CN116829177A

CN116829177A - Effective neutralizing antibodies against SARS-CoV-2 and production and use thereof

Info

Publication number: CN116829177A
Application number: CN202180059673.2A
Authority: CN
Inventors: 何大一; 黄耀星; 刘立鸿; M·S·奈尔; 王鹏飞; 于健; Y·罗
Original assignee: Columbia University in the City of New York
Current assignee: Columbia University in the City of New York
Priority date: 2020-05-20
Filing date: 2021-05-20
Publication date: 2023-09-29

Abstract

The application described herein relates to potent monoclonal and bispecific antibodies capable of neutralizing SARS-CoV-2 virus and methods of producing said antibodies.

Description

Effective neutralizing antibodies against SARS-CoV-2 and production and use thereof

The present application claims the benefit and priority of U.S. provisional patent application number 63/027,935 filed on month 5 and day 20 of 2020, U.S. provisional patent application number 63/032,518 filed on month 5 and day 29 of 2020, U.S. provisional patent application number 63/039,977 filed on month 6 and day 16 of 2020, U.S. provisional patent application number 63/063,106 filed on month 8 and day 7 of 2020, U.S. provisional patent application number 63/123,767 filed on month 12 and day 10 of 2020, U.S. provisional patent application number 63/060,116 filed on month 8 and day 2 of 2020, U.S. provisional patent application number 63/117,908 filed on month 11 and day 24 of 2021, U.S. provisional patent application number 63/165,729 filed on month 3 and 24 of 2020, each of which is incorporated herein by reference in its entirety.

All patents, patent applications, and publications cited herein are incorporated by reference in their entirety. The disclosures of these publications are incorporated herein by reference in their entireties.

This patent disclosure contains material that is subject to copyright protection. The copyright owner has no objection to the facsimile reproduction by anyone of the patent document or the patent disclosure, as it appears in the patent and trademark office patent file or records, but otherwise reserves all copyright rights whatsoever.

Background

Coronavirus disease (covd-19) is an infectious disease caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). This disease affects multiple organs of the body, including the lungs. There is an urgent need to develop therapies against covd-19.

Disclosure of Invention

In certain aspects, the invention provides a bispecific molecule comprising a first polypeptide chain and a second polypeptide chain, wherein: the first polypeptide chain comprises: a first light chain comprising a variable domain and a constant domain; a first heavy chain comprising a variable domain and a constant domain; the second polypeptide chain comprises: a second light chain comprising a variable domain and a constant domain; and a second heavy chain comprising a variable domain and a constant domain.

In some embodiments, the first polypeptide chain and the second polypeptide chain are covalently bonded to each other. In some embodiments, the first heavy chain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 59 or at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 63. In some embodiments, the first light chain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 60 or at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 64. In some embodiments, the second heavy chain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 61 or at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 65. In some embodiments, the second light chain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 62 or at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 66.

In some embodiments, the first heavy chain comprises SEQ ID NO 59 or SEQ ID NO 63. In some embodiments, the first light chain comprises SEQ ID NO. 60 or SEQ ID NO. 64. In some embodiments, the second heavy chain comprises SEQ ID NO. 61 or SEQ ID NO. 65. In some embodiments, the second light chain comprises SEQ ID NO. 62 or SEQ ID NO. 66.

In some embodiments, the variable domain of the first heavy chain comprises CDR1, CDR2 and CDR3 of SEQ ID NO:59 or CDR1, CDR2 and CDR3 of SEQ ID NO:63. In some embodiments, the variable domain of the first light chain comprises CDR1, CDR2 and CDR3 of SEQ ID NO:60 or CDR1, CDR2 and CDR3 of SEQ ID NO:64. In some embodiments, the variable domain of the second heavy chain comprises CDR1, CDR2 and CDR3 of SEQ ID NO:61 or CDR1, CDR2 and CDR3 of SEQ ID NO:65. In some embodiments, the variable domain of the second light chain comprises CDR1, CDR2 and CDR3 of SEQ ID NO:62 or CDR1, CDR2 and CDR3 of SEQ ID NO:66.

In some embodiments, the variable domain of the first heavy chain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 13 or at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 27. In some embodiments, the variable domain of the first light chain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 14 or a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 28. In some embodiments, the variable domain of the second heavy chain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 27 or at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 13. In some embodiments, the variable domain of the second light chain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 28 or a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 14.

In some embodiments, the variable domain of the first heavy chain comprises SEQ ID NO. 13 or SEQ ID NO. 27. In some embodiments, the variable domain of the first light chain comprises SEQ ID NO. 14 or SEQ ID NO. 28. In some embodiments, the variable domain of the second heavy chain comprises SEQ ID NO 27 or SEQ ID NO 13. In some embodiments, the variable domain of the second light chain comprises SEQ ID NO. 28 or SEQ ID NO. 14.

In some embodiments, the molecule comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID NO 59, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID NO, a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 98%, 99% identical to SEQ ID NO 62, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID NO. In some embodiments, the molecule comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 63, a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 98%, 99% identical to SEQ ID No. 66, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID NO. In some embodiments, the molecule comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 13, a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 14, a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 98%, 99% identical to SEQ ID No. 27, and a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID NO.

In some embodiments, the molecule comprises SEQ ID NO 59, SEQ ID NO 60, SEQ ID NO 61, and SEQ ID NO 62. In some embodiments, the molecule comprises SEQ ID NO:63, SEQ ID NO:64, SEQ ID NO:65, and SEQ ID NO:66. In some embodiments, the molecule comprises SEQ ID NO. 13, SEQ ID NO. 14, SEQ ID NO. 27 and SEQ ID NO. 28.

In some embodiments, the molecule further comprises one or more disulfide bonds. In some embodiments, the molecule further comprises one or more linker sequences. In some embodiments, the molecule is capable of neutralizing coronavirus. In some embodiments, the coronavirus is a SARS-CoV-2 strain. In some embodiments, the molecule specifically recognizes a first epitope and a second epitope on SARS-CoV-2 particle. In some embodiments, the first epitope comprises at least a portion of a Receptor Binding Domain (RBD) on the S protein of SARS-CoV-2 particle. In some embodiments, the second epitope comprises at least a portion of an N-terminal domain (NTD) on an S protein of a SARS-CoV-2 particle.

In certain aspects, the invention provides a method of treating a covd-19 infection in a subject in need thereof, the method comprising administering to the subject a composition comprising a bispecific molecule, wherein the bispecific molecule comprises a first polypeptide chain and a second polypeptide chain, wherein: the first polypeptide chain comprises: a first light chain comprising a variable domain and a constant domain; a first heavy chain comprising a variable domain and a constant domain; the second polypeptide chain comprises: a second light chain comprising a variable domain and a constant domain; and a second heavy chain comprising a variable domain and a constant domain.

In some embodiments, the first heavy chain comprises SEQ ID NO 59 or SEQ ID NO 63. In some embodiments, the first light chain comprises SEQ ID NO. 60 or SEQ ID NO. 64. In some embodiments, the second heavy chain comprises SEQ ID NO. 61 or SEQ ID NO. 65. In some embodiments, the second light chain comprises SEQ ID NO. 62 or SEQ ID NO. 66. In some embodiments, the variable domain of the first heavy chain comprises CDR1, CDR2 and CDR3 of SEQ ID NO:59 or CDR1, CDR2 and CDR3 of SEQ ID NO:63. In some embodiments, the variable domain of the first light chain comprises CDR1, CDR2 and CDR3 of SEQ ID NO:60 or CDR1, CDR2 and CDR3 of SEQ ID NO:64. In some embodiments, the variable domain of the second heavy chain comprises CDR1, CDR2 and CDR3 of SEQ ID NO:61 or CDR1, CDR2 and CDR3 of SEQ ID NO:65. In some embodiments, the variable domain of the second light chain comprises CDR1, CDR2 and CDR3 of SEQ ID NO:62 or CDR1, CDR2 and CDR3 of SEQ ID NO:66.

In some embodiments, the molecule further comprises one or more disulfide bonds. In some embodiments, the molecule further comprises one or more linker sequences. In some embodiments, the molecule is capable of neutralizing coronavirus. In some embodiments, the coronavirus is a SARS-CoV-2 strain. In some embodiments, the molecule specifically recognizes a first epitope and a second epitope on SARS-CoV-2 particle. In some embodiments, the first epitope comprises at least a portion of a Receptor Binding Domain (RBD) on a spike protein of a SARS-CoV-2 particle. In some embodiments, the second epitope comprises at least a portion of an N-terminal domain (NTD) on a spike protein of a SARS-CoV-2 particle. In some embodiments, the composition is administered in combination with a second therapeutic agent.

In certain aspects, the invention provides a method of preventing a covd-19 infection in a subject in need thereof, the method comprising administering to the subject a composition comprising a bispecific molecule, wherein the bispecific molecule comprises a first polypeptide chain and a second polypeptide chain, wherein: the first polypeptide chain comprises: a first light chain comprising a variable domain and a constant domain; a first heavy chain comprising a variable domain and a constant domain; the second polypeptide chain comprises: a second light chain comprising a variable domain and a constant domain; and a second heavy chain comprising a variable domain and a constant domain.

In some embodiments, the molecule further comprises one or more disulfide bonds. In some embodiments, the molecule further comprises one or more linker sequences. In some embodiments, the molecule is capable of neutralizing coronavirus. In some embodiments, the coronavirus is a SARS-CoV-2 strain. In some embodiments, the molecule specifically recognizes a first epitope and a second epitope on SARS-CoV-2 particle. In some embodiments, the first epitope comprises at least a portion of a Receptor Binding Domain (RBD) on the S protein of SARS-CoV-2 particle. In some embodiments, the second epitope comprises at least a portion of an N-terminal domain (NTD) on an S protein of a SARS-CoV-2 particle. In some embodiments, the composition is administered in combination with a second therapeutic agent.

In certain aspects, the invention provides a synthetic monoclonal antibody, or functional fragment thereof, comprising a heavy chain having a variable domain and a constant domain and a light chain having a variable domain and a constant domain, wherein the antibody binds an antigen on a coronavirus particle.

In some embodiments, the heavy chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 15. In some embodiments, the light chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 16. In some embodiments, the heavy chain variable domain comprises SEQ ID NO. 15. In some embodiments, the light chain variable domain comprises SEQ ID NO. 16.

In some embodiments, the antibody comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 15 and a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 16. In some embodiments, the antibody comprises SEQ ID NO. 15 and SEQ ID NO. 16.

In some embodiments, the heavy chain variable domain comprises the CDR1, CDR2 and CDR3 regions of SEQ ID NO 15. In some embodiments, the light chain variable domain comprises the CDR1, CDR2 and CDR3 regions of SEQ ID NO. 16.

In some embodiments, the antibody neutralizes coronavirus. In some embodiments, the coronavirus is SARS-CoV-2. In some embodiments, the antibody specifically binds to an epitope on the surface of the coronavirus. In some embodiments, the epitope comprises at least a portion of a coronavirus spike protein. In some embodiments, at least a portion of the coronavirus spike protein comprises a Receptor Binding Domain (RBD) of the spike protein. In some embodiments, the coronavirus is a SARS-CoV-2 coronavirus.

In certain aspects, the invention provides a method of treating a covd-19 infection in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of a synthetic monoclonal antibody, or a functional fragment thereof, comprising a heavy chain having a variable domain and a constant domain and a light chain having a variable domain and a constant domain, wherein the antibody binds an antigen on a coronavirus particle.

In some embodiments, the antibody neutralizes coronavirus. In some embodiments, the coronavirus is SARS-CoV-2. In some embodiments, the antibody neutralizes coronavirus. In some embodiments, the coronavirus is SARS-CoV-2. In some embodiments, the antibody specifically binds to an epitope on the surface of the coronavirus. In some embodiments, the epitope comprises at least a portion of a coronavirus spike protein. In some embodiments, at least a portion of the coronavirus spike protein comprises a receptor binding domain of the spike protein (receptor binding domain, RBD). In some embodiments, the coronavirus is a SARS-CoV-2 coronavirus. In some embodiments, the antibody is administered in combination with a second agent.

In certain aspects, the invention provides a method of preventing a covd-19 infection in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of a synthetic monoclonal antibody, or a functional fragment thereof, comprising a heavy chain having a variable domain and a constant domain and a light chain having a variable domain and a constant domain, wherein the antibody binds an antigen on a coronavirus particle.

In some embodiments, the antibody neutralizes coronavirus. In some embodiments, the coronavirus is SARS-CoV-2. In some embodiments, the antibody neutralizes coronavirus. In some embodiments, the coronavirus is SARS-CoV-2. In some embodiments, the antibody specifically binds to an epitope on the surface of the coronavirus. In some embodiments, the epitope comprises at least a portion of a coronavirus spike protein. In some embodiments, at least a portion of the coronavirus spike protein comprises a Receptor Binding Domain (RBD) of the spike protein. In some embodiments, the coronavirus is a SARS-CoV-2 coronavirus. In some embodiments, the antibody is administered in combination with a second agent.

In some embodiments, the heavy chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 3, a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 7, a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 11, at least 60%, 62%, 65%, 67%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 17, at least 60%, 62%, 67%, 70%, 72%, 75%, 77%, 80%, 98%, 95%, 98%, 99% identical to SEQ ID No. 9.

In some embodiments, the heavy chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 19, a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 29, at least 60%, 62%, 65%, 67%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 31, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 98%, 95%, 99% identical to SEQ ID No. 25.

In some embodiments, the heavy chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID NO, a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 97%, 98%, 99% identical to SEQ ID NO, or a polypeptide sequence that is at least 60%, 62%, 67%, 70%, 72%, 75%, 77%, 80%, 97%, 98%, 99% identical to SEQ ID NO.

In some embodiments, the light chain variable domain comprises a sequence that hybridizes to SEQ ID NO:4, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID NO:8, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID NO:10, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID NO:12, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 95%, 97%, 98%, 99% identical to SEQ ID NO:18, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98% identical to SEQ ID NO.

In some embodiments, the light chain variable domain comprises a sequence that hybridizes to SEQ ID NO:20, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID NO:22, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID NO:26, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID NO:30, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 95%, 97%, 98%, 99% identical to SEQ ID NO:32, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98% identical to SEQ ID NO.

In some embodiments, the light chain variable domain comprises a sequence that hybridizes to SEQ ID NO:34, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID NO:38, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID NO:40, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID NO:44, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 95%, 97%, 98%, 99% identical to SEQ ID NO:46, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 98%, 99% identical to SEQ ID NO: 46.

In some embodiments, the heavy chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 3, and the light chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 4. In some embodiments, the heavy chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 7, and the light chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 8. In some embodiments, the heavy chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 9, and the light chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 10. In some embodiments, the heavy chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 11, and the light chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 12. In some embodiments, the heavy chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 17, and the light chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 18. In some embodiments, the heavy chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 19, and the light chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 20. In some embodiments, the heavy chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 21, and the light chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 22. In some embodiments, the heavy chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 25, and the light chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 26. In some embodiments, the heavy chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 29, and the light chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 30.

In some embodiments, the heavy chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 31, and the light chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 32. In some embodiments, the heavy chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 33, and the light chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 34. In some embodiments, the heavy chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 37, and the light chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 38. In some embodiments, the heavy chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 39, and the light chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 40. In some embodiments, the heavy chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 43, and the light chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 44. In some embodiments, the heavy chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 45, and the light chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 46.

In some embodiments, the heavy chain variable domain comprises SEQ ID NO. 3 and the light chain variable domain comprises SEQ ID NO. 4. In some embodiments, the heavy chain variable domain comprises SEQ ID NO. 7 and the light chain variable domain comprises SEQ ID NO. 8. In some embodiments, the heavy chain variable domain comprises SEQ ID NO. 9 and the light chain variable domain comprises SEQ ID NO. 10. In some embodiments, the heavy chain variable domain comprises SEQ ID NO. 11 and the light chain variable domain comprises SEQ ID NO. 12. In some embodiments, the heavy chain variable domain comprises SEQ ID NO. 17 and the light chain variable domain comprises SEQ ID NO. 18. In some embodiments, the heavy chain variable domain comprises SEQ ID NO. 19 and the light chain variable domain comprises SEQ ID NO. 20. In some embodiments, the heavy chain variable domain comprises SEQ ID NO. 21 and the light chain variable domain comprises SEQ ID NO. 22. In some embodiments, the heavy chain variable domain comprises SEQ ID NO. 25 and the light chain variable domain comprises SEQ ID NO. 26. In some embodiments, the heavy chain variable domain comprises SEQ ID NO. 29 and the light chain variable domain comprises SEQ ID NO. 30. In some embodiments, the heavy chain variable domain comprises SEQ ID NO. 31 and the light chain variable domain comprises SEQ ID NO. 32. In some embodiments, the heavy chain variable domain comprises SEQ ID NO. 33 and the light chain variable domain comprises SEQ ID NO. 34. In some embodiments, the heavy chain variable domain comprises SEQ ID NO. 37 and the light chain variable domain comprises SEQ ID NO. 38. In some embodiments, the heavy chain variable domain comprises SEQ ID NO:39 and the light chain variable domain comprises SEQ ID NO:40. In some embodiments, the heavy chain variable domain comprises SEQ ID NO. 43 and the light chain variable domain comprises SEQ ID NO. 44. In some embodiments, the heavy chain variable domain comprises SEQ ID NO. 45 and the light chain variable domain comprises SEQ ID NO. 46.

