the HC-CDR2, and the HC-CDR3 of the anti-ICAMl heavy chain variable domain comprise amino acid sequences according to HC-CDR1: SEQ ID NO: 19, 25, 46, 49, 52, 55, 58, 61, 64, 67, 70, 73, 76, 79, 82, 85, 88, 91, 94, 97, 100, 103, 106, or 109; HC-CDR2: SEQ ID NO: 20, 26, 47, 50, 53, 56, 59, 62, 65, 68, 71, 74, 77, 80, 83, 86, 89, 92, 95, 98, 101, 104, or 107; HC-CDR3: SEQ ID NO: 21, 27, 48, 51, 54, 57, 60, 63, 66, 69, 72, 75, 78, 81, 84, 87, 90, 93, 96, 99, 102, 105, 108, or 111; and wherein the CDRs comprise from 0-2 amino acid modification(s) (e.g., 0 or 1 amino acid modification(s)) in at least one of the HC-CDR1, HC-CDR2, or HC-CDR3. In some embodiments, the anti-ICAMl light chain variable domain comprises complementarity determining regions (CDRs): LC-CDR1, LC-CDR2, and LC-CDR3, and wherein the LC-CDR1, the LC-CDR2, and the LC-CDR3 of the anti-ICAMl light chain variable domain comprises amino acid sequences according to LC-CDR1: SEQ ID NO: 22, 28, 112, 115, 118, 121, 124, 127, 130, 133, 136, 139, 142, 145, 148, 151, 154, 157, 160, or 163; LC-CDR2: SEQ ID NO: 23, 29, 110, 113, 116, 119, 122, 125, 128, 131, 134, 137, 140, 143, 146, 149, 152, 155, 158, 161, or 164; LC-CDR3: SEQ ID NO: 24, 30, 114, 117, 120, 123, 126, 129, 132, 134, 138, 141, 144, 147, 150, 153, 156, 159, 162, or 165; and wherein the CDRs comprise from 0-2 amino acid modification(s) (e.g., 0 or 1 amino acid modification(s)) in at least one of the LC-CDR1, LC-CDR2, or LC-CDR3. In some embodiments, the anti-ICAMl light chain variable domain comprises complementarity determining regions (CDRs): LC-CDR1, LC-CDR2, and LC-CDR3, and wherein the LC-CDR1, the LC-CDR2, and the LC-CDR3 of the anti-ICAMl light chain variable domain comprises amino acid sequences according to LC-CDR1: SEQ ID NO: 28; LC-CDR2: SEQ ID NO: 29; LC-CDR3: SEQ ID NO: 30; and wherein the CDRs comprise from 0-2 amino acid modification(s) (e.g., 0 or 1 amino acid modification(s)) in at least one of the LC-CDR1, LC-CDR2, or LC-CDR3. In some embodiments, the HC-CDR1, the HC-CDR2, and the HC-CDR3 of the anti-ICAMl heavy chain variable domain comprises amino acid sequences according to HC-CDR1: SEQ ID NO: 19, 25, 46, 49, 52, 55, 58, 61, 64, 67, 70, 73, 76, 79, 82, 85, 88, 91, 94, 97, 100, 103, 106, or 109; HC-CDR2: SEQ ID NO: 20, 26, 47, 50, 53, 56, 59, 62, 65, 68, 71, 74, 77, 80, 83, 86, 89, 92, 95, 98, 101, 104, or 107; HC-CDR3: SEQ ID NO: 21, 27, 48, 51, 54, 57, 60, 63, 66, 69, 72, 75, 78, 81, 84, 87, 90, 93, 96, 99, 102, 105, 108, or 111 and the LC-CDR1, the LC-CDR2, and the LC- CDR3 of the anti-ICAMl light chain variable domain comprises amino acid sequences according to LC- CDR1: SEQ ID NO: 22, 28, 112, 115, 118, 121, 124, 127, 130, 133, 136, 139, 142, 145, 148, 151, 154, 157, 160, or 163; LC-CDR2: SEQ ID NO: 23, 29, 110, 113, 116, 119, 122, 125, 128, 131, 134, 137, 140, 143, 146, 149, 152, 155, 158, 161, or 164; LC-CDR3: SEQ ID NO: 24, 30, 114, 117, 120, 123, 126, 129, 132, 134, 138, 141, 144, 147, 150, 153, 156, 159, 162, or 165. In some embodiments, the HC-CDR1, the HC-CDR2, and the HC-CDR3 of the anti-ICAMl heavy chain variable domain comprises amino acid sequences according to HC-CDR1: SEQ ID NO: 25; HC-CDR2: SEQ ID NO: 26; HC-CDR3: SEQ ID NO: 27; and the LC-CDR1, the LC-CDR2, and the LC-CDR3 of the anti-ICAMl light chain variable domain comprises amino acid sequences according to LC-CDR1: SEQ ID NO: 28; LC-CDR2: SEQ ID NO: 29; LC-CDR3: SEQ ID NO: 30. In some embodiments, the anti-ICAMl heavy chain comprises an amino acid sequence with at least 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to any one of SEQ ID NOs: 35, 36, 41, 166-187, 206-227. . In some embodiments, the anti -I CAM 1 heavy chain comprises an amino acid sequence with at least 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to any one of SEQ ID NO: C. In some embodiments, the anti-ICAMl light chain comprises an amino acid sequence with at least 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to any one of SEQ ID NOs: 39, 40, 44, 45, 188-205, 228-245. In some embodiments, the anti-ICAMl light chain comprises an amino acid sequence with at least 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to any one of SEQ ID NO: D. In some embodiments, the anti-ICAMl heavy chain comprises an amino acid sequence with at least 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to any one of SEQ ID NOs: 35, 36, 37, 38, 41, 42, 43, 166-187, 206-227 and the anti-ICAMl light chain comprises an amino acid sequence with at least 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to any one of SEQ ID NOs: 39, 40, 44, 45, 188-205, 228-245. In some embodiments, the anti-ICAMl heavy chain comprises an amino acid sequence with at least 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NO: C; and the anti-ICAMl light chain comprises an amino acid sequence with at least 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NO: D. In some embodiments, the antibody, or functional fragment or functional variant thereof, that binds specifically to CD47 comprises the Fab, and the antibody, or functional fragment or functional variant thereof, that binds specifically to ICAM1 comprises the scFv. In some embodiments, the anti-CD47 heavy chain comprises an scFv that comprises an amino acid sequence with at least 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NO: 11. In some embodiments, the anti-CD47 heavy chain comprises an amino acid sequence with at least 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NO: A; and the anti- CD47 light chain comprises an amino acid sequence with at least 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NO: B; and the scFv comprises an amino acid sequence with at least 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NO: 11. In some embodiments, the antibody, or functional fragment or functional variant thereof, that binds specifically to CD47 comprises the scFv, and the antibody, or functional fragment or functional variant thereof, that binds specifically to ICAM1 comprises the Fab. In some embodiments, the anti-ICAMl heavy chain comprises an amino acid sequence with at least 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NO: 251 and the anti-ICAMl light chain comprises an amino acid sequence with at least 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NO: 252; and the scFv comprises an amino acid sequence with at least 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NO: 253. In some embodiments, the anti-ICAMl heavy chain comprises an amino acid sequence with at least 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NO: 254 and the anti-ICAMl light chain comprises an amino acid sequence with at least 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NO: 255; and the scFv comprises an amino acid sequence with at least 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NO: 256. In some embodiments, the multispecific antibody comprises at least 3 CDRs of an anti-CD47 binding domain selected from any one of SEQ ID NOs: 1-6 and from 0-2 amino acid modification(s) (e.g., 0-1 amino acid modification(s)) thereof; and at least 3 CDRs of an anti-ICAMl binding domain selected from any one of SEQ ID NO: 19, 25, 46, 49, 52, 55, 58, 61, 64, 67, 70, 73, 76, 79, 82, 85, 88, 91, 94, 97, 100, 103, 106, 109, 20, 26, 47, 50, 53, 56, 59, 62, 65, 68, 71, 74, 77, 80, 83, 86, 89, 92, 95, 98, 101, 104, 107, 21, 27, 48, 51, 54, 57, 60, 63, 66, 69, 72, 75, 78, 81, 84, 87, 90, 93, 96, 99, 102, 105, 108, 111 and from 0-2 amino acid modification(s) (e.g., 0-1 amino acid modification(s)) thereof. In some embodiments, the multispecific antibody further comprises a fragment crystallizable (Fc) region. In some embodiments, the Fc region comprises an IgG CH2 domain and an IgG CH3 domain. In some embodiments, the Fc region comprises a heterodimeric Fc region. In some embodiments, the Fc region comprises at least one amino acid modification that increases the half-life of the multispecific antibody. In some embodiments, the Fc region comprises at least one amino acid modification that modulates its interaction with an Fc receptor. In some embodiments, the Fc region comprises at least one amino acid modification that increases binding of the Fc region to an Fc receptor. In some embodiments, the Fc region comprises at least one amino acid modification that decreases glycosylation of the Fc region. In some embodiments, the modification is an amino acid substitution, deletion, or addition. In some embodiments, the modification is an amino acid substitution. In some embodiments, the at least one amino acid modification that decreases glycosylation of the Fc region comprises an amino acid substitution at a position corresponding to position N297 of human IgGl, wherein the numbering is according to the EU index of Kabat. In some embodiments, the Fc region is afiicosylated. In some embodiments, the Fc region comprises at least one amino acid modification that increases antibody -dependent cellular cytotoxicity (ADCC). In some embodiments, the modification is an amino acid substitution, deletion, or addition. In some embodiments, the modification is an amino acid substitution. In some embodiments, the Fc region comprises at least one mutation that increases antibody-dependent cellular cytotoxicity (ADCC), wherein the at least one mutation that increases ADCC comprises an amino acid substitution at positions corresponding to positions S239, 1332, and A330 of human IgGl, wherein the amino acid numbering is according to the EU index according to Kabat et al. In some embodiments, the amino acid substitutions are S239D, I332E, and A330L, wherein the amino acid numbering is according to the EU index according to Kabat et al. In some embodiments, further comprising the heterodimeric Fc region, wherein the heterodimeric Fc region comprises a knob chain and a hole chain, forming a knob-into-hole (KIH) structure. In some embodiments, the knob chain comprises an IgGl, IgG2, IgG3, or IgG4 domain. In some embodiments, the knob chain comprises an IgGl domain. In some embodiments, the knob chain comprises an IgG2 domain. In some embodiments, the knob chain comprises an IgG3 domain. In some embodiments, the knob chain comprises an IgG4 domain. In some embodiments, the hole chain comprises an IgGl, IgG2, IgG3, or IgG4 domain. In some embodiments, the hole chain comprises an IgGl domain. In some embodiments, the hole chain comprises an IgG2 domain. In some embodiments, the hole chain comprises an IgG3 domain. In some embodiments, the hole chain comprises an IgG4 domain. In some embodiments, the knob chain comprises an amino acid substitution at a position corresponding to T366 of IgGl, wherein amino acid position numbering is according to the EU index according to Kabat et al. In some embodiments, the T366 substitution comprises a T336W mutation, wherein amino acid position numbering is according to the EU index according to Kabat et al. In some embodiments, the hole chain comprises an amino acid substitution at a position corresponding to T366, L368, or Y407 of IgGl, wherein amino acid position numbering is according to the EU index according to Kabat et al. In some embodiments, the hole chain comprises an amino acid substitution at a position corresponding to T366, L368, and Y407 of IgGl, wherein amino acid position numbering is according to the EU index according to Kabat et al. In some embodiments, the T366, L368, or Y407 amino acid substitutions comprise a T366S, L368A, or Y407V of IgGl, wherein amino acid position numbering is according to the EU index according to Kabat et al. In some embodiments, the T366, L368, and Y407 amino acid substitutions comprises a T366S, L368A, and Y407V of IgGl, wherein amino acid position numbering is according to the EU index according to Kabat et al. In some embodiments, the multispecific antibody binds to a target cell. In some embodiments, the target cell expresses CD47 and ICAM1 . In some embodiments, the target cell expresses a lower level of CD47 relative to ICAM1 on the surface of the target cell. In some embodiments, the target cell is a cancer cell. In some embodiments, the cancer is a hematological malignancy. In some embodiments, the cancer is multiple myeloma. In some embodiments, the hematological malignancy is B cell cancer. In some embodiments, the B cell cancer is leukemia or lymphoma. In some embodiments, the B cell cancer is lymphoma, and wherein the lymphoma is B cell lymphoma. In some embodiments, the hematological malignancy is T cell cancer. In some embodiments, the T cell cancer is cutaneous T cell lyphoma, or anaplastic large cell lymphoma. In some embodiments, the cancer cell is from a solid tumor. In some embodiments, the solid tumor is sarcoma, breast cancer, lung cancer, kidney cancer, melanoma, head and neck cancer, ovarian cancer, liver cancer, bladder cancer, thyroid cancer, or carcinoma. In some embodiments, the solid tumor is lung cancer, and wherein the lung cancer is non-small cell lung cancer. In some embodiments, the multispecific antibody inhibits binding of Signal -regulatory protein alpha (SIRPa) to the target cell as determined by an in vitro competition assay. In some embodiments, the binding of Signal-regulatory protein alpha (SIRPa) to the target cell as determined by an in vitro competition assay is inhibited by the multispecific antibody by at least 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100%. In some embodiments, a concentration of the multispecific antibody required to mediate antibody dependent cellular phagocytosis (ADCP) of the target cell by a macrophage is between O.OlnM - 3nM. In some embodiments, the multispecific antibody induces enhanced antibody-dependent cellular phagocytosis (ADCP) of the target cell by a macrophage as compared to ADCP activity induced of the target cell by a macrophage by a monospecific anti-CD47 antibody. In some embodiments, the multispecific antibody has a higher binding activity for CD47 expressed on a surface of a tumor cell than for CD47 expressed on a surface of a red blood cell or a platelet. In some embodiments, a concentration of the multispecific antibody required for half-maximal binding to a human red blood or a human platelet is greater than 500 nM. In some embodiments, the multispecific antibody induces antibody -dependent cellular cytotoxicity (ADCC) mediated killing of the target cell. In some embodiments, the multispecific antibody induces enhanced antibody-dependent cellular cytotoxicity (ADCC) activity on the target cell as compared to ADCC activity induced on the target cell by a monospecific anti-CD47 antibody or a monospecific anti-ICAMl antibody. In some embodiments, the multispecific antibody induces at least 5%, 10%, 15%, 20%, 25%, 30%, 25%, 40%, 45%, or 50% more ADCC activity of the target cell as compared to an ADCC activity of the target cell that is induced by a monospecific anti-CD47 antibody or a monospecific anti-ICAMl antibody. In some embodiments, the multispecific antibody induces complement-dependent cytotoxicity (CDC) mediated killing of the target cell. In some embodiments, the multispecific antibody induces at least 5%, 10%, 15%, 20%, 25%, 30%, 25%, 40%, 45%, or 50% more CDC activity of the target cell as compared to a CDC activity of the target cell that is induced by a monospecific anti-CD47 antibody or a monospecific anti-ICAMl antibody. In some embodiments, a concentration of 600 nM of the multispecific antibody does not induce hemolysis of red blood cells in a hemagglutination assay.

[0002] Disclosed herein are nucleic acid molecules encoding the multispecific antibody of any one of the above embodiments. Disclosed herein are vectors comprising the nucleic acid molecules encoding the multispecific antibody of any one of the above embodiments.

[0003] Disclosed herein are pharmaceutical compositions comprising the multispecific antibody of any one of the above embodiments. In some embodiments, the pharmaceutical composition further comprises a pharmaceutically acceptable carrier, an excipient, or any combinations thereof.

[0004] Disclosed herein are kits that comprise at least one of the multispecific antibody of any one of the above embodiments, the vector of any one of the above embodiments, the nucleic acid of any one of the above embodiments, or the pharmaceutical composition of any one of the above embodiments.

[0005] Disclosed herein are methods of treating a subject having cancer, the method comprising: administering to the subject the multispecific antibody of any one of the above embodiments. In some embodiments, the cancer comprises cancer cells that express CD47 and ICAM1. In some embodiments, the ratio of CD47 to ICAM1 on the surface of the cancer cell is at least about 10: 1, 5: 1, 2.5: 1, 2.0: 1, 1.5: 1, 1: 1, 1: 1.5, 1:2.0, 1:2.5, 1:5, 1: 10, 1: 15, 1:20, 1:50, 1: 100, or 1:200. In some embodiments, the cancer cells that express CD47 and ICAM1 are lysed. In some embodiments, the multispecific antibody induces antibody-dependent cellular cytotoxicity (ADCC) mediated killing of the cancer cells that express CD47 and/or ICAM1. In some embodiments, the multispecific antibody induces complement-dependent cytotoxicity (CDC) mediated killing of the cancer cells that express CD47 and/or ICAM1. In some embodiments, the multispecific antibody induces antibody-dependent cellular phagocytosis (ADCP) of the cancer cells that express CD47 and/or ICAM1. In some embodiments, the cancer is a hematological malignancy. In some embodiments, the cancer is multiple myeloma. In some embodiments, the cancer is B cell cancer. In some embodiments, the cancer is leukemia or lymphoma. In some embodiments, the cancer is lymphoma, and wherein the lymphoma is B-cell lymphoma. In some embodiments, the hematological malignancy is T cell cancer. In some embodiments, the T cell cancer is cutaneous T cell lyphoma, or anaplastic large cell lymphoma. In some embodiments, the cancer is a solid tumor. In some embodiments, the solid tumor is sarcoma, breast cancer, lung cancer, kidney cancer, melanoma, head and neck cancer, ovarian cancer, liver cancer, bladder cancer, thyroid cancer, or carcinoma.In some embodiments, the solid tumor is lung cancer, and wherein the lung cancer is non-small cell lung cancer. In some embodiments, the method further comprises administering to the subject an anti -cancer agent. In some embodiments, the anti-cancer agent is a chemotherapeutic agent or a biologic agent. In some embodiments, the administering is sufficient to reduce or eliminate the cancer as compared to a comparable method lacking the administering. In some embodiments, the reduction is at least about 1 fold, 5 fold, 10 fold, 20 fold, 40 fold, 60 fold, 80 fold, or up to about 100 fold. In some embodiments, the cancer is metastatic.

BRIEF DESCRIPTION OF THE DRAWINGS

[0006] The novel features of the invention are set forth with particularity in the appended claims. A better understanding of the features and advantages of the present invention will be obtained by reference to the following detailed description that sets forth illustrative embodiments, in which the principles of the invention are utilized, and the accompanying drawings of which:

[0007] Figure 1 illustrates a three-chain knob-into-hole anti-CD47 and anti-ICAMl bispecific antibody.

[0008] Figure 2A shows ELISA binding on huCD47 of anti-CD47/ICAMl bispecific antibody (“CD47

X ICAM1”) as compared to control. Figure 2B shows ELISA binding on huICAMl of anti- CD47/ICAM1 bispecific antibody (“CD47 X ICAM1”) as compared to various controls.

[0009] Figure 3A shows flow cytometry binding on huCD47 over-expressing CHO cells of anti- CD47/ICAM1 bispecific antibody (“CD47 X ICAM1”) as compared to various controls. Figure 3B shows flow cytometry binding on cynoCD47 over-expressing CHO cells of anti-CD47/ICAMl bispecific antibody (“CD47 X ICAM1”) as compared to various controls.

[0010] Figure 4 shows a SIRPa blocking assay utilizing Raji cell cultured with anti-CD47/ICAMl bispecific antibody (“CD47 X ICAM1” or VIR47.V8/ICAML245) as compared to various controls.

[0011] Figure 5 shows Antibody Dependent Cellular Phagocytosis (ADCP) of Raji cells as cultured with anti-CD47/ICAMl bispecific antibody (“CD47 X ICAM1” or VIR47.V8/ICAM1.245) as compared to various controls. [0012] Figure 6 shows Antibody-dependent cellular cytotoxicity (ADCC) of HCC-44 cells cultured with anti-CD47/ICAMl bispecific antibody (“CD47 X ICAM1”) as compared to various controls.

[0013] Figure 7A shows FACS binding of anti-CD47/ICAMl bispecific antibody (“CD47 X ICAM1”) on human RBC cells as compared to various controls. Figure 7B shows binding of anti-CD47/ICAMl bispecific antibody (“CD47 X ICAM1”) on human platelet cells as compared to various controls.

[0014] Figure 8 shows hemagglutination of human red blood cells by the indicated bispecific, anti- CD47/ICAM1 bispecific antibody (“CD47 X ICAM1”) as compared to various controls.

[0015] Figure 9 shows an in vivo Raji CDX mouse model of mice administered 0.5 mg/kg or 3mg/kg of anti-CD47/ICAMl bispecific antibody (“CD47 X ICAM1”) as compared to various controls.

[0016] Figures 10A-10B illustrate potent, selective SIRPa blocking activity of anti-CD47/ICAMl bispecific antibody (VIR47.V8/ICAM1.U3) on KYSE-150 (ICAMl^med CD47^med) (Figure 10A) cells, but not on KYSE-30 (ICAMl^nu11 CD47^med) cells (Figure 10B), whereas anti-CD47 antibody VIR47.V8 didn’t show the selectivity.

[0017] Figure 11 shows FACS binding of anti-CD47/ICAMl bispecific antibody (VIR47.V8/ICAM1.U3) on human red blood cells (RBCs).

[0018] Figure 12 shows efficacy of anti-CD47/ICAMl bispecific antibody (VIR47.V8/ICAM1.U3) in in vivo Raji model.

[0019] Figures 13 shows efficacy of anti-CD47/ICAMl bispecific antibody (VIR47.V8/ICAM1.U3) in in vivo HCC44 NSCLC/Kras G12C model.

DETAILED DESCRIPTION

[0020] Cluster of differentiation 47 (CD47), also known as integrin-associated protein (IAP), is a ~50 kDa immunoglobulin superfamily membrane glycoprotein that is overexpressed in numerous blood cancers and solid tumors. High CD47 expression often correlates with more aggressive diseases and poorer clinical outcomes.

[0021] CD47 on the surface of CD47+ cells interacts with signal regulatory protein alpha (SIRPa) expressed on cells of the innate and adaptive immune systems, such as macrophages and dendritic cells. This interaction sends a “don’t eat me” signal that inhibits phagocytosis, thereby allowing CD47+ cells to evade immune surveillance. These data suggest that CD47 may serve as an immune checkpoint and that blocking the CD47-SIRPa interaction could have therapeutic value by switching off the “don’t eat me” signal. Thus, blocking CD47 has emerged as a promising therapeutic strategy with numerous studies showing that interrupting the CD47-SIRPa signaling pathway promotes anti-tumor activity against human cancers in vitro and in vivo.

[0022] Several anti-CD47 monoclonal antibodies (mAbs) have been shown to increase phagocytosis of acute myeloid leukemia cells, non-Hodgkin’s lymphoma cells, breast cancer cells, and ovarian cancer cells. In clinical studies, CD47 mAbs enhanced the anti-tumor activity of other therapeutic antibodies. At least six anti-CD47 mAbs and three SIRPa fusion proteins are in active phase I or II clinical trials for the treatment of human hematological malignancies and solid tumors. [0023] The efficacy of anti-CD47 mAbs is limited by their interactions with red blood cells (RBCs), which also express CD47. RBCs act as a sink to sequester anti-CD47 antibodies, thereby preventing them from binding to malignant CD47-expressing (CD47+) cells. Furthermore, anti-CD47 mAb binding to RBCs leads to hemagglutination and lysis of the RBCs, resulting in anemia. Thus, there is a need for improved methods of treating malignant diseases mediated by CD47+ cells with reduced off-tumor effects.

[0024] Intercellular Adhesion Molecule l(ICAMl) also known as CD54 is a protein that in humans is encoded by the ICAM1 gene. This gene encodes a cell surface glycoprotein which is typically expressed on endothelial cells and cells of the immune system. ICAM1 is involved in lymphoid trafficking and has been shown to be upregulated in several types of cancers.

[0025] The present disclosure is based, at least in part, on the discovery that multispecific antibodies comprising a CD47 binding domain and an ICAM1 binding domain can overcome the problems of monospecific bivalent anti-CD47antibodies by providing an increased local concentration of the anti- CD47 antibody to relevant cell populations (ICAM1+ (high)/CD47+); thereby increasing the potency and killing of target cancer cells.

[0026] Disclosed herein, are multispecific antibodies, comprising a CD47 binding domain and an ICAM1 binding domain. In some embodiments, the multispecific antibodies are bispecific, trispecific, or tetraspecific. In some embodiments, the multispecific antibodies are bispecific. In some embodiments, the multispecific antibodies are bivalent, trivalent, or tetravalent. In some embodiments, the multispecific antibodies are bivalent. In some embodiments, the multispecific antibody comprises an IgGl, IgG2, IgG3, or IgG4 domain. In some embodiments, the multispecific antibody comprises an IgGl domain. In some embodiments, the multispecific antibody comprises an IgG2 domain. In some embodiments, the multispecific antibody comprises an IgG3 domain. In some embodiments, the multispecific antibody comprises an IgG4 domain.

Definitions

[0027] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as is commonly understood by one of skill in the art to which the claimed subject matter belongs. It is to be understood that the foregoing general description and the following detailed description are exemplary and explanatory only and are not restrictive of any subject matter claimed. The section headings used herein are for organizational purposes only and are not to be construed as limiting the subject matter described. In this application, the use of the singular includes the plural unless specifically stated otherwise. It is noted that, as used in the specification and the appended claims, the singular forms “a,” “an” and “the” include plural referents unless the context clearly dictates otherwise. In this application, the use of “or” means “and/or” unless stated otherwise. Furthermore, use of the term “including” as well as other forms, such as “include”, “includes,” and “included,” is not limiting.

[0028] As used herein, ranges and amounts can be expressed as “about” a particular value or range. About also includes the exact amount. Hence “about 5 pL” means “about 5 pL" and also “5 pL.” Generally, the term “about” includes an amount that would be expected to be within experimental error. Unless otherwise indicated, all numbers expressing quantities of ingredients, reaction conditions, and so forth used in the specification and claims are to be understood as being modified in all instances by the term “about.” Accordingly, unless indicated to the contrary, the numerical parameters set forth in the present specification and attached claims are approximations that can vary depending upon the desired properties sought to be obtained by the present application. Generally the term “about,” as used herein when referring to a measurable value such as an amount of weight, time, dose, etc. is meant to encompass in one example variations of ± 20% or ± 10%, in another example ± 5%, in another example ± 1%, and in yet another example ± 0.1% from the specified amount, as such variations are appropriate to perform the disclosed method.

[0029] The section headings used herein are for organizational purposes only and are not to be construed as limiting the subject matter described.

[0030] “Antibodies” and “immunoglobulins” (Igs) are glycoproteins having the same structural characteristics. The terms are used synonymously. In some instances, the antigen specificity of the immunoglobulin is known.

[0031] The term “antibody” is used in the broadest sense and covers fully assembled antibodies, antibody fragments that can bind antigen (e.g., Fab, F(ab’)2, Fv, single chain antibodies, diabodies, antibody chimeras, hybrid antibodies, bispecific antibodies, and the like), and recombinant peptides comprising the forgoing.

[0032] The terms “monoclonal antibody” and “mAb” as used herein refer to an antibody obtained from a substantially homogeneous population of antibodies, i.e., the individual antibodies comprising the population are identical except for possible naturally occurring mutations that may be present in minor amounts.

[0033] “Native antibodies” and “native immunoglobulins” are usually heterotetrameric glycoproteins of about 150,000 Daltons, composed of two identical light (L) chains and two identical heavy (H) chains. Each light chain is linked to a heavy chain by one covalent disulfide bond, while the number of disulfide linkages varies among the heavy chains of different immunoglobulin isotypes. Each heavy and light chain also has regularly spaced intrachain disulfide bridges. Each heavy chain has at one end a variable domain (VH) followed by a number of constant domains. Each light chain has a variable domain at one end (VL) and a constant domain at its other end; the constant domain of the light chain is aligned with the first constant domain of the heavy chain, and the light chain variable domain is aligned with the variable domain of the heavy chain. Particular amino acid residues are believed to form an interface between the light and heavy-chain variable domains.

[0034] The term “variable” refers to the fact that certain portions of the variable domains differ extensively in sequence among antibodies. Variable regions confer antigen-binding specificity. However, the variability is not evenly distributed throughout the variable domains of antibodies. It is concentrated in three segments called complementarity determining regions (CDRs) or hypervariable regions, both in the light chain and the heavy-chain variable domains. The more highly conserved portions of variable domains are celled in the framework (FR) regions. The variable domains of native heavy and light chains each comprise four FR regions, largely adopting a [3-pleated-sheet configuration, connected by three CDRs, which form loops connecting, and in some cases forming part of, the P-pleated- sheet structure. The CDRs in each chain are held together in close proximity by the FR regions and, with the CDRs from the other chain, contribute to the formation of the antigen-binding site of antibodies (see, Kabat et al. (1991) NIH PubL. No. 91-3242, Vol. I, pages 647-669). The constant domains are not involved directly in binding an antibody to an antigen, but exhibit various effector functions, such as Fc receptor (FcR) binding, participation of the antibody in antibody -dependent cellular cytotoxicity, initiation of complement dependent cytotoxicity, and mast cell degranulation.

[0035] The term “hypervariable region,” when used herein, refers to the amino acid residues of an antibody that are responsible for antigen -binding. The hypervariable region comprises amino acid residues from a “complementarily determining region” or “CDR” (i.e., residues 24-34 (LI), 50-56 (L2), and 89-97 (L3) in the light-chain variable domain and 31-35 (Hl), 50-65 (H2), and 95-102 (H3) in the heavy -chain variable domain; Kabat et al. (1991) Sequences of Proteins of Immunological Interest, 5^th Ed. Public Health Service, National Institute of Health, Bethesda, Md.) and/or those residues from a “hypervariable loop” (i.e., residues 26-32 (LI), 50-52 (L2), and 91-96 (L3) in the light-chain variable domain and (Hl), 53-55 (H2), and 96-101 (13) in the heavy chain variable domain; Clothia and Lesk, (1987) J. Mol. Biol., 196:901-917). “Framework” or “FR” residues are those variable domain residues other than the hypervariable region residues, as herein deemed.