In some embodiments, the heavy chain variable domain comprises the CDR1, CDR2 and CDR3 regions of SEQ ID NO 3; CDR1, CDR2 and CDR3 regions of SEQ ID No. 7; CDR1, CDR2 and CDR3 regions of SEQ ID No. 9; CDR1, CDR2 and CDR3 regions of SEQ ID No. 11; or CDR1, CDR2 and CDR3 regions of SEQ ID NO 17. In some embodiments, the heavy chain variable domain comprises the CDR1, CDR2 and CDR3 regions of SEQ ID NO 19; CDR1, CDR2 and CDR3 regions of SEQ ID No. 21; CDR1, CDR2 and CDR3 regions of SEQ ID No. 25; CDR1, CDR2 and CDR3 regions of SEQ ID No. 29; or CDR1, CDR2 and CDR3 regions of SEQ ID NO. 31. In some embodiments, the heavy chain variable domain comprises the CDR1, CDR2 and CDR3 regions of SEQ ID NO 33; CDR1, CDR2 and CDR3 regions of SEQ ID No. 37; CDR1, CDR2 and CDR3 regions of SEQ ID No. 39; CDR1, CDR2 and CDR3 regions of SEQ ID No. 43; or CDR1, CDR2 and CDR3 regions of SEQ ID NO 45. In some embodiments, the light chain variable domain comprises the CDR1, CDR2 and CDR3 regions of SEQ ID NO. 4; CDR1, CDR2 and CDR3 regions of SEQ ID No. 8; CDR1, CDR2 and CDR3 regions of SEQ ID No. 10; CDR1, CDR2 and CDR3 regions of SEQ ID No. 12; or CDR1, CDR2 and CDR3 regions of SEQ ID NO. 18. In some embodiments, the light chain variable domain comprises the CDR1, CDR2 and CDR3 regions of SEQ ID NO. 20; CDR1, CDR2 and CDR3 regions of SEQ ID No. 22; CDR1, CDR2 and CDR3 regions of SEQ ID No. 26; CDR1, CDR2 and CDR3 regions of SEQ ID No. 30; or CDR1, CDR2 and CDR3 regions of SEQ ID NO. 32. In some embodiments, the light chain variable domain comprises the CDR1, CDR2 and CDR3 regions of SEQ ID NO 34; CDR1, CDR2 and CDR3 regions of SEQ ID No. 38; CDR1, CDR2 and CDR3 regions of SEQ ID No. 40; CDR1, CDR2 and CDR3 regions of SEQ ID No. 44; or CDR1, CDR2 and CDR3 regions of SEQ ID NO 46.

In some embodiments, the antibody neutralizes coronavirus. In some embodiments, the coronavirus is a SARS-CoV-2 virus. In some embodiments, the coronavirus is a SARS-CoV-2 variant. In some embodiments, the variant plant is selected from the group consisting of WA1, b.1.1.7, B1.526, B1.351, P1, B1.352, and b.1.427.

In some embodiments, the antibody specifically binds to an epitope on the surface of the coronavirus. In some embodiments, the epitope comprises at least a portion of a coronavirus spike protein. In some embodiments, at least a portion of the coronavirus spike protein comprises a Receptor Binding Domain (RBD) of the spike protein. In some embodiments, at least a portion of the coronavirus spike protein comprises an N-terminal domain (NTD). In some embodiments, the coronavirus is a SARS-CoV-2 coronavirus.

In certain aspects, the invention provides a method of treating or preventing a covd-19 infection in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of a synthetic monoclonal antibody, or a functional fragment thereof, comprising a heavy chain having a variable domain and a constant domain and a light chain having a variable domain and a constant domain, wherein the antibody binds an antigen on a coronavirus particle.

In some embodiments, the light chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 4, a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 8, a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 12, at least 60%, 62%, 65%, 67%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 18, at least 60%, 62%, 67%, 70%, 72%, 75%, 77%, 80%, 98%, 99% identical to SEQ ID No. 10.

In some embodiments, the light chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 20, a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 22, a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 30, at least 60%, 62%, 65%, 67%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 32, at least 60%, 62%, 67%, 70%, 72%, 75%, 77%, 80%, 98%, 99% identical to SEQ ID NO.

In some embodiments, the heavy chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 3, and the light chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 4. In some embodiments, the heavy chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 7, and the light chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 8. In some embodiments, the heavy chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 9, and the light chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 10. In some embodiments, the heavy chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 11, and the light chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 12. In some embodiments, the heavy chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 17, and the light chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 18. In some embodiments, the heavy chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 19, and the light chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 20.

In some embodiments, the heavy chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 21, and the light chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 22. In some embodiments, the heavy chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 25, and the light chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 26. In some embodiments, the heavy chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 29, and the light chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 30. In some embodiments, the heavy chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 31, and the light chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 32. In some embodiments, the heavy chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 33, and the light chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 34. In some embodiments, the heavy chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 37, and the light chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 38.

In some embodiments, the heavy chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 39, and the light chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 40. In some embodiments, the heavy chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 43, and the light chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 44. In some embodiments, the heavy chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 45, and the light chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 46.

In some embodiments, the antibody specifically binds to an epitope on the surface of the coronavirus. In some embodiments, the epitope comprises at least a portion of a coronavirus spike protein. In some embodiments, at least a portion of the coronavirus spike protein comprises a Receptor Binding Domain (RBD) of the spike protein. In some embodiments, at least a portion of the coronavirus spike protein comprises an N-terminal domain (NTD) of the spike protein. In some embodiments, the coronavirus is a SARS-CoV-2 coronavirus. In some embodiments, the composition is administered in combination with a second therapeutic agent.

In some embodiments, the first polypeptide chain and the second polypeptide chain are covalently bonded to each other. In some embodiments, the first heavy chain comprises a sequence identical to SEQ ID NO:51, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to a polypeptide sequence of SEQ ID NO:55, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to a polypeptide sequence of SEQ ID NO: 67%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical polypeptide sequences to at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical polypeptide sequences to at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 95%, 97%, 98%, 99% identical polypeptide sequences to at least 60%, 62%, 65%, 82%, 98%, 99% identical polypeptide sequences to at least 60%, 83%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical polypeptide sequences to at least 60% of SEQ ID NO 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical polypeptide sequences to those of SEQ ID NO:91 at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to a polypeptide sequence of SEQ ID NO:95, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to a polypeptide sequence of SEQ ID NO:99 at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical polypeptide sequence, at least 60%, 119%, 62%, 65%, 67%, 70%, 72%, 75% identical polypeptide sequence, at least 60%, 62%, 65%, 67%, 70%, 75% identical polypeptide sequence to SEQ ID NO 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical polypeptide sequences to SEQ ID NO:123, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to a polypeptide sequence of SEQ ID NO:127 at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to a polypeptide sequence of SEQ ID NO:131 at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID NO:151 at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90% identical to SEQ ID NO:151 92%, 95%, 97%, 98%, 99% identical polypeptide sequence to SEQ ID NO:155, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to a polypeptide sequence of SEQ ID NO:159 at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to a polypeptide sequence of SEQ ID NO:163 is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to the polypeptide sequence of at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 95%, 97%, 98%, 99% identical to the polypeptide sequence of at least 60%, 62%, 65%, 80%, 95%, 98%, 99% identical to the polypeptide sequence of SEQ ID NO of at least 179 is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 98%, 99%, or a polypeptide sequence at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID NO 183.

In some embodiments, the first light chain comprises a sequence identical to SEQ ID NO:52, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to a polypeptide sequence of SEQ ID NO:56 at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to a polypeptide sequence of SEQ ID NO:68 at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical polypeptide sequence to at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 98%, 99% identical polypeptide sequence to at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 95%, 97%, 98%, 99% identical polypeptide sequence to at least 84%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical polypeptide sequence to at least 60%, 88% of SEQ ID NO 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical polypeptide sequences to those of SEQ ID NO:92, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to a polypeptide sequence of SEQ ID NO:96, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to a polypeptide sequence of SEQ ID NO:100, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to a polypeptide sequence at least 60%, 62%, 65%, 67%, 70%, 75%, 90%, 92%, 95%, 98%, 99% identical to a polypeptide sequence at least 120, 62%, 65%, 67%, 70%, 72%, 75% to a polypeptide sequence of SEQ ID NO 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical polypeptide sequences to SEQ ID NO:124 at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to a polypeptide sequence of SEQ ID NO:128 at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to a polypeptide sequence of SEQ ID NO:132, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID NO:140, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID NO:144, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID NO:148, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID NO:152, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90% identical to SEQ ID NO:152, 92%, 95%, 97%, 98%, 99% identical polypeptide sequence to SEQ ID NO:156, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to a polypeptide sequence of SEQ ID NO:160, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to a polypeptide sequence of SEQ ID NO:164 at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical polypeptide sequence to SEQ ID NO 168 at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical polypeptide sequence to SEQ ID NO 172 at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical polypeptide sequence to SEQ ID NO 176 at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 95%, 97%, 98%, 99% identical polypeptide sequence to SEQ ID NO 180 at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, or a polypeptide sequence which is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID NO 184.

In some embodiments, the second heavy chain comprises a sequence identical to SEQ ID NO:53 at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to a polypeptide sequence of SEQ ID NO:57, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to a polypeptide sequence of SEQ ID NO:69 at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical polypeptide sequence to at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical polypeptide sequence to at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 95%, 97%, 98%, 99% identical polypeptide sequence to at least 60%, 85%, 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical polypeptide sequence to at least 60% of SEQ ID NO, 81, 62%, 75%, 72%, 75%, 90%, 80%, 90%, 98%, 99% 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical polypeptide sequences to those of SEQ ID NO:93 at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to a polypeptide sequence of SEQ ID NO:97 at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to a polypeptide sequence of SEQ ID NO:101 at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical polypeptide sequence, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical polypeptide sequence to SEQ ID NO 105, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 90%, 92%, 95%, 97%, 98%, 99% identical polypeptide sequence to SEQ ID NO 121 at least 60%, 62%, 65%, 67%, 70%, 72%, 75% amino acid sequence to SEQ ID NO, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical polypeptide sequences to SEQ ID NO:125 at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to a polypeptide sequence of SEQ ID NO:129, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to a polypeptide sequence of SEQ ID NO:133, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID NO:141, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID NO:149, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID NO:153, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90% identical to SEQ ID NO:153 92%, 95%, 97%, 98%, 99% identical polypeptide sequence to SEQ ID NO:157 at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to the polypeptide sequence of SEQ ID NO:161, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to a polypeptide sequence of SEQ ID NO:165 at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical polypeptide sequence to SEQ ID NO 173 at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical polypeptide sequence to SEQ ID NO 177 at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical polypeptide sequence to SEQ ID NO 181 at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, or a polypeptide sequence at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID NO 185.

In some embodiments, the second light chain comprises a sequence identical to SEQ ID NO:54, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to a polypeptide sequence of SEQ ID NO:58, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to a polypeptide sequence of SEQ ID NO:70, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical polypeptide sequence to at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical polypeptide sequence to at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 95%, 97%, 98%, 99% identical polypeptide sequence to at least 60%, 62%, 65%, 82%, 98%, 99% identical polypeptide sequence to at least 60%, 86%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical polypeptide sequence to at least 60% of SEQ ID NO 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical polypeptide sequences to those of SEQ ID NO:94, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to a polypeptide sequence of SEQ ID NO:98, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to a polypeptide sequence of SEQ ID NO:102 at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical polypeptide sequence, at least 60%, 122%, 62%, 65%, 67%, 70%, 72%, 75% identical polypeptide sequence, 118 at least 60%, 62%, 65%, 67%, 70%, 75%, 99% identical polypeptide sequence to SEQ ID NO, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical polypeptide sequences to SEQ ID NO:126 at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to a polypeptide sequence of SEQ ID NO:130, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to a polypeptide sequence of SEQ ID NO:134 at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical polypeptide sequence to at least 60%, 154%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90% identical polypeptide sequence to at least 60%, 154% to SEQ ID NO, 92%, 95%, 97%, 98%, 99% identical polypeptide sequence to SEQ ID NO:158, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to a polypeptide sequence of SEQ ID NO:162, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to a polypeptide sequence of SEQ ID NO:166 at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical polypeptide sequence to at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical polypeptide sequence to at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 95%, 98%, 99% identical polypeptide sequence to at least 60%, 182%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 98%, 99% identical polypeptide sequence to SEQ ID NO, or a polypeptide sequence at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID NO 186.

In some embodiments, the variable domain of the first heavy chain comprises a sequence that hybridizes to SEQ ID NO:15, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to a polypeptide sequence of SEQ ID NO:27, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to a polypeptide sequence of SEQ ID NO:35 at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical polypeptide sequence to at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 99% identical polypeptide sequence to at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 95%, 97%, 98%, 99% identical polypeptide sequence to at least 60%, 62%, 65%, 80%, 82%, 98%, 99% identical polypeptide sequence to SEQ ID NO 19 at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical polypeptide sequence to SEQ ID NO, a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 13, a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 21, or a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 41.

In some embodiments, the variable domain of the first light chain comprises a sequence that hybridizes to SEQ ID NO:16, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to a polypeptide sequence of SEQ ID NO:28, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to a polypeptide sequence of SEQ ID NO:36, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical polypeptide sequence, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 92%, 95%, 97%, 98%, 99% identical polypeptide sequence of SEQ ID NO, at least 20, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical polypeptide sequence of SEQ ID NO, a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 14, a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 22, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 42.

In some embodiments, the variable domain of the second heavy chain comprises a sequence that hybridizes to SEQ ID NO:27, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to a polypeptide sequence of SEQ ID NO:15, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to a polypeptide sequence of SEQ ID NO:5 at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical polypeptide sequence to at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 99% identical polypeptide sequence to at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 95%, 97%, 98%, 99% identical polypeptide sequence to at least 60%, 29%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 98%, 99% identical polypeptide sequence to SEQ ID NO, and SEQ ID NO:35, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to a polypeptide sequence of SEQ ID NO:23, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to a polypeptide sequence of SEQ ID NO:25, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID NO:37, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID NO:39, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID NO:43, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 95%, 97%, 98%, 99% identical to SEQ ID NO:9, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID NO.

In some embodiments, the variable domain of the second light chain comprises a sequence that hybridizes to SEQ ID NO:28, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to a polypeptide sequence of SEQ ID NO:16, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to a polypeptide sequence of SEQ ID NO:6 at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical polypeptide sequence to at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 99% identical polypeptide sequence to at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 95%, 97%, 98%, 99% identical polypeptide sequence to at least 60%, 62%, 65%, 82%, 98%, 99% identical polypeptide sequence to SEQ ID NO 30 at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical polypeptide sequence to SEQ ID NO, and SEQ ID NO:36, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to a polypeptide sequence of SEQ ID NO:24, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to a polypeptide sequence of SEQ ID NO:26, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical polypeptide sequence to SEQ ID NO, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 95%, 98%, 99% identical polypeptide sequence to SEQ ID NO, or at least 10, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 98%, 99% identical polypeptide sequence to SEQ ID NO.

In some embodiments, the variable domain of the first heavy chain comprises CDR1, CDR2 and CDR3 of SEQ ID NO. 51; CDR1, CDR2 and CDR3 of SEQ ID NO. 55; CDR1, CDR2 and CDR3 of SEQ ID NO 67; CDR1, CDR2 and CDR3 of SEQ ID NO 71; CDR1, CDR2 and CDR3 of SEQ ID NO 75; CDR1, CDR2 and CDR3 of SEQ ID NO 79; CDR1, CDR2 and CDR3 of SEQ ID NO 83; CDR1, CDR2 and CDR3 of SEQ ID NO 87; CDR1, CDR2 and CDR3 of SEQ ID NO. 91; CDR1, CDR2 and CDR3 of SEQ ID NO 95; CDR1, CDR2 and CDR3 of SEQ ID No. 99; CDR1, CDR2 and CDR3 of SEQ ID NO. 103; CDR1, CDR2 and CDR3 of SEQ ID NO 107; CDR1, CDR2 and CDR3 of SEQ ID NO 115; CDR1, CDR2 and CDR3 of SEQ ID No. 119; CDR1, CDR2 and CDR3 of SEQ ID NO. 123; CDR1, CDR2 and CDR3 of SEQ ID NO 127; CDR1, CDR2 and CDR3 of SEQ ID NO. 131; CDR1, CDR2 and CDR3 of SEQ ID NO 139; CDR1, CDR2 and CDR3 of SEQ ID NO 143; CDR1, CDR2 and CDR3 of SEQ ID No. 147; CDR1, CDR2 and CDR3 of SEQ ID NO 151; CDR1, CDR2 and CDR3 of SEQ ID NO 155; CDR1, CDR2 and CDR3 of SEQ ID NO 159; CDR1, CDR2 and CDR3 of SEQ ID NO 163; CDR1, CDR2 and CDR3 of SEQ ID NO 167; CDR1, CDR2 and CDR3 of SEQ ID NO. 171; CDR1, CDR2 and CDR3 of SEQ ID NO 175; CDR1, CDR2 and CDR3 of SEQ ID NO 179; or CDR1, CDR2 and CDR3 of SEQ ID NO 183.

In some embodiments, the variable domain of the first light chain comprises CDR1, CDR2 and CDR3 of SEQ ID NO. 52; CDR1, CDR2 and CDR3 of SEQ ID NO. 56; CDR1, CDR2 and CDR3 of SEQ ID NO. 68; CDR1, CDR2 and CDR3 of SEQ ID NO 72; CDR1, CDR2 and CDR3 of SEQ ID NO 76; CDR1, CDR2 and CDR3 of SEQ ID NO 80; CDR1, CDR2 and CDR3 of SEQ ID NO 84; CDR1, CDR2 and CDR3 of SEQ ID NO 88; CDR1, CDR2 and CDR3 of SEQ ID NO. 92; CDR1, CDR2 and CDR3 of SEQ ID No. 96; CDR1, CDR2 and CDR3 of SEQ ID NO. 100; CDR1, CDR2 and CDR3 of SEQ ID No. 104; CDR1, CDR2 and CDR3 of SEQ ID NO 108; CDR1, CDR2 and CDR3 of SEQ ID NO. 116; CDR1, CDR2 and CDR3 of SEQ ID NO. 120; CDR1, CDR2 and CDR3 of SEQ ID NO 124; CDR1, CDR2 and CDR3 of SEQ ID NO 128; CDR1, CDR2 and CDR3 of SEQ ID NO. 132; CDR1, CDR2 and CDR3 of SEQ ID NO. 140; CDR1, CDR2 and CDR3 of SEQ ID NO. 144; CDR1, CDR2 and CDR3 of SEQ ID NO. 148; CDR1, CDR2 and CDR3 of SEQ ID NO 152; CDR1, CDR2 and CDR3 of SEQ ID NO 156; CDR1, CDR2 and CDR3 of SEQ ID NO. 160; CDR1, CDR2 and CDR3 of SEQ ID NO. 164; CDR1, CDR2 and CDR3 of SEQ ID No. 168; CDR1, CDR2 and CDR3 of SEQ ID NO 172; CDR1, CDR2 and CDR3 of SEQ ID NO. 176; CDR1, CDR2 and CDR3 of SEQ ID NO 180; or CDR1, CDR2 and CDR3 of SEQ ID NO 184.

In some embodiments, the variable domain of the second heavy chain comprises CDR1, CDR2 and CDR3 of SEQ ID NO 53; CDR1, CDR2 and CDR3 of SEQ ID NO 57; CDR1, CDR2 and CDR3 of SEQ ID NO 69; CDR1, CDR2 and CDR3 of SEQ ID NO 73; CDR1, CDR2 and CDR3 of SEQ ID NO 77; CDR1, CDR2 and CDR3 of SEQ ID NO 81; CDR1, CDR2 and CDR3 of SEQ ID NO 85; CDR1, CDR2 and CDR3 of SEQ ID NO 89; CDR1, CDR2 and CDR3 of SEQ ID NO. 93; CDR1, CDR2 and CDR3 of SEQ ID NO 97; CDR1, CDR2 and CDR3 of SEQ ID NO 101; CDR1, CDR2 and CDR3 of SEQ ID NO 105; CDR1, CDR2 and CDR3 of SEQ ID NO 109; CDR1, CDR2 and CDR3 of SEQ ID NO 117; CDR1, CDR2 and CDR3 of SEQ ID NO. 121; CDR1, CDR2 and CDR3 of SEQ ID No. 125; CDR1, CDR2 and CDR3 of SEQ ID No. 129; CDR1, CDR2 and CDR3 of SEQ ID NO 133; CDR1, CDR2 and CDR3 of SEQ ID NO 141; CDR1, CDR2 and CDR3 of SEQ ID NO. 145; CDR1, CDR2 and CDR3 of SEQ ID NO 149; CDR1, CDR2 and CDR3 of SEQ ID NO 153; CDR1, CDR2 and CDR3 of SEQ ID NO. 157; CDR1, CDR2 and CDR3 of SEQ ID NO 161; CDR1, CDR2 and CDR3 of SEQ ID NO. 165; CDR1, CDR2 and CDR3 of SEQ ID NO 169; CDR1, CDR2 and CDR3 of SEQ ID NO 173; CDR1, CDR2 and CDR3 of SEQ ID NO 177; CDR1, CDR2 and CDR3 of SEQ ID NO 181; or CDR1, CDR2 and CDR3 of SEQ ID NO 185.

In some embodiments, the variable domain of the second light chain comprises CDR1, CDR2 and CDR3 of SEQ ID NO. 54; CDR1, CDR2 and CDR3 of SEQ ID NO 58; CDR1, CDR2 and CDR3 of SEQ ID NO 70; CDR1, CDR2 and CDR3 of SEQ ID NO 74; CDR1, CDR2 and CDR3 of SEQ ID NO. 78; CDR1, CDR2 and CDR3 of SEQ ID No. 82; CDR1, CDR2 and CDR3 of SEQ ID NO 86; CDR1, CDR2 and CDR3 of SEQ ID NO. 90; CDR1, CDR2 and CDR3 of SEQ ID No. 94; CDR1, CDR2 and CDR3 of SEQ ID NO 98; CDR1, CDR2 and CDR3 of SEQ ID No. 102; CDR1, CDR2 and CDR3 of SEQ ID NO 106; CDR1, CDR2 and CDR3 of SEQ ID NO. 110; CDR1, CDR2 and CDR3 of SEQ ID NO 118; CDR1, CDR2 and CDR3 of SEQ ID NO. 122; CDR1, CDR2 and CDR3 of SEQ ID NO. 126; CDR1, CDR2 and CDR3 of SEQ ID NO. 130; CDR1, CDR2 and CDR3 of SEQ ID No. 134; CDR1, CDR2 and CDR3 of SEQ ID NO. 142; CDR1, CDR2 and CDR3 of SEQ ID NO 146; CDR1, CDR2 and CDR3 of SEQ ID NO. 150; CDR1, CDR2 and CDR3 of SEQ ID NO 154; CDR1, CDR2 and CDR3 of SEQ ID NO 158; CDR1, CDR2 and CDR3 of SEQ ID NO. 162; CDR1, CDR2 and CDR3 of SEQ ID NO 166; CDR1, CDR2 and CDR3 of SEQ ID NO. 170; CDR1, CDR2 and CDR3 of SEQ ID NO 174; CDR1, CDR2 and CDR3 of SEQ ID NO 178; CDR1, CDR2 and CDR3 of SEQ ID NO 182; or CDR1, CDR2 and CDR3 of SEQ ID NO. 186.

In some embodiments, the molecule comprises SEQ ID NO:51, SEQ ID NO:52, SEQ ID NO:53, and SEQ ID NO:54. In some embodiments, the molecule comprises SEQ ID NO:55, SEQ ID NO:56, SEQ ID NO:57, and SEQ ID NO:58. In some embodiments, the molecule comprises SEQ ID NO:67, SEQ ID NO:68, SEQ ID NO:69, and SEQ ID NO:70. In some embodiments, the molecule comprises SEQ ID NO:71, SEQ ID NO:72, SEQ ID NO:73, and SEQ ID NO:74. In some embodiments, the molecule comprises SEQ ID NO:75, SEQ ID NO:76, SEQ ID NO:77, and SEQ ID NO:78. In some embodiments, the molecule comprises SEQ ID NO:79, SEQ ID NO:80, SEQ ID NO:81, and SEQ ID NO:82. In some embodiments, the molecule comprises SEQ ID NO:83, SEQ ID NO:84, SEQ ID NO:85, and SEQ ID NO:86. In some embodiments, the molecule comprises SEQ ID NO 87, SEQ ID NO 88, SEQ ID NO 89, and SEQ ID NO 90. In some embodiments, the molecule comprises SEQ ID NO. 91, SEQ ID NO. 92, SEQ ID NO. 93 and SEQ ID NO. 94. In some embodiments, the molecule comprises SEQ ID NO 95, SEQ ID NO 96, SEQ ID NO 97, and SEQ ID NO 98. In some embodiments, the molecule comprises SEQ ID NO 99, SEQ ID NO 100, SEQ ID NO 101 and SEQ ID NO 102. In some embodiments, the molecule comprises SEQ ID NO 103,SEQ ID NO:104,SEQ ID NO:105 and SEQ ID NO 106. In some embodiments, the molecule comprises SEQ ID NO:107, SEQ ID NO:108, SEQ ID NO:109, and SEQ ID NO:110. In some embodiments, the molecule comprises SEQ ID NO:115, SEQ ID NO:116, SEQ ID NO:117, and SEQ ID NO:118. In some embodiments, the molecule comprises SEQ ID NO:119, SEQ ID NO:120, SEQ ID NO:121, and SEQ ID NO:122.

In some embodiments, the molecule comprises SEQ ID NO:123, SEQ ID NO:124, SEQ ID NO:125, and SEQ ID NO:126. In some embodiments, the molecule comprises SEQ ID NO:127, SEQ ID NO:128, SEQ ID NO:129, and SEQ ID NO:130. In some embodiments, the molecule comprises SEQ ID NO:131, SEQ ID NO:132, SEQ ID NO:133, and SEQ ID NO:133. In some embodiments, the molecule comprises SEQ ID NO. 139, SEQ ID NO. 140, SEQ ID NO. 141 and SEQ ID NO. 142. In some embodiments, the molecule comprises SEQ ID NO:143, SEQ ID NO:144, SEQ ID NO:145, and SEQ ID NO:146. In some embodiments, the molecule comprises SEQ ID NO:147, SEQ ID NO:148, SEQ ID NO:149, and SEQ ID NO:150. In some embodiments, the molecule comprises SEQ ID NO:151, SEQ ID NO:152, SEQ ID NO:153, and SEQ ID NO:154. In some embodiments, the molecule comprises SEQ ID NO:155, SEQ ID NO:156, SEQ ID NO:157, and SEQ ID NO:158. In some embodiments, the molecule comprises SEQ ID NO 159, SEQ ID NO 160, SEQ ID NO 161 and SEQ ID NO 162. In some embodiments, the molecule comprises SEQ ID NO. 163, SEQ ID NO. 164, SEQ ID NO. 165 and SEQ ID NO. 166. In some embodiments, the molecule comprises SEQ ID NO 167, SEQ ID NO 168, SEQ ID NO 169, and SEQ ID NO 170. In some embodiments, the molecule comprises SEQ ID NO:171, SEQ ID NO:172, SEQ ID NO:173, and SEQ ID NO:174. In some embodiments, the molecule comprises SEQ ID NO:175, SEQ ID NO:176, SEQ ID NO:177, and SEQ ID NO:178. In some embodiments, the molecule comprises SEQ ID NO:179, SEQ ID NO:180, SEQ ID NO:18, and SEQ ID NO:182. In some embodiments, the molecule comprises SEQ ID NO. 183, SEQ ID NO. 184, SEQ ID NO. 185 and SEQ ID NO. 186.

In some embodiments, the molecule further comprises one or more disulfide bonds. In some embodiments, the molecule further comprises one or more linker sequences. In some embodiments, the molecule is capable of neutralizing coronavirus. In some embodiments, the coronavirus is a SARS-CoV-2 virus. In some embodiments, the SARS-CoV-2 virus strain is selected from the group consisting of WA1, B.1.1.7, B1.526, B1.351, P1, B1.352, and B.1.427.

In some embodiments, the molecule specifically recognizes a first epitope and a second epitope on SARS-CoV-2 particle. In some embodiments, the first epitope comprises at least a portion of a Receptor Binding Domain (RBD) on a spike protein of a SARS-CoV-2 particle. In some embodiments, the second epitope comprises at least a portion of an N-terminal domain (NTD) on a spike protein of a SARS-CoV-2 particle.

In certain aspects, the invention provides a method of treating or preventing a covd-19 infection in a subject in need thereof, the method comprising administering to the subject a composition comprising a bispecific molecule comprising a first polypeptide chain and a second polypeptide chain, wherein: the first polypeptide chain comprises: a first light chain comprising a variable domain and a constant domain; a first heavy chain comprising a variable domain and a constant domain; the second polypeptide chain comprises: a second light chain comprising a variable domain and a constant domain; and a second heavy chain comprising a variable domain and a constant domain.

In some embodiments, the second light chain comprises a sequence identical to SEQ ID NO:54, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to a polypeptide sequence of SEQ ID NO:58, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to a polypeptide sequence of SEQ ID NO:70, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical polypeptide sequence to at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical polypeptide sequence to at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 95%, 97%, 98%, 99% identical polypeptide sequence to at least 60%, 62%, 65%, 82%, 98%, 99% identical polypeptide sequence to at least 60%, 86%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical polypeptide sequence to at least 60% of SEQ ID NO 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical polypeptide sequences to those of SEQ ID NO:94, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to a polypeptide sequence of SEQ ID NO:98, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to a polypeptide sequence of SEQ ID NO:102 at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical polypeptide sequence, at least 60%, 122%, 62%, 65%, 67%, 70%, 72%, 75% identical polypeptide sequence, 118 at least 60%, 62%, 65%, 67%, 70%, 75%, 99% identical polypeptide sequence to SEQ ID NO, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical polypeptide sequences to SEQ ID NO:126 at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to a polypeptide sequence of SEQ ID NO:130, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to a polypeptide sequence of SEQ ID NO:134 at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical polypeptide sequence to at least 60%, 154%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90% identical polypeptide sequence to at least 60%, 154% to SEQ ID NO, 92%, 95%, 97%, 98%, 99% identical polypeptide sequence to SEQ ID NO:158, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to a polypeptide sequence of SEQ ID NO:162, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to a polypeptide sequence of SEQ ID NO:166 at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical polypeptide sequence to at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical polypeptide sequence to at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 95%, 98%, 99% identical polypeptide sequence to at least 60%, 182%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 98%, 99% identical polypeptide sequence to SEQ ID NO, a polypeptide sequence which is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 186.

In some embodiments, the variable domain of the first heavy chain comprises CDR1, CDR2 and CDR3 of SEQ ID NO. 51; CDR1, CDR2 and CDR3 of SEQ ID NO. 55; CDR1, CDR2 and CDR3 of SEQ ID NO 67; CDR1, CDR2 and CDR3 of SEQ ID NO 71; CDR1, CDR2 and CDR3 of SEQ ID NO 75; CDR1, CDR2 and CDR3 of SEQ ID NO 79; CDR1, CDR2 and CDR3 of SEQ ID NO 83; CDR1, CDR2 and CDR3 of SEQ ID NO 87; CDR1, CDR2 and CDR3 of SEQ ID NO. 91; CDR1, CDR2 and CDR3 of SEQ ID NO 95; CDR1, CDR2 and CDR3 of SEQ ID No. 99; CDR1, CDR2 and CDR3 of SEQ ID NO 103; CDR1, CDR2 and CDR3 of SEQ ID NO 107; CDR1, CDR2 and CDR3 of SEQ ID NO 115; CDR1, CDR2 and CDR3 of SEQ ID No. 119; CDR1, CDR2 and CDR3 of SEQ ID NO. 123; CDR1, CDR2 and CDR3 of SEQ ID NO 127; CDR1, CDR2 and CDR3 of SEQ ID NO. 131; CDR1, CDR2 and CDR3 of SEQ ID NO 139; CDR1, CDR2 and CDR3 of SEQ ID NO 143; CDR1, CDR2 and CDR3 of SEQ ID No. 147; CDR1, CDR2 and CDR3 of SEQ ID NO 151; CDR1, CDR2 and CDR3 of SEQ ID NO 155; CDR1, CDR2 and CDR3 of SEQ ID NO 159; CDR1, CDR2 and CDR3 of SEQ ID NO 163; CDR1, CDR2 and CDR3 of SEQ ID NO 167; CDR1, CDR2 and CDR3 of SEQ ID NO. 171; CDR1, CDR2 and CDR3 of SEQ ID NO 175; CDR1, CDR2 and CDR3 of SEQ ID NO 179; or CDR1, CDR2 and CDR3 of SEQ ID NO 183.