[0036] “Antibody fragments” comprise a portion of an intact antibody, preferably the antigen-binding or variable region of the intact antibody. Examples of antibody fragments include Fab, Fab, F(ab’)2, and Fv fragments; diabodies; linear antibodies (Zapata et al. (1995) Protein Eng. 10: 1057-1062); single-chain antibody molecules; and multispecific antibodies formed from antibody fragments. Papain digestion of antibodies produces two identical antigen-binding fragments, called “Fab” fragments, each with a single antigen-binding site, and a residual “Fc” fragment, whose name reflects its ability to crystallize readily. Pepsin treatment yields an F(ab’)2 fragment that has two antigen-combining sites and is still capable of cross-linking antigen.

[0037] ‘ ‘Fv” is the minimum antibody fragment that contains a complete antigen recognition and binding site. This region consists of a dimer of one heavy- and one light-chain variable domain in tight, non- covalent association. It is in this configuration that the three CDRs of each variable domain interact to define an antigen-binding site on the surface of the VH-VL dimer. Collectively, the six CDRs confer antigen-binding specificity to the antibody. However, even a single variable domain (or half of an Fv comprising only three CDRs specific for an antigen) has the ability to recognize and bind antigen, although at a lower affinity than the entire binding site.

[0038] The Fab fragment also contains the constant domain of the light chain and the first constant domain (CHI) of the heavy chain. Fab fragments differ from Fab’ fragments by the addition of a few residues at the carboxy terminus of the heavy chain CHI domain including one or more cysteines from the antibody hinge region. Fab’-SH is the designation herein for Fab’ in which the cysteine residue(s) of the constant domains bear a free thiol group. Fab’ fragments are produced by reducing the F(ab’)2 fragment’s heavy chain disulfide bridge. Other chemical couplings of antibody fragments are also known. [0039] The “light chains” of antibodies (immunoglobulins) from any vertebrate species can be assigned to one of two clearly distinct types, called kappa (K) and lambda (X), based on the amino acid sequences of their constant domains.

[0040] Depending on the amino acid sequence of the constant domain of their heavy chains, immunoglobulins can be assigned to different classes. There are five major classes of human immunoglobulins: IgA, IgD, IgE, IgG, IgM, and IgY, and several of these may be further divided into subclasses (isotypes), e.g., IgGl, IgG2, IgG3, IgG4, IgAl, and IgA2. The heavy -chain constant domains that correspond to the different classes of immunoglobulins are called alpha, delta, epsilon, gamma, and mu, respectively. The subunit structures and three-dimensional configurations of different classes of immunoglobulins are well known. Different isotypes have different effector functions. For example, human IgGl and IgG3 isotypes have ADCC (antibody dependent cell-mediated cytotoxicity) activity. [0041] The CDR sequence(s) for the antibodies disclosed herein, or the anti-CD47 or anti-ICAMl binding domain sequences disclosed herein, may be defined or determined according to (i) the Kabat numbering system (Kabat et al. (1971) Ann. NY Acad. Sci. 190:382-391 and, Kabat et al. (1991) Sequences of Proteins of Immunological Interest Fifth Edition, U.S. Department of Health and Human Services, NIH Publication No. 91-3242); or (ii) the Chothia numbering scheme, which will be referred to herein as the “Chothia CDRs” (see, e.g., Chothia and Lesk, 1987, J. Mol. Biol., 196:901-917; Al- Lazikani et al., 1997, J. Mol. Biol., 273 :927-948; Chothia et al., 1992, J. Mol. Biol., 227:799-817; Tramontane A et al. , 1990, J. Mol. Biol. 215(1): 175-82; and U.S. Patent No. 7,709,226); or (iii) the ImMunoGeneTics (IMGT) numbering system, for example, as described in Lefranc, M.-P., 1999, The Immunologist, 7: 132-136 and Lefranc, M.-P. et al, 1999, Nucleic Acids Res., 27:209-212 (“IMGT CDRs”); or (iv) MacCallum et al, 1996, J. Mol. Biol., 262:732-745. See also, e.g., Martin, A., “Protein Sequence and Structure Analysis of Antibody Variable Domains,” in Antibody Engineering, Kontermann and Diibel, eds., Chapter 31, pp. 422-439, Springer- Verlag, Berlin (2001).

[0042] With respect to the Kabat numbering system, CDRs within an antibody heavy chain molecule are typically present at amino acid positions 31 to 35, which optionally can include one or two additional amino acids, following 35 (referred to in the Kabat numbering scheme as 35 A and 35B) (CDR1), amino acid positions 50 to 65 (CDR2), and amino acid positions 95 to 102 (CDR3). Using the Kabat numbering system, CDRs within an antibody light chain molecule are typically present at amino acid positions 24 to 34 (CDR1), amino acid positions 50 to 56 (CDR2), and amino acid positions 89 to 97 (CDR3). As is well known to those of skill in the art, using the Kabat numbering system, the actual linear amino acid sequence of the antibody variable domain can contain fewer or additional amino acids due to a shortening or lengthening of a FR and/or CDR and, as such, an amino acid’s Kabat number is not necessarily the same as its linear amino acid number. [0043] The term “chimeric” antibody refers to an antibody in which a portion of the heavy and/or light chain is derived from a particular source or species, while the remainder of the heavy and/or light chain is derived from a different source or species.

[0044] The term “human antibody” or “humanized antibody”, as used herein, is intended to include antibodies having variable and constant regions derived from human germ line immunoglobulin sequences. Human antibodies are well-known in the state of the art (van Dijk, M.A., and van de Winkel, J.G., Curr. Opin. Chem. Biol. 5 (2001) 368-374). In some instances, human antibodies are also produced in transgenic animals (e.g., mice) that are capable, upon immunization, of producing a full repertoire or a selection of human antibodies in the absence of endogenous immunoglobulin production. Transfer of the human germ-line immunoglobulin gene array in such germ-line mutant mice will result in the production of human antibodies upon antigen challenge (see, e.g., Jakobovits, A., et al, Proc. Natl. Acad. Sci. USA 90 (1993) 2551-2555; Jakobovits, A., et al, Nature 362 (1993) 255-258; Bruggemann, M., et al, Year Immunol. 7 (1993) 33-40). In additional instances, human antibodies are also produced in phage display libraries (Hoogenboom, H.R., and Winter, G., J. Mol. Biol. 227 (1992) 381-388; Marks, J.D., et al, J. Mol. Biol. 222 (1991) 581-597). The techniques of Cole et al. and Boemer et al. are also available for the preparation of human monoclonal antibodies (Cole, et al., Monoclonal Antibodies and Cancer Therapy, Alan R. Liss, p. 77 (1985); and Boemer, P., et al, J. Immunol. 147 (1991) 86-95). [0045] The term “recombinant human antibody”, as used herein, is intended to include all human antibodies that are prepared, expressed, created or isolated by recombinant means, such as antibodies isolated from a host cell such as a NSO or CHO cell or from an animal (e.g. a mouse) that is transgenic for human immunoglobulin genes or antibodies expressed using a recombinant expression vector transfected into a host cell. Such recombinant human antibodies have variable and constant regions in a rearranged form. In some cases, the recombinant human antibodies have been subjected to in vivo somatic hypermutation. Thus, the amino acid sequences of the VH and VL regions of the recombinant antibodies are sequences that, while derived from and related to human germ line VH and VL sequences, may not naturally exist within the human antibody germ line repertoire in vivo.

[0046] The term “valent” as used herein denotes the presence of a specified number of binding sites in an antigen binding molecule. As such, the terms “bivalent”, “tetravalent”, and “hexavalent” denote the presence of two binding sites, four binding sites, and six binding sites, respectively, in an antigen binding molecule. The bispecific antibodies according to the invention are at least “bivalent” and may be “trivalent” or “multivalent” (e.g. “tetravalent” or “hexavalent”). In a particular aspect, the antibodies of the present invention have two or more binding sites and are bispecific. That is, the antibodies may be bispecific even in cases where there are more than two binding sites (i.e. that the antibody is trivalent or multivalent). In particular, the invention relates to bispecific bivalent antibodies, having one binding site for each antigen they specifically bind to.

[0047] The term “bispecific” means that the antibody is able to specifically bind to two distinct antigenic determinants, for example two binding sites each formed by a pair of an antibody heavy chain variable domain (VH) and an antibody light chain variable domain (VL) binding to different antigens . Such a bispecific antibody is an 1+1 format. Other bispecific antibody formats are 2+1 formats (comprising two binding sites for a first antigen or epitope and one binding site for a second antigen or epitope) or 2+2 formats (comprising two binding sites for a first antigen or epitope and two binding sites for a second antigen or epitope). Typically, a bispecific antibody comprises two antigen binding sites, each of which is specific for a different antigenic determinant. The term “multispecific” means that the antibody is able to specifically bind to two or more distinct antigenic determinants for example two binding sites each formed by a pair of an antibody heavy chain variable domain (VH) and an antibody light chain variable domain (VL) binding to different antigens.

[0048] The terms “individual(s)”, “subject(s)” and “patient(s)” are used interchangeably herein and refer to any mammal. In some embodiments, the mammal is a human. In some embodiments, the mammal is a non-human. None of the terms require or are limited to situations characterized by the supervision (e.g. constant or intermittent) of a health care worker (e.g. a doctor, a registered nurse, a nurse practitioner, a physician’s assistant, an orderly or a hospice worker).

[0049] As used herein, the term “percent (%) amino acid sequence identity” with respect to a sequence is defined as the percentage of amino acid residues in a candidate sequence that are identical with the amino acid residues in the specific sequence, after aligning the sequences and introducing gaps, if necessary, to achieve the maximum percent sequence identity, and not considering any conservative substitutions as part of the sequence identity. Alignment for purposes of determining percent amino acid sequence identity can be achieved in various ways that are within the skill in the art, for instance, using publicly available computer software such as EMBOSS MATCHER, EMBOSS WATER, EMBOSS STRETCHER, EMBOSS NEEDLE, EMBOSS LALIGN, BLAST, BLAST-2, ALIGN or Megalign (DNASTAR) software. Those skilled in the art can determine appropriate parameters for measuring alignment, including any algorithms needed to achieve maximal alignment over the full length of the sequences being compared.

[0050] In situations where ALIGN -2 is employed for amino acid sequence comparisons, the % amino acid sequence identity of a given amino acid sequence A to, with, or against a given amino acid sequence B (which can alternatively be phrased as a given amino acid sequence A that has or comprises a certain % amino acid sequence identity to, with, or against a given amino acid sequence B) is calculated as follows: 100 times the fraction X/Y, where X is the number of amino acid residues scored as identical matches by the sequence alignment program ALIGN-2 in that program’s alignment of A and B, and where Y is the total number of amino acid residues in B. It will be appreciated that where the length of amino acid sequence A is not equal to the length of amino acid sequence B, the % amino acid sequence identity of A to B will not equal the % amino acid sequence identity of B to A. Unless specifically stated otherwise, all % amino acid sequence identity values used herein are obtained as described in the immediately preceding paragraph using the ALIGN-2 computer program.

[0051] The terms “cancer” and “tumor” are used interchangeably herein, encompass all types of oncogenic processes and/or cancerous growths. In embodiments, cancer includes primary tumors as well as metastatic tissues or malignantly transformed cells, tissues, or organs. In embodiments, cancer encompasses all histopathologies and stages, e.g., stages of invasiveness/severity, of a cancer. In embodiments, cancer includes relapsed and/or resistant cancer.

[0052] As used herein, “treatment” (and grammatical variations thereof such as “treat” or “treating”) refers to clinical intervention in an attempt to alter the natural course of the individual being treated, and can be performed either for prophylaxis or during the course of clinical pathology. Desirable effects of treatment include, but are not limited to, preventing occurrence or recurrence of disease, alleviation of symptoms, diminishment of any direct or indirect pathological consequences of the disease, preventing metastasis, decreasing the rate of disease progression, amelioration or palliation of the disease state, and remission or improved prognosis. In some embodiments, the molecules of the invention are used to delay development of a disease or to slow the progression of a disease.

[0053] As used herein, the terms “Antibody-dependent cellular cytotoxicity” and “ADCC” refer to a cell- mediated reaction in which non-specific cytotoxic cells (e.g., Natural Killer (NK) cells, neutrophils, and macrophages) recognize bound antibody on a target cell and subsequently cause lysis of the target cell.

CD47 Binding Domains

[0054] In some embodiments, disclosed herein is a multispecific antibody that comprises an anti-CD47 binding domain. In some embodiments, the CD47 binding domain comprises an antibody or antigen binding fragment or variant thereof. In some embodiments, the antibody or antigen binding fragment or variant thereof is a monoclonal antibody. In some embodiments, the antibody or antigen binding fragment or variant thereof is a human antibody, a murine antibody, a humanized antibody, or a chimeric antibody. In some embodiments, the CD47 binding domain comprises a monovalent Fab, a bivalent Fab’2, a singlechain variable fragment (scFv), or functional fragment or variant thereof. In some embodiments, the CD47 binding domain comprises an IgGl, IgG2, IgG3, or IgG4 domain. In some embodiments, the CD47 binding domain comprises an IgGl domain. In some embodiments, the CD47 binding domain comprises an IgG2 domain. In some embodiments, the CD47 binding domain comprises an IgG3 domain. In some embodiments, the CD47 binding domain comprises an IgG4 domain.

[0055] In some embodiments, the antibody, or functional fragment or functional variant thereof that binds specifically to CD47 comprises an anti-CD47 heavy chain and an anti-CD47 light chain.

[0056] In some embodiments, the anti-CD47 heavy chain comprises an anti-CD47 heavy chain variable domain. In some embodiments, the anti-CD47 heavy chain variable domain comprises a variable domain of an IgGl, IgG2, IgG3, or IgG4 heavy chain. In some embodiments, the anti-CD47 light chain comprises an anti-CD47 light chain variable domain. In some embodiments, the anti-CD47 light chain variable domain comprises a variable domain of a Kappa or Lambda light chain.

[0057] In some embodiments, the anti-CD47 heavy chain variable domain comprises the variable domain of an IgGl heavy chain and the anti-CD47 light chain variable domain comprises the variable domain of a Kappa or Lambda light chain. In some embodiments, the anti-CD47 heavy chain variable domain comprises the variable domain of an IgG2 heavy chain and the anti-CD47 light chain variable domain comprises the variable domain of a Kappa or Lambda light chain. In some embodiments, the anti-CD47 heavy chain variable domain comprises the variable domain of an IgG3 heavy chain and the anti-CD47 light chain variable domain comprises the variable domain of a Kappa or Lambda light chain. In some embodiments, the anti-CD47 heavy chain variable domain comprises the variable domain of an IgG4 heavy chain and the anti-CD47 light chain variable domain comprises the variable domain of a Kappa or Lambda light chain.

[0058] In some embodiments, the anti-CD47 heavy chain variable domain comprises the variable domain of an IgGl heavy chain and the anti-CD47 light chain variable domain comprises the variable domain of a Kappa light chain. In some embodiments, the anti-CD47 heavy chain variable domain comprises the variable domain of an IgG2 heavy chain and the anti-CD47 light chain variable domain comprises the variable domain of a Kappa light chain. In some embodiments, the anti-CD47 heavy chain variable domain comprises the variable domain of an IgG3 heavy chain and the anti-CD47 light chain variable domain comprises the variable domain of a Kappa light chain. In some embodiments, the anti-CD47 heavy chain variable domain comprises the variable domain of an IgG4 heavy chain and the anti-CD47 light chain variable domain comprises the variable domain of a Kappa light chain.

[0059] In some embodiments, the anti-CD47 heavy chain variable domain comprises the variable domain of an IgGl heavy chain and the anti-CD47 light chain variable domain comprises the variable domain of a Lambda light chain. In some embodiments, the anti-CD47 heavy chain variable domain comprises the variable domain of an IgG2 heavy chain and the anti-CD47 light chain variable domain comprises the variable domain of a Lambda light chain. In some embodiments, the anti-CD47 heavy chain variable domain comprises the variable domain of an IgG3 heavy chain and the anti-CD47 light chain variable domain comprises the variable domain of a Lambda light chain. In some embodiments, the anti-CD47 heavy chain variable domain comprises the variable domain of an IgG4 heavy chain and the anti-CD47 light chain variable domain comprises the variable domain of a Lambda light chain.

[0060] In some embodiments, the antibody, or functional fragment or functional variant thereof, that binds specifically to CD47 comprises a single-chain variable fragment (scFv) or an antigen-binding fragment (Fab). In some embodiments, the antibody, or functional fragment or functional variant thereof, that binds specifically to CD47 comprises a single-chain variable fragment. In some embodiments, the antibody, or functional fragment or functional variant thereof, that binds specifically to CD47 comprises an antigen-binding fragment (Fab).

[0061] In some embodiments, the anti-CD47 heavy chain variable domain comprises complementarity determining regions (CDRs): HC-CDR1, HC-CDR2, and HC-CDR3, and wherein the HC-CDR1, the HC- CDR2, and the HC-CDR3 of the anti-CD47 heavy chain variable domain comprise amino acid sequences according to HC-CDR1: SEQ ID NO: 1; HC-CDR2: SEQ ID NO: 2; HC-CDR3: SEQ ID NO: 3; and wherein the CDRs comprise from 0-2 amino acid modification(s) (e.g., 0 or 1 amino acid modification(s)) in at least one of the HC-CDR1, HC-CDR2, or HC-CDR3. In some embodiments, the HC-CDR1, the HC- CDR2, and the HC-CDR3 of the anti-CD47 heavy chain variable domain comprises amino acid sequences according to HC-CDR1: SEQ ID NO: 1; HC-CDR2: SEQ ID NO: 2; and HC-CDR3: SEQ ID NO: 3. [0062] In some embodiments, the anti-CD47 light chain variable domain comprises complementarity determining regions (CDRs): LC-CDR1, LC-CDR2, and LC-CDR3, and wherein the LC-CDR1, the LC- CDR2, and the LC-CDR3 of the anti-CD47 light chain variable domain comprises amino acid sequences according to LC-CDR1: SEQ ID NO: 4; LC-CDR2: SEQ ID NO: 5; LC-CDR3: SEQ ID NO: 6; and wherein the CDRs comprise from 0-2 amino acid modification(s) (e.g., 0 or 1 amino acid modification(s)) in at least one of the LC-CDR1, LC-CDR2, or LC-CDR3. In some embodiments, the anti-CD47 light chain variable domain comprises complementarity determining regions (CDRs): LC-CDR1, LC-CDR2, and LC-CDR3, and wherein the LC-CDR1, the LC-CDR2, and the LC-CDR3 of the anti-CD47 light chain variable domain comprises amino acid sequences according to LC-CDR1: SEQ ID NO: 4; LC-CDR2: SEQ ID NO: 5; and LC-CDR3: SEQ ID NO: 6.

[0063] In some embodiments, the anti-CD47 heavy chain variable domain comprises complementarity determining regions (CDRs): HC-CDR1, HC-CDR2, and HC-CDR3, and wherein the HC-CDR1, the HC- CDR2, and the HC-CDR3 of the anti-CD47 heavy chain variable domain comprise amino acid sequences according to HC-CDR1: SEQ ID NO: 1; HC-CDR2: SEQ ID NO: 2; HC-CDR3: SEQ ID NO: 3; and wherein the CDRs comprise from 0-2 amino acid modification(s) (e.g., 0 or 1 amino acid modification(s)) in at least one of the HC-CDR1, HC-CDR2, or HC-CDR3, and the anti-CD47 light chain variable domain comprises complementarity determining regions (CDRs): LC-CDR1, LC-CDR2, and LC-CDR3, and wherein the LC-CDR1, the LC-CDR2, and the LC-CDR3 of the anti-CD47 light chain variable domain comprises amino acid sequences according to LC-CDR1: SEQ ID NO: 4; LC-CDR2: SEQ ID NO: 5; LC- CDR3: SEQ ID NO: 6; and wherein the CDRs comprise from 0-2 amino acid modification(s) (e.g., 0 or 1 amino acid modification(s)) in at least one of the LC-CDR1, LC-CDR2, or LC-CDR3. In some embodiments, the HC-CDR1, the HC-CDR2, and the HC-CDR3 of the anti-CD47 heavy chain variable domain comprises amino acid sequences according to HC-CDR1: SEQ ID NO: 1; HC-CDR2: SEQ ID NO: 2; HC-CDR3: SEQ ID NO: 3 and the LC-CDR1, the LC-CDR2, and the LC-CDR3 of the anti-CD47 light chain variable domain comprises amino acid sequences according to LC-CDR1: SEQ ID NO: 4;

LC-CDR2: SEQ ID NO: 5; and LC-CDR3: SEQ ID NO: 6.

[0064] In some embodiments, the anti-CD47 heavy chain comprises an amino acid sequence with at least 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NO: 7.

[0065] In some embodiments, the anti-CD47 heavy chain comprises an amino acid sequence of at least 400 consecutive amino acid residues of SEQ ID NO: 7. In some embodiments, the anti-CD47 heavy chain comprises an amino acid sequence of at least 420 consecutive amino acid residues of SEQ ID NO: 7. In some embodiments, the anti-CD47 heavy chain comprises an amino acid sequence of at least 430 consecutive amino acid residues of SEQ ID NO: 7. In some embodiments, the anti-CD47 heavy chain comprises an amino acid sequence of at least 440 consecutive amino acid residues of SEQ ID NO: 7. In some embodiments, the anti-CD47 heavy chain comprises an amino acid sequence of at least 445 consecutive amino acid residues of SEQ ID NO: 7.

[0066] In some embodiments, the anti-CD47 heavy chain comprises an amino acid sequence of at least 400 consecutive amino acid residues of SEQ ID NO: 7 and has at least 95% sequence identity to the at least 400 consecutive amino acid residues of SEQ ID NO: 7. In some embodiments, the anti-CD47 heavy chain comprises an amino acid sequence of at least 420 consecutive amino acid residues of SEQ ID NO: 7 and has at least 95% sequence identity to the at least 420 consecutive amino acid residues of SEQ ID NO: 7. In some embodiments, the anti-CD47 heavy chain comprises an amino acid sequence of at least 430 consecutive amino acid residues of SEQ ID NO: 7 and has at least 95% sequence identity to the at least 430 consecutive amino acid residues of SEQ ID NO: 7. In some embodiments, the anti-CD47 heavy chain comprises an amino acid sequence of at least 440 consecutive amino acid residues of SEQ ID NO: 7 and has at least 95% sequence identity to the at least 440 consecutive amino acid residues of SEQ ID NO: 7. In some embodiments, the anti-CD47 heavy chain comprises an amino acid sequence of at least 445 consecutive amino acid residues of SEQ ID NO: 7 and has at least 95% sequence identity to the at least 445 consecutive amino acid residues of SEQ ID NO: 7.

[0067] In some embodiments, the anti-CD47 light chain comprises an amino acid sequence with at least 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NOs: 9.

[0068] In some embodiments, the anti-CD47 light chain comprises an amino acid sequence of at least 180 consecutive amino acid residues of SEQ ID NO: 9. In some embodiments, the anti-CD47 light chain comprises an amino acid sequence of at least 190 consecutive amino acid residues of SEQ ID NO: 9. In some embodiments, the anti-CD47 light chain comprises an amino acid sequence of at least 200 consecutive amino acid residues of SEQ ID NO: 9. In some embodiments, the anti-CD47 light chain comprises an amino acid sequence of at least 205 consecutive amino acid residues of SEQ ID NO: 9. In some embodiments, the anti-CD47 light chain comprises an amino acid sequence of at least 210 consecutive amino acid residues of SEQ ID NO: 9.

[0069] In some embodiments, the anti-CD47 light chain comprises an amino acid sequence of at least 180 consecutive amino acid residues of SEQ ID NO: 9 and has at least 95% sequence identity to the at least 180 consecutive amino acid residues of SEQ ID NO: 9. In some embodiments, the anti-CD47 light chain comprises an amino acid sequence of at least 190 consecutive amino acid residues of SEQ ID NO: 9 and has at least 95% sequence identity to the at least 190 consecutive amino acid residues of SEQ ID NO: 9. In some embodiments, the anti-CD47 light chain comprises an amino acid sequence of at least 200 consecutive amino acid residues of SEQ ID NO: 9 and has at least 95% sequence identity to the at least 200 consecutive amino acid residues of SEQ ID NO: 9. In some embodiments, the anti-CD47 light chain comprises an amino acid sequence of at least 205 consecutive amino acid residues of SEQ ID NO: 9 and has at least 95% sequence identity to the at least 205 consecutive amino acid residues of SEQ ID NO: 9. In some embodiments, the anti-CD47 light chain comprises an amino acid sequence of at least 210 consecutive amino acid residues of SEQ ID NO: 9 and has at least 95% sequence identity to the at least 210 consecutive amino acid residues of SEQ ID NO: 9.

[0070] In some embodiments, the anti-CD47 heavy chain comprises an amino acid sequence with at least 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NOs: 7 and the anti-CD47 light chain comprises an amino acid sequence with at least 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NOs: 9.

[0071] In some embodiments, the anti-CD47 heavy chain comprises an amino acid sequence with at least 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NO: A.

[0072] In some embodiments, the anti-CD47 light chain comprises an amino acid sequence with at least 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NO: B.

[0073] In some embodiments, the anti-CD47 heavy chain comprises an amino acid sequence with at least 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NO: A and the anti-CD47 light chain comprises an amino acid sequence with at least 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NOs: B.

[0074] In some embodiments, the antibody, or functional fragment or functional variant thereof, that binds specifically to CD47 comprises the scFv. In some embodiments, the scFv that binds specifically to CD47 comprises an amino acid sequence with at least 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NO: 11. [0075] In some embodiments, the scFv that binds specifically to CD47 comprises an amino acid sequence of at least 440 consecutive amino acid residues of SEQ ID NO: 11. In some embodiments, the scFv that binds specifically to CD47 comprises an amino acid sequence of at least 450 consecutive amino acid residues of SEQ ID NO: 11. In some embodiments, the scFv that binds specifically to CD47 comprises an amino acid sequence of at least 460 consecutive amino acid residues of SEQ ID NO: 11. In some embodiments, the scFv that binds specifically to CD47 comprises an amino acid sequence of at least 465 consecutive amino acid residues of SEQ ID NO: 11. In some embodiments, the scFv that binds specifically to CD47 comprises an amino acid sequence of at least 470 consecutive amino acid residues of SEQ ID NO: 11. In some embodiments, the scFv that binds specifically to CD47 comprises an amino acid sequence of at least 475 consecutive amino acid residues of SEQ ID NO: 11.

[0076] In some embodiments, the scFv that binds specifically to CD47 comprises an amino acid sequence of at least 440 consecutive amino acid residues of SEQ ID NO: 11 and has at least 95% sequence identity to the at least 440 consecutive amino acid residues of SEQ ID NO: 11. In some embodiments, the scFv that binds specifically to CD47 comprises an amino acid sequence of at least 450 consecutive amino acid residues of SEQ ID NO: 11 and has at least 95% sequence identity to the at least 450 consecutive amino acid residues of SEQ ID NO: 11. In some embodiments, the scFv that binds specifically to CD47 comprises an amino acid sequence of at least 460 consecutive amino acid residues of SEQ ID NO: 11 and has at least 95% sequence identity to the at least 460 consecutive amino acid residues of SEQ ID NO: 11. In some embodiments, the scFv that binds specifically to CD47 comprises an amino acid sequence of at least 465 consecutive amino acid residues of SEQ ID NO: 11 and has at least 95% sequence identity to the at least 465 consecutive amino acid residues of SEQ ID NO: 11. In some embodiments, the scFv that binds specifically to CD47 comprises an amino acid sequence of at least 470 consecutive amino acid residues of SEQ ID NO: 11 and has at least 95% sequence identity to the at least 470 consecutive amino acid residues of SEQ ID NO: 11. In some embodiments, the scFv that binds specifically to CD47 comprises an amino acid sequence of at least 475 consecutive amino acid residues of SEQ ID NO: 11 and has at least 95% sequence identity to the at least 475 consecutive amino acid residues of SEQ ID NO: 11.

Table 1. CD47 Binding Domain Amino Acid and Nucleotide Sequences; the CDR sequences have been determined according to Kabat numbering

ICAM1 Binding Domains

[0077] In some embodiments, disclosed herein is a multispecific antibody that comprises an anti-ICAMl binding domain. In some embodiments, the ICAM1 binding domain comprises an antibody or antigen binding fragment or variant thereof. In some embodiments, the antibody or antigen binding fragment or variant thereof is a monoclonal antibody. In some embodiments, the antibody or antigen binding fragment or variant thereof is a human antibody, a murine antibody, a humanized antibody, or a chimeric antibody. In some embodiments, the ICAM1 binding domain comprises a monovalent Fab, a bivalent Fab’2, a single-chain variable fragment (scFv), or functional fragment or variant thereof.

[0078] In some embodiments, the ICAM1 binding domain comprises an IgGl, IgG2, IgG3, or IgG4 domain. In some embodiments, the ICAM1 binding domain comprises an IgGl domain. In some embodiments, the ICAM1 binding domain comprises an IgG2 domain. In some embodiments, the ICAM1 binding domain comprises an IgG3 domain. In some embodiments, the ICAM1 binding domain comprises an IgG4 domain.

[0079] In some embodiments, the antibody, or functional fragment or functional variant thereof that binds specifically to ICAM1 comprises an anti-ICAMl heavy chain and an anti-ICAMl light chain. [0080] In some embodiments, the anti-ICAMl heavy chain comprises an anti-ICAMl heavy chain variable domain. In some embodiments, the anti-ICAMl heavy chain variable domain comprises a variable domain of an IgGl, IgG2, IgG3, or IgG4 heavy chain. In some embodiments, the anti-ICAMl light chain comprises an anti-ICAMl light chain variable domain. In some embodiments, the anti-ICAMl light chain variable domain comprises a variable domain of a Kappa or Lambda light chain. In some embodiments, the anti-ICAMl heavy chain variable domain comprises the variable domain of an IgGl heavy chain and the anti-ICAMl light chain variable domain comprises the variable domain of a Kappa or Lambda light chain. In some embodiments, the anti-ICAMl heavy chain variable domain comprises the variable domain of an IgG2 heavy chain and the anti-ICAMl light chain variable domain comprises the variable domain of a Kappa or Lambda light chain. In some embodiments, the anti-ICAMl heavy chain variable domain comprises the variable domain of an IgG3 heavy chain and the anti-ICAMl light chain variable domain comprises the variable domain of a Kappa or Lambda light chain. In some embodiments, the anti-ICAMl heavy chain variable domain comprises the variable domain of an IgG4 heavy chain and the anti-ICAMl light chain variable domain comprises the variable domain of a Kappa or Lambda light chain.