In some embodiments, the molecule comprises SEQ ID NO:51, SEQ ID NO:52, SEQ ID NO:53, and SEQ ID NO:54. In some embodiments, the molecule comprises SEQ ID NO:55, SEQ ID NO:56, SEQ ID NO:57, and SEQ ID NO:58. In some embodiments, the molecule comprises SEQ ID NO:67, SEQ ID NO:68, SEQ ID NO:69, and SEQ ID NO:70. In some embodiments, the molecule comprises SEQ ID NO:71, SEQ ID NO:72, SEQ ID NO:73, and SEQ ID NO:74. In some embodiments, the molecule comprises SEQ ID NO:75, SEQ ID NO:76, SEQ ID NO:77, and SEQ ID NO:78. In some embodiments, the molecule comprises SEQ ID NO:79, SEQ ID NO:80, SEQ ID NO:81, and SEQ ID NO:82. In some embodiments, the molecule comprises SEQ ID NO:83, SEQ ID NO:84, SEQ ID NO:85, and SEQ ID NO:86. In some embodiments, the molecule comprises SEQ ID NO 87, SEQ ID NO 88, SEQ ID NO 89, and SEQ ID NO 90. In some embodiments, the molecule comprises SEQ ID NO. 91, SEQ ID NO. 92, SEQ ID NO. 93 and SEQ ID NO. 94. In some embodiments, the molecule comprises SEQ ID NO 95, SEQ ID NO 96, SEQ ID NO 97, and SEQ ID NO 98. In some embodiments, the molecule comprises SEQ ID NO 99, SEQ ID NO 100, SEQ ID NO 101 and SEQ ID NO 102. In some embodiments, the molecule comprises SEQ ID NO:103, SEQ ID NO:104, SEQ ID NO:105, and SEQ ID NO:106. In some embodiments, the molecule comprises SEQ ID NO:107, SEQ ID NO:108, SEQ ID NO:109, and SEQ ID NO:110. In some embodiments, the molecule comprises SEQ ID NO:115, SEQ ID NO:116, SEQ ID NO:117, and SEQ ID NO:118.

In some embodiments, the molecule comprises SEQ ID NO:119, SEQ ID NO:120, SEQ ID NO:121, and SEQ ID NO:122. In some embodiments, the molecule comprises SEQ ID NO:123, SEQ ID NO:124, SEQ ID NO:125, and SEQ ID NO:126. In some embodiments, the molecule comprises SEQ ID NO:127, SEQ ID NO:128, SEQ ID NO:129, and SEQ ID NO:130. In some embodiments, the molecule comprises SEQ ID NO:131, SEQ ID NO:132, SEQ ID NO:133, and SEQ ID NO:133. In some embodiments, the molecule comprises SEQ ID NO. 139, SEQ ID NO. 140, SEQ ID NO. 141 and SEQ ID NO. 142. In some embodiments, the molecule comprises SEQ ID NO:143, SEQ ID NO:144, SEQ ID NO:145, and SEQ ID NO:146. In some embodiments, the molecule comprises SEQ ID NO:147, SEQ ID NO:148, SEQ ID NO:149, and SEQ ID NO:150. In some embodiments, the molecule comprises SEQ ID NO. 151, SEQ ID NO. 152, SEQ ID NO. 153 and SEQ ID NO. 154. In some embodiments, the molecule comprises SEQ ID NO:155, SEQ ID NO:156, SEQ ID NO:157, and SEQ ID NO:158. In some embodiments, the molecule comprises SEQ ID NO 159, SEQ ID NO 160, SEQ ID NO 161 and SEQ ID NO 162. In some embodiments, the molecule comprises SEQ ID NO. 163, SEQ ID NO. 164, SEQ ID NO. 165 and SEQ ID NO. 166. In some embodiments, the molecule comprises SEQ ID NO 167, SEQ ID NO 168, SEQ ID NO 169, and SEQ ID NO 170. In some embodiments, the molecule comprises SEQ ID NO:171, SEQ ID NO:172, SEQ ID NO:173, and SEQ ID NO:174. In some embodiments, the molecule comprises SEQ ID NO:175, SEQ ID NO:176, SEQ ID NO:177, and SEQ ID NO:178. In some embodiments, the molecule comprises SEQ ID NO:179, SEQ ID NO:180, SEQ ID NO:181 and SEQ ID NO:182. In some embodiments, the molecule comprises SEQ ID NO. 183, SEQ ID NO. 184, SEQ ID NO. 185 and SEQ ID NO. 186.

In some embodiments, the molecule further comprises one or more disulfide bonds. In some embodiments, the molecule further comprises one or more linker sequences. In some embodiments, the molecule is capable of neutralizing coronavirus. In some embodiments, the coronavirus is a SARS-CoV-2 virus. In some embodiments, the SARS-CoV-2 virus strain is selected from the group consisting of WA1, B.1.1.7, B1.526, B1.351, P1, B1.352, and B.1.427. In some embodiments, the molecule specifically recognizes a first epitope and a second epitope on SARS-CoV-2 particle. In some embodiments, the first epitope comprises at least a portion of a Receptor Binding Domain (RBD) on a spike protein of a SARS-CoV-2 particle. In some embodiments, the second epitope comprises at least a portion of an N-terminal domain (NTD) on a spike protein of a SARS-CoV-2 particle. In some embodiments, the composition is administered in combination with a second therapeutic agent.

In some embodiments, the heavy chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 5. In some embodiments, the heavy chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 41. In some embodiments, the heavy chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 35. In some embodiments, the heavy chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 1. In some embodiments, the heavy chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 23. In some embodiments, the heavy chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 13.

In some embodiments, the heavy chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 27. In some embodiments, the light chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 6. In some embodiments, the light chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 42. In some embodiments, the light chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 36. In some embodiments, the light chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 2.

In some embodiments, the light chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 24. In some embodiments, the light chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 14. In some embodiments, the light chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 28. In some embodiments, the heavy chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 5, and the light chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 6. In some embodiments, the heavy chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 41, and the light chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 42. In some embodiments, the heavy chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 35, and the light chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 36.

In some embodiments, the heavy chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 1, and the light chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 2. In some embodiments, the heavy chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 23, and the light chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 24. In some embodiments, the heavy chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 13, and the light chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 14. In some embodiments, the heavy chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 27, and the light chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 28.

In some embodiments, the heavy chain variable domain comprises SEQ ID NO. 5 and the light chain variable domain comprises SEQ ID NO. 6. In some embodiments, the heavy chain variable domain comprises SEQ ID NO. 41 and the light chain variable domain comprises SEQ ID NO. 42. In some embodiments, the heavy chain variable domain comprises SEQ ID NO. 35 and the light chain variable domain comprises SEQ ID NO. 36. In some embodiments, the heavy chain variable domain comprises SEQ ID NO. 1 and the light chain variable domain comprises SEQ ID NO. 2. In some embodiments, the heavy chain variable domain comprises SEQ ID NO. 23 and the light chain variable domain comprises SEQ ID NO. 24. In some embodiments, the heavy chain variable domain comprises SEQ ID NO. 13 and the light chain variable domain comprises SEQ ID NO. 14. In some embodiments, the heavy chain variable domain comprises SEQ ID NO. 27 and the light chain variable domain comprises SEQ ID NO. 28. In some embodiments, the heavy chain variable domain comprises the CDR1, CDR2 and CDR3 regions of SEQ ID NO. 5. In some embodiments, the heavy chain variable domain comprises the CDR1, CDR2 and CDR3 regions of SEQ ID NO 41. In some embodiments, the heavy chain variable domain comprises the CDR1, CDR2 and CDR3 regions of SEQ ID NO. 1.

In some embodiments, the heavy chain variable domain comprises the CDR1, CDR2 and CDR3 regions of SEQ ID NO. 23. In some embodiments, the heavy chain variable domain comprises the CDR1, CDR2 and CDR3 regions of SEQ ID NO. 13. In some embodiments, the heavy chain variable domain comprises the CDR1, CDR2 and CDR3 regions of SEQ ID NO 27.

In some embodiments, the light chain variable domain comprises the CDR1, CDR2 and CDR3 regions of SEQ ID NO. 6. In some embodiments, the light chain variable domain comprises the CDR1, CDR2 and CDR3 regions of SEQ ID NO. 42. In some embodiments, the light chain variable domain comprises the CDR1, CDR2 and CDR3 regions of SEQ ID NO. 36. In some embodiments, the light chain variable domain comprises the CDR1, CDR2 and CDR3 regions of SEQ ID NO. 2. In some embodiments, the light chain variable domain comprises the CDR1, CDR2 and CDR3 regions of SEQ ID NO. 24. In some embodiments, the light chain variable domain comprises the CDR1, CDR2 and CDR3 regions of SEQ ID NO. 14. In some embodiments, the light chain variable domain comprises the CDR1, CDR2 and CDR3 regions of SEQ ID NO. 28. In some embodiments, the heavy chain variable domain comprises the CDR1, CDR2 and CDR3 regions of SEQ ID NO. 5 and the light chain variable domain comprises the CDR1, CDR2 and CDR3 regions of SEQ ID NO. 6. In some embodiments, the heavy chain variable domain comprises the CDR1, CDR2 and CDR3 regions of SEQ ID NO. 41 and the light chain variable domain comprises the CDR1, CDR2 and CDR3 regions of SEQ ID NO. 42. In some embodiments, the heavy chain variable domain comprises the CDR1, CDR2 and CDR3 regions of SEQ ID NO. 35 and the light chain variable domain comprises the CDR1, CDR2 and CDR3 regions of SEQ ID NO. 36. In some embodiments, the heavy chain variable domain comprises the CDR1, CDR2 and CDR3 regions of SEQ ID NO. 1 and the light chain variable domain comprises the CDR1, CDR2 and CDR3 regions of SEQ ID NO. 2.

In some embodiments, the heavy chain variable domain comprises the CDR1, CDR2 and CDR3 regions of SEQ ID NO. 23 and the light chain variable domain comprises the CDR1, CDR2 and CDR3 regions of SEQ ID NO. 24. In some embodiments, the heavy chain variable domain comprises the CDR1, CDR2 and CDR3 regions of SEQ ID NO. 13 and the light chain variable domain comprises the CDR1, CDR2 and CDR3 regions of SEQ ID NO. 14. In some embodiments, the heavy chain variable domain comprises the CDR1, CDR2 and CDR3 regions of SEQ ID NO. 27 and the light chain variable domain comprises the CDR1, CDR2 and CDR3 regions of SEQ ID NO. 28.

In some embodiments, the antibody neutralizes coronavirus. In some embodiments, the coronavirus is a SARS-CoV-2 virus. In some embodiments, the coronavirus is a SARS-CoV-2 variant. In some embodiments, the variant plant is selected from the group consisting of WA1, b.1.1.7, B1.526, B1.351, P1, B1.352, and b.1.427. In some embodiments, the antibody specifically binds to an epitope on the surface of the coronavirus. In some embodiments, the epitope comprises at least a portion of a coronavirus spike protein. In some embodiments, at least a portion of the coronavirus spike protein comprises a Receptor Binding Domain (RBD) of the spike protein. In some embodiments, at least a portion of the coronavirus spike protein comprises an N-terminal domain (NTD) of the spike protein. In some embodiments, the coronavirus is a SARS-CoV-2 coronavirus.

In some embodiments, the heavy chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 5. In some embodiments, the heavy chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 41. In some embodiments, the heavy chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 35. In some embodiments, the heavy chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 1. In some embodiments, the heavy chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 23. In some embodiments, the heavy chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 13. In some embodiments, the heavy chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 27. In some embodiments, the light chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 6.

In some embodiments, the light chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 42. In some embodiments, the light chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 36. In some embodiments, the light chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 2. In some embodiments, the light chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 24. In some embodiments, the light chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 14. In some embodiments, the light chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 28.

In some embodiments, the heavy chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 5, and the light chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 6. In some embodiments, the heavy chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 41, and the light chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 42. In some embodiments, the heavy chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 35, and the light chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 36. In some embodiments, the heavy chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 1, and the light chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 2. In some embodiments, the heavy chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 23, and the light chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 24.

In some embodiments, the heavy chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 13, and the light chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 14. In some embodiments, the heavy chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 27, and the light chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 28.

In some embodiments, the heavy chain variable domain comprises SEQ ID NO. 5 and the light chain variable domain comprises SEQ ID NO. 6. In some embodiments, the heavy chain variable domain comprises SEQ ID NO. 41 and the light chain variable domain comprises SEQ ID NO. 42. In some embodiments, the heavy chain variable domain comprises SEQ ID NO:35 and the light chain variable domain comprises SEQ ID NO:36. In some embodiments, the heavy chain variable domain comprises SEQ ID NO. 1 and the light chain variable domain comprises SEQ ID NO. 2. In some embodiments, the heavy chain variable domain comprises SEQ ID NO. 23 and the light chain variable domain comprises SEQ ID NO. 24. In some embodiments, the heavy chain variable domain comprises SEQ ID NO. 13 and the light chain variable domain comprises SEQ ID NO. 14.

In some embodiments, the heavy chain variable domain comprises SEQ ID NO. 27 and the light chain variable domain comprises SEQ ID NO. 28. In some embodiments, the heavy chain variable domain comprises the CDR1, CDR2 and CDR3 regions of SEQ ID NO. 5. In some embodiments, the heavy chain variable domain comprises the CDR1, CDR2 and CDR3 regions of SEQ ID NO 41. In some embodiments, the heavy chain variable domain comprises the CDR1, CDR2 and CDR3 regions of SEQ ID NO. 1. In some embodiments, the heavy chain variable domain comprises the CDR1, CDR2 and CDR3 regions of SEQ ID NO. 23. In some embodiments, the heavy chain variable domain comprises the CDR1, CDR2 and CDR3 regions of SEQ ID NO. 13. In some embodiments, the heavy chain variable domain comprises the CDR1, CDR2 and CDR3 regions of SEQ ID NO 27. In some embodiments, the light chain variable domain comprises the CDR1, CDR2 and CDR3 regions of SEQ ID NO. 6. In some embodiments, the light chain variable domain comprises the CDR1, CDR2 and CDR3 regions of SEQ ID NO. 42. In some embodiments, the light chain variable domain comprises the CDR1, CDR2 and CDR3 regions of SEQ ID NO. 36. In some embodiments, the light chain variable domain comprises the CDR1, CDR2 and CDR3 regions of SEQ ID NO. 2. In some embodiments, the light chain variable domain comprises the CDR1, CDR2 and CDR3 regions of SEQ ID NO. 24.

In some embodiments, the light chain variable domain comprises the CDR1, CDR2 and CDR3 regions of SEQ ID NO. 14. In some embodiments, the light chain variable domain comprises the CDR1, CDR2 and CDR3 regions of SEQ ID NO. 28. In some embodiments, the heavy chain variable domain comprises the CDR1, CDR2 and CDR3 regions of SEQ ID NO. 5 and the light chain variable domain comprises the CDR1, CDR2 and CDR3 regions of SEQ ID NO. 6. In some embodiments, the heavy chain variable domain comprises the CDR1, CDR2 and CDR3 regions of SEQ ID NO. 41 and the light chain variable domain comprises the CDR1, CDR2 and CDR3 regions of SEQ ID NO. 42. In some embodiments, the heavy chain variable domain comprises the CDR1, CDR2 and CDR3 regions of SEQ ID NO. 35 and the light chain variable domain comprises the CDR1, CDR2 and CDR3 regions of SEQ ID NO. 36. In some embodiments, the heavy chain variable domain comprises the CDR1, CDR2 and CDR3 regions of SEQ ID NO. 1 and the light chain variable domain comprises the CDR1, CDR2 and CDR3 regions of SEQ ID NO. 2. In some embodiments, the heavy chain variable domain comprises the CDR1, CDR2 and CDR3 regions of SEQ ID NO. 23 and the light chain variable domain comprises the CDR1, CDR2 and CDR3 regions of SEQ ID NO. 24. In some embodiments, the heavy chain variable domain comprises the CDR1, CDR2 and CDR3 regions of SEQ ID NO. 13 and the light chain variable domain comprises the CDR1, CDR2 and CDR3 regions of SEQ ID NO. 14. In some embodiments, the heavy chain variable domain comprises the CDR1, CDR2 and CDR3 regions of SEQ ID NO. 27 and the light chain variable domain comprises the CDR1, CDR2 and CDR3 regions of SEQ ID NO. 28.

In some embodiments, the antibody neutralizes coronavirus. In some embodiments, the coronavirus is a SARS-CoV-2 virus. In some embodiments, the coronavirus is a SARS-CoV-2 variant. In some embodiments, the variant plant is selected from the group consisting of WA1, b.1.1.7, B1.526, B1.351, P1, B1.352, and b.1.427. In some embodiments, the antibody specifically binds to an epitope on the surface of the coronavirus. In some embodiments, the epitope comprises at least a portion of a coronavirus spike protein. In some embodiments, at least a portion of the coronavirus spike protein comprises a Receptor Binding Domain (RBD) of the spike protein. In some embodiments, at least a portion of the coronavirus spike protein comprises an N-terminal domain (NTD) of the spike protein. In some embodiments, the coronavirus is a SARS-CoV-2 coronavirus. In some embodiments, the antibody is administered in combination with a second agent.

In certain aspects, the invention provides a bispecific molecule comprising a first polypeptide chain and a second polypeptide chain, wherein: the first polypeptide chain comprises: a first light chain comprising a variable domain and a constant domain; a first heavy chain comprising a variable domain and a constant domain; the second polypeptide chain comprises: a second light chain comprising a variable domain and a constant domain; and a second heavy chain comprising a variable domain and a constant domain. In some embodiments, the first polypeptide chain and the second polypeptide chain are covalently bonded to each other.

In some embodiments, the first heavy chain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID NO, a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 72%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID NO, 211, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 98%, 99% identical to SEQ ID NO. In some embodiments, the first light chain comprises a sequence identical to SEQ ID NO:48 at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical polypeptide sequence, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical polypeptide sequence to SEQ ID NO 112, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical polypeptide sequence to SEQ ID NO 188 at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 95%, 97%, 98%, 99% identical polypeptide sequence to SEQ ID NO 212 at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 98%, 99% identical polypeptide sequence to SEQ ID NO.

In some embodiments, the second heavy chain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 49, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 113, a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 189, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 213, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 92%, 98%, 99% identical to SEQ ID No. 213. In some embodiments, the second light chain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 50, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 114, a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 190, at least 60%, 62%, 65%, 67%, 70%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 214, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 98%, 99% identical to SEQ ID NO. In some embodiments, the variable domain of the first heavy chain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 35, a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 13.

In some embodiments, the variable domain of the first light chain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 36, a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 14. In some embodiments, the variable domain of the second heavy chain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 13, a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 43, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 43. In some embodiments, the variable domain of the second light chain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 46, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 44.

In some embodiments, the variable domain of the first heavy chain comprises CDR1, CDR2 and CDR3 of SEQ ID NO 47; CDR1, CDR2 and CDR3 of SEQ ID NO 111; CDR1, CDR2 and CDR3 of SEQ ID NO. 135; CDR1, CDR2 and CDR3 of SEQ ID NO 187; or CDR1, CDR2 and CDR3 of SEQ ID NO 211. In some embodiments, the variable domain of the first light chain comprises CDR1, CDR2 and CDR3 of SEQ ID NO. 48; CDR1, CDR2 and CDR3 of SEQ ID NO 112; CDR1, CDR2 and CDR3 of SEQ ID NO 136; CDR1, CDR2 and CDR3 of SEQ ID NO. 188; or CDR1, CDR2 and CDR3 of SEQ ID NO. 212. In some embodiments, the variable domain of the second heavy chain comprises CDR1, CDR2 and CDR3 of SEQ ID NO. 49; CDR1, CDR2 and CDR3 of SEQ ID NO 113; CDR1, CDR2 and CDR3 of SEQ ID NO 137; CDR1, CDR2 and CDR3 of SEQ ID NO 189; or CDR1, CDR2 and CDR3 of SEQ ID NO 213. In some embodiments, the variable domain of the second light chain comprises CDR1, CDR2 and CDR3 of SEQ ID NO. 50; CDR1, CDR2 and CDR3 of SEQ ID NO. 114; CDR1, CDR2 and CDR3 of SEQ ID NO 138; CDR1, CDR2 and CDR3 of SEQ ID NO. 190; or CDR1, CDR2 and CDR3 of SEQ ID NO 214.

In some embodiments, the molecule comprises SEQ ID NO:47, SEQ ID NO:48, SEQ ID NO:49, and SEQ ID NO:50. In some embodiments, the molecule comprises SEQ ID NO:111, SEQ ID NO:112, SEQ ID NO:113, and SEQ ID NO:114. In some embodiments, the molecule comprises SEQ ID NO:135, SEQ ID NO:137, and SEQ ID NO:138. In some embodiments, the molecule comprises SEQ ID NO. 187, SEQ ID NO. 188, SEQ ID NO. 189 and SEQ ID NO. 190. In some embodiments, the molecule comprises SEQ ID NO:211, SEQ ID NO:212, SEQ ID NO:213, and SEQ ID NO:214.

In some embodiments, the molecule further comprises one or more disulfide bonds. In some embodiments, the molecule further comprises one or more linker sequences. In some embodiments, the molecule is capable of neutralizing coronavirus. In some embodiments, the coronavirus is a SARS-CoV-2 virus. In some embodiments, the SARS-CoV-2 virus strain is selected from the group consisting of WA1, B.1.1.7, B1.526, B1.351, P1, B1.352, and B.1.427. In some embodiments, the molecule specifically recognizes a first epitope and a second epitope on SARS-CoV-2 particle. In some embodiments, the first epitope comprises at least a portion of a Receptor Binding Domain (RBD) on a spike protein of a SARS-CoV-2 particle. In some embodiments, the second epitope comprises at least a portion of an N-terminal domain (NTD) on a spike protein of a SARS-CoV-2 particle.

In certain aspects, the invention provides a method of treating or preventing a covd-19 infection in a subject in need thereof, the method comprising administering to the subject a composition comprising a bispecific molecule comprising a first polypeptide chain and a second polypeptide chain, wherein: the first polypeptide chain comprises: a first light chain comprising a variable domain and a constant domain; a first heavy chain comprising a variable domain and a constant domain; the second polypeptide chain comprises: a second light chain comprising a variable domain and a constant domain; and a second heavy chain comprising a variable domain and a constant domain. In some embodiments, the first polypeptide chain and the second polypeptide chain are covalently bonded to each other.

In some embodiments, the first heavy chain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID NO, a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 72%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID NO, 211, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 98%, 99% identical to SEQ ID NO.

In some embodiments, the first light chain comprises a sequence identical to SEQ ID NO:48 at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical polypeptide sequence, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical polypeptide sequence to SEQ ID NO 112, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical polypeptide sequence to SEQ ID NO 188 at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 95%, 97%, 98%, 99% identical polypeptide sequence to SEQ ID NO 212 at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 98%, 99% identical polypeptide sequence to SEQ ID NO.

In some embodiments, the second heavy chain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 49, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 113, a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 189, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 213, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 92%, 98%, 99% identical to SEQ ID No. 213.

In some embodiments, the second light chain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 50, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 114, a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 190, at least 60%, 62%, 65%, 67%, 70%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 214, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 98%, 99% identical to SEQ ID NO.

In some embodiments, the variable domain of the first heavy chain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 35, a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 13. In some embodiments, the variable domain of the first light chain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 36, a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 14. In some embodiments, the variable domain of the second heavy chain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 13, a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 43, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 43.

In some embodiments, the variable domain of the second light chain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 46, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 44.

In some embodiments, the variable domain of the first heavy chain comprises CDR1, CDR2 and CDR3 of SEQ ID NO 47; CDR1, CDR2 and CDR3 of SEQ ID NO 111; CDR1, CDR2 and CDR3 of SEQ ID NO. 135; CDR1, CDR2 and CDR3 of SEQ ID NO 187; or CDR1, CDR2 and CDR3 of SEQ ID NO 211. In some embodiments, the variable domain of the first light chain comprises CDR1, CDR2 and CDR3 of SEQ ID NO. 48; CDR1, CDR2 and CDR3 of SEQ ID NO 112; CDR1, CDR2 and CDR3 of SEQ ID NO 136; CDR1, CDR2 and CDR3 of SEQ ID NO. 188; or CDR1, CDR2 and CDR3 of SEQ ID NO. 212. In some embodiments, the variable domain of the second heavy chain comprises CDR1, CDR2 and CDR3 of SEQ ID NO. 49; CDR1, CDR2 and CDR3 of SEQ ID NO 113; CDR1, CDR2 and CDR3 of SEQ ID NO 137; CDR1, CDR2 and CDR3 of SEQ ID NO 189; or CDR1, CDR2 and CDR3 of SEQ ID NO 213. In some embodiments, the variable domain of the second light chain comprises CDR1, CDR2 and CDR3 of SEQ ID NO. 50; CDR1, CDR2 and CDR3 of SEQ ID NO. 114; CDR1, CDR2 and CDR3 of SEQ ID NO 138; CDR1, CDR2 and CDR3 of SEQ ID NO. 190; or CDR1, CDR2 and CDR3 of SEQ ID NO 214. In some embodiments, the molecule comprises SEQ ID NO:47, SEQ ID NO:48, SEQ ID NO:49, and SEQ ID NO:50. In some embodiments, the molecule comprises SEQ ID NO:111, SEQ ID NO:112, SEQ ID NO:113, and SEQ ID NO:114. In some embodiments, the molecule comprises SEQ ID NO:135, SEQ ID NO:137, and SEQ ID NO:138. In some embodiments, the molecule comprises SEQ ID NO. 187, SEQ ID NO. 188, SEQ ID NO. 189 and SEQ ID NO. 190. In some embodiments, the molecule comprises SEQ ID NO:211, SEQ ID NO:212, SEQ ID NO:213, and SEQ ID NO:214.

In some embodiments, the molecule specifically recognizes a first epitope and a second epitope on SARS-CoV-2 particle. In some embodiments, the first epitope comprises at least a portion of a Receptor Binding Domain (RBD) on a spike protein of a SARS-CoV-2 particle. In some embodiments, the second epitope comprises at least a portion of an N-terminal domain (NTD) on a spike protein of a SARS-CoV-2 particle. In some embodiments, the composition is administered in combination with a second therapeutic agent.

Brief description of the drawings

The patent or application contains at least one original drawing in color. To meet the requirements of PCT patent application, many of the figures provided herein are black and white representations of images originally created in color.

FIGS. 1A-C show images of coronaviruses. FIG. 1A shows a schematic of SARS-CoV-2 virus particles. FIG. 1B shows the spike protein trimer structure. Fig. 1C shows an electron micrograph of coronavirus particles. Arrows indicate viral spike proteins on the viral membrane.

FIGS. 2A-G show individual antibody responses to SARS-CoV-2 virus. FIGS. 2A-F illustrate ELISA binding assays in which multiple plasma samples of patients with COVID-19 (represented by lines of different colors) were tested for their ability to bind to SARS-CoV-2 nucleocapsid antigen (FIGS. 2A and D), SARS-CoV-2 envelope trimer antigen (FIGS. 2B and E) and SARS-CoV envelope trimer antigen (FIGS. 2C and F). Figures 2A-C show responses to critically ill patients. Figures 2D-F show responses to non-critically ill patients. FIG. 2G shows that there is a higher SARS-CoV-2 antibody response in the plasma sample of the severe patient with COVID-19 than the non-severe patient with COVID-19.

FIGS. 3A-J show antibody responses effective in neutralizing SARS-CoV-2. Figures 3A-F show individual neutralization assays for both critically ill patients (figures 3A-C) and non-critically ill patients (figures 3D-F). FIGS. 3A and D show neutralization of SARS-CoV-2 pseudovirus. FIGS. 3B and E show neutralization of SARS-CoV pseudovirus. FIGS. 3C and F show neutralization of SARS-CoV-2 live virus. Fig. 3G shows the infection dose (ID ₅₀ ) Or generating a plot of the estimated number of virus particles required for 50% infection, from figures 3A-F. FIGS. 3H-J show ICs obtained by pseudovirus neutralization assay ₅₀ Value and live virus neutralization IC ₅₀ (FIG. 3H), half maximal plasma dilution (FIG. 3I) and 1:400OD ₄₅₀ (FIG. 3J) correlation.

FIGS. 4A-C show schematic diagrams for identifying potent neutralizing antibodies against COVID-19. Fig. 4A shows a schematic diagram of the production of effective neutralizing antibodies. FIG. 4B shows memory B cells sorted from healthy individuals and from patients with COVID-19. Figure 4C shows a table of seven patient donors and the number of antibodies isolated from each donor.

FIGS. 5A-D show binding and neutralization assays of mAbs against SARS-CoV-2 virus. FIG. 5A shows a binding assay of synthetic antibodies to SARS-CoV-2 spike protein trimer. FIG. 5B shows a binding assay of synthetic antibodies to the SARS-CoV-2 receptor binding domain. FIG. 5C shows a neutralization assay of synthetic antibodies with SARS-CoV-2 pseudovirus. FIG. 5D shows a neutralization assay of synthetic antibodies with SARS-CoV-2 live virus.

FIGS. 6A-F show epitope mapping of SARS-CoV-2 specific monoclonal antibodies as determined by competition ELISA. FIGS. 6A-C show the binding affinity of a single synthetic mAb to a Receptor Binding Domain (RBD), wherein antibodies Nos. 2-4, 2-15 and 2-38 specifically bind to RBDs. FIGS. 6D-F show the binding affinity of a single synthetic mAb to the Receptor Binding Domain (RBD), wherein Nos. 2-51, 4-8 and 4-32 show no binding affinity to RBD.

FIGS. 7A-E show Surface Plasmon Resonance (SPR) assays for mAb binding to spike protein trimers. Each panel represents the binding affinity and binding kinetics of the indicated monoclonal antibodies (i.e., 2-4, 2-15, 2-38, etc.) for SARS-CoV-2 envelope trimer. The color bars represent the different dilutions of the corresponding antibodies tested. FIG. 7A shows mAb 2-4. FIG. 7B shows mAb 2-15. FIG. 7C shows mAb2-38. FIG. 7D shows mAb 2-51. FIG. 7E shows mAb 4-8.

FIGS. 8A-D show a preliminary screen of mAbs that bind to and neutralize SARS-CoV-2 virus. Figure 8A shows mabs binding to spike trimer proteins. Fig. 8B shows mabs binding to the Receptor Binding Domain (RBD). FIG. 8C shows an IC infected with a pseudovirus ₅₀ Values. Fig. 8D shows the% inhibition values of live virus infection.

FIG. 9 shows a summary of SARS-CoV-2 specific mAb screening. The amount of most effective neutralizing mAb: 17.

FIGS. 10A-I show that binding of mAbs to spike protein trimers, RBDs and N-terminal domains (NTDs) is effectively neutralized. Fig. 10A, D and G show binding to the spike protein trimer. Fig. 10B, E and H show the combination with RBD. Fig. 10C, F and I show the binding to NTD. Figures 10A-C show binding of mabs that bind efficiently to RBD. Figures 10D-F show binding of mabs that bind efficiently to NTD. Fig. 10G-I show binding of mabs that bind to epitopes other than RBD or NTD.

FIGS. 11A-F show SARS-CoV-2 neutralization activity of selected mAbs. Fig. 11A shows the neutralizing activity of RBD-targeted mabs to neutralize pseudovirus infection. Fig. 11B shows the neutralizing activity of NTD-targeted mabs to neutralize pseudovirus infection. Fig. 11C shows neutralizing activity of mabs targeting epitopes other than RBD or NTD to neutralize pseudovirus infection. Fig. 11D shows the neutralizing activity of RBD-targeted mabs to neutralize live virus infection. Fig. 11E shows the neutralizing activity of NTD-targeted mabs to neutralize live virus infection. Fig. 11F shows neutralizing activity of mabs targeting epitopes other than RBD or NTD to neutralize live virus infection.

Fig. 12A-B show the CryoEM structure of RBD-targeting monoclonal antibodies. Fig. 12A shows a side view of the mAb. Fig. 12B shows a top view of the mAb.

Fig. 13A-B show the CryoEM structure of monoclonal antibodies targeting NTD. Fig. 13A shows a side view of a mAb. Fig. 13B shows a top view of the mAb.

FIG. 14 shows three SARS-CoV-2 neutralization epitope clusters identified on spike protein trimers.