[0081] In some embodiments, the antibody, or functional fragment or functional variant thereof, that binds specifically to ICAM1 comprises a single-chain variable fragment (scFv) or an antigen-binding fragment (Fab). In some embodiments, the antibody, or functional fragment or functional variant thereof, that binds specifically to ICAM1 comprises a single-chain variable fragment (scFv). In some embodiments, the antibody, or functional fragment or functional variant thereof, that binds specifically to ICAM1 comprises an antigen-binding fragment (Fab).

[0082] In some embodiments, the anti-ICAMl heavy chain variable domain comprises complementarity determining regions (CDRs): HC-CDR1, HC-CDR2, and HC-CDR3, and wherein the HC-CDR1, the HC- CDR2, and the HC-CDR3 of the anti-ICAMl heavy chain variable domain comprise amino acid sequences according to HC-CDR1: SEQ ID NO: 19, 25, 46, 49, 52, 55, 58, 61, 64, 67, 70, 73, 76, 79, 82, 85, 88, 91, 94, 97, 100, 103, 106, or 109; HC-CDR2: SEQ ID NO: 20, 26, 47, 50, 53, 56, 59, 62, 65, 68, 71, 74, 77, 80, 83, 86, 89, 92, 95, 98, 101, 104, or 107; HC-CDR3: SEQ ID NO: 21, 27, 48, 51, 54, 57, 60, 63, 66, 69, 72, 75, 78, 81, 84, 87, 90, 93, 96, 99, 102, 105, 108, or 111; and wherein the CDRs comprise from 0-2 amino acid modification(s) (e.g., 0 or 1 amino acid modification(s)) in at least one of the HC- CDR1, HC-CDR2, or HC-CDR3. In some embodiments, the anti-ICAMl heavy chain variable domain comprises complementarity determining regions (CDRs): HC-CDR1, HC-CDR2, and HC-CDR3, and wherein the HC-CDR1, the HC-CDR2, and the HC-CDR3 of the anti-ICAMl heavy chain variable domain comprise amino acid sequences according to HC-CDR1: SEQ ID NO: 25; HC-CDR2: SEQ ID NO: 26; HC-CDR3: SEQ ID NO: 27; and wherein the CDRs comprise from 0-2 amino acid modification(s) (e.g., 0 or 1 amino acid modification(s)) in at least one of the HC-CDR1, HC-CDR2, or HC-CDR3.

[0083] In some embodiments, the anti-ICAMl light chain variable domain comprises complementarity determining regions (CDRs): LC-CDR1, LC-CDR2, and LC-CDR3, and wherein the LC-CDR1, the LC- CDR2, and the LC-CDR3 of the anti-ICAMl light chain variable domain comprises amino acid sequences according to LC-CDR1: SEQ ID NO: 22, 28, 112, 115, 118, 121, 124, 127, 130, 133, 136, 139, 142, 145, 148, 151, 154, 157, 160, or 163; LC-CDR2: SEQ ID NO: 23, 29, 110, 113, 116, 119, 122, 125, 128, 131, 134, 137, 140, 143, 146, 149, 152, 155, 158, 161, or 164; LC-CDR3: SEQ ID NO: 24, 30, 114, 117, 120, 123, 126, 129, 132, 134, 138, 141, 144, 147, 150, 153, 156, 159, 162, or 165; and wherein the CDRs comprise from 0-2 amino acid modification(s) (e.g., 0 or 1 amino acid modification(s)) in at least one of the LC-CDR1, LC-CDR2, or LC-CDR3.

[0084] In some embodiments, the HC-CDR1, the HC-CDR2, and the HC-CDR3 of the anti-ICAMl heavy chain variable domain comprises amino acid sequences according to HC-CDR1: SEQ ID NO: 19, 25, 46, 49, 52, 55, 58, 61, 64, 67, 70, 73, 76, 79, 82, 85, 88, 91, 94, 97, 100, 103, 106, or 109; HC-CDR2: SEQ ID NO: 20, 26, 47, 50, 53, 56, 59, 62, 65, 68, 71, 74, 77, 80, 83, 86, 89, 92, 95, 98, 101, 104, or 107; HC-CDR3: SEQ ID NO: 21, 27, 48, 51, 54, 57, 60, 63, 66, 69, 72, 75, 78, 81, 84, 87, 90, 93, 96, 99, 102, 105, 108, or 111 and the LC-CDR1, the LC-CDR2, and the LC-CDR3 of the anti-ICAMl light chain variable domain comprises amino acid sequences according to LC-CDR1: SEQ ID NO: 22, 28, 112, 115, 118, 121, 124, 127, 130, 133, 136, 139, 142, 145, 148, 151, 154, 157, 160, or 163; LC-CDR2: SEQ ID NO: 23, 29, 110, 113, 116, 119, 122, 125, 128, 131, 134, 137, 140, 143, 146, 149, 152, 155, 158, 161, or 164; LC-CDR3: SEQ ID NO: 24, 30, 114, 117, 120, 123, 126, 129, 132, 134, 138, 141, 144, 147, 150, 153, 156, 159, 162, or 165.

[0085] In some embodiments, the anti-ICAMl heavy chain variable domain comprises complementarity determining regions (CDRs): HC-CDR1, HC-CDR2, and HC-CDR3, and wherein the HC-CDR1, the HC- CDR2, and the HC-CDR3 of the anti-ICAMl heavy chain variable domain comprise amino acid sequences according to HC-CDR1: SEQ ID NO: 19; HC-CDR2: SEQ ID NO: 20; HC-CDR3: SEQ ID NO: 21; and wherein the CDRs comprise from 0-2 amino acid modification(s) (e.g., 0 or 1 amino acid modification(s)) in at least one of the HC-CDR1, HC-CDR2, or HC-CDR3. In some embodiments, the HC-CDR1, the HC-CDR2, and the HC-CDR3 of the anti-ICAMl heavy chain variable domain comprises amino acid sequences according to HC-CDR1: SEQ ID NO: 19; HC-CDR2: SEQ ID NO: 20; and HC- CDR3: SEQ ID NO: 21.

[0086] In some embodiments, the anti-ICAMl light chain variable domain comprises complementarity determining regions (CDRs): LC-CDR1, LC-CDR2, and LC-CDR3, and wherein the LC-CDR1, the LC- CDR2, and the LC-CDR3 of the anti-ICAMl light chain variable domain comprises amino acid sequences according to LC-CDR1: SEQ ID NO: 22; LC-CDR2: SEQ ID NO: 23; LC-CDR3: SEQ ID NO: 24; and wherein the CDRs comprise from 0-2 amino acid modification(s) (e.g., 0 or 1 amino acid modification(s)) in at least one of the LC-CDR1, LC-CDR2, or LC-CDR3. In some embodiments, the anti-ICAMl light chain variable domain comprises complementarity determining regions (CDRs): LC-CDR1, LC-CDR2, and LC-CDR3, and wherein the LC-CDR1, the LC-CDR2, and the LC-CDR3 of the anti-ICAMl light chain variable domain comprises amino acid sequences according to LC-CDR1: SEQ ID NO: 22; LC- CDR2: SEQ ID NO: 23; and LC-CDR3: SEQ ID NO: 24. [0087] In some embodiments, the anti -I CAM 1 heavy chain variable domain comprises complementarity determining regions (CDRs): HC-CDR1, HC-CDR2, and HC-CDR3, and wherein the HC-CDR1, the HC- CDR2, and the HC-CDR3 of the anti-ICAMl heavy chain variable domain comprise amino acid sequences according to HC-CDR1: SEQ ID NO: 19; HC-CDR2: SEQ ID NO: 20; HC-CDR3: SEQ ID NO: 21; and wherein the CDRs comprise from 0-2 amino acid modification(s) (e.g., 0 or 1 amino acid modification(s)) in at least one of the HC-CDR1, HC-CDR2, or HC-CDR3, and the anti-ICAMl light chain variable domain comprises complementarity determining regions (CDRs): LC-CDR1, LC-CDR2, and LC-CDR3, and wherein the LC-CDR1, the LC-CDR2, and the LC-CDR3 of the anti-ICAMl light chain variable domain comprises amino acid sequences according to LC-CDR1: SEQ ID NO: 22; LC- CDR2: SEQ ID NO:: 23; LC-CDR3: SEQ ID NO: 24; and wherein the CDRs comprise from 0-2 amino acid modification(s) (e.g., 0 or 1 amino acid modification(s)) in at least one of the LC-CDR1, LC-CDR2, or LC-CDR3. In some embodiments, the HC-CDR1, the HC-CDR2, and the HC-CDR3 of the anti- ICAMl heavy chain variable domain comprises amino acid sequences according to HC-CDR1: SEQ ID NO: 19; HC-CDR2: SEQ ID NO: 20; HC-CDR3: SEQ ID NO: 21 and the LC-CDR1, the LC-CDR2, and the LC-CDR3 of the anti-ICAMl light chain variable domain comprises amino acid sequences according to LC-CDR1: SEQ ID NO: 22; LC-CDR2: SEQ ID NO: 23; and LC-CDR3: SEQ ID NO: 24.

[0088] In some embodiments, the anti-ICAMl heavy chain variable domain comprises complementarity determining regions (CDRs): HC-CDR1, HC-CDR2, and HC-CDR3, and wherein the HC-CDR1, the HC- CDR2, and the HC-CDR3 of the anti-ICAMl heavy chain variable domain comprise amino acid sequences according to HC-CDR1: SEQ ID NO: 25; HC-CDR2: SEQ ID NO: 26; HC-CDR3: SEQ ID NO: 27; and wherein the CDRs comprise from 0-2 amino acid modification(s) (e.g., 0 or 1 amino acid modification(s)) in at least one of the HC-CDR1, HC-CDR2, or HC-CDR3. In some embodiments, the HC-CDR1, the HC-CDR2, and the HC-CDR3 of the anti-ICAMl heavy chain variable domain comprises amino acid sequences according to HC-CDR1: SEQ ID NO: 25; HC-CDR2: SEQ ID NO: 26; and HC- CDR3: SEQ ID NO: 27.

[0089] In some embodiments, the anti-ICAMl light chain variable domain comprises complementarity determining regions (CDRs): LC-CDR1, LC-CDR2, and LC-CDR3, and wherein the LC-CDR1, the LC- CDR2, and the LC-CDR3 of the anti-ICAMl light chain variable domain comprises amino acid sequences according to LC-CDR1: SEQ ID NO: 28; LC-CDR2: SEQ ID NO: 29; LC-CDR3: SEQ ID NO: 30; and wherein the CDRs comprise from 0-2 amino acid modification(s) (e.g., 0 or 1 amino acid modification(s)) in at least one of the LC-CDR1, LC-CDR2, or LC-CDR3. In some embodiments, the anti-ICAMl light chain variable domain comprises complementarity determining regions (CDRs): LC-CDR1, LC-CDR2, and LC-CDR3, and wherein the LC-CDR1, the LC-CDR2, and the LC-CDR3 of the anti-ICAMl light chain variable domain comprises amino acid sequences according to LC-CDR1: SEQ ID NO: 28; LC- CDR2: SEQ ID NO: 29; and LC-CDR3: SEQ ID NO: 30.

[0090] In some embodiments, the anti-ICAMl heavy chain variable domain comprises complementarity determining regions (CDRs): HC-CDR1, HC-CDR2, and HC-CDR3, and wherein the HC-CDR1, the HC- CDR2, and the HC-CDR3 of the anti-ICAMl heavy chain variable domain comprise amino acid sequences according to HC-CDR1: SEQ ID NO: 25; HC-CDR2: SEQ ID NO: 26; HC-CDR3: SEQ ID NO: 27; and wherein the CDRs comprise from 0-2 amino acid modification(s) (e.g., 0 or 1 amino acid modification(s)) in at least one of the HC-CDR1, HC-CDR2, or HC-CDR3, and the anti-ICAMl light chain variable domain comprises complementarity determining regions (CDRs): LC-CDR1, LC-CDR2, and LC-CDR3, and wherein the LC-CDR1, the LC-CDR2, and the LC-CDR3 of the anti-ICAMl light chain variable domain comprises amino acid sequences according to LC-CDR1: SEQ ID NO: 28; LC- CDR2: SEQ ID NO:: 29; LC-CDR3: SEQ ID NO: 30; and wherein the CDRs comprise from 0-2 amino acid modification(s) (e.g., 0 or 1 amino acid modification(s)) in at least one of the LC-CDR1, LC-CDR2, or LC-CDR3. In some embodiments, the HC-CDR1, the HC-CDR2, and the HC-CDR3 of the anti- ICAMl heavy chain variable domain comprises amino acid sequences according to HC-CDR1: SEQ ID NO: 25; HC-CDR2: SEQ ID NO: 26; HC-CDR3: SEQ ID NO: 27 and the LC-CDR1, the LC-CDR2, and the LC-CDR3 of the anti-ICAMl light chain variable domain comprises amino acid sequences according to LC-CDR1: SEQ ID NO: 28; LC-CDR2: SEQ ID NO: 29; and LC-CDR3: SEQ ID NO: 30.

[0091] In some embodiments, the anti-ICAMl heavy chain comprises an amino acid sequence with at least 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NO: 31.

[0092] In some embodiments, the anti-ICAMl heavy chain comprises an amino acid sequence with at least 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to of SEQ ID NO: C.

[0093] In some embodiments, the anti-ICAMl heavy chain comprises an amino acid sequence of at least 400 consecutive amino acid residues of SEQ ID NO: 31. In some embodiments, the anti-ICAMl heavy chain comprises an amino acid sequence of at least 420 consecutive amino acid residues of SEQ ID NO: 31. In some embodiments, the anti-ICAMl heavy chain comprises an amino acid sequence of at least 430 consecutive amino acid residues of SEQ ID NO: 31. In some embodiments, the anti-ICAMl heavy chain comprises an amino acid sequence of at least 440 consecutive amino acid residues of SEQ ID NO: 31. In some embodiments, the anti-ICAMl heavy chain comprises an amino acid sequence of at least 445 consecutive amino acid residues of SEQ ID NO: 31.

[0094] In some embodiments, the anti-ICAMl heavy chain comprises an amino acid sequence of at least 400 consecutive amino acid residues of SEQ ID NO: 31 and has at least 95% sequence identity to the at least 400 consecutive amino acid residues of SEQ ID NO: 31. In some embodiments, the anti-ICAMl heavy chain comprises an amino acid sequence of at least 420 consecutive amino acid residues of SEQ ID NO: 31 and has at least 95% sequence identity to the at least 420 consecutive amino acid residues of SEQ ID NO: 31. In some embodiments, the anti-ICAMl heavy chain comprises an amino acid sequence of at least 430 consecutive amino acid residues of SEQ ID NO: 31 and has at least 95% sequence identity to the at least 430 consecutive amino acid residues of SEQ ID NO: 31. In some embodiments, the anti-ICAMl heavy chain comprises an amino acid sequence of at least 440 consecutive amino acid residues of SEQ ID NO: 31 and has at least 95% sequence identity to the at least 440 consecutive amino acid residues of SEQ ID NO: 31. In some embodiments, the anti-ICAMl heavy chain comprises an amino acid sequence of at least 445 consecutive amino acid residues of SEQ ID NO: 31 and has at least 95% sequence identity to the at least 445 consecutive amino acid residues of SEQ ID NO: 31.

[0095] In some embodiments, the anti -I CAM 1 light chain comprises an amino acid sequence with at least 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NOs: 33.

[0096] In some embodiments, the anti -I CAM 1 light chain comprises an amino acid sequence with at least 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NO: D.

[0097] In some embodiments, the anti -I CAM 1 light chain comprises an amino acid sequence of at least 180 consecutive amino acid residues of SEQ ID NO: 33. In some embodiments, the anti-ICAMl light chain comprises an amino acid sequence of at least 190 consecutive amino acid residues of SEQ ID NO: 33. In some embodiments, the anti-ICAMl light chain comprises an amino acid sequence of at least 200 consecutive amino acid residues of SEQ ID NO: 33. In some embodiments, the anti-ICAMl light chain comprises an amino acid sequence of at least 205 consecutive amino acid residues of SEQ ID NO: 33. In some embodiments, the anti-ICAMl light chain comprises an amino acid sequence of at least 210 consecutive amino acid residues of SEQ ID NO: 33.

[0098] In some embodiments, the anti-ICAMl light chain comprises an amino acid sequence of at least 180 consecutive amino acid residues of SEQ ID NO: 33 and has at least 95% sequence identity to the at least 180 consecutive amino acid residues of SEQ ID NO: 33. In some embodiments, the anti-ICAMl light chain comprises an amino acid sequence of at least 190 consecutive amino acid residues of SEQ ID NO: 33 and has at least 95% sequence identity to the at least 190 consecutive amino acid residues of SEQ ID NO: 33. In some embodiments, the anti-ICAMl light chain comprises an amino acid sequence of at least 200 consecutive amino acid residues of SEQ ID NO: 33 and has at least 95% sequence identity to the at least 200 consecutive amino acid residues of SEQ ID NO: 33. In some embodiments, the anti-ICAMl light chain comprises an amino acid sequence of at least 205 consecutive amino acid residues of SEQ ID NO: 33 and has at least 95% sequence identity to the at least 205 consecutive amino acid residues of SEQ ID NO: 33. In some embodiments, the anti-ICAMl light chain comprises an amino acid sequence of at least 210 consecutive amino acid residues of SEQ ID NO: 33 and has at least 95% sequence identity to the at least 210 consecutive amino acid residues of SEQ ID NO: 33.

[0099] In some embodiments, the anti-ICAMl heavy chain comprises an amino acid sequence with at least 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NOs: 31 and the anti-ICAMl light chain comprises an amino acid sequence with at least 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NOs: 33.

[0100] In some embodiments, the anti-ICAMl heavy chain comprises an amino acid sequence with at least 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NO: 35. [0101] In some embodiments, the anti -I CAM 1 heavy chain comprises an amino acid sequence of at least 400 consecutive amino acid residues of SEQ ID NO: 35. In some embodiments, the anti-ICAMl heavy chain comprises an amino acid sequence of at least 420 consecutive amino acid residues of SEQ ID NO:

35. In some embodiments, the anti-ICAMl heavy chain comprises an amino acid sequence of at least 430 consecutive amino acid residues of SEQ ID NO: 35. In some embodiments, the anti-ICAMl heavy chain comprises an amino acid sequence of at least 440 consecutive amino acid residues of SEQ ID NO: 35. In some embodiments, the anti-ICAMl heavy chain comprises an amino acid sequence of at least 445 consecutive amino acid residues of SEQ ID NO: 35.

[0102] In some embodiments, the anti-ICAMl heavy chain comprises an amino acid sequence of at least 400 consecutive amino acid residues of SEQ ID NO: 35 and has at least 95% sequence identity to the at least 400 consecutive amino acid residues of SEQ ID NO: 35. In some embodiments, the anti-ICAMl heavy chain comprises an amino acid sequence of at least 420 consecutive amino acid residues of SEQ ID NO: 35 and has at least 95% sequence identity to the at least 420 consecutive amino acid residues of SEQ ID NO: 35. In some embodiments, the anti-ICAMl heavy chain comprises an amino acid sequence of at least 430 consecutive amino acid residues of SEQ ID NO: 35 and has at least 95% sequence identity to the at least 430 consecutive amino acid residues of SEQ ID NO: 35. In some embodiments, the anti-ICAMl heavy chain comprises an amino acid sequence of at least 440 consecutive amino acid residues of SEQ ID NO: 35 and has at least 95% sequence identity to the at least 440 consecutive amino acid residues of SEQ ID NO: 35. In some embodiments, the anti-ICAMl heavy chain comprises an amino acid sequence of at least 445 consecutive amino acid residues of SEQ ID NO: 35 and has at least 95% sequence identity to the at least 445 consecutive amino acid residues of SEQ ID NO: 35.

[0103] In some embodiments, the anti-ICAMl heavy chain comprises an amino acid sequence with at least 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NO: 36.

[0104] In some embodiments, the anti-ICAMl heavy chain comprises an amino acid sequence of at least 400 consecutive amino acid residues of SEQ ID NO: 36. In some embodiments, the anti-ICAMl heavy chain comprises an amino acid sequence of at least 420 consecutive amino acid residues of SEQ ID NO:

36. In some embodiments, the anti-ICAMl heavy chain comprises an amino acid sequence of at least 430 consecutive amino acid residues of SEQ ID NO: 36. In some embodiments, the anti-ICAMl heavy chain comprises an amino acid sequence of at least 440 consecutive amino acid residues of SEQ ID NO: 36. In some embodiments, the anti-ICAMl heavy chain comprises an amino acid sequence of at least 445 consecutive amino acid residues of SEQ ID NO: 36.

[0105] In some embodiments, the anti-ICAMl heavy chain comprises an amino acid sequence of at least 400 consecutive amino acid residues of SEQ ID NO: 36 and has at least 95% sequence identity to the at least 400 consecutive amino acid residues of SEQ ID NO: 36. In some embodiments, the anti-ICAMl heavy chain comprises an amino acid sequence of at least 420 consecutive amino acid residues of SEQ ID NO: 36 and has at least 95% sequence identity to the at least 420 consecutive amino acid residues of SEQ ID NO: 36. In some embodiments, the anti-ICAMl heavy chain comprises an amino acid sequence of at least 430 consecutive amino acid residues of SEQ ID NO: 36 and has at least 95% sequence identity to the at least 430 consecutive amino acid residues of SEQ ID NO: 36. In some embodiments, the anti-ICAMl heavy chain comprises an amino acid sequence of at least 440 consecutive amino acid residues of SEQ ID NO: 36 and has at least 95% sequence identity to the at least 440 consecutive amino acid residues of SEQ ID NO: 36. In some embodiments, the anti-ICAMl heavy chain comprises an amino acid sequence of at least 445 consecutive amino acid residues of SEQ ID NO: 36 and has at least 95% sequence identity to the at least 445 consecutive amino acid residues of SEQ ID NO: 36.

[0106] In some embodiments, the anti-ICAMl light chain comprises an amino acid sequence with at least 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NOs: 39.

[0107] In some embodiments, the anti-ICAMl light chain comprises an amino acid sequence of at least 180 consecutive amino acid residues of SEQ ID NO: 39. In some embodiments, the anti-ICAMl light chain comprises an amino acid sequence of at least 190 consecutive amino acid residues of SEQ ID NO: 39. In some embodiments, the anti-ICAMl light chain comprises an amino acid sequence of at least 200 consecutive amino acid residues of SEQ ID NO: 39. In some embodiments, the anti-ICAMl light chain comprises an amino acid sequence of at least 205 consecutive amino acid residues of SEQ ID NO: 39. In some embodiments, the anti-ICAMl light chain comprises an amino acid sequence of at least 210 consecutive amino acid residues of SEQ ID NO: 39.

[0108] In some embodiments, the anti-ICAMl light chain comprises an amino acid sequence of at least 180 consecutive amino acid residues of SEQ ID NO: 39 and has at least 95% sequence identity to the at least 180 consecutive amino acid residues of SEQ ID NO: 39. In some embodiments, the anti-ICAMl light chain comprises an amino acid sequence of at least 190 consecutive amino acid residues of SEQ ID NO: 39 and has at least 95% sequence identity to the at least 190 consecutive amino acid residues of SEQ ID NO: 39. In some embodiments, the anti-ICAMl light chain comprises an amino acid sequence of at least 200 consecutive amino acid residues of SEQ ID NO: 39 and has at least 95% sequence identity to the at least 200 consecutive amino acid residues of SEQ ID NO: 39. In some embodiments, the anti-ICAMl light chain comprises an amino acid sequence of at least 205 consecutive amino acid residues of SEQ ID NO: 39 and has at least 95% sequence identity to the at least 205 consecutive amino acid residues of SEQ ID NO: 39. In some embodiments, the anti-ICAMl light chain comprises an amino acid sequence of at least 210 consecutive amino acid residues of SEQ ID NO: 39 and has at least 95% sequence identity to the at least 210 consecutive amino acid residues of SEQ ID NO: 39.

[0109] In some embodiments, the anti-ICAMl heavy chain comprises an amino acid sequence with at least 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NOs: 35 or 36 and the anti-ICAMl light chain comprises an amino acid sequence with at least 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NOs: 39. [0110] In some embodiments, the anti-ICAMl heavy chain comprises an amino acid sequence with at least 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NO: 41.

[oni] In some embodiments, the anti-ICAMl heavy chain comprises an amino acid sequence of at least 400 consecutive amino acid residues of SEQ ID NO: 41. In some embodiments, the anti-ICAMl heavy chain comprises an amino acid sequence of at least 420 consecutive amino acid residues of SEQ ID NO: 41. In some embodiments, the anti-ICAMl heavy chain comprises an amino acid sequence of at least 430 consecutive amino acid residues of SEQ ID NO: 41. In some embodiments, the anti-ICAMl heavy chain comprises an amino acid sequence of at least 440 consecutive amino acid residues of SEQ ID NO: 41. In some embodiments, the anti-ICAMl heavy chain comprises an amino acid sequence of at least 445 consecutive amino acid residues of SEQ ID NO: 41.

[0112] In some embodiments, the anti-ICAMl heavy chain comprises an amino acid sequence of at least 400 consecutive amino acid residues of SEQ ID NO: 41 and has at least 95% sequence identity to the at least 400 consecutive amino acid residues of SEQ ID NO: 41. In some embodiments, the anti-ICAMl heavy chain comprises an amino acid sequence of at least 420 consecutive amino acid residues of SEQ ID NO: 41 and has at least 95% sequence identity to the at least 420 consecutive amino acid residues of SEQ ID NO: 41. In some embodiments, the anti-ICAMl heavy chain comprises an amino acid sequence of at least 430 consecutive amino acid residues of SEQ ID NO: 41 and has at least 95% sequence identity to the at least 430 consecutive amino acid residues of SEQ ID NO: 41. In some embodiments, the anti-ICAMl heavy chain comprises an amino acid sequence of at least 440 consecutive amino acid residues of SEQ ID NO: 41 and has at least 95% sequence identity to the at least 440 consecutive amino acid residues of SEQ ID NO: 41. In some embodiments, the anti-ICAMl heavy chain comprises an amino acid sequence of at least 445 consecutive amino acid residues of SEQ ID NO: 41 and has at least 95% sequence identity to the at least 445 consecutive amino acid residues of SEQ ID NO: 41.

[0113] In some embodiments, the anti-ICAMl heavy chain comprises an amino acid sequence with at least 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NO: 260.

[0114] In some embodiments, the anti-ICAMl heavy chain comprises an amino acid sequence of at least 400 consecutive amino acid residues of SEQ ID NO: 260. In some embodiments, the anti-ICAMl heavy chain comprises an amino acid sequence of at least 420 consecutive amino acid residues of SEQ ID NO: 260. In some embodiments, the anti-ICAMl heavy chain comprises an amino acid sequence of at least 430 consecutive amino acid residues of SEQ ID NO: 260. In some embodiments, the anti-ICAMl heavy chain comprises an amino acid sequence of at least 440 consecutive amino acid residues of SEQ ID NO: 260. In some embodiments, the anti-ICAMl heavy chain comprises an amino acid sequence of at least 445 consecutive amino acid residues of SEQ ID NO: 260.

[0115] In some embodiments, the anti-ICAMl heavy chain comprises an amino acid sequence of at least 400 consecutive amino acid residues of SEQ ID NO: 260 and has at least 95% sequence identity to the at least 400 consecutive amino acid residues of SEQ ID NO: 260. In some embodiments, the anti-ICAMl heavy chain comprises an amino acid sequence of at least 420 consecutive amino acid residues of SEQ ID NO: 260 and has at least 95% sequence identity to the at least 420 consecutive amino acid residues of SEQ ID NO: 260. In some embodiments, the anti-ICAMl heavy chain comprises an amino acid sequence of at least 430 consecutive amino acid residues of SEQ ID NO: 260 and has at least 95% sequence identity to the at least 430 consecutive amino acid residues of SEQ ID NO: 260. In some embodiments, the anti-ICAMl heavy chain comprises an amino acid sequence of at least 440 consecutive amino acid residues of SEQ ID NO: 260 and has at least 95% sequence identity to the at least 440 consecutive amino acid residues of SEQ ID NO: 260. In some embodiments, the anti-ICAMl heavy chain comprises an amino acid sequence of at least 445 consecutive amino acid residues of SEQ ID NO: 260 and has at least 95% sequence identity to the at least 445 consecutive amino acid residues of SEQ ID NO: 260.

[0116] In some embodiments, the anti-ICAMl light chain comprises an amino acid sequence with at least 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NOs: 44.

[0117] In some embodiments, the anti-ICAMl light chain comprises an amino acid sequence of at least 180 consecutive amino acid residues of SEQ ID NO: 44. In some embodiments, the anti-ICAMl light chain comprises an amino acid sequence of at least 190 consecutive amino acid residues of SEQ ID NO: 39. In some embodiments, the anti-ICAMl light chain comprises an amino acid sequence of at least 200 consecutive amino acid residues of SEQ ID NO: 44. In some embodiments, the anti-ICAMl light chain comprises an amino acid sequence of at least 205 consecutive amino acid residues of SEQ ID NO: 44. In some embodiments, the anti-ICAMl light chain comprises an amino acid sequence of at least 210 consecutive amino acid residues of SEQ ID NO: 44.