FIGS. 15A-E show isolation of SARS-CoV-2mAb from critically infected patients. FIG. 15A shows the plasma neutralization profile of 40 patients on SARS-CoV-2 pseudovirus (highlighted are the 5 best neutralizers selected). All 252 parts of the transfection supernatant were screened for binding to S trimer and RBD, and for neutralization of SARS-CoV-2 pseudovirus and live virus. For pseudovirus neutralization, 50% inhibition dilutions (IC 50) of each supernatant are plotted. For live virus, a semi-quantitative representation of inhibition at 1:50 dilution is plotted, the neutralization level ranged from (-) (none to (+++) and (3) completely neutralizing). The effective antibodies identified later are marked with vertical lines and at the bottom. Antibodies from each patient were shown graphically or stained as shown in fig. 15A. Fig. 15B shows s-trimer binding. Fig. 15C shows RBD binding. Fig. 15D shows pseudovirus neutralization. Figure 15E shows live virus neutralization.

FIGS. 16A-O show characterization of SARS-CoV-2 effective neutralizing mAb. FIGS. 16A, D and G show the binding spectra of 19 purified potent neutralizing mAbs against SARS-CoV-2S trimer (left). FIG. 16B, FIG. E and FIG. H show the binding spectra (middle) of 19 purified potent neutralizing mAbs against SARS-CoV-2 RBD. FIG. 16C, FIG. F and FIG. I show the binding spectra of 19 purified potent neutralizing mAbs against SARS-CoV-2NTD (right). Note that mAb 2-30 binds a variety of proteins at high concentrations. FIG. 16J shows pseudo-virus neutralization against SARS-CoV-2 RBD. FIG. 16K shows pseudovirus neutralization against SARS-CoV-2 NTD. FIG. 16L shows pseudovirus neutralization against other epitopes on SARS-CoV-2 virus particles. FIG. 16M shows live virus neutralization against SARS-CoV-2 RBD. FIG. 16N shows live virus neutralization against SARS-CoV-2 NTD. FIG. 16O shows live virus neutralization against other epitopes on SARS-CoV-2 virus particles. The neutralization profile was for 19 purified mabs. The triplicate of the single repeat and neutralization of the binding experiments are expressed as mean ± Standard Error (SEM).

Figures 17A-D show epitope mapping of selected neutralizing and non-neutralizing mabs. Competition results of non-RBD binders (fig. 17A) and RBD binders (fig. 17B) in blocking ACE2 or biotinylated mAb binding to S trimer. In addition, the ability of each mAb to bind NTD and RBD is shown. The numbers in each box show the area under each competition curve (AUC) tested by ELISA. +/-means binding/non-binding of mAb to protein. Fig. 17C shows a wien (Venn) plot interpretation of the results of fig. 17B. Fig. 17D shows a wien (Venn) plot interpretation of the results of fig. 17B.

Fig. 18A-D show Cryo-EM reconstruction of Fab-spike complexes and visualization of neutralizing epitopes on spike surfaces. FIG. 18A shows Cryo-EM 3D reconstruction of antibody 2-4 complexed with S trimer, with total resolution ofDensity is colored according to the spike domain, RBD is green, NTD is orange, and the other areas are gray. FIG. 18B shows Cryo-EM reconstruction of antibody 4-8 complexed with S trimer (banding pattern, staining as in a); the total resolution is->The RBD is in a "fully down" configuration. The resolution of antibody density is limited by molecular motion. Although the binding of Fab to NTD and the location of antibodies are clear, the identity of the heavy and light chains is not yet established. FIG. 18C shows->Cryo-EM reconstruction of antibody 2-43 complexed with S trimer at resolution reveals quaternary epitopes that involve RBD of one subunit and NTD of the next subunit. Fig. 18D shows the mapping of the wien (Venn) plot of fig. 17B on the surface of viral spikes.

Fig. 19 shows patient information. Abbreviations: ARDS, acute respiratory distress syndrome; MV, mechanical ventilation; hsCRP, high sensitivity C reactive protein, upper limit of normal value (ULN) >10mg/L; ESR, erythrocyte sedimentation rate, uln=20 mm/hr; interleukin 6, uln=5 pg/mL; ferritin, uln=150 ng/mL; d-dimer quantification uln=0.8 μg/mL FEU.

Figure 20 shows a summary of mAb screening.

Figure 21 shows Cryo-EM data acquisition, refinement and validation statistics.

FIGS. 22A-C show SARS-CoV-2S trimer-specific antibody isolation strategy. FIG. 22A shows a summary of the isolation of S-trimer specific mAbs from memory B cells in the blood of infected patients. FIG. 22B shows the results of sorting S trimer-specific memory B cells using flow cytometry. Fig. 22C shows an enlarged view of the S-trimer positive memory B cell group for each patient. The inset numbers represent the absolute number and percentage of S trimer-specific memory B cells isolated from each case.

FIGS. 23A-G show the genetic profile of SARS-CoV-2 specific antibody repertoire. Fig. 23A shows that most of the 121 trimer S-specific antibodies are of IgG isotype. Fig. 23B shows that heavy chains kappa (fig. 23C) and lambda (fig. 23D) light chains can be equally used. Compared to the IgG repertoire of healthy human donors, IGHV3-30 and IGKV3-20 genes were overexpressed in the heavy and light chain repertoires, respectively (β2-test, p < 0.05). Figure 23E shows that the use of the igh j6 gene in antigen-specific antibodies is significantly higher (β2-assay, p < 0.05). Figure 23F shows that the CDRH3 length of the antigen-specific antibodies was significantly longer than that of healthy donors (Kolmogorov-Smirnov test), p=0.014. Figure 23G shows that V region nucleotide somatic hypermutation levels were significantly lower for heavy and light chains than for healthy donor antibodies (Kolmogorov-Smirnov test, p < 0.001).

FIG. 24 shows a best-fit pseudo-virus neutralization curve for 130 samples positive in at least one of the screens shown in FIGS. 15B-E. 18 parts of the transfection supernatant showing significantly better efficacy are highlighted in color, while the other transfection supernatants with non-or weak neutralization activity are shown in gray. Another supernatant (patient 1) was originally omitted from the pseudovirus screen, but was later found to be an effective neutralizing mAb (1-87), also highlighted.

Figure 25 shows the pseudo-virus neutralization profile of 12 purified mabs that bind strongly to S trimer but have weaker or no virus neutralization activity. Four mabs with weak neutralizing activity against SARS-CoV-2 pseudovirus are shown in solid lines, the remaining 8 non-neutralizing mabs are shown in dashed lines (dashed lines).

FIGS. 26A-C show that monoclonal Ab2-43 binding to the S trimer expressed on the surface of an Expi293 cell can be competed by mAbs against RBD, but only minimally by mAbs against the NTD region. FIG. 26A shows the results of 2-43 and protein. Fig. 26B shows the results for mabs against NTD. Fig. 26C shows the results for mabs against RBD.

FIGS. 27A-F show Cryo-EM data processing for antibodies 2-4 complexed with S-trimer. Fig. 27A shows representative micrographs and CTFs of the micrographs. Fig. 27B shows a representative class 2D average. Fig. 27C shows the resolution of a correspondence map with C3 symmetry calculated from 3 DFSC. Fig. 27D shows the global local resolution calculated by crysparc at FSC cutoff of 0.5. Representative densities of Fab 2-4 and RBD junctions, showing CRR H3, L3 and L3 (fig. 27E), and N-linked glycosylation at CDR H2 and ASN58 (fig. 27F).

FIGS. 28A-E show Fab 2-4 binding. FIG. 28A shows the binding sites of Fab 2-4 to RBD. FIG. 28B shows antibodies 2-4, S309 ⁸ And BD-23 ⁹ At the site of trimeric CoV-2 spike. Figure 28C shows somatic hypermutations found only in the heavy chain of antibodies 2-4, shown in brown. The mutation a60T produced NxT sequence leading to N58 glycosylation. FIG. 28D shows antibody 2-4 complexed with S trimer. CDR loops are shown in color, 751 and side chains show residues for interaction. FIG. 28E shows antibody BD-23 ⁹ Compounding with S trimer.

FIGS. 29A-F show Cryo-EM data processing for antibody 4-8 complexed with S trimer. Fig. 29A shows representative micrographs and CTFs of the micrographs. Fig. 29B shows a representative class 2D average. FIG. 29C shows the resolution of spikes in the downward conformation of the RBD complexed with Fab 4-8. FIG. 29D shows the resolution of spikes in the upward conformation of RBD complexed with Fab 4-8. FIG. 29E shows the local resolution of spikes in the downward conformation of RBD complexed with Fab 4-8 at a FSC cutoff value of 0.5. Two thresholds are shown. FIG. 29F shows the local resolution of spikes in the up-directed conformation of RBD complexed with Fab 4-8 at a FSC cutoff value of 0.5. Two thresholds are shown.

FIG. 30 shows a 3D reconstruction of neutralizing antibody 4-8 targeting NTD complexed with SARS-CoV-2 spike trimer having a 1-RBD-up conformation.

FIGS. 31A-D show Cryo-EM data processing for antibody 2-43 complexed with S trimer. Fig. 31A shows representative micrographs and CTFs of the micrographs. Fig. 31B shows a representative class 2D average. FIG. 31C shows the resolution of Fab 2-43 complexed with S trimer. Fig. 31D shows the global local resolution calculated by crysparc at FSC cutoff (cutoff) of 0.5.

FIGS. 32A-B show monoclonal antibodies 2-15mut. FIG. 32A shows virus neutralization using 2-15mut antibodies. FIG. 32B shows the efficacy of 2-15mut antibodies.

FIGS. 33A-B show selection mutants of the 4-8 monoclonal antibodies. FIG. 33A shows neutralization of antibodies 4-8 (39/51) and 4-8 (39/51/57). FIG. 33B shows the efficacy of antibodies 4-8 (39/51) and 4-8 (39/51/57).

FIG. 34 shows the IC of selected monoclonal antibody mutants ₅₀ And IC ₉₀ Values.

FIG. 35 shows that antibody 2-36 is a potent neutralizing agent against SARS-CoV pseudovirus.

FIGS. 36A-B show ELISA binding of antibodies 2-36 to SARS-CoV-2 spike trimer (FIG. 36A) and SARS-CoV spike trimer (FIG. 36B). Antibody CR3022 with cross-activity was previously reported as a control.

FIGS. 37A-D show the binding affinities of antibodies 2-36 and antibodies 2-4 for SARS-CoV-2 spike trimer and SARS-CoV spike trimer, as determined by SPR. Antibody 2-36 was demonstrated to bind to SARS-CoV-2 (FIG. 37A) and SARS-CoV (FIG. 37B) spikes, but with higher affinity for SARS-CoV-2 spikes. As a control, SARS-CoV-2 specific antibody 2-4 binds only to SARS-CoV-2 spike (FIG. 37C) and not to SARS-CoV spike (FIG. 37D).

Fig. 38A-B show the neutralization assay results. The antibodies 2-36 neutralized both SARS-CoV-2 (FIG. 38A) and SARS-CoV (FIG. 38B) compared to the control antibody CR3022 which neutralized SARS-CoV (FIG. 38B) but only very weakly neutralized SARS-CoV-2 (FIG. 38A) and compared to the SARS-CoV-2 specific antibody 2-4 which neutralized only SARS-CoV-2 (FIG. 38A).

FIGS. 39A-B show the neutralization of SARS-like coronavirus with antibodies 2-36 using hACE 2. Fig. 39A shows the lineage of coronaviruses, including several coronaviruses from bat and pangolin scales. FIG. 39B shows that antibody 2-36 is capable of neutralizing SARS-CoV and SARS-CoV-2 and its related lineage viruses, but that antibody 2-36 is not capable of neutralizing MERS-CoV and 229E, indicating its breadth.

FIGS. 40A-D show cryo-EM structures of antibody 2-36 complexed with SARS-CoV-2. FIG. 40A shows the cryo-EM structure of 2-36 complexed with SARS-CoV-2 spike trimer, and FIG. 40B further details how 2-36 interacts with spike at the CDR level. Figure 40C shows that antibody 2-36 binds to one side of RBD and competes with ACE2 for binding to RBD, although its epitope does not overlap with the ACE2 binding site. Figure 40D depicts that spike proteins are highly conserved.

FIG. 41 shows a schematic of an engineered bispecific antibody that binds the binding specificity of two neutralizing antibodies into one.

FIG. 42 shows that the 2-17/1-57 bispecific antibody against SARS-CoV-2 does not enhance efficacy compared to the corresponding single arm counterpart. Ab X and Ab Y are single arm independent controls. X=pgdm 1400 (HIV neutralization Ab). Y=a32 (HIV neutralization Ab).

FIG. 43 shows the effective neutralization of bispecific antibodies 2-17/2-7 against SARS-CoV-2 as compared to the corresponding single arm counterparts. Ab X and Ab Y are single arm independent controls. X=pgdm 1400 (HIV neutralization Ab). Y=a32 (HIV neutralization Ab).

FIG. 44 shows the effective neutralization of bispecific antibodies 2-17/2-15 against SARS-CoV-2 as compared to the corresponding single arm counterparts. Ab X and Ab Y are single arm independent controls. X=pgdm 1400 (HIV neutralization Ab). Y=a32 (HIV neutralization Ab).

FIG. 45 shows the effective neutralization of bispecific antibody 2-17/2-30 against SARS-CoV-2 as compared to the corresponding single arm counterpart. Ab X and Ab Y are single arm independent controls. X=pgdm 1400 (HIV neutralization Ab). Y=a32 (HIV neutralization Ab).

FIG. 46 shows the effective neutralization of bispecific antibody 2-17/4-20 against SARS-CoV-2 as compared to the corresponding single arm counterpart. Ab X and Ab Y are single arm independent controls. X=pgdm 1400 (HIV neutralization Ab). Y=a32 (HIV neutralization Ab).

FIG. 47 shows the effective neutralization of bispecific antibody 5-24/1-57 against SARS-CoV-2 as compared to the corresponding single arm counterpart. Ab X and Ab Y are single arm independent controls. X=pgdm 1400 (HIV neutralization Ab). Y=a32 (HIV neutralization Ab).

FIG. 48 shows the effective neutralization of bispecific antibodies 5-24/2-15 against SARS-CoV-2 as compared to the corresponding single arm counterparts. Ab X and Ab Y are single arm independent controls. X=pgdm 1400 (HIV neutralization Ab). Y=a32 (HIV neutralization Ab).

FIG. 49 shows that bispecific antibody 5-24/4-20 against SARS-CoV-2 does not enhance efficacy (IC 50) as compared to the corresponding single arm counterpart. Ab X and Ab Y are single arm independent controls. X=pgdm 1400 (HIV neutralization Ab). Y=a32 (HIV neutralization Ab).

FIG. 50 shows the effective neutralization of bispecific antibody 1-20/1-68 against SARS-CoV-2 as compared to the corresponding single arm counterpart. Ab X and Ab Y are single arm independent controls. X=pgdm 1400 (HIV neutralization Ab). Y=a32 (HIV neutralization Ab).

FIG. 51 shows the effective neutralization of bispecific antibodies 1-20/2-17 against SARS-CoV-2 as compared to the corresponding single arm counterparts. Ab X and Ab Y are single arm independent controls. X=pgdm 1400 (HIV neutralization Ab). Y=a32 (HIV neutralization Ab).

FIG. 52 shows the effective neutralization of bispecific antibody 1-20/4-18 against SARS-CoV-2 as compared to the corresponding single arm counterpart. Ab X and Ab Y are single arm independent controls. X=pgdm 1400 (HIV neutralization Ab). Y=a32 (HIV neutralization Ab).

FIG. 53 shows the effective neutralization of bispecific antibodies 1-20/5-24 against SARS-CoV-2 as compared to the corresponding single arm counterparts. Ab X and Ab Y are single arm independent controls. X=pgdm 1400 (HIV neutralization Ab). Y=a32 (HIV neutralization Ab).

FIG. 54 shows effective neutralization against SARS-CoV-2. Bispecific antibodies with 2-17 as a component were more effective than the single arm control (2-17/A32, A32 being a control antibody that was inactive against SARS-CoV-2), indicating that both arms contribute to the neutralizing activity of the bispecific molecule.

FIG. 55 shows the effective neutralization of SARS-CoV-2 with a bispecific antibody containing 5-24 as a component. In antibody 5-24/A32, A32 was a control antibody that was inactive against SARS-CoV-2, indicating that both arms contribute to the neutralizing activity of the bispecific molecule.

FIG. 56 shows the effective neutralization of SARS-CoV-2 with bispecific antibodies containing 1-20 as components. In antibodies 1-20/A32, A32 was a control antibody that was inactive against SARS-CoV-2, indicating that both arms contribute to the neutralizing activity of the bispecific molecule.

FIGS. 57A-E show bispecific antibodies with 1-20 (RBD) as parent antibodies. FIG. 57A shows virus neutralization with bispecific antibodies 1-20/5-24, 1-20/4-18, 1-20/2-17 and 1-20/1-68. FIG. 57B shows virus neutralization of bispecific antibodies 1-20/5-24 as compared to control antibodies. FIG. 57C shows virus neutralization of bispecific antibodies 1-20/4-18 as compared to control antibodies. FIG. 57D shows virus neutralization of bispecific antibodies 1-20/2-17 as compared to control antibodies. FIG. 57E shows virus neutralization of bispecific antibody 1-20/1-68 as compared to control antibody.

FIG. 58 shows the efficacy of bispecific antibodies with 1-20 (RBD) as parent antibodies.

FIGS. 59A-G show bispecific antibodies with 2-17 (NTD) as the parent antibody. FIG. 59A shows virus neutralization with bispecific antibodies 2-17/2-7, 2-17/1-57, 2-36/2-17, 2-17/4-20, 2-17/2-30 and 2-17/2-15. FIG. 59B shows virus neutralization of bispecific antibody 2-17/2-7 as compared to control antibody. FIG. 59C shows virus neutralization of bispecific antibody 2-17/1-57 as compared to control antibody. FIG. 59D shows virus neutralization of bispecific antibody 2-17/2-15 as compared to control antibody. FIG. 59E shows virus neutralization of bispecific antibody 2-17/4-20 as compared to control antibody. FIG. 59F shows virus neutralization of bispecific antibody 2-17/2-30 as compared to control antibody. FIG. 59G shows virus neutralization of bispecific antibody 2-36/2-17 as compared to control antibody.

FIG. 60 shows the efficacy of bispecific antibodies with 2-17 (NTD) as the parent antibody.

FIGS. 61A-D show bispecific antibodies with 5-24 (NTD) as the parent antibody. FIG. 61A shows virus neutralization with bispecific antibodies 5-24/4-20, 5-24/1-57, 5-24/2-15 and 1-20/5-24. FIG. 61B shows virus neutralization of bispecific antibody 5-24/2-15 as compared to control antibody. FIG. 61C shows virus neutralization of bispecific antibody 5-24/1-57 as compared to control antibody. FIG. 61D shows virus neutralization of bispecific antibody 5-24/4-20 as compared to control antibody.

FIG. 62 shows the efficacy of bispecific antibodies with 5-24 (NTD) as the parent antibody.

FIGS. 63A-D show bispecific antibodies with 2-36 (RBD) as parent antibodies. FIG. 63A shows virus neutralization with bispecific antibodies 2-36/1-68, 2-36/4-18 and 2-36/2-17. FIG. 63B shows virus neutralization of bispecific antibody 2-36/2-17 as compared to control antibody. FIG. 63C shows virus neutralization of bispecific antibody 2-36/1-68 as compared to control antibody. FIG. 63D shows virus neutralization of bispecific antibody 2-36/4-18 as compared to control antibody.

FIG. 64 shows the efficacy of bispecific antibodies with 2-36 (RBD) as parent antibody.

FIGS. 65A-C show bispecific antibodies with 2-30 (RBD) as parent antibodies. FIG. 65A shows virus neutralization of bispecific antibodies 2-30/4-18 and 2-30/1-68. FIG. 65B shows virus neutralization of bispecific antibody 2-30/1-68 as compared to control antibody. FIG. 65C shows virus neutralization of bispecific antibody 2-30/4-18 as compared to control antibody.

FIG. 66 shows the efficacy of bispecific antibodies with 2-30 (RBD) as parent antibody.

FIGS. 67A-D show comparative evaluation of bispecific antibody 2-17/2-7. FIG. 67A shows the efficacy of a 2-17/2-7 bispecific antibody compared to an antibody in which one arm is similar to the single parent component of the bispecific antibody and the other arm is an unrelated control. It also compares the efficacy of bispecific antibodies to a combination of two single arm IgG controls. FIG. 67B shows virus neutralization with a combination of parent antibodies and each parent antibody. FIG. 67C shows virus neutralization of bispecific antibody 2-17/2-7 as compared to the combination of parent antibodies. Figure 67D shows the efficacy of the combined parent monoclonal antibodies each tested in half the amount of the corresponding bispecific antibody. Since each monoclonal antibody has two identical arms, whereas each monoclonal antibody of each bispecific antibody has only one arm, testing in half amounts allows the same concentration of each monoclonal antibody arm to be compared to the bispecific antibody and monoclonal antibody combination.

FIGS. 68A-D show comparative evaluations of bispecific antibodies 1-20/4-18. FIG. 68A compares the efficacy of 1-20/4-18 bispecific antibodies with antibodies in which one arm is similar to the single parent component of the bispecific antibody and the other arm is an unrelated control. It also compares the efficacy of bispecific antibodies to a combination of two single arm IgG controls. FIG. 68B shows virus neutralization with a combination of parent antibodies and each parent antibody. FIG. 68C shows virus neutralization of bispecific antibodies 1-20/4-18 as compared to the combination of parent antibodies. Figure 68D shows the efficacy of the combined parent monoclonal antibodies each tested in half the amount of the corresponding bispecific antibody.

FIGS. 69A-D show comparative evaluations of bispecific antibodies 1-20/5-24. FIG. 69A compares the efficacy of 1-20/5-24 bispecific antibodies with antibodies in which one arm is similar to the single parent component of the bispecific antibody and the other arm is an unrelated control. It also compares the efficacy of bispecific antibodies to a combination of two single arm IgG controls. FIG. 69B shows virus neutralization with a combination of parent antibodies and each parent antibody. FIG. 69C shows virus neutralization of bispecific antibodies 1-20/5-24 as compared to the combination of parent antibodies. Figure 69D shows the efficacy of the combined parent monoclonal antibodies each tested in half the amount of the corresponding bispecific antibody.

FIGS. 70A-D show comparative evaluations of bispecific antibody 2-17/4-20. FIG. 70A compares the efficacy of 2-17/4-20 bispecific antibodies to antibodies in which one arm is similar to the single parent component of the bispecific antibody and the other arm is an unrelated control. It also compares the efficacy of bispecific antibodies to a combination of two single arm IgG controls. FIG. 70B shows virus neutralization with a combination of parent antibodies and each parent antibody. FIG. 70C shows virus neutralization of bispecific antibody 2-17/4-20 as compared to the combination of parent antibodies. Figure 70D shows the efficacy of the combined parent monoclonal antibodies each tested in half the amount of the corresponding bispecific antibody.

FIGS. 71A-B show the in vitro efficacy of bispecific antibodies against SARS-CoV-2 real virus strain USA/WA 1. Fig. 71A shows an IC ₉₀ Neutralization of ten bispecific antibodies at a concentration between 0.004 μg/ml and 0.025 μg/ml. Fig. 71B shows an IC ₉₀ The concentration was between 0.026. Mu.g/ml and 0.Neutralization of nine bispecific antibodies between 046 μg/ml. True viruses are another term of use for "infectious" viruses. The term may also refer to "live viruses". The first strain isolated in the United states by USA/WA1 strain WAs supplied by the BEI resource center.

FIG. 72 shows the minimum to maximum IC ₉₀ The concentration provides in vitro efficacy of the bispecific antibody. The highlighted antibodies were tested against the control.

FIGS. 73A-B show a display IC ₉₀ And IC ₅₀ In vitro potency of bispecific antibody at concentration. FIG. 73A shows an IC showing a threshold value relative to 0.05 μg/ml ₉₀ And IC ₅₀ In vitro potency of bispecific antibody at concentration. FIG. 73B shows an IC showing a threshold value relative to 0.01 μg/ml ₉₀ And IC ₅₀ In vitro potency of bispecific antibody at concentration.

FIGS. 74A-D show comparative evaluations of bispecific antibodies 1-57/1-68. FIG. 74A shows virus neutralization of bispecific antibodies 1-57/1-68 compared to two control bispecific antibodies (each antibody comprising a parent antibody arm and a control antibody arm), and compared to a mixture of two control bispecific antibodies. AbX is an irrelevant control antibody arm, where AbX is PGDM1400. FIG. 74B shows virus neutralization with a combination of parent antibodies and each parent antibody. FIG. 74C shows virus neutralization of bispecific antibody 1-57/1-68 as compared to the combination of parent antibodies. FIG. 74D shows the efficacy of a mixture of bispecific antibodies 1-57/1-68 and each parent monoclonal antibody, each tested in half the amount of the corresponding bispecific antibody. The comparison data is from a single experiment, while the data in the summary table is the average of multiple replicates.

FIGS. 75A-D show comparative evaluation of bispecific antibody 2-17/2-7. FIG. 75A shows virus neutralization of bispecific antibodies 2-17/2-7 compared to two control bispecific antibodies (each antibody comprising a parent antibody arm and a control antibody arm), and compared to a mixture of two control bispecific antibodies. AbX and AbY are unrelated control antibody arms, where AbX is PGDM1400 and AbY is a32. FIG. 75B shows virus neutralization with a combination of parent antibodies and each parent antibody. FIG. 75C shows virus neutralization of bispecific antibody 2-17/2-7 as compared to the combination of parent antibodies. FIG. 75D shows the efficacy of a mixture of bispecific antibodies 2-17/2-7 and each parent monoclonal antibody, each tested in half the amount of the corresponding bispecific antibody. The comparison data is from a single experiment, while the data in the summary table is the average of multiple replicates.

FIGS. 76A-D show comparative evaluations of bispecific antibodies 1-20/4-18. FIG. 76A shows virus neutralization of bispecific antibodies 1-20/4-18 as compared to two control bispecific antibodies (each antibody comprising a parent antibody arm and a control antibody arm) and as compared to a mixture of two control bispecific antibodies. AbX and AbY are unrelated control antibody arms, where AbX is PGDM1400 and AbY is a32. FIG. 76B shows virus neutralization with a combination of parent antibodies and each parent antibody. FIG. 76C shows virus neutralization of bispecific antibodies 1-20/4-18 as compared to the combination of parent antibodies. FIG. 76D shows the efficacy of a mixture of bispecific antibodies 1-20/4-18 and each parent monoclonal antibody, each tested in half the amount of the corresponding bispecific antibody. The comparison data is from a single experiment, while the data in the summary table is the average of multiple replicates.

FIGS. 77A-D show comparative evaluations of bispecific antibodies 1-20/5-24. FIG. 77A shows virus neutralization of bispecific antibodies 1-20/5-24 compared to two control bispecific antibodies (each antibody comprising a parent antibody arm and a control antibody arm) and compared to a mixture of two control bispecific antibodies. AbX and AbY are unrelated control antibody arms, where AbX is PGDM1400 and AbY is a32. FIG. 77B shows virus neutralization with a combination of parent antibodies and each parent antibody. FIG. 77C shows virus neutralization of bispecific antibodies 1-20/5-24 as compared to the combination of parent antibodies. FIG. 77D shows the efficacy of a mixture of bispecific antibodies 1-20/5-24 and each parent monoclonal antibody, each tested in half the amount of the corresponding bispecific antibody. The comparison data is from a single experiment, while the data in the summary table is the average of multiple replicates.

FIGS. 78A-D show comparative evaluations of bispecific antibodies 2-30/1-68. FIG. 78A shows virus neutralization of bispecific antibodies 2-30/1-68 compared to two control bispecific antibodies (each antibody comprising a parent antibody arm and a control antibody arm) and compared to a mixture of two control bispecific antibodies. AbX is an irrelevant control antibody arm, where AbX is PGDM1400. FIG. 78B shows virus neutralization with a combination of parent antibodies and each parent antibody. FIG. 78C shows virus neutralization of bispecific antibody 2-30/1-68 as compared to the combination of parent antibodies. FIG. 78D shows the efficacy of a mixture of bispecific antibodies 2-30/1-68 and each parent monoclonal antibody, each tested in half the amount of the corresponding bispecific antibody. The comparison data is from a single experiment, while the data in the summary table is the average of multiple replicates.

FIGS. 79A-D show comparative evaluation of bispecific antibodies 2-7/4-8 (39/51). FIG. 79A shows virus neutralization of bispecific antibodies 2-7/4-8 (39/51) compared to two control bispecific antibodies, each comprising a parent antibody arm and a control antibody arm, and compared to a mixture of two control bispecific antibodies. AbX is an irrelevant control antibody arm, where AbX is PGDM1400. FIG. 79B shows virus neutralization with a combination of parent antibodies and each parent antibody. FIG. 79C shows virus neutralization of bispecific antibodies 2-7/4-8 (39/51) as compared to the combination of parent antibodies. FIG. 79D shows the efficacy of a mixture of bispecific antibodies 2-7/4-8 (39/51) and each parent monoclonal antibody, each tested in half the amount of the corresponding bispecific antibody. The comparison data is from a single experiment, while the data in the summary table is the average of multiple replicates.

FIGS. 80A-D show comparative evaluations of bispecific antibodies 1-57/1-87. FIG. 80A shows virus neutralization of bispecific antibodies 1-57/1-87 as compared to two control bispecific antibodies, each comprising a parent antibody arm and a control antibody arm, and as compared to a mixture of two control bispecific antibodies. AbX is an irrelevant control antibody arm, where AbX is PGDM1400. FIG. 80B shows virus neutralization with a combination of parent antibodies and each parent antibody. FIG. 80C shows virus neutralization of bispecific antibodies 1-57/1-87 as compared to the combination of parent antibodies. FIG. 80D shows the efficacy of a mixture of bispecific antibodies 1-57/1-87 and each parent monoclonal antibody, each tested in half the amount of the corresponding bispecific antibody. The comparison data is from a single experiment, while the data in the summary table is the average of multiple replicates.

Figure 81 shows the efficacy of other bispecific antibodies.

FIGS. 82A-B show the neutralizing activity of antibodies specific for worker Cheng Huashuang. FIG. 82A shows the neutralizing activity of bispecific antibodies 2-17/2-7, 1-57/5-7, 5-7/1-57, 2-7/5-7, 5-7/2-7 against wild type virus (WA 1) at various antibody concentrations. FIG. 82B shows the antibody efficacy (IC 50 and IC 90) of bispecific antibodies 2-17/2-7, 1-57/5-7, 5-7/1-57, 2-7/5-7, 5-7/2-7 against wild type virus (WA 1).

FIGS. 83A-B show 2-7/5-7 bispecific and affinity antibody activity. FIG. 83A shows the neutralizing activity of bispecific antibody 2-7/5-7, parent antibodies 2-7, 5-7, and combinations of 2-7 and 5-7 parent antibodies against wild type virus (WA 1) at various antibody concentrations. FIG. 83B shows the IC of bispecific antibody 2-7/5-7, parent antibodies 2-7, 5-7, and combinations of parent antibodies 2-7 and 5-7 ₅₀ Concentration.

FIGS. 84A-B show the activity of bispecific antibodies 2-7/5-7 against SARS-CoV-2 wild-type (WA 1) and variant B1.1.7 (UK), B1.526 (New York), B1.351 (south Africa) and P.1 (Brazil). FIG. 84A shows the neutralizing activity of bispecific antibody 2-7/5-7 against various SARS-CoV-2 strains at different antibody concentrations. FIG. 84B shows the IC of bispecific antibody 2-7/5-7 against various SARS-CoV-2 strains ₅₀ And IC ₉₀ Concentration.

FIGS. 85A-D show neutralization of pseudoviruses with bispecific 2-7/5-7 antibodies having mutations associated with SARS-CoV-2 variant. FIG. 85A shows the antibody neutralization profile against a circulating SARS-CoV-2 virus variant. FIG. 85B shows antibody efficacy against circulating SARS-CoV-2 virus variants. FIG. 85C shows an antibody neutralization profile for SARS-CoV-2 variant with high frequency mutations. FIG. 85D shows antibody efficacy against SARS-CoV-2 variant with high frequency mutation.

FIGS. 86A-F show the use of bispecific antibodies 2-7/5-7 to neutralize pseudoviruses with SARS-CoV-2 mutation corresponding to the "related variant (variant of concern)". Fig. 86A shows the antibody neutralization profile against b.1.1.7 (uk virus variant) with NTD mutations. Figure 86B shows antibody efficacy against b.1.1.7 (uk virus variant) with NTD mutations. Fig. 86C shows an antibody neutralization profile for b.1.352 (SA) virus variants with NTD mutations. Figure 86D shows antibody efficacy against b.1.352 (SA) virus variants with NTD mutations. Figure 86E shows an antibody neutralization profile for p.1 (BZ) virus variants with NTD mutations. Figure 86F shows antibody efficacy against p.1 (BZ) virus variants with NTD mutations.