[0118] In some embodiments, the anti-ICAMl light chain comprises an amino acid sequence of at least 180 consecutive amino acid residues of SEQ ID NO: 44 and has at least 95% sequence identity to the at least 180 consecutive amino acid residues of SEQ ID NO: 44. In some embodiments, the anti-ICAMl light chain comprises an amino acid sequence of at least 190 consecutive amino acid residues of SEQ ID NO: 44 and has at least 95% sequence identity to the at least 190 consecutive amino acid residues of SEQ ID NO: 44. In some embodiments, the anti-ICAMl light chain comprises an amino acid sequence of at least 200 consecutive amino acid residues of SEQ ID NO: 44 and has at least 95% sequence identity to the at least 200 consecutive amino acid residues of SEQ ID NO: 44. In some embodiments, the anti-ICAMl light chain comprises an amino acid sequence of at least 205 consecutive amino acid residues of SEQ ID NO: 44 and has at least 95% sequence identity to the at least 205 consecutive amino acid residues of SEQ ID NO: 44. In some embodiments, the anti-ICAMl light chain comprises an amino acid sequence of at least 210 consecutive amino acid residues of SEQ ID NO: 44 and has at least 95% sequence identity to the at least 210 consecutive amino acid residues of SEQ ID NO: 44.

[0119] In some embodiments, the anti-ICAMl heavy chain comprises an amino acid sequence with at least 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NOs: 41 or 260 and the anti-ICAMl light chain comprises an amino acid sequence with at least 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%,

89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NOs: 44.

[0120] In some embodiments, the anti -I CAM 1 heavy chain comprises an amino acid sequence with at least 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%_;

96%, 97%, 98%, 99%, or 100% identity to any amino acid sequence of Table 2.

[0121] In some embodiments, the anti -I CAM 1 light chain comprises an amino acid sequence with at least 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%_;

96%, 97%, 98%, 99%, or 100% identity to any amino acid sequence of Table 2.

[0122] In some embodiments, the anti -I CAM 1 heavy chain comprises an amino acid sequence with at least 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%_;

96%, 97%, 98%, 99%, or 100% identity to SEQ ID NO: C; and the anti-ICAMl light chain comprises an amino acid sequence with at least 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%,

91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NO: D.

Table 2. ICAM1 Binding Domain Amino Acid and Nucleotide Sequences; the CDR sequences have been determined according to Kabat numbering

Multispecific Antibodies that Bind to CD47 and ICAM1

[0123] In some embodiments, the antibody, or functional fragment or functional variant thereof, that binds specifically to CD47 comprises the Fab, and the antibody, or functional fragment or functional variant thereof, that binds specifically to ICAM1 comprises the scFv.

[0124] In some embodiments, the antibody, or functional fragment or functional variant thereof, that binds specifically to CD47 comprises the scFv, and the antibody, or functional fragment or functional variant thereof, that binds specifically to ICAM1 comprises the Fab. In some embodiments, the anti- ICAM1 heavy chain comprises an amino acid sequence with at least 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NOs: 251 and the anti-ICAMl light chain comprises an amino acid sequence with at least 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NOs: 252; and the scFv comprises an amino acid sequence with at least 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NOs: 253. In some embodiments, the anti- ICAMl heavy chain comprises an amino acid sequence with at least 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NOs: 254 and the anti-ICAMl light chain comprises an amino acid sequence with at least 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NOs: 255; and the scFv comprises an amino acid sequence with at least 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NOs: 256.

[0125] In some embodiments, the multispecific antibody comprises at least 3 CDRs of an anti-CD47 binding domain selected from any one of SEQ ID NOs: 1-6 and from 0-2 amino acid modification(s) (e.g., 0-1 amino acid modification(s)) thereof; and at least 3 CDRs of an anti-ICAMl binding domain selected from any one of SEQ ID NO: 19, 25, 46, 49, 52, 55, 58, 61, 64, 67, 70, 73, 76, 79, 82, 85, 88, 91, 94, 97, 100, 103, 106, 109, 20, 26, 47, 50, 53, 56, 59, 62, 65, 68, 71, 74, 77, 80, 83, 86, 89, 92, 95, 98, 101, 104, 107, 21, 27, 48, 51, 54, 57, 60, 63, 66, 69, 72, 75, 78, 81, 84, 87, 90, 93, 96, 99, 102, 105, 108, 111 and from 0-2 amino acid modification(s) (e.g., 0-1 amino acid modification(s)) thereof.

Table 3. Exemplary Amino Acid Sequences of Multispecific Antibodies that bind to CD47 and ICAM1; the CDR sequences (bold and underlined) have been determined according to Kabat numbering

[0126] In some embodiments, the multispecific antibody further comprises a fragment crystallizable (Fc) region. In some embodiments, the Fc region comprises an IgG CH2 domain and an IgG CH3 domain. In some embodiments, the Fc region comprises a heterodimeric Fc region. In some embodiments, the heterodimeric Fc region comprises a(n) (e.g., human) IgGl, IgG2, IgG3, or IgG4 domain. In some embodiments, the heterodimeric Fc region comprises a(n) (e.g., human) IgGl domain. In some embodiments, the heterodimeric Fc region comprises a(n) (e.g., human) IgG2 domain. In some embodiments, the heterodimeric Fc region comprises a(n) (e.g., human) IgG3 domain. In some embodiments, the heterodimeric Fc region comprises a(n) (e.g., human) IgG4 domain.

[0127] In some embodiments, the Fc region comprises at least one amino acid modification that increases the half-life of the multispecific antibody. In some embodiments, the Fc region comprises at least one amino acid modification that modulates its interaction with an Fc receptor. In some embodiments, the Fc region comprises at least one amino acid modification that increases binding of the Fc region to an Fc receptor. In some embodiments, the Fc region comprises at least one amino acid modification that decreases glycosylation of the Fc region. In some embodiments, the modification is an amino acid substitution, deletion, or addition. In some embodiments, the modification is an amino acid substitution. In some embodiments, the at least one amino acid modification that decreases glycosylation of the Fc region comprises an amino acid substitution at a position corresponding to position N297 of human IgGl, wherein the numbering is according to the EU index of Kabat. In some embodiments, the Fc region is afiicosylated.

[0128] In some embodiments, the Fc region comprises at least one amino acid modification that increases antibody-dependent cellular cytotoxicity (ADCC). In some embodiments, the modification is an amino acid substitution, deletion, or addition. In some embodiments, the modification is an amino acid substitution. In some embodiments, the Fc region comprises at least one mutation that increases antibodydependent cellular cytotoxicity (ADCC), wherein the at least one mutation that increases ADCC comprises an amino acid substitution at positions corresponding to positions S239, 1332, and A330 of human IgGl, wherein the amino acid numbering is according to the EU index according to Kabat et al. In some embodiments, the amino acid substitutions are S239D, I332E, and A330L, wherein the amino acid numbering is according to the EU index according to Kabat et al.

[0129] In some embodiments, the heterodimeric Fc region, wherein the heterodimeric Fc region comprises a knob chain and a hole chain, forming a knob-into-hole (KIH) structure. In some embodiments, the knob chain comprises a(n) (e.g., human) IgGl, IgG2, IgG3, or IgG4 domain. In some embodiments, the knob chain comprises a(n) (e.g., human) IgGl domain. In some embodiments, the knob chain comprises a(n) (e.g., human) IgG2 domain. In some embodiments, the knob chain comprises a(n) (e.g., human) IgG3 domain. In some embodiments, the knob chain comprises a(n) (e.g., human) IgG4 domain. In some embodiments, the hole chain comprises a(n) (e.g., human) IgGl, IgG2, IgG3, or IgG4 domain. In some embodiments, the hole chain comprises a(n) (e.g., human) IgGl domain. In some embodiments, the hole chain comprises a(n) (e.g., human) IgG2 domain. In some embodiments, the hole chain comprises a(n) (e.g., human) IgG3 domain. In some embodiments, the hole chain comprises a(n) (e.g., human) IgG4 domain. In some embodiments, the knob chain comprises an amino acid substitution at a position corresponding to T366 of IgGl, wherein amino acid position numbering is according to the EU index according to Kabat et al. In some embodiments, the T366 substitution comprises a T336W mutation, wherein amino acid position numbering is according to the EU index according to Kabat et al. In some embodiments, the hole chain comprises an amino acid substitution at a position corresponding to T366, L368, or Y407 of IgGl, wherein amino acid position numbering is according to the EU index according to Kabat et al. In some embodiments, the hole chain comprises an amino acid substitution at a position corresponding to T366, L368, and Y407 of IgGl, wherein amino acid position numbering is according to the EU index according to Kabat et al. In some embodiments, the T366, L368, or Y407 amino acid substitutions comprise a T366S, L368A, or Y407V of IgGl, wherein amino acid position numbering is according to the EU index according to Kabat et al. In some embodiments, the T366, L368, and Y407 amino acid substitutions comprises a T366S, L368A, and Y407V of IgGl, wherein amino acid position numbering is according to the EU index according to Kabat et al.

[0130] In some embodiments, the multispecific antibody binds to a target cell. In some embodiments, the target cell expresses CD47 and ICAM1. In some embodiments, the target cell expresses a lower level of CD47 relative to ICAM1 on the surface of the target cell. In some embodiments, the ratio of CD47 to ICAM1 on the surface of the target cell is at least about 10: 1, 5: 1, 2.5: 1, 2.0: 1, 1.5: 1, 1: 1, 1: 1.5, 1:2.0, 1:2.5, 1:5, 1: 10, 1: 15, 1:20, 1:50, 1: 100, or 1:200.

[0131] In some embodiments, the target cell is a cancer cell. In some embodiments, the cancer is a hematological malignancy. In some embodiments, the cancer is multiple myeloma. In some embodiments, the hematological malignancy is B cell cancer. In some embodiments, the B cell cancer is leukemia or lymphoma. In some embodiments, the B cell cancer is lymphoma, and wherein the lymphoma is B cell lymphoma. In some embodiments, the hematological malignancy is T cell cancer. In some embodiments, the T cell cancer is cutaneous T cell lyphoma, or anaplastic large cell lymphoma. In some embodiments, the cancer cell is from a solid tumor. In some embodiments, the solid tumor is sarcoma, breast cancer, lung cancer, kidney cancer, melanoma, head and neck cancer, ovarian cancer, liver cancer, bladder cancer, thyroid cancer, or carcinoma. In some embodiments, the solid tumor is lung cancer, and wherein the lung cancer is non-small cell lung cancer.

[0132] In some embodiments, the multispecific antibody binds to a cancer cell that expresses CD47 and ICAM1 on the surface, and wherein the ratio of CD47 to ICAM1 on the surface of the target cell is at least about 10: 1, 5: 1, 2.5: 1, 2.0: 1, 1.5: 1, 1: 1, 1: 1.5, 1:2.0, 1:2.5, 1:5, 1: 10, 1: 15, 1:20, 1:50, 1: 100, or 1:200. [0133] In some embodiments, the multispecific antibody inhibits binding of Signal-regulatory protein alpha (SIRPa) to the target cell as determined by an in vitro competition assay. In some embodiments, the binding of Signal-regulatory protein alpha (SIRPa) to the target cell as determined by an in vitro competition assay is inhibited by the multispecific antibody by at least 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100%.

[0134] In some embodiments, a concentration of the multispecific antibody required to mediate antibody dependent cellular phagocytosis (ADCP) of the target cell is between 0.0 InM - 3nM, 0.05nM - 5nM, 0.5nM - 2nM, or 0.0 InM - lOnM. In some embodiments, a concentration of the multispecific antibody required to mediate antibody -dependent cellular phagocytosis (ADCP) of the target cell is between 0.0 InM - 3nM. In some embodiments, the multispecific antibody induces enhanced antibody -dependent cellular phagocytosis (ADCP) of the target cell by a macrophage as compared to ADCP activity induced of the target cell by a macrophage by a monospecific anti-CD47 antibody.

[0135] In some embodiments, the multispecific antibody has a higher binding activity for CD47 expressed on a surface of a tumor cell than for CD47 expressed on a surface of a red blood cell or a platelet.

[0136] In some embodiments, a concentration of the multispecific antibody required for half-maximal binding to a human red blood or a human platelet is greater than 500 nM. In some embodiments, a concentration of the multispecific antibody required for half-maximal binding to a human red blood or a human platelet is greater than 600 nM, 700nM, 800nM, or 900 nM. In some embodiments, a concentration of the multispecific antibody required for half-maximal binding to a human red blood or a human platelet is greater than IpM.

[0137] In some embodiments, the multispecific antibody induces antibody -dependent cellular cytotoxicity (ADCC) mediated killing of the target cell. In some embodiments, the multispecific antibody induces enhanced antibody -dependent cellular cytotoxicity (ADCC) activity on the target cell as compared to ADCC activity induced on the target cell by a monospecific anti-CD47 antibody or a monospecific anti-ICAMl antibody. In some embodiments, the multispecific antibody induces at least 5%, 10%, 15%, 20%, 25%, 30%, 25%, 40%, 45%, or 50% more ADCC activity of the target cell as compared to an ADCC activity of the target cell that is induced by a monospecific anti-CD47 antibody or a monospecific anti-ICAMl antibody.

[0138] In some embodiments, the multispecific antibody induces complement-dependent cytotoxicity (CDC) mediated killing of the target cell. In some embodiments, the multispecific antibody induces at least 5%, 10%, 15%, 20%, 25%, 30%, 25%, 40%, 45%, or 50% more CDC activity of the target cell as compared to a CDC activity of the target cell that is induced by a monospecific anti-CD47 antibody or a monospecific anti-ICAMl antibody.

[0139] In some embodiments, a concentration of 600 nM of the multispecific antibody does not induce hemolysis of red blood cells in a hemagglutination assay. Polynucleotides Encoding Multispecific Antibodies that Bind to CD47 and ICAM1

[0140] Disclosed herein, in some embodiments, are isolated recombinant nucleic acid molecules encoding multispecific antibody polypeptide or polypeptide complexes that comprise a CD47 binding domain and an ICAM1 binding domain. Disclosed herein, are isolated recombinant nucleic acid molecules encoding polypeptide sequences of Tables 1-3.

[0141] Disclosed herein are isolated recombinant nucleic acid molecules encoding polypeptide sequences comprising amino acid sequences of SEQ ID NOs: 251, 252, and 253, or amino acid sequences that have at least 90%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NOs: 251, 252, and 253.

[0142] Disclosed herein are isolated recombinant nucleic acid molecules encoding polypeptide sequences comprising amino acid sequences of SEQ ID NOs: 254, 255, and 256, or amino acid sequences that have at least 90%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NOs: 254, 255, and 256.

Pharmaceutical Compositions

[0143] Disclosed herein, in some embodiments, are pharmaceutical compositions comprising: (a) the multispecific antibodies that bind to CD47 and ICAM1 as disclosed herein; and (b) a pharmaceutically acceptable excipient.

[0144] In some embodiments, the pharmaceutical composition comprises (a) polypeptide sequences comprising amino acid sequences of SEQ ID NOs: 251, 252, and 253, or amino acid sequences that have at least 90%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NOs: 251, 252, and 253, and (b) a pharmaceutically acceptable excipient.

[0145] In some embodiments, the pharmaceutical composition comprises (a) polypeptide sequences comprising amino acid sequences of SEQ ID NOs: 254, 255, and 256 or amino acid sequences that have at least 90%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NOs: 254, 255, and 256, and (b) a pharmaceutically acceptable excipient.

[0146] In some embodiments, the multispecific antibody further comprises a detectable label, a therapeutic agent, or a pharmacokinetic modifying moiety. In some embodiments, the detectable label comprises a fluorescent label, a radiolabel, an enzyme, a nucleic acid probe, or a contrast agent.

[0147] For administration to a subject, the multispecific antibody as disclosed herein, may be provided in a pharmaceutical composition together with one or more pharmaceutically acceptable carriers or excipients. The term “pharmaceutically acceptable carrier” includes, but is not limited to, any carrier that does not interfere with the effectiveness of the biological activity of the ingredients and that is not toxic to the patient to whom it is administered. Examples of suitable pharmaceutical carriers are well known in the art and include phosphate buffered saline solutions, water, emulsions, such as oil/water emulsions, various types of wetting agents, sterile solutions etc. Such carriers can be formulated by conventional methods and can be administered to the subject at a suitable dose. Preferably, the compositions are sterile. These compositions may also contain adjuvants such as preservative, emulsifying agents and dispersing agents. Prevention of the action of microorganisms may be ensured by the inclusion of various antibacterial and antifungal agents. [0148] The pharmaceutical composition may be in any suitable form, (depending upon the desired method of administration). It may be provided in unit dosage form, may be provided in a sealed container and may be provided as part of a kit. Such a kit may include instructions for use. It may include a plurality of said unit dosage forms.

[0149] The pharmaceutical composition may be adapted for administration by any appropriate route, including a parenteral (e.g., subcutaneous, intramuscular, or intravenous) route. Such compositions may be prepared by any method known in the art of pharmacy, for example by mixing the active ingredient with the carrier(s) or excipient(s) under sterile conditions.

[0150] Dosages of the substances of the present disclosure can vary between wide limits, depending upon the disease or disorder to be treated, the age and condition of the individual to be treated, etc. and a physician will ultimately determine appropriate dosages to be used.

Production of Multispecific Antibodies that bind to CD47 and ICAM1

[0151] In some embodiments, polypeptides described herein (e.g., antibodies and its binding fragments) are produced using any method known in the art to be useful for the synthesis of polypeptides (e.g., antibodies), in particular, by chemical synthesis or by recombinant expression, and are preferably produced by recombinant expression techniques.

[0152] In some instances, an antibody or its binding fragment thereof is expressed recombinantly, and the nucleic acid encoding the antibody or its binding fragment is assembled from chemically synthesized oligonucleotides (e.g., as described in Kutmeier et al., 1994, BioTechniques 17:242), which involves the synthesis of overlapping oligonucleotides containing portions of the sequence encoding the antibody, annealing and ligation of those oligonucleotides, and then amplification of the ligated oligonucleotides by PCR.

[0153] Alternatively, a nucleic acid molecule encoding an antibody is optionally generated from a suitable source (e.g., an antibody cDNA library, or cDNA library generated from any tissue or cells expressing the immunoglobulin) by PCR amplification using synthetic primers hybridizable to the 3' and 5' ends of the sequence or by cloning using an oligonucleotide probe specific for the particular gene sequence.

[0154] In some instances, an antibody or its binding is optionally generated by immunizing an animal, such as a mouse, to generate polyclonal antibodies or, more preferably, by generating monoclonal antibodies, e.g., as described by Kohler and Milstein (1975, Nature 256:495-497) or, as described by Kozbor et al. (1983, Immunology Today 4:72) or Cole et al. (1985 in Monoclonal Antibodies and Cancer Therapy, Alan R. Liss, Inc., pp. 77-96). Alternatively, a clone encoding at least the Fab portion of the antibody is optionally obtained by screening Fab expression libraries (e.g., as described in Huse et al., 1989, Science 246: 1275-1281) for clones of Fab fragments that bind the specific antigen or by screening antibody libraries (See, e.g., Clackson et al., 1991, Nature 352:624; Hane et al., 1997 Proc. Natl. Acad. Sci. USA 94:4937).

[0155] In some embodiments, techniques developed for the production of “chimeric antibodies” (Morrison et al., 1984, Proc. Natl. Acad. Sci. 81:851-855; Neuberger et al., 1984, Nature 312:604-608; Takeda et al., 1985, Nature 314:452-454) by splicing genes from a mouse antibody molecule of appropriate antigen specificity together with genes from a human antibody molecule of appropriate biological activity are used. A chimeric antibody is a molecule in which different portions are derived from different animal species, such as those having a variable region derived from a murine monoclonal antibody and a human immunoglobulin constant region.

[0156] In some embodiments, techniques described for the production of single chain antibodies (U.S. Pat. No. 4,694,778; Bird, 1988, Science 242:423-42; Huston et al., 1988, Proc. Natl. Acad. Sci. USA 85:5879-5883; and Ward et al., 1989, Nature 334:544-54) are adapted to produce single chain antibodies. Single chain antibodies are formed by linking the heavy and light chain fragments of the Fv region via an amino acid bridge, resulting in a single chain polypeptide. Techniques for the assembly of functional Fv fragments in E. coli are also optionally used (Skerra et al., 1988, Science 242: 1038-1041).

[0157] In some embodiments, an expression vector comprising the nucleotide sequence of an antibody or the nucleotide sequence of an antibody is transferred to a host cell by conventional techniques (e.g., electroporation, liposomal transfection, and calcium phosphate precipitation), and the transfected cells are then cultured by conventional techniques to produce the antibody. In specific embodiments, the expression of the antibody is regulated by a constitutive, an inducible or a tissue, specific promoter.

[0158] In some embodiments, a variety of host-expression vector systems is utilized to express an antibody, or its binding fragment described herein. Such host-expression systems represent vehicles by which the coding sequences of the antibody is produced and subsequently purified, but also represent cells that are, when transformed or transfected with the appropriate nucleotide coding sequences, express an antibody or its binding fragment in situ. These include, but are not limited to, microorganisms such as bacteria (e.g., E. coli and B. subtilis) transformed with recombinant bacteriophage DNA, plasmid DNA or cosmid DNA expression vectors containing an antibody or its binding fragment coding sequences; yeast (e.g., Saccharomyces Pichia) transformed with recombinant yeast expression vectors containing an antibody or its binding fragment coding sequences; insect cell systems infected with recombinant virus expression vectors (e.g., baculovirus) containing an antibody or its binding fragment coding sequences; plant cell systems infected with recombinant virus expression vectors (e.g., cauliflower mosaic virus (CaMV) and tobacco mosaic virus (TMV)) or transformed with recombinant plasmid expression vectors (e.g., Ti plasmid) containing an antibody or its binding fragment coding sequences; or mammalian cell systems (e.g., COS, CHO, BH, 293, 293T, 3T3 cells) harboring recombinant expression constructs containing promoters derived from the genome of mammalian cells (e.g., metallothionein promoter) or from mammalian viruses (e.g. the adenovirus late promoter; the vaccinia virus 7.5K promoter).

[0159] For long-term, high-yield production of recombinant proteins, stable expression is preferred. In some instances, cell lines that stably express an antibody are optionally engineered. Rather than using expression vectors that contain viral origins of replication, host cells are transformed with DNA controlled by appropriate expression control elements (e.g., promoter, enhancer, sequences, transcription terminators, polyadenylation sites, etc.), and a selectable marker. Following the introduction of the foreign DNA, engineered cells are then allowed to grow for 1 -2 days in an enriched media, and then are switched to a selective media. The selectable marker in the recombinant plasmid confers resistance to the selection and allows cells to stably integrate the plasmid into their chromosomes and grow to form foci that in turn are cloned and expanded into cell lines. This method can advantageously be used to engineer cell lines which express the antibody or its binding fragments.

[0160] In some instances, a number of selection systems are used, including but not limited to the herpes simplex virus thymidine kinase (Wigler et al., 1977, Cell 11:223), hypoxanthine -guanine phosphoribosyltransferase (Szybalska & Szybalski, 192, Proc. Natl. Acad. Sci. USA 48:202), and adenine phosphoribosyltransferase (Lowy et al., 1980, Cell 22:817) genes are employed in tk-, hgprt- or aprt- cells, respectively. Also, antimetabolite resistance are used as the basis of selection for the following genes: dhfr, which confers resistance to methotrexate (Wigler et al., 1980, Proc. Natl. Acad. Sci. USA 77:357; O’Hare et al., 1981, Proc. Natl. Acad. Sci. USA 78: 1527); gpt, which confers resistance to mycophenolic acid (Mulligan & Berg, 1981, Proc. Natl. Acad. Sci. USA 78:2072); neo, which confers resistance to the aminoglycoside G-418 (Clinical Pharmacy 12:488-505; Wu and Wu, 1991, Biotherapy 3:87-95; Tolstoshev, 1993, Ann. Rev. Pharmacol. Toxicol. 32:573-596; Mulligan, 1993, Science 260:926- 932; and Morgan and Anderson, 1993, Ann. Rev. Biochem. 62: 191-217; May 1993, TIB TECH 11(5): 155-215) and hygro, which confers resistance to hygromycin (Santerre et al., 1984, Gene 30: 147). Methods commonly known in the art of recombinant DNA technology which can be used are described in Ausubel et al. (eds., 1993, Current Protocols in Molecular Biology, John Wiley & Sons, NY; Kriegler, 1990, Gene Transfer and Expression, A Laboratory Manual, Stockton Press, NY; and in Chapters 12 and 13, Dracopoli et al. (eds), 1994, Current Protocols in Human Genetics, John Wiley & Sons, NY.;

Colberre-Garapin et al., 1981, J. Mol. Biol. 150: 1).

[0161] In some instances, the expression levels of an antibody are increased by vector amplification (for a review, see Bebbington and Hentschel, the use of vectors based on gene amplification for the expression of cloned genes in mammalian cells in DNA cloning, Vol. 3. (Academic Press, New York, 1987)). When a marker in the vector system expressing an antibody is amplifiable, an increase in the level of inhibitor present in culture of host cell will increase the number of copies of the marker gene. Since the amplified region is associated with the nucleotide sequence of the antibody, production of the antibody will also increase (Crouse et al., 1983, Mol. Cell Biol. 3:257).

[0162] In some instances, any method known in the art for purification of an antibody is used, for example, by chromatography (e.g., ion exchange, affinity, particularly by affinity for the specific antigen after Protein A, and sizing column chromatography), centrifugation, differential solubility, or by any other standard technique for the purification of proteins.

Expression Vectors

[0163] In some embodiments, vectors include any suitable vectors derived from either a eukaryotic or prokaryotic sources. In some cases, vectors are obtained from bacteria (e.g. E. coli), insects, yeast (e.g. Pichia pastoris), algae, or mammalian sources. Exemplary bacterial vectors include pACYC177, pASK75, pBAD vector series, pBADM vector series, pET vector series, pETM vector series, pGEX vector series, pHAT, pHAT2, pMal-c2, pMal-p2, pQE vector series, pRSET A, pRSET B, pRSET C, pTrcHis2 series, pZA31-Luc, pZE21-MCS-l, pFLAG ATS, pFLAG CTS, pFLAG MAC, pFLAG Shift-12c, pTAC-MAT- 1, pFLAG CTC, or pTAC-MAT-2.

[0164] Exemplary insect vectors include pFastBacl, pFastBac DUAL, pFastBac ET, pFastBac HTa, pFastBac HTb, pFastBac HTc, pFastBac M30a, pFastBact M30b, pFastBac, M30c, pVL1392, pVL1393, pVL1393 MIO, pVL1393 Ml 1, pVL1393 M12, FLAG vectors such as pPolh-FLAGl or pPolh-MAT 2, or MAT vectors such as pPolh-MATl, or pPolh-MAT2.

[0165] In some cases, yeast vectors include Gateway® pDEST™ 14 vector, Gateway® pDEST™ 15 vector, Gateway® pDEST™ 17 vector, Gateway® pDEST™ 24 vector, Gateway® pYES-DEST52 vector, pBAD-DEST49 Gateway® destination vector, pAO815 Pichia vector, pFLDl Pichi pastoris vector, pGAPZA,B, & C Pichia pastoris vector, pPIC3.5K Pichia vector, pPIC6 A, B, & C Pichia vector, pPIC9K Pichia vector, pTEFl/Zeo, pYES2 yeast vector, pYES2/CT yeast vector, pYES2/NT A, B, & C yeast vector, or pYES3/CT yeast vector.

[0166] Exemplary algae vectors include pChlamy-4 vector or MCS vector.

[0167] Examples of mammalian vectors include transient expression vectors or stable expression vectors. Mammalian transient expression vectors may include pRK5, p3xFLAG-CMV 8, pFLAG-Myc-CMV 19, pFLAG-Myc-CMV 23, pFLAG-CMV 2, pFLAG-CMV 6a,b,c, pFLAG-CMV 5.1, pFLAG-CMV 5a,b,c, p3xFLAG-CMV 7.1, pFLAG-CMV 20, p3xFLAG-Myc-CMV 24, pCMV-FLAG-MATl, pCMV-FLAG- MAT2, pBICEP-CMV 3, or pBICEP-CMV 4. Mammalian stable expression vector may include pFLAG- CMV 3, p3xFLAG-CMV 9, p3xFLAG-CMV 13, pFLAG-Myc-CMV 21, p3xFLAG-Myc-CMV 25, pFLAG-CMV 4, p3xFLAG-CMV 10, p3xFLAG-CMV 14, pFLAG-Myc-CMV 22, p3xFLAG-Myc-CMV 26, pBICEP-CMV 1, or pBICEP-CMV 2.

[0168] In some instances, a cell-free system is a mixture of cytoplasmic and/or nuclear components from a cell and is used for in vitro nucleic acid synthesis. In some cases, a cell-free system utilizes either prokaryotic cell components or eukaryotic cell components. Sometimes, a nucleic acid synthesis is obtained in a cell-free system based on for example Drosophila cell, Xenopus egg, or HeLa cells.

Exemplary cell-free systems include, but are not limited to, E. coli S30 Extract system, E. coli T7 S30 system, or PURExpress®.

Host Cells

[0169] In some embodiments, a host cell includes any suitable cell such as a naturally derived cell or a genetically modified cell. In some instances, a host cell is a production host cell. In some instances, a host cell is a eukaryotic cell. In other instances, a host cell is a prokaryotic cell. In some cases, a eukaryotic cell includes fungi (e.g., yeast cells), animal cell or plant cell. In some cases, a prokaryotic cell is a bacterial cell. Examples of bacterial cell include gram-positive bacteria or gram-negative bacteria. Sometimes the gram-negative bacteria is anaerobic, rod-shaped, or both.

[0170] In some instances, gram-positive bacteria include Actinobacteria, Firmicutes or Tenericutes. In some cases, gram-negative bacteria include Aquificae, Deinococcus-Thermus, Fibrobacteres- Chlorobi/Bacteroidetes (FCB group), Fusobacteria, Gemmatimonadetes, Nitrospirae, Planctomycetes- Verrucomicrobia/ Chlamydiae (PVC group), Proteobacteria, Spirochaetes or Synergistetes. Other bacteria can be Acidobacteria, Chloroflexi, Chrysiogenetes, Cyanobacteria, Deferribacteres, Dictyoglomi, Thermodesulfobacteria or Thermotogae. A bacterial cell can be Escherichia coli, Clostridium botulinum, or Coli bacilli.

[0171] Exemplary prokaryotic host cells include, but are not limited to, BL21, Maehl™, DH10B™, TOPIO, DH5a, DHIOBac™, OmniMax™, MegaX™, DH12S™, INV110, TOP10F’, INVaF, TOP10/P3, ccdB Survival, PIR1, PIR2, Stbl2™, Stbl3™, or Stbl4™.

[0172] In some instances, animal cells include a cell from a vertebrate or from an invertebrate. In some cases, an animal cell includes a cell from a marine invertebrate, fish, insects, amphibian, reptile, or mammal. In some cases, a fungus cell includes a yeast cell, such as brewer’s yeast, baker’s yeast, or wine yeast.

[0173] Fungi include ascomycetes such as yeast, mold, filamentous fungi, basidiomycetes, or zygomycetes. In some instances, yeast includes Ascomycota or Basidiomycota. In some cases, Ascomycota includes Saccharomycotina (true yeasts, e.g. Saccharomyces cerevisiae (baker’s yeast)) or Taphrinomycotina (e.g. Schizosaccharomycetes (fission yeasts)). In some cases, Basidiomycota includes Agaricomycotina (e.g. Tremellomycetes) or Pucciniomycotina (e.g. Microbotryomycetes).

[0174] Exemplary yeast or filamentous fungi include, for example, the genus: Saccharomyces, Schizosaccharomyces, Candida, Pichia, Hansenula, Kluyveromyces, Zygosaccharomyces, Yarrowia, Trichosporon, Rhodosporidi, Aspergillus, Fusarium, or Trichoderma. Exemplary yeast or filamentous fungi include, for example, the species: Saccharomyces cerevisiae, Schizosaccharomyces pombe, Candida utilis, Candida boidini, Candida albicans, Candida tropicalis, Candida stellatoidea, Candida glabrata, Candida krusei, Candida parapsilosis, Candida guilliermondii, Candida viswanathii, Candida lusitaniae, Rhodotorula mucilaginosa, Pichia metanolica, Pichia angusta, Pichia pastoris, Pichia anomala, Hansenula polymorpha, Kluyveromyces lactis, Zygosaccharomyces rouxii, Yarrowia lipolytica, Trichosporon pullulans, Rhodosporidium toru-Aspergillus niger, Aspergillus nidulans, Aspergillus awamori, Aspergillus oryzae, Trichoderma reesei, Yarrowia lipolytica, Brettanomyces bruxellensis, Candida 58xpress58, Schizosaccharomyces pombe, Torulaspora delbrueckii, Zygosaccharomyces bailii, Cryptococcus neoformans, Cryptococcus gattii, or Saccharomyces boulardii.

[0175] Exemplary yeast host cells include, but are not limited to, Pichia pastoris yeast strains such as GS115, KM71H, SMD1168, SMD1168H, and X-33; and Saccharomyces cerevisiae yeast strain such as INVScl.

[0176] In some instances, additional animal cells include cells obtained from a mollusk, arthropod, annelid or sponge. In some cases, an additional animal cell is a mammalian cell, e.g., from a primate, ape, equine, bovine, porcine, canine, feline or rodent. In some cases, a rodent includes mouse, rat, hamster, gerbil, hamster, chinchilla, fancy rat, or guinea pig.

[0177] Exemplary mammalian host cells include, but are not limited to, 293A cell line, 293FT cell line, 293F cells , 293 H cells, CHO DG44 cells, CHO-S cells, CHO-K1 cells, FUT8 KO CHOK1, Expi293F™ cells, Flp-In™ T-REx™ 293 cell line, Flp-In™-293 cell line, Flp-In™-3T3 cell line, Flp-In™-BHK cell line, Flp-In™-CHO cell line, Flp-In™-CV-l cell line, Flp-In™-Jurkat cell line, FreeStyle™ 293-F cells, FreeStyle™ CHO-S cells, GripTite™ 293 MSR cell line, GS-CHO cell line, HepaRG™ cells, T-REx™ Jurkat cell line, Per.C6 cells, T-REx™-293 cell line, T-REx™-CHO cell line, and T-REx™-HeLa cell line.

[0178] In some instances, a mammalian host cell is a stable cell line, or a cell line that has incorporated a genetic material of interest into its own genome and has the capability to express the product of the genetic material after many generations of cell division. In some cases, a mammalian host cell is a transient cell line, or a cell line that has not incorporated a genetic material of interest into its own genome and does not have the capability to express the product of the genetic material after many generations of cell division.

[0179] Exemplary insect host cells include, but are not limited to, Drosophila S2 cells, Sf9 cells, Sf21 cells, High Five™ cells, and 59xpress+® cells.

[0180] In some instances, plant cells include a cell from algae. Exemplary insect cell lines include, but are not limited to, strains from Chlamydomonas reinhardtii 137c, or Synechococcus elongatus PPC 7942. Articles of Manufacture

[0181] In another aspect of the invention, an article of manufacture containing materials useful for the treatment, prevention and/or diagnosis of the disorders described above is provided. The article of manufacture comprises a container and a label or package insert on or associated with the container. Suitable containers include, for example, bottles, vials, syringes, IV solution bags, etc. The containers may be formed from a variety of materials such as glass or plastic. The container holds a composition which is by itself or combined with another composition effective for treating, preventing and/or diagnosing the condition and may have a sterile access port (for example the container may be an intravenous solution bag or a vial having a stopper that is pierceable by a hypodermic injection needle). At least one active agent in the composition is a bispecific antibody comprising a first antigen-binding site that specifically binds to CD47 and a second antigen -binding site that specifically binds to ICAM1 as defined herein before.

[0182] The label or package insert indicates that the composition is used for treating the condition of choice. Moreover, the article of manufacture may comprise (a) a first container with a composition contained therein, wherein the composition comprises the bispecific antibody of the invention; and (b) a second container with a composition contained therein, wherein the composition comprises a further cytotoxic or otherwise therapeutic agent. The article of manufacture in this embodiment of the invention may further comprise a package insert indicating that the compositions can be used to treat a particular condition.

[0183] Alternatively, or additionally, the article of manufacture may further comprise a second (or third) container comprising a pharmaceutically-acceptable buffer, such as bacteriostatic water for injection (BWFI), phosphate-buffered saline, Ringer’s solution and dextrose solution. It may further include other materials desirable from a commercial and user standpoint, including other buffers, diluents, filters, needles, and syringes. Methods of Use

[0184] In some embodiments, are methods of treating cancer in a subject in need thereof comprising administering to the subject a multispecific antibody as disclosed herein that binds to CD47 and ICAM1. In some embodiments, the cancer comprises cancer cells that express CD47 and ICAM1. In some embodiments, the cancer cells express more ICAM1 protein molecules on its cell surface than CD47 protein molecules on its cell surface. In some embodiments, the ratio of CD47 protein molecules to ICAM1 protein molecules on its cell surface is at least about 1: 1, 1: 1.5, 1:2.0, 1:2.5, 1:5, 1: 10, 1: 15, 1:20, 1:50, 1: 100, or 1:200. In some embodiments, the multispecific antibody induces the cancer cells that express CD47 and ICAM1 to be lysed. In some embodiments, the multispecific antibody induces antibody-dependent cellular cytotoxicity (ADCC) mediated killing of the cancer cells that express CD47 and ICAM1. In some embodiments, the multispecific antibody induces complement-dependent cytotoxicity (CDC) mediated killing of the cancer cells that express CD47 and ICAM1. In some embodiments, the multispecific antibody induces antibody-dependent cellular phagocytosis (ADCP) of the cancer cells that express CD47 and ICAM1. In some embodiments, the cancer is a hematological malignancy. In some embodiments, the cancer is multiple myeloma. In some embodiments, the cancer is B cell cancer. In some embodiments, the cancer is leukemia or lymphoma. In some embodiments, the cancer is lymphoma. In some embodiments, the lymphoma is B-cell lymphoma. In some embodiments, the hematological malignancy is T cell cancer. In some embodiments, the T cell cancer is cutaneous T cell lyphoma, or anaplastic large cell lymphoma. In some embodiments, the cancer is a solid tumor. In some embodiments, the solid tumor is sarcoma, breast cancer, lung cancer, kidney cancer, melanoma, head and neck cancer, ovarian cancer, liver cancer, bladder cancer, thyroid cancer, or carcinoma. In some embodiments, the solid tumor is lung cancer. In some embodiments, the lung cancer is non-small cell lung cancer. In some embodiments, the method further comprises administering to the subject an anti -cancer agent. In some embodiments, the anti-cancer agent is a chemotherapeutic agent or a biologic agent. In some embodiments, the administering is sufficient to reduce or eliminate the cancer as compared to a comparable method lacking the administering. In some embodiments, the reduction is at least about 1- fold, 5 fold, 10 fold, 20 fold, 40 fold, 60 fold, 80 fold, or up to about 100 fold.

[0185] While preferred embodiments of the present invention have been shown and described herein, it will be obvious to those skilled in the art that such embodiments are provided by way of example only. Numerous variations, changes, and substitutions will now occur to those skilled in the art without departing from the invention. It should be understood that various alternatives to the embodiments of the invention described herein may be employed in practicing the invention. It is intended that the following claims define the scope of the invention and that methods and structures within the scope of these claims and their equivalents be covered thereby.

List of Embodiments

[0186] The following list of embodiments of the invention are to be considered as disclosing various features of the invention, which features can be considered to be specific to the particular embodiment under which they are discussed, or which are combinable with the various other features as listed in other embodiments. Thus, simply because a feature is discussed under one particular embodiment does not necessarily limit the use of that feature to that embodiment.

[0187] Embodiment 1. A multispecific antibody comprising a CD47 binding domain and an Intercellular Adhesion Molecule 1 (ICAM1) binding domain.

[0188] Embodiment 2. The multispecific antibody of Embodiment 1, wherein the multispecific antibody is bispecific, trispecific, or tetraspecific.

[0189] Embodiment 3. The multispecific antibody of Embodiment 2, wherein the multispecific antibody is bispecific.

[0190] Embodiment 4. The multispecific antibody of Embodiment 1, wherein the multispecific antibody is bivalent, trivalent, or tetravalent.

[0191] Embodiment 5. The multispecific antibody of Embodiment 4, wherein the multispecific antibody is bivalent.

[0192] Embodiment 6. The multispecific antibody of any one of Embodiments 1-5, wherein the CD47 binding domain comprises an antibody, or functional fragment or functional variant thereof, that binds specifically to CD47.

[0193] Embodiment 7. The multispecific antibody of Embodiment 6, wherein the antibody, or functional fragment or functional variant thereof that binds specifically to CD47 comprises an anti-CD47 heavy chain and an anti-CD47 light chain.

[0194] Embodiment 8. The multispecific antibody of Embodiment 7, wherein the anti-CD47 heavy chain comprises an anti-CD47 heavy chain variable domain.

[0195] Embodiment 9. The multispecific antibody of Embodiment 7, wherein the anti-CD47 heavy chain variable domain comprises a variable domain of an IgGl, IgG2, IgG3, or IgG4 heavy chain.

[0196] Embodiment 10. The multispecific antibody of any one of Embodiments 7-9, wherein the anti- CD47 light chain comprises an anti-CD47 light chain variable domain.

[0197] Embodiment 11. The multispecific antibody of Embodiment 10, wherein the anti-CD47 light chain variable domain comprises a variable domain of a Kappa or Lambda light chain.

[0198] Embodiment 12. The multispecific antibody of Embodiments 8-11, wherein the anti-CD47 heavy chain variable domain comprises the variable domain of an IgGl heavy chain and the anti-CD47 light chain variable domain comprises the variable domain of a Kappa or Lambda light chain.

[0199] Embodiment 13. The multispecific antibody according to any one of Embodiments 6-12, wherein the antibody, or functional fragment or functional variant thereof, that binds specifically to CD47 comprises a single-chain variable fragment (scFv) or an antigen-binding fragment (Fab).

[0200] Embodiment 14. The multispecific antibody of Embodiment 8 or 9, wherein the anti-CD47 heavy chain variable domain comprises complementarity determining regions (CDRs): HC-CDR1, HC-CDR2, and HC-CDR3, and wherein the HC-CDR1, the HC-CDR2, and the HC-CDR3 of the anti-CD47 heavy chain variable domain comprise amino acid sequences according to HC-CDR1: SEQ ID NO: 1; HC- CDR2: SEQ ID NO: 2; HC-CDR3: SEQ ID NO: 3; and wherein the CDRs comprise from 0-2 amino acid modification(s) in at least one of the HC-CDR1, HC-CDR2, or HC-CDR3. [0201] Embodiment 15. The multispecific antibody of Embodiment 8 or 9, wherein the anti-CD47 heavy chain variable domain comprises complementarity determining regions (CDRs): HC-CDR1, HC-CDR2, and HC-CDR3, and wherein the HC-CDR1, the HC-CDR2, and the HC-CDR3 of the anti-CD47 heavy chain variable domain comprise amino acid sequences according to HC-CDR1: SEQ ID NO: 1; HC- CDR2: SEQ ID NO: 2; HC-CDR3: SEQ ID NO: 3; and wherein the CDRs comprise from 0-1 amino acid modification(s) in at least one of the HC-CDR1, HC-CDR2, or HC-CDR3.

[0202] Embodiment 16. The multispecific antibody of Embodiment 8 or 9, wherein the anti-CD47 heavy chain variable domain comprises complementarity determining regions (CDRs): HC-CDR1, HC-CDR2, and HC-CDR3, and wherein the HC-CDR1, the HC-CDR2, and the HC-CDR3 of the anti-CD47 heavy chain variable domain comprise amino acid sequences according to HC-CDR1: SEQ ID NO: 1; HC- CDR2: SEQ ID NO: 2; HC-CDR3: SEQ ID NO: 3.

[0203] Embodiment 17. The multispecific antibody of Embodiment 10 or 11, wherein the anti-CD47 light chain variable domain comprises complementarity determining regions (CDRs): LC-CDR1, LC- CDR2, and LC-CDR3, and wherein the LC-CDR1, the LC-CDR2, and the LC-CDR3 of the anti-CD47 light chain variable domain comprises amino acid sequences according to LC-CDR1: SEQ ID NO: 4; LC-CDR2: SEQ ID NO: 5; LC-CDR3: SEQ ID NO: 6; and wherein the CDRs comprise from 0-2 amino acid modification(s) in at least one of the LC-CDR1, LC-CDR2, or LC-CDR3.

[0204] Embodiment 18. The multispecific antibody of Embodiment 10 or 11, wherein the anti-CD47 light chain variable domain comprises complementarity determining regions (CDRs): LC-CDR1, LC- CDR2, and LC-CDR3, and wherein the LC-CDR1, the LC-CDR2, and the LC-CDR3 of the anti-CD47 light chain variable domain comprises amino acid sequences according to LC-CDR1: SEQ ID NO: 4; LC-CDR2: SEQ ID NO: 5; LC-CDR3: SEQ ID NO: 6; and wherein the CDRs comprise from 0-1 amino acid modification(s) in at least one of the LC-CDR1, LC-CDR2, or LC-CDR3.

[0205] Embodiment 19. The multispecific antibody of Embodiment 10 or 11, wherein the anti-CD47 light chain variable domain comprises complementarity determining regions (CDRs): LC-CDR1, LC- CDR2, and LC-CDR3, and wherein the LC-CDR1, the LC-CDR2, and the LC-CDR3 of the anti-CD47 light chain variable domain comprises amino acid sequences according to LC-CDR1: SEQ ID NO: 4; LC-CDR2: SEQ ID NO: 5; LC-CDR3: SEQ ID NO: 6.

[0206] Embodiment 20. The multispecific antibody of Embodiment 7, wherein the HC-CDR1, the HC- CDR2, and the HC-CDR3 of the anti-CD47 heavy chain variable domain comprises amino acid sequences according to HC-CDR1: SEQ ID NO: 1; HC-CDR2: SEQ ID NO: 2; HC-CDR3: SEQ ID NO: 3 and the LC-CDR1, the LC-CDR2, and the LC-CDR3 of the anti-CD47 light chain variable domain comprises amino acid sequences according to LC-CDR1: SEQ ID NO: 4; LC-CDR2: SEQ ID NO: 5; LC-CDR3: SEQ ID NO: 6; wherein the CDRs comprise from 0-2 amino acid modification(s) in at least one of the HC-CDR1, HC-CDR2, or HC-CDR3; and wherein the CDRs comprise from 0-2 amino acid modification(s) in at least one of the LC-CDR1, LC-CDR2, or LC-CDR3

[0207] Embodiment 21. The multispecific antibody of Embodiment 7, wherein the HC-CDR1, the HC- CDR2, and the HC-CDR3 of the anti-CD47 heavy chain variable domain comprises amino acid sequences according to HC-CDR1: SEQ ID NO: 1; HC-CDR2: SEQ ID NO: 2; HC-CDR3: SEQ ID NO: 3 and the LC-CDR1, the LC-CDR2, and the LC-CDR3 of the anti-CD47 light chain variable domain comprises amino acid sequences according to LC-CDR1: SEQ ID NO: 4; LC-CDR2: SEQ ID NO: 5; LC-CDR3: SEQ ID NO: 6; wherein the CDRs comprise from 0-1 amino acid modification(s) in at least one of the HC-CDR1, HC-CDR2, or HC-CDR3; and wherein the CDRs comprise from 0-1 amino acid modification(s) in at least one of the LC-CDR1, LC-CDR2, or LC-CDR3

[0208] Embodiment 22. The multispecific antibody of Embodiment 7, wherein the HC-CDR1, the HC- CDR2, and the HC-CDR3 of the anti-CD47 heavy chain variable domain comprises amino acid sequences according to HC-CDR1: SEQ ID NO: 1; HC-CDR2: SEQ ID NO: 2; HC-CDR3: SEQ ID NO: 3 and the LC-CDR1, the LC-CDR2, and the LC-CDR3 of the anti-CD47 light chain variable domain comprises amino acid sequences according to LC-CDR1: SEQ ID NO: 4; LC-CDR2: SEQ ID NO: 5; LC-CDR3: SEQ ID NO: 6.

[0209] Embodiment 23. The multispecific antibody of Embodiment 7, wherein the antibody, or functional fragment or functional variant thereof that binds specifically to CD47 comprises the scFv, and the antibody, or functional fragment or functional variant thereof, that binds specifically to ICAM1 comprises the Fab, and wherein the HC-CDR1, the HC-CDR2, and the HC-CDR3 of the anti-CD47 heavy chain variable domain comprises amino acid sequences according to HC-CDR1: SEQ ID NO: 1; HC- CDR2: SEQ ID NO: 2; HC-CDR3: SEQ ID NO: 3 and the LC-CDR1, the LC-CDR2, and the LC-CDR3 of the anti-CD47 light chain variable domain comprises amino acid sequences according to LC-CDR1: SEQ ID NO: 4; LC-CDR2: SEQ ID NO: 5; LC-CDR3: SEQ ID NO: 6.

[0210] Embodiment 24. The multispecific antibody of Embodiment 7, wherein the antibody, or functional fragment or functional variant thereof that binds specifically to CD47 comprises the Fab, and the antibody, or functional fragment or functional variant thereof, that binds specifically to ICAM1 comprises the scFv, and wherein the HC-CDR1, the HC-CDR2, and the HC-CDR3 of the anti-CD47 heavy chain variable domain comprises amino acid sequences according to HC-CDR1: SEQ ID NO: 1; HC-CDR2: SEQ ID NO: 2; HC-CDR3: SEQ ID NO: 3 and the LC-CDR1, the LC-CDR2, and the LC- CDR3 of the anti-CD47 light chain variable domain comprises amino acid sequences according to LC- CDR1: SEQ ID NO: 4; LC-CDR2: SEQ ID NO: 5; LC-CDR3: SEQ ID NO: 6.

[0211] Embodiment 25. The multispecific antibody of Embodiment 8 or 9, wherein the anti-CD47 heavy chain comprises an amino acid sequence with at least 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to of SEQ ID NO: A.

[0212] Embodiment 26. The multispecific antibody of Embodiment 8 or 9, wherein the anti-CD47 heavy chain comprises an amino acid sequence with at least 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to of SEQ ID NO: 7.

[0213] Embodiment 27. The multispecific antibody of Embodiment 10 or 11, wherein the anti-CD47 light chain comprises an amino acid sequence with at least 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID

NO: B.

[0214] Embodiment 28. The multispecific antibody of Embodiment 7, wherein the anti-CD47 heavy chain comprises an amino acid sequence with at least 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%,

88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NO: A; and the anti-CD47 light chain comprises an amino acid sequence with at least 75%, 80%, 81%, 82%,

83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or

100% identity to SEQ ID NO: B.

[0215] Embodiment 29. The multispecific antibody of Embodiment 13, wherein the anti-CD47 heavy chain variable domain comprises a single-chain variable fragment (scFv) comprising an amino acid sequence with at least 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NOM E

[0216] Embodiment 30. The multispecific antibody of Embodiment 10 or 11, wherein the anti-CD47 light chain comprises an amino acid sequence with at least 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NO: 9.

[0217] Embodiment 31. The multispecific antibody of Embodiment 7, wherein the anti-CD47 heavy chain comprises an amino acid sequence with at least 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NO: 7 and the anti-CD47 light chain comprises an amino acid sequence with at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NO: 9.

[0218] Embodiment 32. The multispecific antibody of any one of Embodiments 1-28, wherein the ICAM1 binding domain comprises an antibody, or functional fragment or functional variant thereof, that binds specifically to ICAM1.

[0219] Embodiment 33. The multispecific antibody of any one of Embodiments 1-32, wherein the antibody, or functional fragment or functional variant thereof that binds specifically to ICAM1 comprises an anti-ICAMl heavy chain and an anti-ICAMl light chain.

[0220] Embodiment 34. The multispecific antibody of Embodiment 33, wherein the anti-ICAMl heavy chain comprises an anti-ICAMl heavy chain variable domain.

[0221] Embodiment 35. The multispecific antibody of Embodiment 34, wherein the anti-ICAMl heavy chain variable domain comprises a variable domain of an IgGl, IgG2, IgG3, or IgG4 heavy chain.

[0222] Embodiment 36. The multispecific antibody of any one of Embodiments 33-35, wherein the anti- ICAMl light chain comprises an anti-ICAMl light chain variable domain.

[0223] Embodiment 37. The multispecific antibody of Embodiment 36, wherein the anti-ICAMl light chain variable domain comprises a variable domain of a Kappa or Lambda light chain.

[0224] Embodiment 38. The multispecific antibody of any one of Embodiments 28-37, wherein the anti- ICAMl heavy chain variable domain comprises the variable domain of an IgGl heavy chain and the anti- ICAMl light chain variable domain comprises the variable domain of a Kappa or Lambda light chain. [0225] Embodiment 39. The multispecific antibody according to any of Embodiments 1-32, wherein the antibody, or functional fragment or functional variant thereof, that binds specifically to ICAM1 comprises a single-chain variable fragment (scFv) or an antigen-binding fragment (Fab).

[0226] Embodiment 40. The multispecific antibody of Embodiment 34 or 35, wherein the anti-ICAMl heavy chain variable domain comprises complementarity determining regions (CDRs): HC-CDR1, HC- CDR2, and HC-CDR3, and wherein the HC-CDR1, the HC-CDR2, and the HC-CDR3 of the anti-ICAMl heavy chain variable domain comprise amino acid sequences according to HC-CDR1: SEQ ID NO: 19, 25, 46, 49, 52, 55, 58, 61, 64, 67, 70, 73, 76, 79, 82, 85, 88, 91, 94, 97, 100, 103, 106, or 109; HC-CDR2: SEQ ID NO: 20, 26, 47, 50, 53, 56, 59, 62, 65, 68, 71, 74, 77, 80, 83, 86, 89, 92, 95, 98, 101, 104, or 107; HC-CDR3: SEQ ID NO: 21, 27, 48, 51, 54, 57, 60, 63, 66, 69, 72, 75, 78, 81, 84, 87, 90, 93, 96, 99, 102, 105, 108, or 111; and wherein the CDRs comprise from 0-2 amino acid modification(s) in at least one of the HC-CDR1, HC-CDR2, or HC-CDR3.

[0227] Embodiment 41. The multispecific antibody of Embodiment 34 or 35, wherein the anti-ICAMl heavy chain variable domain comprises complementarity determining regions (CDRs): HC-CDR1, HC- CDR2, and HC-CDR3, and wherein the HC-CDR1, the HC-CDR2, and the HC-CDR3 of the anti-ICAMl heavy chain variable domain comprise amino acid sequences according to HC-CDR1: SEQ ID NO: 25; HC-CDR2: SEQ ID NO: 26; HC-CDR3: SEQ ID NO: 27; and wherein the CDRs comprise from 0-2 amino acid modification(s) in at least one of the HC-CDR1, HC-CDR2, or HC-CDR3.

[0228] Embodiment 42. The multispecific antibody of Embodiment 34 or 35, wherein the anti-ICAMl heavy chain variable domain comprises complementarity determining regions (CDRs): HC-CDR1, HC- CDR2, and HC-CDR3, and wherein the HC-CDR1, the HC-CDR2, and the HC-CDR3 of the anti-ICAMl heavy chain variable domain comprise amino acid sequences according to HC-CDR1: SEQ ID NO: 25; HC-CDR2: SEQ ID NO: 26; HC-CDR3: SEQ ID NO: 27; and wherein the CDRs comprise from 0-1 amino acid modification(s) in at least one of the HC-CDR1, HC-CDR2, or HC-CDR3.

[0229] Embodiment 43. The multispecific antibody of Embodiment 34 or 35, wherein the anti-ICAMl heavy chain variable domain comprises complementarity determining regions (CDRs): HC-CDR1, HC- CDR2, and HC-CDR3, and wherein the HC-CDR1, the HC-CDR2, and the HC-CDR3 of the anti-ICAMl heavy chain variable domain comprise amino acid sequences according to HC-CDR1: SEQ ID NO: 25; HC-CDR2: SEQ ID NO: 26; HC-CDR3: SEQ ID NO: 27.

[0230] Embodiment 44. The multispecific antibody of Embodiment 36 or 37, wherein the anti-ICAMl light chain variable domain comprises complementarity determining regions (CDRs): LC-CDR1, LC- CDR2, and LC-CDR3, and wherein the LC-CDR1, the LC-CDR2, and the LC-CDR3 of the anti-ICAMl light chain variable domain comprises amino acid sequences according to LC-CDR1: SEQ ID NO: 22, 28, 112, 115, 118, 121, 124, 127, 130, 133, 136, 139, 142, 145, 148, 151, 154, 157, 160, or 163; LC- CDR2: SEQ ID NO: 23, 29, 110, 113, 116, 119, 122, 125, 128, 131, 134, 137, 140, 143, 146, 149, 152, 155, 158, 161, or 164; LC-CDR3: SEQ ID NO: 24, 30, 114, 117, 120, 123, 126, 129, 132, 134, 138, 141, 144, 147, 150, 153, 156, 159, 162, or 165; and wherein the CDRs comprise from 0-2 amino acid modification(s) in at least one of the LC-CDR1, LC-CDR2, or LC-CDR3. [0231] Embodiment 45. The multispecific antibody of Embodiment 36 or 37, wherein the anti-ICAMl light chain variable domain comprises complementarity determining regions (CDRs): LC-CDR1, LC- CDR2, and LC-CDR3, and wherein the LC-CDR1, the LC-CDR2, and the LC-CDR3 of the anti-ICAMl light chain variable domain comprises amino acid sequences according to LC-CDR1: SEQ ID NO: 28; LC-CDR2: SEQ ID NO: 29; LC-CDR3: SEQ ID NO: 30; and wherein the CDRs comprise from 0-2 amino acid modification(s) in at least one of the LC-CDR1, LC-CDR2, or LC-CDR3.

[0232] Embodiment 46. The multispecific antibody of Embodiment 36 or 37, wherein the anti-ICAMl light chain variable domain comprises complementarity determining regions (CDRs): LC-CDR1, LC- CDR2, and LC-CDR3, and wherein the LC-CDR1, the LC-CDR2, and the LC-CDR3 of the anti-ICAMl light chain variable domain comprises amino acid sequences according to LC-CDR1: SEQ ID NO: 28; LC-CDR2: SEQ ID NO: 29; LC-CDR3: SEQ ID NO: 30; and wherein the CDRs comprise from 0-1 amino acid modification(s) in at least one of the LC-CDR1, LC-CDR2, or LC-CDR3.

[0233] Embodiment 47. The multispecific antibody of Embodiment 36 or 37, wherein the anti-ICAMl light chain variable domain comprises complementarity determining regions (CDRs): LC-CDR1, LC- CDR2, and LC-CDR3, and wherein the LC-CDR1, the LC-CDR2, and the LC-CDR3 of the anti-ICAMl light chain variable domain comprises amino acid sequences according to LC-CDR1: SEQ ID NO: 28; LC-CDR2: SEQ ID NO: 29; LC-CDR3: SEQ ID NO: 30.

[0234] Embodiment 48. The multispecific antibody of Embodiment 33, wherein the HC-CDR1, the HC- CDR2, and the HC-CDR3 of the anti-ICAMl heavy chain variable domain comprises amino acid sequences according to HC-CDR1: SEQ ID NO: 19, 25, 46, 49, 52, 55, 58, 61, 64, 67, 70, 73, 76, 79, 82, 85, 88, 91, 94, 97, 100, 103, 106, or 109; HC-CDR2: SEQ ID NO: 20, 26, 47, 50, 53, 56, 59, 62, 65, 68, 71, 74, 77, 80, 83, 86, 89, 92, 95, 98, 101, 104, or 107; HC-CDR3: SEQ ID NO: 21, 27, 48, 51, 54, 57, 60, 63, 66, 69, 72, 75, 78, 81, 84, 87, 90, 93, 96, 99, 102, 105, 108, or 111 and the LC-CDRl, the LC-CDR2, and the LC-CDR3 of the anti-ICAMl light chain variable domain comprises amino acid sequences according to LC-CDR1: SEQ ID NO: 22, 28, 112, 115, 118, 121, 124, 127, 130, 133, 136, 139, 142, 145, 148, 151, 154, 157, 160, or 163; LC-CDR2: SEQ ID NO: 23, 29, 110, 113, 116, 119, 122, 125, 128, 131, 134, 137, 140, 143, 146, 149, 152, 155, 158, 161, or 164; LC-CDR3: SEQ ID NO: 24, 30, 114, 117, 120, 123, 126, 129, 132, 134, 138, 141, 144, 147, 150, 153, 156, 159, 162, or 165.

[0235] Embodiment 49. The multispecific antibody of Embodiment 33, wherein the HC-CDR1, the HC- CDR2, and the HC-CDR3 of the anti-ICAMl heavy chain variable domain comprises amino acid sequences according to HC-CDR1: SEQ ID NO: 25; HC-CDR2: SEQ ID NO: 26; HC-CDR3: SEQ ID NO: 27; and the LC-CDR1, the LC-CDR2, and the LC-CDR3 of the anti-ICAMl light chain variable domain comprises amino acid sequences according to LC-CDR1: SEQ ID NO: 28; LC-CDR2: SEQ ID NO: 29; LC-CDR3: SEQ ID NO: 30; wherein the CDRs comprise from 0-2 amino acid modification(s) in at least one of the HC-CDR1, HC-CDR2, or HC-CDR3; and wherein the CDRs comprise from 0-2 amino acid modification(s) in at least one of the LC-CDR1, LC-CDR2, or LC-CDR3.

[0236] Embodiment 50. The multispecific antibody of Embodiment 33, wherein the HC-CDR1, the HC- CDR2, and the HC-CDR3 of the anti-ICAMl heavy chain variable domain comprises amino acid sequences according to HC-CDR1: SEQ ID NO: 25; HC-CDR2: SEQ ID NO: 26; HC-CDR3: SEQ ID NO: 27; and the LC-CDR1, the LC-CDR2, and the LC-CDR3 of the anti-ICAMl light chain variable domain comprises amino acid sequences according to LC-CDR1: SEQ ID NO: 28; LC-CDR2: SEQ ID NO: 29; LC-CDR3: SEQ ID NO: 30; wherein the CDRs comprise from 0-1 amino acid modification(s) in at least one of the HC-CDR1, HC-CDR2, or HC-CDR3; and wherein the CDRs comprise from 0-1 amino acid modification(s) in at least one of the LC-CDR1, LC-CDR2, or LC-CDR3.

[0237] Embodiment 51. The multispecific antibody of Embodiment 33, wherein the HC-CDR1, the HC- CDR2, and the HC-CDR3 of the anti-ICAMl heavy chain variable domain comprises amino acid sequences according to HC-CDR1: SEQ ID NO: 25; HC-CDR2: SEQ ID NO: 26; HC-CDR3: SEQ ID NO: 27; and the LC-CDR1, the LC-CDR2, and the LC-CDR3 of the anti-ICAMl light chain variable domain comprises amino acid sequences according to LC-CDR1: SEQ ID NO: 28; LC-CDR2: SEQ ID NO: 29; LC-CDR3: SEQ ID NO: 30.

[0238] Embodiment 52. The multispecific antibody of Embodiment 33, wherein the anti-ICAMl heavy chain comprises an amino acid sequence with at least 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to any one of SEQ ID NOs: 35, 36, 41, 166-187, 206-227.

[0239] Embodiment 53. The multispecific antibody of Embodiment 33, wherein the anti-ICAMl heavy chain comprises an amino acid sequence with at least 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to of SEQ ID NO: C.

[0240] Embodiment 54. The multispecific antibody of Embodiment 33, wherein the anti-ICAMl light chain comprises an amino acid sequence with at least 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%,

88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to any one of SEQ

ID NOs: 39, 40, 44, 45, 188-205, 228-245.

[0241] Embodiment 55. The multispecific antibody of Embodiment 33, wherein the anti-ICAMl light chain comprises an amino acid sequence with at least 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%,

88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NO: D.

[0242] Embodiment 56. The multispecific antibody of Embodiment 33, wherein the anti-ICAMl heavy chain comprises an amino acid sequence with at least 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%,

88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity at least 75%, 80%,

81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%,

99%, or 100% identity to any one of SEQ ID NOs: 35, 36, 37, 38, 41, 42, 43, 166-187, 206-227 and the anti-ICAMl light chain comprises an amino acid sequence with at least 75%, 80%, 81%, 82%, 83%, 84%,

85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to any one of SEQ ID NOs: 39, 40, 44, 45, 188-205, 228-245.

[0243] Embodiment 57. The multispecific antibody of Embodiment 33, wherein the anti-ICAMl heavy chain comprises an amino acid sequence with at least 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%,

88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NO: C; and the anti-ICAMl light chain comprises an amino acid sequence with at least 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NO: D.

[0244] Embodiment 58. The multispecific antibody of Embodiment 33, wherein the anti-ICAMl heavy chain comprises an amino acid sequence with at least 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NO: 41. [0245] Embodiment 59. The multispecific antibody of Embodiment 33, wherein the anti-ICAMl light chain comprises an amino acid sequence with at least 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NO: 44. [0246] Embodiment 60. The multispecific antibody of Embodiment 33, wherein the anti-ICAMl heavy chain comprises an amino acid sequence with at least 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NO: 41; and wherein the anti-ICAMl light chain comprises an amino acid sequence with at least 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NO: 44.

[0247] Embodiment 61. The multispecific antibody of any one of Embodiments 13-60, wherein the antibody, or functional fragment or functional variant thereof, that binds specifically to CD47 comprises the Fab, and the antibody, or functional fragment or functional variant thereof, that binds specifically to ICAM1 comprises the scFv.

[0248] Embodiment 63. The multispecific antibody of Embodiment 61, wherein the anti-CD47 heavy chain comprises an amino acid sequence with at least 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NO: A, and the anti-CD47 light chain comprises an amino acid sequence with at least 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NO: B; and the scFv comprises an amino acid sequence with at least 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NO: 11.

[0249] Embodiment 64. The multispecific antibody of any one of Embodiments 13-60, wherein the anti- CD47 heavy chain comprises an scFv that comprises an amino acid sequence with at least 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NO: 11.

[0250] Embodiment 65. The multispecific antibody of any one of Embodiments 13-60, wherein the antibody, or functional fragment or functional variant thereof, that binds specifically to CD47 comprises the scFv, and the antibody, or functional fragment or functional variant thereof, that binds specifically to ICAM1 comprises the Fab.

[0251] Embodiment 66. The multispecific antibody of Embodiment 65, wherein the anti-ICAMl heavy chain comprises an amino acid sequence with at least 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NO: 251 and the anti-ICAMl light chain comprises an amino acid sequence with at least 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NO: 252; and the scFv comprises an amino acid sequence with at least 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NO: 253.

[0252] Embodiment 67. The multispecific antibody of Embodiment 65, wherein the anti-ICAMl heavy chain comprises an amino acid sequence with at least 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NO: 254 and the anti-ICAMl light chain comprises an amino acid sequence with at least 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NO: 255.

[0253] Embodiment 68. The multispecific antibody of Embodiment 65, wherein the anti-ICAMl heavy chain comprises an amino acid sequence with at least 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NO: 254 and the anti-ICAMl light chain comprises an amino acid sequence with at least 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NO: 255; and the scFv comprises an amino acid sequence with at least 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NO: 256.

[0254] Embodiment 70. The multispecific antibody of any one of Embodiments 1-69, wherein the multispecific antibody comprises at least 3 CDRs of an anti-CD47 binding domain selected from any one of SEQ ID NO: 1-6 and from 0-2 amino acid modification(s) thereof; and at least 3 CDRs of an anti- ICAMl binding domain selected from any one of SEQ ID NO: 19, 25, 46, 49, 52, 55, 58, 61, 64, 67, 70, 73, 76, 79, 82, 85, 88, 91, 94, 97, 100, 103, 106, 109, 20, 26, 47, 50, 53, 56, 59, 62, 65, 68, 71, 74, 77, 80, 83, 86, 89, 92, 95, 98, 101, 104, 107, 21, 27, 48, 51, 54, 57, 60, 63, 66, 69, 72, 75, 78, 81, 84, 87, 90, 93, 96, 99, 102, 105, 108, 111 and from 0-2 amino acid modification(s) thereof.

[0255] Embodiment 71. The multispecific antibody of any one of Embodiments 1 -69, wherein the multispecific antibody comprises at least 3 CDRs of an anti-CD47 binding domain selected from any one of SEQ ID NOs: 1-6, and from 0-2 amino acid modification(s) thereof; and at least 3 CDRs of an anti- ICAMl binding domain selected from any one of SEQ ID NO: 25-30, and from 0-2 amino acid modification(s) thereof.

[0256] Embodiment 72. The multispecific antibody of any one of Embodiments 1-69, wherein the multispecific antibody comprises at least 3 CDRs of an anti-CD47 binding domain selected from any one of SEQ ID NOs: 1-6, and from 0-1 amino acid modification(s) thereof; and at least 3 CDRs of an anti- ICAMl binding domain selected from any one of SEQ ID NO: 25-30, and from 0-1 amino acid modification(s) thereof.

[0257] Embodiment 73. The multispecific antibody of any one of Embodiments 1-69, wherein the multispecific antibody comprises at least 3 CDRs of an anti-CD47 binding domain selected from any one of SEQ ID NOs: 1-6; and at least 3 CDRs of an anti-ICAMl binding domain selected from any one of SEQ ID NO: 25-30.

[0258] Embodiment 74. The multispecific antibody of any one of Embodiments 1-69, wherein the multispecific antibody comprises 6 CDRs of an anti-CD47 binding domain having the sequences of SEQ ID NOs: 1-6, and from 0-2 amino acid modification(s) thereof; and 6 CDRs of an anti-ICAMl binding domain having the sequences of SEQ ID NO: 25-30, and from 0-2 amino acid modification(s) thereof. [0259] Embodiment 75. The multispecific antibody of any one of Embodiments 1-69, wherein the multispecific antibody comprises 6 CDRs of an anti-CD47 binding domain having the sequences of SEQ ID NOs: 1-6, and from 0-1 amino acid modification(s) thereof; and 6 CDRs of an anti-ICAMl binding domain having the sequences of SEQ ID NO: 25-30, and from 0-1 amino acid modification(s) thereof. [0260] Embodiment 76. The multispecific antibody of any one of Embodiments 1-69, wherein the multispecific antibody comprises 6 CDRs of an anti-CD47 binding domain having the sequences of SEQ ID NOs: 1-6; and 6 CDRs of an anti-ICAMl binding domain having the sequences of SEQ ID NO: 25-30. [0261] Embodiment 77. The multispecific antibody of any one of Embodiments 1-69, wherein the antibody, or functional fragment or functional variant thereof that binds specifically to CD47 comprises the scFv, and the antibody, or functional fragment or functional variant thereof, that binds specifically to ICAM1 comprises the Fab, and wherein the HC-CDR1, the HC-CDR2, and the HC-CDR3 of the anti- CD47 heavy chain variable domain comprises amino acid sequences according to HC-CDR1: SEQ ID NO: 1; HC-CDR2: SEQ ID NO: 2; HC-CDR3: SEQ ID NO: 3 and the LC-CDR1, the LC-CDR2, and the LC-CDR3 of the anti-CD47 light chain variable domain comprises amino acid sequences according to LC-CDR1: SEQ ID NO: 4; LC-CDR2: SEQ ID NO: 5; LC-CDR3: SEQ ID NO: 6, and wherein the HC- CDR1, the HC-CDR2, and the HC-CDR3 of the anti-ICAMl heavy chain variable domain comprises amino acid sequences according to HC-CDR1: SEQ ID NO: 25; HC-CDR2: SEQ ID NO: 26; HC-CDR3: SEQ ID NO: 27; and the LC-CDR1, the LC-CDR2, and the LC-CDR3 of the anti-ICAMl light chain variable domain comprises amino acid sequences according to LC-CDR1: SEQ ID NO: 28; LC-CDR2: SEQ ID NO: 29; LC-CDR3: SEQ ID NO: 30

[0262] Embodiment 78. The multispecific antibody of any one of Embodiments 1-69, wherein the antibody, or functional fragment or functional variant thereof that binds specifically to CD47 comprises the Fab, and the antibody, or functional fragment or functional variant thereof, that binds specifically to ICAM1 comprises the scFv, and wherein the HC-CDR1, the HC-CDR2, and the HC-CDR3 of the anti- CD47 heavy chain variable domain comprises amino acid sequences according to HC-CDR1: SEQ ID NO: 1; HC-CDR2: SEQ ID NO: 2; HC-CDR3: SEQ ID NO: 3 and the LC-CDR1, the LC-CDR2, and the LC-CDR3 of the anti-CD47 light chain variable domain comprises amino acid sequences according to LC-CDR1: SEQ ID NO: 4; LC-CDR2: SEQ ID NO: 5; LC-CDR3: SEQ ID NO: 6; and wherein the HC- CDR1, the HC-CDR2, and the HC-CDR3 of the anti-ICAMl heavy chain variable domain comprises amino acid sequences according to HC-CDR1: SEQ ID NO: 25; HC-CDR2: SEQ ID NO: 26; HC-CDR3: SEQ ID NO: 27; and the LC-CDR1, the LC-CDR2, and the LC-CDR3 of the anti-ICAMl light chain variable domain comprises amino acid sequences according to LC-CDR1: SEQ ID NO: 28; LC-CDR2: SEQ ID NO: 29; LC-CDR3: SEQ ID NO: 30.

[0263] Embodiment 79. The multispecific antibody of any one of Embodiments 1-78, further comprising a fragment crystallizable (Fc) region.

[0264] Embodiment 80. The multispecific antibody of Embodiment 79, wherein the Fc region comprises an IgG CH2 domain and an IgG CH3 domain.

[0265] Embodiment 81. The multispecific antibody of Embodiment 79 or 80, wherein the Fc region comprises a heterodimeric Fc region.

[0266] Embodiment 82. The multispecific antibody of any one of Embodiments 79-81, wherein the Fc region comprises at least one amino acid modification that increases the half-life of the multispecific antibody.

[0267] Embodiment 83. The multispecific antibody of any one of Embodiments 79-82, wherein the Fc region comprises at least one amino acid modification that modulates its interaction with an Fc receptor. [0268] Embodiment 84. The multispecific antibody of any one of Embodiments 79-83, wherein the Fc region comprises at least one amino acid modification that increases binding of the Fc region to an Fc receptor.

[0269] Embodiment 85. The multispecific antibody of any one of Embodiments 79-84, wherein the Fc region comprises at least one amino acid modification that decreases glycosylation of the Fc region. [0270] Embodiment 86. The multispecific antibody of Embodiment 85, wherein the modification is an amino acid substitution, deletion, or addition.

[0271] Embodiment 87. The multispecific antibody of Embodiment 86, wherein the modification is an amino acid substitution.

[0272] Embodiment 88. The multispecific antibody of Embodiment 87, wherein the at least one amino acid modification that decreases glycosylation of the Fc region comprises an amino acid substitution at a position corresponding to position N297 of human IgGl, wherein the numbering is according to the EU index of Kabat.

[0273] Embodiment 89. The multispecific antibody of any one of Embodiments 79-88, wherein the Fc region is afucosylated.

[0274] Embodiment 90. The multispecific antibody of any one of Embodiments 79-89, wherein the Fc region comprises at least one amino acid modification that increases antibody -dependent cellular cytotoxicity (ADCC).

[0275] Embodiment 91. The multispecific antibody of Embodiment 90, wherein the modification is an amino acid substitution, deletion, or addition.

[0276] Embodiment 92. The multispecific antibody of Embodiment 91, wherein the modification is an amino acid substitution.

[0277] Embodiment 93. The multispecific antibody of any one of Embodiments 79-92, wherein the Fc region comprises at least one mutation that increases antibody -dependent cellular cytotoxicity (ADCC), wherein the at least one mutation that increases ADCC comprises an amino acid substitution at positions corresponding to positions S239, 1332, and A330 of human IgGl, wherein the amino acid numbering is according to the EU index according to Kabat et al.

[0278] Embodiment 94. The multispecific antibody of Embodiment 93, wherein the amino acid substitutions are S239D, I332E, and A330L, wherein the amino acid numbering is according to the EU index according to Kabat et al.

[0279] Embodiment 95. The multispecific antibody of any one of Embodiments 81 -94, comprising the heterodimeric Fc region, wherein the heterodimeric Fc region comprises a knob chain and a hole chain, forming a knob-into-hole (KIH) structure.

[0280] Embodiment 96. The multispecific antibody of Embodiment 95, wherein the knob chain comprises an amino acid substitution at a position corresponding to T366 of IgGl, wherein amino acid position numbering is according to the EU index according to Kabat et al.

[0281] Embodiment 97. The multispecific antibody of Embodiment 96, wherein the T366 substitution comprises a T336W mutation, wherein amino acid position numbering is according to the EU index according to Kabat et al.

[0282] Embodiment 98. The multispecific antibody of any one of Embodiments 95-97, wherein the hole chain comprises an amino acid substitution at a position corresponding to T366, L368, or Y407 of IgGl, wherein amino acid position numbering is according to the EU index according to Kabat et al.

[0283] Embodiment 99. The multispecific antibody of Embodiment 98, wherein the hole chain comprises an amino acid substitution at a position corresponding to T366, L368, and Y407 of IgGl, wherein amino acid position numbering is according to the EU index according to Kabat et al.

[0284] Embodiment 100. The multispecific antibody of Embodiment 99, wherein the T366, L368, or Y407 amino acid substitutions comprise a T366S, L368A, or Y407V of IgGl, wherein amino acid position numbering is according to the EU index according to Kabat et al.

[0285] Embodiment 101. The multispecific antibody of Embodiment 100, wherein the T366, L368, and Y407 amino acid substitutions comprises a T366S, L368A, and Y407V of IgGl, wherein amino acid position numbering is according to the EU index according to Kabat et al.

[0286] Embodiment 102. The multispecific antibody of any one of Embodiments 1-101, wherein the multispecific antibody binds to a target cell.

[0287] Embodiment 103. The multispecific antibody of Embodiment 102, wherein the target cell expresses CD47 and ICAM1.

[0288] Embodiment 104. The multispecific antibody of Embodiment 103, wherein the target cell expresses a lower level of CD47 relative to ICAM1 on the surface of the target cell.

[0289] Embodiment 105. The multispecific antibody of Embodiment 104, wherein the ratio of CD47 to ICAM1 on the surface of the target cell is at least about 10:1, 5:1, 2.5: 1, 2.0: 1, 1.5: 1, 1: 1, 1: 1.5, 1:2.0, 1:2.5, 1:5, 1: 10, 1:15, 1:20, 1:50, 1: 100, or 1:200.

[0290] Embodiment 106. The multispecific antibody of any one of Embodiments 102-105, wherein the target cell is a cancer cell. [0291] Embodiment 107. The multispecific antibody of Embodiment 106, wherein the cancer is a hematological malignancy or multiple myeloma.

[0292] Embodiment 108. The multispecific antibody of Embodiment 107, wherein the hematological malignancy is B cell cancer or T cell cancer (such as cutaneous T cell lyphoma, or anaplastic large cell lymphoma).

[0293] Embodiment 109. The multispecific antibody of Embodiment 108, wherein the B cell cancer is leukemia or lymphoma.

[0294] Embodiment 110. The multispecific antibody of Embodiment 109, wherein the B cell cancer is lymphoma, and wherein the lymphoma is B cell lymphoma.

[0295] Embodiment 111. The multispecific antibody of Embodiment 106, wherein the cancer cell is from a solid tumor.

[0296] Embodiment 112. The multispecific antibody of Embodiment 111, wherein the solid tumor is sarcoma, breast cancer, lung cancer, kidney cancer, melanoma, head and neck cancer, ovarian cancer, liver cancer, bladder cancer, thyroid cancer, or carcinoma.

[0297] Embodiment 113. The multispecific antibody of Embodiment 111, wherein the solid tumor is lung cancer, and wherein the lung cancer is non-small cell lung cancer.

[0298] Embodiment 114. The multispecific antibody of any one of Embodiments 102-113, wherein the multispecific antibody binds to a cancer cell that expresses CD47 and ICAM1 on the surface, and wherein the ratio of CD47 to ICAM1 on the surface of the target cell is at least about 10: 1, 5: 1, 2.5: 1, 2.0: 1, 1.5: 1, 1: 1, 1: 1.5, 1:2.0, 1:2.5, 1:5, 1: 10, 1: 15, 1:20, 1:50, 1 : 100, or 1:200.

[0299] Embodiment 115. The multispecific antibody of any one of Embodiments 1-114, wherein the multispecific antibody inhibits binding of Signal-regulatory protein alpha (SIRPa) to the target cell as determined by an in vitro competition assay.

[0300] Embodiment 116. The multispecific antibody of Embodiment 115, wherein the binding of Signal- regulatory protein alpha (SIRPa) to the target cell as determined by an in vitro competition assay is inhibited by the multispecific antibody by at least 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100%.

[0301] Embodiment 117. The multispecific antibody of Embodiment 116, wherein a concentration of the multispecific antibody required to mediate antibody -dependent cellular phagocytosis (ADCP) of the target cell by a macrophage is from 0.0 InM to 3nM.

[0302] Embodiment 118. The multispecific antibody of Embodiment 116 or 117, wherein the multispecific antibody induces enhanced antibody-dependent cellular phagocytosis (ADCP) of the target cell by a macrophage as compared to ADCP activity induced of the target cell by a macrophage by a monospecific anti-CD47 antibody.

[0303] Embodiment 119. The multispecific antibody of any one of Embodiments 102-118, wherein the multispecific antibody has a higher binding activity for CD47 expressed on a surface of a tumor cell than for CD47 expressed on a surface of a red blood cell or a platelet. [0304] Embodiment 120. The multispecific antibody of any one of Embodiments 102-119, wherein a concentration of the multispecific antibody required for half-maximal binding to a human red blood or a human platelet is greater than 500 nM.

[0305] Embodiment 121. The multispecific antibody of any one of Embodiments 102-120, wherein the multispecific antibody induces antibody -dependent cellular cytotoxicity (ADCC) mediated killing of the target cell.

[0306] Embodiment 122. The multispecific antibody of Embodiment 121, wherein the multispecific antibody induces enhanced antibody-dependent cellular cytotoxicity (ADCC) activity on the target cell as compared to ADCC activity induced on the target cell by a monospecific anti-CD47 antibody or a monospecific anti-ICAMl antibody.

[0307] Embodiment 123. The multispecific antibody of Embodiment 122, wherein the multispecific antibody induces at least 5%, 10%, 15%, 20%, 25%, 30%, 25%, 40%, 45%, or 50% more ADCC activity of the target cell as compared to an ADCC activity of the target cell that is induced by a monospecific anti-CD47 antibody or a monospecific anti-ICAMl antibody.

[0308] Embodiment 124. The multispecific antibody of any one of Embodiments 102-123, wherein the multispecific antibody induces complement-dependent cytotoxicity (CDC) mediated killing of the target cell.

[0309] Embodiment 125. The multispecific antibody of Embodiment 124, wherein the multispecific antibody induces at least 5%, 10%, 15%, 20%, 25%, 30%, 25%, 40%, 45%, or 50% more CDC activity of the target cell as compared to a CDC activity of the target cell that is induced by a monospecific anti- CD47 antibody or a monospecific anti-ICAMl antibody.

[0310] Embodiment 126. The multispecific antibody of any one of Embodiments 102-125, wherein a concentration of 600 nM of the multispecific antibody does not induce hemolysis of red blood cells in a hemagglutination assay.

[0311] Embodiment 127. A nucleic acid molecule encoding the multispecific antibody of any one of Embodiments 1-126.

[0312] Embodiment 128. A vector comprising the nucleic acid molecule of Embodiment 127.

[0313] Embodiment 129. A pharmaceutical composition comprising the multispecific antibody of any one of Embodiments 1-126.

[0314] Embodiment 130. The pharmaceutical composition of Embodiment 129, further comprising a pharmaceutically acceptable carrier, an excipient, or any combinations thereof.

[0315] Embodiment 131. A method of treating a subject having cancer, the method comprising: administering to the subject the multispecific antibody of any one of Embodiments 1-126 or the pharmaceutical composition of Embodiment 129 or 130.

[0316] Embodiment 132. The method of Embodiment 131, wherein the cancer comprises cancer cells that express CD47 and ICAM1. [0317] Embodiment 133. The method of Embodiment 132, wherein the ratio of CD47 to ICAM1 on the surface of the cancer cell is at least about 10: 1, 5: 1, 2.5: 1, 2.0: 1, 1.5: 1, 1: 1, 1: 1.5, 1:2.0, 1:2.5, 1:5, 1: 10, 1: 15, 1:20, 1:50, 1: 100, or 1:200.

[0318] Embodiment 134. The method of Embodiment 132 or 133, wherein the cancer cells that express CD47 and ICAM1 are lysed.

[0319] Embodiment 135. The method of any one of Embodiments 131-134, wherein the multispecific antibody induces antibody-dependent cellular cytotoxicity (ADCC) mediated killing of the cancer cells that express CD47 and/or ICAM1.

[0320] Embodiment 136. The method of any one of Embodiments 131-135, wherein the multispecific antibody induces complement-dependent cytotoxicity (CDC) mediated killing of the cancer cells that express CD47 and/or ICAM1.

[0321] Embodiment 137. The method of any one of Embodiments 131-136, wherein the multispecific antibody induces antibody-dependent cellular phagocytosis (ADCP) of the cancer cells that express CD47 and/or ICAM1.

[0322] Embodiment 138. The method of any one of Embodiments 131-137, wherein the cancer is a hematological malignancy or multiple myeloma.

[0323] Embodiment 139. The method of Embodiment 138, wherein the cancer is B cell cancer or T cell cancer (such as cutaneous T cell lyphoma, or anaplastic large cell lymphoma).

[0324] Embodiment 140. The method of Embodiment 139, wherein the cancer is leukemia or lymphoma. [0325] Embodiment 141. The method of Embodiment 140, wherein the cancer is lymphoma, and wherein the lymphoma is B-cell lymphoma.

[0326] Embodiment 142. The method of any one of Embodiments 131-137, wherein the cancer is a solid tumor.

[0327] Embodiment 143. The method of Embodiment 142, wherein the solid tumor is sarcoma, breast cancer, lung cancer, kidney cancer, melanoma, head and neck cancer, ovarian cancer, liver cancer, bladder cancer, thyroid cancer, or carcinoma.

[0328] Embodiment 144. The method of Embodiment 143, wherein the solid tumor is lung cancer, and wherein the lung cancer is non-small cell lung cancer.

[0329] Embodiment 145. The method of any one of Embodiments 131-144, further comprising administering to the subject an anti -cancer agent.

[0330] Embodiment 146. The method of Embodiment 145, wherein the anti -cancer agent is a chemotherapeutic agent or a biologic agent.

[0331] Embodiment 147. The method of any one of Embodiments 146, wherein the administering is sufficient to reduce or eliminate the cancer as compared to a comparable method lacking the administering.

[0332] Embodiment 148. The method of Embodiment 147, wherein the reduction is at least about 1 fold, 5 fold, 10 fold, 20 fold, 40 fold, 60 fold, 80 fold, or up to about 100 fold. [0333] Embodiment 149. The method of any one of Embodiments 131-148, wherein the cancer is metastatic.

[0334] Embodiment 150. A kit that comprises at least one of:

(a) the multispecific antibody of any one of Embodiments 1-126;

(b) the vector of Embodiment 128;

(c) the nucleic acid molecule of Embodiment 127; or

(d) the pharmaceutical composition of Embodiment 129 or 130.

[0335] Embodiment 151. A multispecific antibody of any one of Embodiments 1-126, a nucleic acid molecule of Embodiment 127, a vector of Embodiment 128, or a pharmaceutical composition of Embodiment 129 or 130, for use in a method for the treatment of cancer in a subject in need thereof, the method comprising administering the multispecific antibody, the nucleic acid molecule, the vector, or the pharmaceutical composition to the subject.

EXAMPLES

Example 1: Generation of Human/Cyno CD47

[0336] DNA encoding the extracellular domains (ECD) of human CD47 (19aa-139aa) and cyno CD47 (4aa-126aa) were cloned into pRK5 (ATCC Cat#209784). The resulting constructs with C-terminal 6xHis tags were transfected into Expi293F cells. After 72 hours, CD47-expressing cells were harvested by centrifugation for 5 minutes at 2000 rpm at 4 °C. The supernatant was collected. Ni-NTA (Qiagen, Cat# 30410) resin was pre -equilibrated with buffer A (137 mM NaCl, 2.7 mM KC1, lOmM Na2HPO4, 2mM KH2PO4, pH 7.4) and incubated with the supernatant for 2 hours at 4 °C on a rotator. The resin was filled in the Ni Sepharose excel (GE, Cat# GE17371201) and washed with buffer A until no signal (OD595, about 20-30 column volumes (CV) was observed by Coomassie-Brilliant Blue G-250. The target protein was eluted using buffer B (137 mM NaCl, 2.7 mM KC1, 10 mM Na2HPC>4, 2mM KH2PO4, pH 7.4, 250 mM imidazole) for 3 column volumes (CV). The SuperdexTX 200 increase column (GE, Cat# GE28-9909-44) was pre-equilibrated by buffer A, then the eluate was loaded onto the column. The column was washed with buffer A and fractions were collected. Different fractions were resolved on a 12% SDS-PAGE and desired fractions were combined and neutralized with buffer C (137 mM NaCl, 2.7 mM KC1, 10 mM Na2HPC>4, 2mM KH2PO4, pH 7.4). The target protein was concentrated by using an ultra-filtration tube (Amicon, Cat#42409) with a molecular cutoff of 30 kDa, then aliquoted and snap frozen using liquid N2 and stored at -80 °C.

Example 2: Generation of Anti-CD47/ICAM1 Bispecific Antibodies

[0337] The anti-CD47 benchmark antibody BMK-1 is based on 5F9.G4 sequence from Gilead/Forty Seven, the CD47 BMK-2 is based on TTI-621 sequence from Pfizer/Trillium.

[0338] Anti-CD47/ICAM1 bispecific antibodies with different binding stoichiometry and geometry were designed and generated using VH and VL sequences from the anti-CD47 antibody VIR47.V8 and anti- ICAM1 antibody sequences. The corresponding heavy chain (HC) and light chain (LC) DNAs were synthesized and cloned into the pRK5 mammalian expression vector (ATCC). Each HC and LC pair were then co-transfected in CHO cells. The conditioned medium was harvested by centrifugation (4 °C, 4000rpm for 40min), then fdtered to remove cell debris. The clarified medium was loaded onto MabSelect SuRe column (GE, 17-5438) which was pre-equilibrated with Buffer A (25mM Tris, 150mM NaCl, pH 8.0). The column was washed sequentially with 5 column volume of Buffer A, then 30 column volume of Buffer B (Buffer A + 0.1% Triton X100 + 0. 1% Triton XI 14), then 15 column volume of Buffer A. The antibodies were eluted with Buffer C (lOOmM sodium citrate, 150mM NaCl, pH 3.0) and neutralized immediately with Buffer D (200 mM Arginine, 137 mM Succinic acid, pH 5.0). The final product was dialyzed against the buffer 20 mM His, 5% sucrose, pH 5.5, concentrated and filtrated through MILLEX- MP 0.22um (MILLIPORE). Figure 1 illustrates the bispecific formats that were generated for evaluation. Example 3: Evaluating Target Antigen Binding Activity of the Anti-CD47/ICAM1 Bispecific Antibody

Antibody Binding to Antigen Measured by ELISA

[0339] A 96-well plate was coated overnight at 4 °C with 1 pg/ml recombinant huCD47 or huICAMl . After washing 3 times, the plate was blocked with 300 pl 1% BSA in PBST at 37 °C for 1 hour. Serially diluted antibodies were added and incubated at 37 °C for 1 hour. The plate was then washed 4 times with PBST and incubated with 1:5000 diluted peroxidase labeled goat anti-human IgG (Fab specific) secondary antibody (Sigma, Cat# A0293) for 1 hour at 37 °C. The plate was washed again 4 times with PBST, incubated with 3,3',5,5'-tetramethylbenzidine (TMB) substrate for 15 min at room temperature, terminated with IN HC1, and then read at 450 nm. Figure 2A and Figure 2B showed the ELISA binding results of anti-CD47/ICAMl bispecific antibody on CD47 and ICAM1, respectively. The anti- CD47/ICAM1 bispecific antibody (“CD47 X ICAM1”) showed binding on huCD47 and huICAMl in a dose dependent manner, weaker than its parental anti-CD47 mab(VIR47.V8) or anti-ICAMl reference since it was only one arm binding.

Antibody Binding to Cells as Measured by Flow Cytometry

[0340] Antibody binding to CD47 over-expressing CHO cell line was quantified by flow cytometry. Harvested cells were centrifuged at 2000 rpm for 5 min, resuspended in 10 - 15 ml ice-cold culture medium, and then counted. Cells were resuspended in blocking buffer (PBS plus 2% FBS) at a concentration of 3x 10⁶ cells/mL. 100 pL of the cell suspension was dispensed into each well of a 96-well plate. Purified antibodies were diluted to the desired concentrations with blocking buffer and 100 pL of diluted antibodies were added to the well and incubated for 1 hour at 4 °C. The cells were then washed 3 times with PBS plus 2% FBS. After the third wash, the cells were resuspended in 100 pL 1:500 diluted Alexa Fluor 488 labeled Mouse anti -Human IgGl Fc secondary antibody (Invitrogen, Cat#: A 10631) and incubated for 1 hour at 4 °C in the dark. The cells were then washed 3 times with 200 pL PBS by centrifuging at 2000 rpm for 5 min. After the last wash, the cells were resuspended in 300 pL cold PBS and analyzed on a FACSVerse™ (BD Biosciences) flow cytometer. Figure 3A and Figure 3B showed the FACS binding results of anti-CD47/ICAMl bispecific antibody on huCD47 and cynoCD47 overexpressing CHO cell lines, respectively, in a dose dependent manner. Example 4: Inhibition of SIRPa Binding to CD47+ Cells with anti-CD47/ICAMl bispecific antibody (“CD47 X ICAM1”)

[0341] Tumor cells were harvested, centrifuged, and then resuspended in FACS buffer (PBS plus 2% FBS) at a concentration of 2x 10⁶ cells/mL. 100 pL of the cell suspension was dispensed into each well of a 96-well plate. The plate was centrifuged for 5 min at 300g, and the supernatants were discarded. The cells were incubated with 50 pl per well of serially diluted bispecific or bivalent anti-CD47 antibodies and a constant amount of SIRPa-mIgG2a fusion protein (0.2 pg/ml for Raji cells) in FACS buffer for Ih at 4°C. Then, the plates were washed twice with FACS buffer and incubated for 1 hour at 4°C in the dark with 100 pL of Alexa Fluor 488 donkey anti-Mouse IgG(H+L) secondary antibody (Invitrogen, Cat#A21202, 1: 1000). After washing twice with FACS buffer, the plates were resuspended with 300 pL FACS buffer and analyzed by flow cytometry. Figure 4 showed the SIRPa blocking results for anti- CD47/ICAM1 bispecific antibody, with the guide ICAM1, the bispecific “CD47 X ICAM1” and VIR47.V8/ICAM1.245 (corresponding to the construct “VIR47.V8.knob/ICAMl.245.hole” having sequences of SEQ ID NOs. 251-253 detailed in Table 3 hereinabove) showed superior SIRPa blocking activity to its parental anti-CD47 mab (VIR47.V8) on an ICAM1+/CD47+ Raji cell line.

Example 5: Antibody-Dependent Cellular Phagocytosis with anti-CD47/ICAMl bispecific antibody (“CD47 X ICAM1” and VIR47.V8/ICAM1.245)

[0342] The bispecific antibodies were tested in an antibody-dependent cellular phagocytosis (ADCP) assay. Peripheral blood mononuclear cells (PBMCs) were isolated from human donors. Monocytes were enriched using a Human Monocyte Enrichment Kit without CD 16 depletion (STEMCELL, Cat # 19058). Isolated monocytes were differentiated into macrophages by culturing monocytes in complete culture media (RPMI 1640 + 10% FBS) with 20 ng/ml of human Macrophage Colony-Stimulating Factor (M- CSF, Peprotech, Cat #: 3-25-10). The media was changed every three days. After 7 days of culturing in M-CSF containing culture media, macrophages were collected and counted. Target tumor cells were collected and washed with D-PBS two times to remove remaining FBS. The washed tumor cells were resuspended in PBS at a cell concentration of 5 - 10x 10⁶/mL. Cancer cells were stained with CFSE (ebiosciences, Cat#: 65-0580-84) at a final concentration of 3 pM and mixed immediately. The cells were stained in the dark at room temperature for 10 minutes. The staining was terminated by adding 4-5 volumes of cold complete media and incubating on ice for 5 minutes. Stained cells were washed three times with RPMI 1640 + 10% FBS. Cells were resuspended in 1 ml RPMI 1640 + 10% FBS and counted, then adjusted the cell numbers to 3x 10⁵ cells/mL. 50 pl cells were seeded into a 96 well deep U-plate (Axygen, Cat #: P-DW-20-C) wherein each well contained 1.5 x 104 cells. 50 pl of diluted antibodies were added to each well. 100 pl of macrophages (1.5 x 104 cells) were added to each well and incubated at 37 °C, 5% CO2 for 1.5 hours. After incubation, cells were washed with 2 ml of 2% FBS in D-PBS once. 100 pl of diluted Fc blocker (Human TruStain FcX (Fc Receptor Blocking Solution), Biolegend Cat #: 422302)) was added and the cells were incubated at room temperature for 10 minutes. 20 pl of diluted anti-human CD1 lb antibody was added to each well and incubated for 30 minutes at 4°C in the dark. Cells were washed with 2% FBS-D-PBS once. Phagocytosis was detected in a flow cytometer by the appearance of CFSE/CD1 lb double positive cells indicative of macrophages that engulfed the tumor cells.

[0343] In Figure 5., Bispecific antibody “CD47 X ICAMl”and VIR47.V8.knob/ICAM1.245.hole both showed superior ADCP activity to its parental anti-CD47 mab (VIR47.V8) on an ICAM1+/CD47+ Raji cell line.

Example 6: Antibody-Dependent Cellular Cytotoxicity with anti-CD47/ICAMl bispecific antibody (“CD47 X ICAM1”)

The ability of anti-CD47 antibodies to mediate antibody-dependent cellular cytotoxicity (ADCC) was tested on HCC44 cancer cells, which express high levels of CD47 and ICAM1 . The cells were washed once with balanced salt solution or culture medium and cell numbers were adjusted to IxlO⁶ cells/ml. 2 pL of BATDA fluorescence enhancing ligand (Perkin Elmer, Cat# C 136-100) was then added to each mb of cells and incubated for 20 min at 37 °C in a cell incubator. After incubation, cells were centrifuged, culture medium was aspirated. The labeled cells were washed 4 times with PBS. After the final wash, cells were re-suspended in culture medium and adjusted to 5 x 10⁴ cell /ml. 200 pL cell suspension was then added to each well of the 96-well plate to make the cell number per well to lx 10⁴. Background release was determined by withdrawing an aliquot of the labeled target cells, centrifuge and supernatant was transferred into an empty well. The reading was background release. IxlO⁴ labeled target cells were transferred to sterile 96-well assay plate. Antibodies were serially diluted with RPMI-1640 containing 10% FBS. 50 pL of serially-diluted antibodies were added to assay plate containing target cell and incubated at 37 °C, 5% CO2 for 5-10min. Effector cells NK92/CD 16a 176V were harvested and suspended in RPMI-1640 containing 10% FBS. 50 ul/well effector cells were added to each well of assay plate at different E:T ratio. Set up controls: target spontaneous (target cell+100 pL medium); target maximum (target cell+100 pL medium+10 pL lysis buffer); background (100 pL the labeled target cell supernatant and 100 pL dilution medium). The plates were incubated in a humidified 5 % CO2 atmosphere at 37 °C for 2 hours. At the end of incubation, 10 pL of Lysis Buffer (Perkin Elmer, Cat# 4005-0010) was added to the maximum release well. The plates were centrifuged for 5 min at 500g. 20 pL of the supernatant from each well was transferred to a flat-bottom detection plate. 200 pL of Europium Solution (Perkin Elmer, Cat# Cl 35 -100) was then added to each well of the detection plate.

The plate was shaken at 250 rpm for 15 min at room temperature and the fluorescence was then measured in a time-resolved fluorometer within 5 hrs., Figure 6. Bispecific antibody “CD47 X ICAM1” showed superior ADCC activity to ICAM1 reference or VIR47.V8 bivalent antibody on HCC44 human non-small lung cancer cells.

Example 7: Antibody Binding to Red Blood Cells/Platelet and Hemagglutination with anti- CD47/ICAM1 bispecific antibody (“CD47 X ICAM1”)

[0344] The RBC binding assay was performed by spinning down fresh human whole blood at 200g for 10 minutes. Collected RBCs were washed twice with PBS and counted using flow cytometry. I x lO⁶ cells were dispensed into each well of a 96 well culture plate. Serially diluted anti-CD47 antibodies were added and incubated for 1 hour at 4 °C. Cells were washed with FACS buffer (PBS + 2% FBS) twice. Secondary antibody (Alexa Fluor® 488 Goat Anti-Human IgG (H+L)) was added and incubated for 1 hour at 4 °C. Cells were washed twice and resuspended in 200 pl of FACS buffer and analyzed by flow cytometry.

[0345] In Figure 7A and 7B bispecific antibody “CD47 X ICAM1” showed much less RBC and platelet binding, respectively, as compared to the CD47 BMK-1 benchmark antibody and its parental VIR47.V8 bivalent antibody. Furthermore, hemagglutination was not detected for “CD47 X ICAM1,” even at the highest tested concentration (lOOug/ml).

[0346] The hemagglutination assay was performed by diluting human red blood cells (RBCs) and incubating RBCs at 37°C for 2 hours with a titration of CD47 antibodies (from 100 pg/ml) in a round bottom 96 well plate. Hemagglutination is demonstrated by the presence of crosslinked RBCs, which appear as a haze because they do not settle to the bottom of the well, in contrast to non-hemagglutinated RBCs.

[0347] In Figure 8. Bispecific antibody “CD47 X ICAM1” didn’t show any hemagglutination, control BMK-1 showed dose dependent hemagglutination under the same condition.

Example 8: In Vivo Efficacy Study with anti-CD47/ICAMl bispecific antibody (“CD47 X ICAM1”) [0348] 10 x 10" Raji cells with Matrigel were injected subcutaneously (sc) into the flank of 6-8-week-old SCID mice. Tumors were measured every 2-3 days using a digital caliper and volumes were calculated using the formula (width x length x height x Pi)/6. Mice were randomized to different groups for treatment when the tumor volume reached to 100-150 mm3, mice were treated with 3mg/kg or 0.5mg/kg of antibodies intravenously twice a week for 3 weeks. Tumor volumes were estimated twice weekly. Tumor volumes were monitored until they reached the maximum volume of approximately 2,500 mm³ or maximum permissible markers of discomfort in the mice were reached (i.e., mouse discomfort or body weight loss reached maximum allowable levels), at which time the mice were sacrificed.

[0349] In Figure 9, bispecific antibody “CD47 X ICAM1” at 0.5mg/kg and 3mg/kg and VIR47.V8 at 0.5mg/kg showed significant inhibition of tumor growth, superior to bench mark antibody BMK-1, and VIR47.V8 at 3mg/kg showed tumor growth inhibition completely,

Example 9: Inhibition of SIPRa Binding to CD47+ tumor Cells with anti-CD47/ICAMl bispecific antibody (VIR47.V8/ICAM1.U3)

[0350] Inhibition of SIPRa binding to CD47+ or CD47+/ICAM1 tumor cells with anti-CD47/ICAMl bispecific antibody were performed according to the methods described in Example 4. Figure 10A showed SIPRa blocking data obtained on KYSE-150 (ICAMl^med CD47^med) cells; and Figure 10B showed SIPRa blocking data obtained on KYSE-30 (ICAMl^nu11 CD47^med) cells. As shown in Figures 10A-10B, anti-CD47 mAbs showed similar SIRPa blocking activity on ICAM1+/CD47+ and CD47+ only tumor cells without selectivity; whereas anti-CD47/ICAMl bispecific antibody (VIR47.V8/ICAM1.U3) (corresponding to the construct “ICAM1 ,U3.knob/VIR47.V8.Hole” having sequences of SEQ ID NOs. 254-256 detailed in Table 3 hereinabove) showed potent SIRPa blocking activity on ICAM1+/CD47+ tumor cells, but not on CD47+ only tumor cells. Example 10: FACS binding to human Red Blood Cells with anti-CD47/ICAMl bispecific antibody (VIR47.V8/ICAM1.U3)

[0351] The FACS binding assay was performed according to the methods described in Example 7. As illustrated in Figure 11, anti-CD47/ICAMl bispecific antibody (VIR47.V8/ICAM1.U3) (corresponding to the construct “ICAMl.U3.knob/VIR47.V8.Hole” having sequences of SEQ ID NOs. 254-256 detailed in Table 3 hereinabove) showed reduced RBC binding compared with BMK-1 and its bivalent anti-CD47 antibody VIR47.V8 significantly.

Example 13: In Vivo Efficacy Study in Raji Model

[0352] In vivo efficacy study in Raji model was performed according to the methods described in Example 8.

[0353] In Figure 12, anti-CD47/ICAMl bispecific antibody (VIR47.V8/ICAM1.U3) (corresponding to the construct “ICAMl.U3.knob/VIR47.V8.Hole” having sequences of SEQ ID NOs. 254-256 detailed in Table 3 hereinabove) showed significant tumor growth inhibition at 0.3mg/kg and complete tumor inhibition at 1 mg/kg treatment in Raji model, whereas Lenalidomide 50mg/kg didn’t show any tumor inhibition in the same model.

Example 14: In Vivo Efficacy Study in HCC44 Model (NSCLC/Kras G12C)

[0354] Female Balb/c nude mice of 6-8 weeks in age (Shanghai Lingchang Biotechnology) were inoculated subcutaneously with 5xl0⁶ HCC44 (NSCLC/Kras G12C) tumor cells in 0.2 mL of PBS supplemented with Matrigel ( 1 : 1) for tumor development. Tumor volume was measured using a caliper device and calculated with the following formula: Tumor volume = (length x width ²) / 2. Once the average tumor volume reached approximately 150-200 mm³, the animals were randomized to 6 animals per group and started treatment. Test antibodies were administered through bolus tail vein injection at a dose of 10 mg/kg or 20 mg/kg twice a week for 5 consecutive weeks. Tumor volume was measured twice weekly. Animals were humanely euthanized if their tumor volumes exceeded 3000 mm³ or if they experienced over 20 % of body weight loss.

[0355] The results are summarized in FIG.13. Treatment with bispecific antibody VIR47.V8/ICAM1.U3 (corresponding to the construct “ICAMl.U3.knob/VIR47.V8.Hole” having sequences of SEQ ID NOs. 254-256 detailed in Table 3 hereinabove) showed significant tumor regression whereas treatment with its anti-CD47 bivalent antibody VIR47.V8 only showed partial tumor growth inhibition. These data demonstrates the superior in vivo anti-tumor activity of VIR47.V8/ICAM1.U3 bispecific antibody over its anti- CD47 bivalent antibody VIR47.V8 against ICAM1 and CD47 expressing lung cancer.

[0356] While the foregoing disclosure has been described in some detail for purposes of clarity and understanding, it will be clear to one skilled in the art from a reading of this disclosure that various changes in form and detail can be made without departing from the true scope of the disclosure. For example, all the techniques and apparatus described above can be used in various combinations. All publications, patents, patent applications, and/or other documents cited in this application are incorporated by reference in their entirety for all purposes to the same extent as if each individual publication, patent, patent application, and/or other document were individually and separately indicated to be incorporated by reference for all purposes.

Claims

CLAIMS WHAT IS CLAIMED IS:

1. A multispecific antibody comprising a CD47 binding domain and an Intercellular Adhesion Molecule 1 (ICAM1) binding domain.

2. The multispecific antibody of claim 1, wherein the multispecific antibody is bispecific, trispecific, or tetraspecific.

3. The multispecific antibody of claim 2, wherein the multispecific antibody is bispecific.

4. The multispecific antibody of claim 1, wherein the multispecific antibody is bivalent, trivalent, or tetravalent.

5. The multispecific antibody of claim 4, wherein the multispecific antibody is bivalent.

6. The multispecific antibody of claim 1, wherein the CD47 binding domain comprises an antibody, or functional fragment or functional variant thereof, that binds specifically to CD47.

7. The multispecific antibody of claim 6, wherein the antibody, or functional fragment or functional variant thereof that binds specifically to CD47 comprises an anti-CD47 heavy chain and an anti-CD47 light chain.

8. The multispecific antibody of claim 7, wherein the anti-CD47 heavy chain comprises an anti- CD47 heavy chain variable domain.

9. The multispecific antibody of claim 7, wherein the anti-CD47 heavy chain variable domain comprises a variable domain of an IgGl, IgG2, IgG3, or IgG4 heavy chain.

10. The multispecific antibody of claim 7, wherein the anti-CD47 light chain comprises an anti- CD47 light chain variable domain.

11. The multispecific antibody of claim 10, wherein the anti-CD47 light chain variable domain comprises a variable domain of a Kappa or Lambda light chain.

12. The multispecific antibody of claim 8, wherein the anti-CD47 heavy chain variable domain comprises the variable domain of an IgGl heavy chain and the anti-CD47 light chain variable domain comprises the variable domain of a Lambda light chain.

13. The multispecific antibody of claim 8, wherein the anti-CD47 heavy chain variable domain comprises the variable domain of an IgGl heavy chain and the anti-CD47 light chain variable domain comprises the variable domain of a Kappa light chain.

14. The multispecific antibody according to claim 6, wherein the antibody, or functional fragment or functional variant thereof, that binds specifically to CD47 comprises a single-chain variable fragment (scfv) or an antigen-binding fragment (fab).

15. The multispecific antibody of claim 8, wherein the anti-CD47 heavy chain variable domain comprises complementarity determining regions (CDRs): HC-CDR1, HC-CDR2, and HC-CDR3, and wherein the HC-CDR1, the HC-CDR2, and the HC-CDR3 of the anti-CD47 heavy chain variable domain comprise amino acid sequences according to HC-CDR1: SEQ ID NO: 1; HC-CDR2: SEQ ID NO: 2; HC- CDR3: SEQ ID NO: 3; and wherein the CDRs comprise from 0-2 amino acid modification(s) in at least one of the HC-CDR1, HC-CDR2, or HC-CDR3.

-83-

16. The multispecific antibody of claim 8, wherein the anti-CD47 heavy chain variable domain comprises complementarity determining regions (CDRs): HC-CDR1, HC-CDR2, and HC-CDR3, and wherein the HC-CDR1, the HC-CDR2, and the HC-CDR3 of the anti-CD47 heavy chain variable domain comprise amino acid sequences according to HC-CDR1: SEQ ID NO: 1; HC-CDR2: SEQ ID NO: 2; HC- CDR3: SEQ ID NO: 3; and wherein the CDRs comprise from 0-1 amino acid modification(s) in at least one of the HC-CDR1, HC-CDR2, or HC-CDR3.

17. The multispecific antibody of claim 8, wherein the anti-CD47 heavy chain variable domain comprises complementarity determining regions (CDRs): HC-CDR1, HC-CDR2, and HC-CDR3, and wherein the HC-CDR1, the HC-CDR2, and the HC-CDR3 of the anti-CD47 heavy chain variable domain comprise amino acid sequences according to HC-CDR1: SEQ ID NO: 1; HC-CDR2: SEQ ID NO: 2; HC- CDR3: SEQ ID NO: 3.

18. The multispecific antibody of claim 10, wherein the anti-CD47 light chain variable domain comprises complementarity determining regions (CDRs): LC-CDR1, LC-CDR2, and LC-CDR3, and wherein the LC-CDR1, the LC-CDR2, and the LC-CDR3 of the anti-CD47 light chain variable domain comprises amino acid sequences according to LC-CDR1: SEQ ID NO: 4; LC-CDR2: SEQ ID NO: 5; LC- CDR3: SEQ ID NO: 6; and wherein the CDRs comprise from 0-2 amino acid modification(s) in at least one of the LC-CDR1, LC-CDR2, or LC-CDR3.

19. The multispecific antibody of claim 10, wherein the anti-CD47 light chain variable domain comprises complementarity determining regions (CDRs): LC-CDR1, LC-CDR2, and LC-CDR3, and wherein the LC-CDR1, the LC-CDR2, and the LC-CDR3 of the anti-CD47 light chain variable domain comprises amino acid sequences according to LC-CDR1: SEQ ID NO: 4; LC-CDR2: SEQ ID NO: 5; LC- CDR3: SEQ ID NO: 6; and wherein the CDRs comprise from 0-1 amino acid modification(s) in at least one of the LC-CDR1, LC-CDR2, or LC-CDR3.

20. The multispecific antibody of claim 10, wherein the anti-CD47 light chain variable domain comprises complementarity determining regions (CDRs): LC-CDR1, LC-CDR2, and LC-CDR3, and wherein the LC-CDR1, the LC-CDR2, and the LC-CDR3 of the anti-CD47 light chain variable domain comprises amino acid sequences according to LC-CDR1: SEQ ID NO: 4; LC-CDR2: SEQ ID NO: 5; LC- CDR3: SEQ ID NO: 6.

21. The multispecific antibody of claim 7, wherein the HC-CDR1, the HC-CDR2, and the HC- CDR3 of the anti-CD47 heavy chain variable domain comprises amino acid sequences according to HC- CDR1: SEQ ID NO: 1; HC-CDR2: SEQ ID NO: 2; HC-CDR3: SEQ ID NO: 3 and the LC-CDR1, the LC-CDR2, and the LC-CDR3 of the anti-CD47 light chain variable domain comprises amino acid sequences according to LC-CDR1: SEQ ID NO: 4; LC-CDR2: SEQ ID NO: 5; LC-CDR3: SEQ ID NO: 6; wherein the CDRs comprise from 0-2 amino acid modification(s) in at least one of the HC-CDR1, HC- CDR2, or HC-CDR3; and wherein the CDRs comprise from 0-2 amino acid modification(s) in at least one of the LC-CDR1, LC-CDR2, or LC-CDR3

22. The multispecific antibody of claim 7, wherein the HC-CDR1, the HC-CDR2, and the HC- CDR3 of the anti-CD47 heavy chain variable domain comprises amino acid sequences according to HC-

-84- CDR1: SEQ ID NO: 1; HC-CDR2: SEQ ID NO: 2; HC-CDR3: SEQ ID NO: 3 and the LC-CDR1, the LC-CDR2, and the LC-CDR3 of the anti-CD47 light chain variable domain comprises amino acid sequences according to LC-CDR1: SEQ ID NO: 4; LC-CDR2: SEQ ID NO: 5; LC-CDR3: SEQ ID NO: 6; wherein the CDRs comprise from 0-1 amino acid modification(s) in at least one of the HC-CDR1, HC- CDR2, or HC-CDR3; and wherein the CDRs comprise from 0-1 amino acid modification(s) in at least one of the LC-CDR1, LC-CDR2, or LC-CDR3

23. The multispecific antibody of claim 7, wherein the HC-CDR1, the HC-CDR2, and the HC- CDR3 of the anti-CD47 heavy chain variable domain comprises amino acid sequences according to HC- CDR1: SEQ ID NO: 1; HC-CDR2: SEQ ID NO: 2; HC-CDR3: SEQ ID NO: 3 and the LC-CDR1, the LC-CDR2, and the LC-CDR3 of the anti-CD47 light chain variable domain comprises amino acid sequences according to LC-CDR1: SEQ ID NO: 4; LC-CDR2: SEQ ID NO: 5; LC-CDR3: SEQ ID NO: 6.

24. The multispecific antibody of claim 7, wherein the antibody, or functional fragment or functional variant thereof that binds specifically to CD47 comprises the scFv, and the antibody, or functional fragment or functional variant thereof, that binds specifically to ICAM1 comprises the Fab, and wherein the HC-CDR1, the HC-CDR2, and the HC-CDR3 of the anti-CD47 heavy chain variable domain comprises amino acid sequences according to HC-CDR1: SEQ ID NO: 1; HC-CDR2: SEQ ID NO: 2; HC-CDR3: SEQ ID NO: 3 and the LC-CDR1, the LC-CDR2, and the LC-CDR3 of the anti-CD47 light chain variable domain comprises amino acid sequences according to LC-CDR1: SEQ ID NO: 4; LC- CDR2: SEQ ID NO: 5; LC-CDR3: SEQ ID NO: 6.

25. The multispecific antibody of claim 7, wherein the antibody, or functional fragment or functional variant thereof that binds specifically to CD47 comprises the Fab, and the antibody, or functional fragment or functional variant thereof, that binds specifically to ICAM1 comprises the scFv, and wherein the HC-CDR1, the HC-CDR2, and the HC-CDR3 of the anti-CD47 heavy chain variable domain comprises amino acid sequences according to HC-CDR1: SEQ ID NO: 1; HC-CDR2: SEQ ID NO: 2; HC-CDR3: SEQ ID NO: 3 and the LC-CDR1, the LC-CDR2, and the LC-CDR3 of the anti-CD47 light chain variable domain comprises amino acid sequences according to LC-CDR1: SEQ ID NO: 4; LC-CDR2: SEQ ID NO: 5; LC-CDR3: SEQ ID NO: 6.

26. The multispecific antibody of claim 8, wherein the anti-CD47 heavy chain comprises an amino acid sequence with at least 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to of SEQ ID NO: A.

27. The multispecific antibody of claim 8, wherein the anti-CD47 heavy chain comprises an amino acid sequence with at least 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to of SEQ ID NO: 7.

28. The multispecific antibody of claim 10, wherein the anti-CD47 light chain comprises an amino acid sequence with at least 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NO: B.

-85-

29. The multispecific antibody of claims 7, wherein the anti-CD47 heavy chain comprises an amino acid sequence with at least 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NO: A; and the anti- CD47 light chain comprises an amino acid sequence with at least 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NO: B.

30. The multispecific antibody of claim 13, wherein the anti-CD47 heavy chain variable domain comprises a single-chain variable fragment (scFv) comprising an amino acid sequence with at least 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NO: 11.

31. The multispecific antibody of claim 10, wherein the anti-CD47 light chain comprises an amino acid sequence with at least 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NO: 9.

32. The multispecific antibody of claims 7, wherein the anti-CD47 heavy chain comprises an amino acid sequence with at least 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NO: 7 and the anti-CD47 light chain comprises an amino acid sequence with at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NO: 9.

33. The multispecific antibody of claim 1, wherein the ICAM1 binding domain comprises an antibody, or functional fragment or functional variant thereof, that binds specifically to ICAM1.

34. The multispecific antibody of claim 33, wherein the antibody, or functional fragment or functional variant thereof that binds specifically to ICAM1 comprises an anti-ICAMl heavy chain and an anti-ICAMl light chain.

35. The multispecific antibody of claim 34, wherein the anti-ICAMl heavy chain comprises an anti-ICAMl heavy chain variable domain.

36. The multispecific antibody of claim 35, wherein the anti-ICAMl heavy chain variable domain comprises a variable domain of an IgGl, IgG2, IgG3, or IgG4 heavy chain.

37. The multispecific antibody of claim 34, wherein the anti-ICAMl light chain comprises an anti-ICAMl light chain variable domain.

38. The multispecific antibody of claim 37, wherein the anti-ICAMl light chain variable domain comprises a variable domain of a Kappa or Lambda light chain.

39. The multispecific antibody of claim 28, wherein the anti-ICAMl heavy chain variable domain comprises the variable domain of an IgGl heavy chain and the anti-ICAMl light chain variable domain comprises the variable domain of a Kappa or Lambda light chain.

40. The multispecific antibody according to claim 33, wherein the antibody, or functional fragment or functional variant thereof, that binds specifically to ICAM1 comprises a single-chain variable fragment (scFv) or an antigen -binding fragment (Fab).

-86-

41. The multispecific antibody of claim 35, wherein the anti -I CAM 1 heavy chain variable domain comprises complementarity determining regions (CDRs): HC-CDR1, HC-CDR2, and HC-CDR3, and wherein the HC-CDR1, the HC-CDR2, and the HC-CDR3 of the anti-ICAMl heavy chain variable domain comprise amino acid sequences according to HC-CDR1: SEQ ID NO: 19, 25, 46, 49, 52, 55, 58, 61, 64, 67, 70, 73, 76, 79, 82, 85, 88, 91, 94, 97, 100, 103, 106, or 109; HC-CDR2: SEQ ID NO: 20, 26, 47, 50, 53, 56, 59, 62, 65, 68, 71, 74, 77, 80, 83, 86, 89, 92, 95, 98, 101, 104, or 107; HC-CDR3: SEQ ID NO: 21, 27, 48, 51, 54, 57, 60, 63, 66, 69, 72, 75, 78, 81, 84, 87, 90, 93, 96, 99, 102, 105, 108, or 111; and wherein the CDRs comprise from 0-2 amino acid modification(s) in at least one of the HC-CDR1, HC- CDR2, or HC-CDR3.

42. The multispecific antibody of claim 35, wherein the anti-ICAMl heavy chain variable domain comprises complementarity determining regions (CDRs): HC-CDR1, HC-CDR2, and HC-CDR3, and wherein the HC-CDR1, the HC-CDR2, and the HC-CDR3 of the anti-ICAMl heavy chain variable domain comprise amino acid sequences according to HC-CDR1: SEQ ID NO: 25; HC-CDR2: SEQ ID NO: 26; HC-CDR3: SEQ ID NO: 27; and wherein the CDRs comprise from 0-2 amino acid modification(s) in at least one of the HC-CDR1, HC-CDR2, or HC-CDR3.

43. The multispecific antibody of claim 35, wherein the anti-ICAMl heavy chain variable domain comprises complementarity determining regions (CDRs): HC-CDR1, HC-CDR2, and HC-CDR3, and wherein the HC-CDR1, the HC-CDR2, and the HC-CDR3 of the anti-ICAMl heavy chain variable domain comprise amino acid sequences according to HC-CDR1: SEQ ID NO: 25; HC-CDR2: SEQ ID NO: 26; HC-CDR3: SEQ ID NO: 27; and wherein the CDRs comprise from 0-1 amino acid modification(s) in at least one of the HC-CDR1, HC-CDR2, or HC-CDR3.

44. The multispecific antibody of claim 35, wherein the anti-ICAMl heavy chain variable domain comprises complementarity determining regions (CDRs): HC-CDR1, HC-CDR2, and HC-CDR3, and wherein the HC-CDR1, the HC-CDR2, and the HC-CDR3 of the anti-ICAMl heavy chain variable domain comprise amino acid sequences according to HC-CDR1: SEQ ID NO: 25; HC-CDR2: SEQ ID NO: 26; HC-CDR3: SEQ ID NO: 27.

45. The multispecific antibody of claim 37, wherein the anti-ICAMl light chain variable domain comprises complementarity determining regions (CDRs): LC-CDR1, LC-CDR2, and LC-CDR3, and wherein the LC-CDR1, the LC-CDR2, and the LC-CDR3 of the anti-ICAMl light chain variable domain comprises amino acid sequences according to LC-CDR1: SEQ ID NO: 22, 28, 112, 115, 118, 121, 124, 127, 130, 133, 136, 139, 142, 145, 148, 151, 154, 157, 160, or 163; LC-CDR2: SEQ ID NO: 23, 29, 110, 113, 116, 119, 122, 125, 128, 131, 134, 137, 140, 143, 146, 149, 152, 155, 158, 161, or 164; LC-CDR3: SEQ ID NO: 24, 30, 114, 117, 120, 123, 126, 129, 132, 134, 138, 141, 144, 147, 150, 153, 156, 159, 162, or 165; and wherein the CDRs comprise from 0-2 amino acid modification(s) in at least one of the LC- CDR1, LC-CDR2, or LC-CDR3.

46. The multispecific antibody of claim 37, wherein the anti-ICAMl light chain variable domain comprises complementarity determining regions (CDRs): LC-CDR1, LC-CDR2, and LC-CDR3, and wherein the LC-CDR1, the LC-CDR2, and the LC-CDR3 of the anti-ICAMl light chain variable domain

-87- comprises amino acid sequences according to LC-CDR1: SEQ ID NO: 28; LC-CDR2: SEQ ID NO: 29; LC-CDR3: SEQ ID NO: 30; and wherein the CDRs comprise from 0-2 amino acid modification(s) in at least one of the LC-CDR1, LC-CDR2, or LC-CDR3.

47. The multispecific antibody of claim 37, wherein the anti -I CAM 1 light chain variable domain comprises complementarity determining regions (CDRs): LC-CDR1, LC-CDR2, and LC-CDR3, and wherein the LC-CDR1, the LC-CDR2, and the LC-CDR3 of the anti-ICAMl light chain variable domain comprises amino acid sequences according to LC-CDR1: SEQ ID NO: 28; LC-CDR2: SEQ ID NO: 29; LC-CDR3: SEQ ID NO: 30; and wherein the CDRs comprise from 0-1 amino acid modification(s) in at least one of the LC-CDR1, LC-CDR2, or LC-CDR3.

48. The multispecific antibody of claim 37, wherein the anti-ICAMl light chain variable domain comprises complementarity determining regions (CDRs): LC-CDR1, LC-CDR2, and LC-CDR3, and wherein the LC-CDR1, the LC-CDR2, and the LC-CDR3 of the anti-ICAMl light chain variable domain comprises amino acid sequences according to LC-CDR1: SEQ ID NO: 28; LC-CDR2: SEQ ID NO: 29; LC-CDR3: SEQ ID NO: 30.

49. The multispecific antibody of claim 34, wherein the HC-CDR1, the HC-CDR2, and the HC- CDR3 of the anti-ICAMl heavy chain variable domain comprises amino acid sequences according to HC-CDR1: SEQ ID NO: 19, 25, 46, 49, 52, 55, 58, 61, 64, 67, 70, 73, 76, 79, 82, 85, 88, 91, 94, 97, 100, 103, 106, or 109; HC-CDR2: SEQ ID NO: 20, 26, 47, 50, 53, 56, 59, 62, 65, 68, 71, 74, 77, 80, 83, 86, 89, 92, 95, 98, 101, 104, or 107; HC-CDR3: SEQ ID NO: 21, 27, 48, 51, 54, 57, 60, 63, 66, 69, 72, 75, 78, 81, 84, 87, 90, 93, 96, 99, 102, 105, 108, or 111 and the LC-CDR1, the LC-CDR2, and the LC-CDR3 of the anti-ICAMl light chain variable domain comprises amino acid sequences according to LC-CDR1: SEQ ID NO: 22, 28, 112, 115, 118, 121, 124, 127, 130, 133, 136, 139, 142, 145, 148, 151, 154, 157, 160, or 163; LC-CDR2: SEQ ID NO: 23, 29, 110, 113, 116, 119, 122, 125, 128, 131, 134, 137, 140, 143, 146, 149, 152, 155, 158, 161, or 164; LC-CDR3: SEQ ID NO: 24, 30, 114, 117, 120, 123, 126, 129, 132, 134, 138, 141, 144, 147, 150, 153, 156, 159, 162, or 165.

50. The multispecific antibody of claim 34, wherein the HC-CDR1, the HC-CDR2, and the HC- CDR3 of the anti-ICAMl heavy chain variable domain comprises amino acid sequences according to HC-CDR1: SEQ ID NO: 25; HC-CDR2: SEQ ID NO: 26; HC-CDR3: SEQ ID NO: 27; and the LC- CDR1, the LC-CDR2, and the LC-CDR3 of the anti-ICAMl light chain variable domain comprises amino acid sequences according to LC-CDR1: SEQ ID NO: 28; LC-CDR2: SEQ ID NO: 29; LC-CDR3: SEQ ID NO: 30; wherein the CDRs comprise from 0-2 amino acid modification(s) in at least one of the HC- CDR1, HC-CDR2, or HC-CDR3; and wherein the CDRs comprise from 0-2 amino acid modification(s) in at least one of the LC-CDR1, LC-CDR2, or LC-CDR3.

51. The multispecific antibody of claim 34, wherein the HC-CDR1, the HC-CDR2, and the HC- CDR3 of the anti-ICAMl heavy chain variable domain comprises amino acid sequences according to HC-CDR1: SEQ ID NO: 25; HC-CDR2: SEQ ID NO: 26; HC-CDR3: SEQ ID NO: 27; and the LC- CDR1, the LC-CDR2, and the LC-CDR3 of the anti-ICAMl light chain variable domain comprises amino acid sequences according to LC-CDR1: SEQ ID NO: 28; LC-CDR2: SEQ ID NO: 29; LC-CDR3: SEQ

-88- ID NO: 30; wherein the CDRs comprise from 0-1 amino acid modification(s) in at least one of the HC- CDR1, HC-CDR2, or HC-CDR3; and wherein the CDRs comprise from 0-1 amino acid modification(s) in at least one of the LC-CDR1, LC-CDR2, or LC-CDR3.

52. The multispecific antibody of claim 34, wherein the HC-CDR1, the HC-CDR2, and the HC- CDR3 of the anti-ICAMl heavy chain variable domain comprises amino acid sequences according to HC-CDR1: SEQ ID NO: 25; HC-CDR2: SEQ ID NO: 26; HC-CDR3: SEQ ID NO: 27; and the LC- CDR1, the LC-CDR2, and the LC-CDR3 of the anti-ICAMl light chain variable domain comprises amino acid sequences according to LC-CDR1: SEQ ID NO: 28; LC-CDR2: SEQ ID NO: 29; LC-CDR3: SEQ ID NO: 30.

53. The multispecific antibody of claim 34, wherein the anti-ICAMl heavy chain comprises an amino acid sequence with at least 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%,

91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to any one of SEQ ID NOs: 35, 36,

41, 166-187, 206-227.

54. The multispecific antibody of claim 34, wherein the anti-ICAMl heavy chain comprises an amino acid sequence with at least 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%,

91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to of SEQ ID NO: C.

55. The multispecific antibody of claim 34, wherein the anti-ICAMl light chain comprises an amino acid sequence with at least 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%,

91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to any one of SEQ ID NOs: 39, 40,

44, 45, 188-205, 228-245.

56. The multispecific antibody of claim 34, wherein the anti-ICAMl light chain comprises an amino acid sequence with at least 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%,

91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NO: D.

57. The multispecific antibody of claim 34, wherein the anti-ICAMl heavy chain comprises an amino acid sequence with at least 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%,

91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity at least 75%, 80%, 81%, 82%, 83%_;

84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to any one of SEQ ID NOs: 35, 36, 37, 38, 41, 42, 43, 166-187, 206-227 and the anti-ICAMl light chain comprises an amino acid sequence with at least 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to any one of SEQ ID NOs: 39, 40, 44, 45, 188-205, 228-245.

58. The multispecific antibody of claim 34, wherein the anti-ICAMl heavy chain comprises an amino acid sequence with at least 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NO: C; and the anti- ICAMl light chain comprises an amino acid sequence with at least 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NO: D.

59. The multispecific antibody of claim 34, wherein the anti-ICAMl heavy chain comprises an amino acid sequence with at least 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NO: 41.

60. The multispecific antibody of claim 34, wherein the anti-ICAMl light chain comprises an amino acid sequence with at least 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NO: 44.

61. The multispecific antibody of claim 34, wherein the anti-ICAMl heavy chain comprises an amino acid sequence with at least 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NO: 41; and wherein the anti-ICAMl light chain comprises an amino acid sequence with at least 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NO: 44.

62. The multispecific antibody of claim 13, wherein the antibody, or functional fragment or functional variant thereof, that binds specifically to CD47 comprises the Fab, and the antibody, or functional fragment or functional variant thereof, that binds specifically to ICAM1 comprises the scFv.

63. The multispecific antibody of claim 62, wherein the anti-CD47 heavy chain comprises an amino acid sequence with at least 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NO: A, and the anti- CD47 light chain comprises an amino acid sequence with at least 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NO: B; and the scFv comprises an amino acid sequence with at least 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NO: 11.

64. The multispecific antibody of claim 13, wherein the anti-CD47 heavy chain comprises an scFv that comprises an amino acid sequence with at least 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NO: 11.

65. The multispecific antibody of claim 13, wherein the antibody, or functional fragment or functional variant thereof, that binds specifically to CD47 comprises the scFv, and the antibody, or functional fragment or functional variant thereof, that binds specifically to ICAM1 comprises the Fab.

66. The multispecific antibody of claim 65, wherein the anti-ICAMl heavy chain comprises an amino acid sequence with at least 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%,

91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NO: 251 and the anti-

ICAM1 light chain comprises an amino acid sequence with at least 75%, 80%, 81%, 82%, 83%, 84%,

85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NO: 252; and the scFv comprises an amino acid sequence with at least 75%, 80%, 81%, 82%,

100% identity to SEQ ID NO: 253.

67. The multispecific antibody of claim 65, wherein the anti -I CAM 1 heavy chain comprises an amino acid sequence with at least 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NO: 254 and the anti- ICAM1 light chain comprises an amino acid sequence with at least 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NO: 255.

68. The multispecific antibody of claim 65, wherein the anti-ICAMl heavy chain comprises an amino acid sequence with at least 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NO: 254 and the anti- ICAMl light chain comprises an amino acid sequence with at least 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NO: 255; and the scFv comprises an amino acid sequence with at least 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NO: 256.

69. The multispecific antibody of claim 1, wherein the multispecific antibody comprises at least 3 CDRs of an anti-CD47 binding domain selected from any one of SEQ ID NO: 1-6 and from 0-2 amino acid modification(s) thereof; and at least 3 CDRs of an anti-ICAMl binding domain selected from any one of SEQ ID NO: 19, 25, 46, 49, 52, 55, 58, 61, 64, 67, 70, 73, 76, 79, 82, 85, 88, 91, 94, 97, 100, 103, 106, 109, 20, 26, 47, 50, 53, 56, 59, 62, 65, 68, 71, 74, 77, 80, 83, 86, 89, 92, 95, 98, 101, 104, 107, 21, 27, 48, 51, 54, 57, 60, 63, 66, 69, 72, 75, 78, 81, 84, 87, 90, 93, 96, 99, 102, 105, 108, 111 and from 0-2 amino acid modification(s) thereof.

70. The multispecific antibody of claim 1, wherein the multispecific antibody comprises at least 3 CDRs of an anti-CD47 binding domain selected from any one of SEQ ID NOs: 1-6, and from 0-2 amino acid modification(s) thereof; and at least 3 CDRs of an anti-ICAMl binding domain selected from any one of SEQ ID NO: 25-30, and from 0-2 amino acid modification(s) thereof.

71. The multispecific antibody of claim 1, wherein the multispecific antibody comprises at least 3 CDRs of an anti-CD47 binding domain selected from any one of SEQ ID NOs: 1-6, and from 0-1 amino acid modification(s) thereof; and at least 3 CDRs of an anti-ICAMl binding domain selected from any one of SEQ ID NO: 25-30, and from 0-1 amino acid modification(s) thereof.

72. The multispecific antibody of claim 1, wherein the multispecific antibody comprises at least 3 CDRs of an anti-CD47 binding domain selected from any one of SEQ ID NOs: 1-6; and at least 3 CDRs of an anti-ICAMl binding domain selected from any one of SEQ ID NO: 25-30.

73. The multispecific antibody of claim 1, wherein the multispecific antibody comprises 6 CDRs of an anti-CD47 binding domain having the sequences of SEQ ID NOs: 1-6, and from 0-2 amino acid modification(s) thereof; and 6 CDRs of an anti-ICAMl binding domain having the sequences of SEQ ID NO: 25-30, and from 0-2 amino acid modification(s) thereof.

74. The multispecific antibody of claim 1, wherein the multispecific antibody comprises 6 CDRs of an anti-CD47 binding domain having the sequences of SEQ ID NOs: 1-6, and from 0-1 amino acid modification(s) thereof; and 6 CDRs of an anti-ICAMl binding domain having the sequences of SEQ ID NO: 25-30, and from 0-1 amino acid modification(s) thereof.

75. The multispecific antibody of claim 1, wherein the multispecific antibody comprises 6 CDRs of an anti-CD47 binding domain having the sequences of SEQ ID NOs: 1-6; and 6 CDRs of an anti- ICAMl binding domain having the sequences of SEQ ID NO: 25-30.

76. The multispecific antibody of claim 1, wherein the antibody, or functional fragment or functional variant thereof that binds specifically to CD47 comprises the scFv, and the antibody, or functional fragment or functional variant thereof, that binds specifically to ICAM1 comprises the Fab, and wherein the HC-CDR1, the HC-CDR2, and the HC-CDR3 of the anti-CD47 heavy chain variable domain comprises amino acid sequences according to HC-CDR1: SEQ ID NO: 1; HC-CDR2: SEQ ID NO: 2; HC-CDR3: SEQ ID NO: 3 and the LC-CDR1, the LC-CDR2, and the LC-CDR3 of the anti-CD47 light chain variable domain comprises amino acid sequences according to LC-CDR1: SEQ ID NO: 4; LC- CDR2: SEQ ID NO: 5; LC-CDR3: SEQ ID NO: 6, and wherein the HC-CDR1, the HC-CDR2, and the HC-CDR3 of the anti-ICAMl heavy chain variable domain comprises amino acid sequences according to HC-CDR1: SEQ ID NO: 25; HC-CDR2: SEQ ID NO: 26; HC-CDR3: SEQ ID NO: 27; and the LC- CDR1, the LC-CDR2, and the LC-CDR3 of the anti-ICAMl light chain variable domain comprises amino acid sequences according to LC-CDR1: SEQ ID NO: 28; LC-CDR2: SEQ ID NO: 29; LC-CDR3: SEQ ID NO: 30

77. The multispecific antibody of claim 1, wherein the antibody, or functional fragment or functional variant thereof that binds specifically to CD47 comprises the Fab, and the antibody, or functional fragment or functional variant thereof, that binds specifically to ICAM1 comprises the scFv, and wherein the HC-CDR1, the HC-CDR2, and the HC-CDR3 of the anti-CD47 heavy chain variable domain comprises amino acid sequences according to HC-CDR1: SEQ ID NO: 1; HC-CDR2: SEQ ID NO: 2; HC-CDR3: SEQ ID NO: 3 and the LC-CDR1, the LC-CDR2, and the LC-CDR3 of the anti-CD47 light chain variable domain comprises amino acid sequences according to LC-CDR1: SEQ ID NO: 4; LC-CDR2: SEQ ID NO: 5; LC-CDR3: SEQ ID NO: 6; and wherein the HC-CDR1, the HC-CDR2, and the HC-CDR3 of the anti-ICAMl heavy chain variable domain comprises amino acid sequences according to HC-CDR1: SEQ ID NO: 25; HC-CDR2: SEQ ID NO: 26; HC-CDR3: SEQ ID NO: 27; and the LC-CDR1, the LC-CDR2, and the LC-CDR3 of the anti-ICAMl light chain variable domain comprises amino acid sequences according to LC-CDR1: SEQ ID NO: 28; LC-CDR2: SEQ ID NO: 29; LC-CDR3: SEQ ID NO: 30.

78. The multispecific antibody of claim 1, further comprising a fragment crystallizable (Fc) region.

79. The multispecific antibody of claim 78, wherein the Fc region comprises an IgG CH2 domain and an IgG CH3 domain.

80. The multispecific antibody of claim 78, wherein the Fc region comprises a heterodimeric Fc region.

81. The multispecific antibody of claim 78, wherein the Fc region comprises at least one amino acid modification that increases the half-life of the multispecific antibody.

82. The multispecific antibody of claim 78, wherein the Fc region comprises at least one amino acid modification that modulates its interaction with an Fc receptor.

83. The multispecific antibody of claim 78, wherein the Fc region comprises at least one amino acid modification that increases binding of the Fc region to an Fc receptor.

84. The multispecific antibody of claim 78, wherein the Fc region comprises at least one amino acid modification that decreases glycosylation of the Fc region.

85. The multispecific antibody of claim 84, wherein the modification is an amino acid substitution, deletion, or addition.

86. The multispecific antibody of claim 85, wherein the modification is an amino acid substitution.

87. The multispecific antibody of claim 86, wherein the at least one amino acid modification that decreases glycosylation of the Fc region comprises an amino acid substitution at a position corresponding to position N297 of human IgGl, wherein the numbering is according to the EU index of Kabat.

88. The multispecific antibody of claim 78, wherein the Fc region is afucosylated.

89. The multispecific antibody of claim 78, wherein the Fc region comprises at least one amino acid modification that increases antibody -dependent cellular cytotoxicity (ADCC).

90. The multispecific antibody of claim 89, wherein the modification is an amino acid substitution, deletion, or addition.

91. The multispecific antibody of claim 90, wherein the modification is an amino acid substitution.

92. The multispecific antibody of claim 78, wherein the Fc region comprises at least one mutation that increases antibody-dependent cellular cytotoxicity (ADCC), wherein the at least one mutation that increases ADCC comprises an amino acid substitution at positions corresponding to positions S239, 1332, and A330 of human IgGl, wherein the amino acid numbering is according to the EU index according to Kabat et al.

93. The multispecific antibody of claim 92, wherein the amino acid substitutions are S239D, I332E, and A330L, wherein the amino acid numbering is according to the EU index according to Kabat et al.

94. The multispecific antibody of claim 80, comprising the heterodimeric Fc region, wherein the heterodimeric Fc region comprises a knob chain and a hole chain, forming a knob-into-hole (KIH) structure.

95. The multispecific antibody of claim 94, wherein the knob chain comprises an IgGl, IgG2, IgG3, or IgG4 domain.

96. The multispecific antibody of claim 94, wherein the hole chain comprises an IgGl, IgG2, IgG3, or IgG4 domain.

-93-

97. The multispecific antibody of claim 94, wherein the knob chain comprises an amino acid substitution at aposition corresponding to T366 of IgGl, wherein amino acid position numbering is according to the EU index according to Kabat et al.

98. The multispecific antibody of claim 97, wherein the T366 substitution comprises a T336W mutation, wherein amino acid position numbering is according to the EU index according to Kabat et al.

99. The multispecific antibody of claim 94, wherein the hole chain comprises an amino acid substitution at aposition corresponding to T366, L368, or Y407 of IgGl, wherein amino acid position numbering is according to the EU index according to Kabat et al.

100. The multispecific antibody of claim 99, wherein the hole chain comprises an amino acid substitution at a position corresponding to T366, L368, and Y407 of IgGl, wherein amino acid position numbering is according to the EU index according to Kabat et al.

101. The multispecific antibody of claim 100, wherein the T366, L368, or Y407 amino acid substitutions comprise a T366S, L368A, or Y407V of IgGl, wherein amino acid position numbering is according to the EU index according to Kabat et al.

102. The multispecific antibody of claim 101, wherein the T366, L368, and Y407 amino acid substitutions comprises a T366S, L368A, and Y407V of IgGl, wherein amino acid position numbering is according to the EU index according to Kabat et al.

103. The multispecific antibody of claim 1, wherein the multispecific antibody binds to a target cell.

104. The multispecific antibody of claim 103, wherein the target cell expresses CD47 and ICAM1.

105. The multispecific antibody of claim 104, wherein the target cell expresses a lower level of CD47 relative to ICAM1 on the surface of the target cell.

106. The multispecific antibody of claim 103, wherein the target cell is a cancer cell.

107. The multispecific antibody of claim 106, wherein the cancer is a hematological malignancy.

108. The multispecific antibody of claim 106, wherein the cancer is multiple myeloma.

109. The multispecific antibody of claim 107, wherein the hematological malignancy is B cell cancer.

110. The multispecific antibody of claim 108, wherein the B cell cancer is leukemia or lymphoma.

111. The multispecific antibody of claim 110, wherein the B cell cancer is lymphoma, and wherein the lymphoma is B cell lymphoma.

112. The multispecific antibody of claim 104, the hematological malignancy is T cell cancer.

113. The multispecific antibody of claim 112, the T cell cancer is cutaneous T cell lyphoma, or anaplastic large cell lymphoma.

114. The multispecific antibody of claim 106, wherein the cancer cell is from a solid tumor.

-94-

115. The multispecific antibody of claim 114, wherein the solid tumor is sarcoma, breast cancer, lung cancer, kidney cancer, melanoma, head and neck cancer, ovarian cancer, liver cancer, bladder cancer, thyroid cancer, or carcinoma.

116. The multispecific antibody of claim 114, wherein the solid tumor is lung cancer, and wherein the lung cancer is non-small cell lung cancer.

117. The multispecific antibody of claim 1, wherein the multispecific antibody inhibits binding of Signal-regulatory protein alpha (SIRPa) to the target cell as determined by an in vitro competition assay.

118. The multispecific antibody of claim 117, wherein the binding of Signal -regulatory protein alpha (SIRPa) to the target cell as determined by an in vitro competition assay is inhibited by the multispecific antibody by at least 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100%.

119. The multispecific antibody of claim 118, wherein a concentration of the multispecific antibody required to mediate antibody-dependent cellular phagocytosis (ADCP) of the target cell by a macrophage is from 0.0 InM to 3nM.

120. The multispecific antibody of claim 118, wherein the multispecific antibody induces enhanced antibody-dependent cellular phagocytosis (ADCP) of the target cell by a macrophage as compared to ADCP activity induced of the target cell by a macrophage by a monospecific anti-CD47 antibody.

121. The multispecific antibody of claim 103, wherein the multispecific antibody has a higher binding activity for CD47 expressed on a surface of a ICAM1 expressing tumor cell than for CD47 expressed on a surface of a red blood cell or a platelet.

122. The multispecific antibody of claim 103, wherein a concentration of the multispecific antibody required for half-maximal binding to a human red blood or a human platelet is greater than 500 nM.

123. The multispecific antibody of claim 103, wherein the multispecific antibody induces antibody-dependent cellular cytotoxicity (ADCC) mediated killing of the target cell.

124. The multispecific antibody of claim 123, wherein the multispecific antibody induces enhanced antibody-dependent cellular cytotoxicity (ADCC) activity on the target cell as compared to ADCC activity induced on the target cell by a monospecific anti-CD47 antibody or a monospecific anti- ICAM1 antibody.

125. The multispecific antibody of claim 124, wherein the multispecific antibody induces at least 5%, 10%, 15%, 20%, 25%, 30%, 25%, 40%, 45%, or 50% more ADCC activity of the target cell as compared to an ADCC activity of the target cell that is induced by a monospecific anti-CD47 antibody or a monospecific anti -I CAM 1 antibody.

126. The multispecific antibody of claim 103, wherein the multispecific antibody induces antibody-dependent cellular phagocytosis (ADCP) mediated killing of the target cell.

127. The multispecific antibody of claim 126, wherein the multispecific antibody induces at least 5%, 10%, 15%, 20%, 25%, 30%, 25%, 40%, 45%, or 50% more ADCP activity of the target cell as

-95- compared to a ADCP activity of the target cell that is induced by a monospecific anti-CD47 antibody or a monospecific anti-ICAMl antibody.

128. The multispecific antibody of claim 103, wherein a concentration of 600 nM of the multispecific antibody does not induce hemolysis of red blood cells in a hemagglutination assay.

129. A nucleic acid molecule encoding the multispecific antibody of claim 1.

130. A vector comprising the nucleic acid molecule of claim 129.

131. A pharmaceutical composition comprising the multispecific antibody of claim 1.

132. The pharmaceutical composition of claim 131, further comprising a pharmaceutically acceptable carrier, an excipient, or any combinations thereof.

133. A method of treating a subject having cancer, the method comprising: administering to the subject the multispecific antibody of claim 1.

134. The method of claim 133, wherein the cancer comprises cancer cells that express CD47 and ICAM1.

135. The method of claim 134, wherein the cancer cells that express CD47 and ICAM1 are lysed.

136. The method of claim 133, wherein the multispecific antibody induces antibody -dependent cellular cytotoxicity (ADCC) mediated killing of the cancer cells that express CD47 and/or ICAM1.

137. The method of claim 133, wherein the multispecific antibody induces antibody -dependent cellular phagocytosis (ADCP) of the cancer cells that express CD47 and/or ICAM1.

138. The method of claim 133, wherein the cancer is a hematological malignancy.

139. The method of claim 138, wherein the cancer is Multiple Myeloma.

140. The method of claim 138, wherein the cancer is B cell cancer.

141. The method of claim 139, wherein the cancer is leukemia or lymphoma.

142. The method of claim 141, wherein the cancer is lymphoma, and wherein the lymphoma is

B-cell lymphoma.

143. The method of claim 138, the hematological malignancy is T cell cancer.

144. The method of claim 143, the T cell cancer is cutaneous T cell lyphoma, or anaplastic large cell lymphoma.

145. The method of claim 133, wherein the cancer is a solid tumor.

146. The method of claim 145, wherein the solid tumor is sarcoma, breast cancer, lung cancer, kidney cancer, melanoma, head and neck cancer, ovarian cancer, liver cancer, bladder cancer, thyroid cancer, or carcinoma.

147. The method of claim 146, wherein the solid tumor is lung cancer, and wherein the lung cancer is non-small cell lung cancer.

148. The method of claim 133, further comprising administering to the subject an anti-cancer agent.

149. The method of claim 148, wherein the anti-cancer agent is a chemotherapeutic agent or a biologic agent.

-96-

150. The method of claim 149, wherein the administering is sufficient to reduce or eliminate the cancer as compared to a comparable method lacking the administering.

151. The method of claim 150, wherein the reduction is at least about 1 fold, 5 fold, 10 fold, 20 fold, 40 fold, 60 fold, 80 fold, or up to about 100 fold.

152. The method of claim 133, wherein the cancer is metastatic.

153. A kit that comprises at least one of:

(a) the multispecific antibody of claim 1 ;

(b) the vector of claim 130;

(c) the nucleic acid molecule of claim 129; or

(d) the pharmaceutical composition of claim 131.

-97-