FIGS. 87A-F show the use of bispecific antibodies 2-7/5-7 to neutralize pseudoviruses with SARS-CoV-2 mutation corresponding to the "related variant". Fig. 87A shows an antibody neutralization profile for a b.1.427 (CA) virus variant having an NTD mutation. Figure 87B shows antibody efficacy against b.1.427 (CA) virus variants with NTD mutations. Fig. 87C shows an antibody neutralization curve for a b.1.526 (NY) variant with NTD mutations. Figure 87D shows antibody efficacy against b.1.526 (NY) variants with NTD mutations. Fig. 87E shows an antibody neutralization profile for NJ variants with NTD mutations. Figure 87F shows antibody efficacy against NJ variants with NTD mutations.

FIG. 88 shows a summary of bispecific antibody 2-7/5-7 activity against variants. IC50 is shown as circles. IC90 is shown as square.

FIGS. 89A-C show the neutralizing activity of bispecific antibody 2-7/5-7 and parent 2-7 and 5-7 monoclonal antibodies against SARS-CoV-2 variant (live virus). FIG. 89A shows the neutralization profile of bispecific antibody 2-7/5-7. FIG. 89B shows the neutralization profile of parent antibodies 2-7. FIG. 89C shows the neutralization profile of parent antibodies 5-7.

FIGS. 90A-D show virus neutralization with monoclonal antibodies 2-36. FIG. 90A shows the screening of transfected supernatants for neutralization activity of SARS-CoV-2 pseudovirus. FIG. 90B shows screening of transfected supernatants for neutralization activity of SARS-CoV pseudovirus. FIG. 90C shows 2-36 neutralization IC50 (. Mu.g/mL) against SARS-CoV-2 Pseudovirus (PV) and Live Virus (LV). FIG. 90D shows 2-36 neutralization IC50 (. Mu.g/mL) against SARS-CoV Pseudovirus (PV) and Live Virus (LV).

FIGS. 91A-B show binding of monoclonal antibodies 2-36 to SARS-CoV-2 (FIG. 91A) and SARS-CoV (FIG. 91B) spikes as determined by ELISA.

FIGS. 92A-H show the binding affinities of monoclonal antibodies 2-36 and 2-4 to SARS-CoV-2 and SARS-CoV viruses as measured by SPR. FIG. 92A shows the binding affinity of 2-36 for SARS-CoV-2 spike protein. FIG. 92B shows the binding affinity of 2-4 for SARS-CoV-2 spike protein. FIG. 92C shows the binding affinity of 2-36 for SARS-CoV spike protein. FIG. 92D shows the binding affinity of 2-4 for SARS-CoV spike protein. FIG. 92E shows the binding affinity of 2-36 for the SARS-CoV-2 Receptor Binding Domain (RBD). FIG. 92F shows the binding affinity of 2-4 for SARS-CoV-2 RBD. FIG. 92G shows the binding affinity of 2-36 for SARS-CoV RBD. FIG. 92H shows the binding affinity of 2-4 for SARS-CoV RBD.

FIGS. 93A-B show binding to SARS-CoV-2 spike protein. FIG. 93A shows that binding of monoclonal antibody 2-36 to SARS-CoV-2 spike protein is inhibited by CR 3022. FIG. 93B shows that monoclonal antibody 2-36 inhibits binding of hACE2 to SARS-CoV-2 spike protein.

FIG. 94 shows a conservation analysis of RBD in SARS CoV-2, SARS CoV-1, bat CoV (Bat Covs) and human CoV (Human Covs) strains. Analysis showed that the 2-36 binding sites are highly conserved; gray is the high conserved region and red is the low conserved region. The 2-36 binding sites are marked in blue.

FIGS. 95A-D show the neutralization of SARS-CoV-2 variants and SARS-like coronaviruses with monoclonal antibodies 2-36 using hACE 2. Fig. 95A shows neutralization of living variants. Fig. 95B shows neutralization of pseudo-variants. FIG. 95C shows a virus' evolutionary tree. Figure 95D shows antibody efficacy against viruses.

FIGS. 96A-D show the neutralization of SARS-like coronavirus with monoclonal antibodies 2-36 using hACE 2. FIG. 96A shows virus neutralization of 2-36 antibodies. Fig. 96B shows virus neutralization of S309 antibody. FIG. 96C shows virus neutralization of COVA1-16 antibodies. Figure 96D shows virus neutralization of CR3022 antibodies.

Figures 97A-D show in vitro selection of 2-36 antibody escape mutations. FIG. 97A shows the evolution of escape mutations during in vitro passage of SARS-CoV-2USA/WA1 in Vero E6 cells in the presence of 2-36. FIG. 97B shows the neutralization activity of 2-36 for virus passaging tests. Fig. 97C shows spike proteins with locally selected mutations. FIG. 97D shows that selected escape mutations were introduced into pseudoviruses, and then 2-36 neutralization activity was tested against these viruses.

Detailed description of the invention

Definition of the definition

The following are definitions of terms used in the present specification. Unless otherwise indicated, the initial definition provided herein for a group or term applies to that group or term throughout this specification, either alone or as part of another group. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art.

The singular forms "a", "an" and "the" include plural referents unless the context clearly dictates otherwise. The use of the terms "a" or "an" when used in conjunction with the term "comprising" in the claims and/or specification may mean "one" or "one" but it is also consistent with the meaning of "one or more", "at least one" and "one or more (one or more than one)".

As used herein, the term "about" is used herein to mean about, roughly, approximately, or within that range. When the term "about" is used in connection with a numerical range, it modifies that range by extending the boundaries above and below the values set forth. Generally, the term "about (about)" is used herein to modify values above and below the stated values to a 20% change either upward or downward (higher or lower).

As used herein, the term "subject" refers to a vertebrate. In one embodiment, the subject is a mammal or a mammalian species. In one embodiment, the subject is a human. In one embodiment, the subject is a healthy adult. In other embodiments, the subject is a non-human vertebrate, including but not limited to a non-human primate, laboratory animal, livestock, racing, domestic animal, and non-domestic animal. In one embodiment, the term "human subjects" refers to a healthy adult population.

As used herein, the term "patient" refers to a human or animal.

The term "mammal" includes, but is not limited to, a human, mouse, rat, guinea pig, dog, cat, horse, cow, pig or non-human primate, such as a monkey, chimpanzee, baboon or rhesus. In one embodiment, the mammal is a human.

As used herein, the term "therapeutically effective (therapeutically effective)" includes preventing and treating a subject suspected of having a viral infection.

In some embodiments, the variable domain of the first heavy chain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 13, or at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 27. In some embodiments, the variable domain of the first light chain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 14, or a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 28. In some embodiments, the variable domain of the second heavy chain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 27, or at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 13. In some embodiments, the variable domain of the second light chain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 28, or a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 14.

In some embodiments, the second light chain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID NO 59, a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID NO 62 at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID NO. In some embodiments, the molecule comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 63, a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 64, a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 97%, 98%, 99% identical to SEQ ID No. 66, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID NO. In some embodiments, the molecule comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 13, a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 14, a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 98%, 99% identical to SEQ ID No. 28, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID NO.

In some embodiments, the first polypeptide chain and the second polypeptide chain are covalently bonded to each other. In some embodiments, the first heavy chain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID NO 59, or at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID NO 63. In some embodiments, the first light chain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 60, or at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 64. In some embodiments, the first heavy chain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 61, or at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 65. In some embodiments, the second light chain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 62, or at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 66.

In some embodiments, the molecule comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID NO 59, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID NO, a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 98%, 99% identical to SEQ ID NO 62, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID NO. In some embodiments, the molecule comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 63, a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 64, a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 97%, 98%, 99% identical to SEQ ID No. 66, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID NO. In some embodiments, the molecule comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 13, a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 14, a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 98%, 99% identical to SEQ ID No. 28, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID NO.

In some embodiments, the variable domain of the first heavy chain comprises CDR1, CDR2 and CDR3 of SEQ ID NO:59 or CDR1, CDR2 and CDR3 of SEQ ID NO:63. In some embodiments, the variable domain of the first light chain comprises CDR1, CDR2 and CDR3 of SEQ ID NO:60 or CDR1, CDR2 and CDR3 of SEQ ID NO:64. In some embodiments, the variable domain of the second heavy chain comprises CDR1, CDR2 and CDR3 of SEQ ID NO. 61 or CDR1, CDR2 and CDR3 of SEQ ID NO. 65. In some embodiments, the variable domain of the second light chain comprises CDR1, CDR2 and CDR3 of SEQ ID NO:62 or CDR1, CDR2 and CDR3 of SEQ ID NO:66.

In some embodiments, the molecule comprises SEQ ID NO 59, SEQ ID NO 60, SEQ ID NO 61, and SEQ ID NO 62. In some embodiments, the molecule comprises SEQ ID NO 63, SEQ ID NO 64, SEQ ID NO 65, and SEQ ID NO 66. In some embodiments, the molecule comprises SEQ ID NO. 13, SEQ ID NO. 14, SEQ ID NO. 27 and SEQ ID NO. 28.

In some embodiments, the antibody comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 15, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 16. In some embodiments, the antibody comprises SEQ ID NO. 15 and SEQ ID NO. 16.

In some embodiments, the heavy chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 19. At least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID NO. 21, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID NO. 29, or at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID NO. 31.

In some embodiments, the heavy chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 3, and the light chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 4. In some embodiments, the heavy chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 7, and the light chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 8, in some embodiments, the heavy chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 10, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to the light chain variable domain. In some embodiments, the heavy chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 11, and the light chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 12. In some embodiments, the heavy chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 17, and the light chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 18. In some embodiments, the heavy chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 19, and the light chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 20. In some embodiments, the heavy chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 21, and the light chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 22. In some embodiments, the heavy chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 25, and the light chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 26. In some embodiments, the heavy chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 29, and the light chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 30.

In some embodiments, the heavy chain variable domain comprises SEQ ID NO. 3 and the light chain variable domain comprises SEQ ID NO. 4. In some embodiments, the heavy chain variable domain comprises SEQ ID NO. 7 and the light chain variable domain comprises SEQ ID NO. 8. In some embodiments, the heavy chain variable domain comprises SEQ ID NO. 9 and the light chain variable domain comprises SEQ ID NO. 10. In some embodiments, the heavy chain variable domain comprises SEQ ID NO. 11 and the light chain variable domain comprises SEQ ID NO. 12. In some embodiments, the heavy chain variable domain comprises SEQ ID NO. 17 and the light chain variable domain comprises SEQ ID NO. 18. In some embodiments, the heavy chain variable domain comprises SEQ ID NO. 19 and the light chain variable domain comprises SEQ ID NO. 20. In some embodiments, the heavy chain variable domain comprises SEQ ID NO. 21 and the light chain variable domain comprises SEQ ID NO. 22. In some embodiments, the heavy chain variable domain comprises SEQ ID NO. 25 and the light chain variable domain comprises SEQ ID NO. 26. In some embodiments, the heavy chain variable domain comprises SEQ ID NO. 29 and the light chain variable domain comprises SEQ ID NO. 30. In some embodiments, the heavy chain variable domain comprises SEQ ID NO. 31 and the light chain variable domain comprises SEQ ID NO. 32. In some embodiments, the heavy chain variable domain comprises SEQ ID NO. 33 and the light chain variable domain comprises SEQ ID NO. 34. In some embodiments, the heavy chain variable domain comprises SEQ ID NO. 37 and the light chain variable domain comprises SEQ ID NO. 38. In some embodiments, the heavy chain variable domain comprises SEQ ID NO 39 and the light chain variable domain comprises SEQ ID NO 40. In some embodiments, the heavy chain variable domain comprises SEQ ID NO. 43 and the light chain variable domain comprises SEQ ID NO. 44. In some embodiments, the heavy chain variable domain comprises SEQ ID NO. 45 and the light chain variable domain comprises SEQ ID NO. 46.

In some embodiments, the molecule comprises SEQ ID NO:51, SEQ ID NO:52, SEQ ID NO:53, and SEQ ID NO:54. In some embodiments, the molecule comprises SEQ ID NO:55, SEQ ID NO:56, SEQ ID NO:57, and SEQ ID NO:58. In some embodiments, the molecule comprises SEQ ID NO:67, SEQ ID NO:68, SEQ ID NO:69, and SEQ ID NO:70. In some embodiments, the molecule comprises SEQ ID NO:71, SEQ ID NO:72, SEQ ID NO:73, and SEQ ID NO:74. In some embodiments, the molecule comprises SEQ ID NO:75, SEQ ID NO:76, SEQ ID NO:77, and SEQ ID NO:78. In some embodiments, the molecule comprises SEQ ID NO:79, SEQ ID NO:80, SEQ ID NO:81, and SEQ ID NO:82. In some embodiments, the molecule comprises SEQ ID NO:83, SEQ ID NO:84, SEQ ID NO:85, and SEQ ID NO:86. In some embodiments, the molecule comprises SEQ ID NO 87, SEQ ID NO 88, SEQ ID NO 89, and SEQ ID NO 90. In some embodiments, the molecule comprises SEQ ID NO. 91, SEQ ID NO. 92, SEQ ID NO. 93 and SEQ ID NO. 94. In some embodiments, the molecule comprises SEQ ID NO 95, SEQ ID NO 96, SEQ ID NO 97, and SEQ ID NO 98. In some embodiments, the molecule comprises SEQ ID NO 99, SEQ ID NO 100, SEQ ID NO 101 and SEQ ID NO 102. In some embodiments, the molecule comprises SEQ ID NO:103, SEQ ID NO:104, SEQ ID NO:105, and SEQ ID NO:106. In some embodiments, the molecule comprises SEQ ID NO:107, SEQ ID NO:108, SEQ ID NO:109, and SEQ ID NO:110. In some embodiments, the molecule comprises SEQ ID NO:115, SEQ ID NO:116, SEQ ID NO:117, and SEQ ID NO:118. In some embodiments, the molecule comprises SEQ ID NO:119, SEQ ID NO:120, SEQ ID NO:121, and SEQ ID NO:122.

In some embodiments, the molecule comprises SEQ ID NO:123, SEQ ID NO:124, SEQ ID NO:125, and SEQ ID NO:126. In some embodiments, the molecule comprises SEQ ID NO:127, SEQ ID NO:128, SEQ ID NO:129, and SEQ ID NO:130. In some embodiments, the molecule comprises SEQ ID NO:131, SEQ ID NO:132, SEQ ID NO:133, and SEQ ID NO:133. In some embodiments, the molecule comprises SEQ ID NO. 139, SEQ ID NO. 140, SEQ ID NO. 141 and SEQ ID NO. 142. In some embodiments, the molecule comprises SEQ ID NO:143, SEQ ID NO:144, SEQ ID NO:145, and SEQ ID NO:146. In some embodiments, the molecule comprises SEQ ID NO:147, SEQ ID NO:148, SEQ ID NO:149, and SEQ ID NO:150. In some embodiments, the molecule comprises SEQ ID NO:151, SEQ ID NO:152, SEQ ID NO:153, and SEQ ID NO:154. In some embodiments, the molecule comprises SEQ ID NO:155, SEQ ID NO:156, SEQ ID NO:157, and SEQ ID NO:158. In some embodiments, the molecule comprises SEQ ID NO 159, SEQ ID NO 160, SEQ ID NO 161 and SEQ ID NO 162. In some embodiments, the molecule comprises SEQ ID NO. 163, SEQ ID NO. 164, SEQ ID NO. 165 and SEQ ID NO. 166. In some embodiments, the molecule comprises SEQ ID NO 167, SEQ ID NO 168, SEQ ID NO 169, and SEQ ID NO 170. In some embodiments, the molecule comprises SEQ ID NO:171, SEQ ID NO:172, SEQ ID NO:173, and SEQ ID NO:174. In some embodiments, the molecule comprises SEQ ID NO:175, SEQ ID NO:176, SEQ ID NO:177, and SEQ ID NO:178. In some embodiments, the molecule comprises SEQ ID NO:179, SEQ ID NO:180, SEQ ID NO:181 and SEQ ID NO:182. In some embodiments, the molecule comprises SEQ ID NO. 183, SEQ ID NO. 184, SEQ ID NO. 185 and SEQ ID NO. 186.

In some embodiments, the molecule comprises SEQ ID NO:119, SEQ ID NO:120, SEQ ID NO:121, and SEQ ID NO:122. In some embodiments, the molecule comprises SEQ ID NO:123, SEQ ID NO:124, SEQ ID NO:125, and SEQ ID NO:126. In some embodiments, the molecule comprises SEQ ID NO:127, SEQ ID NO:128, SEQ ID NO:129, and SEQ ID NO:130. In some embodiments, the molecule comprises SEQ ID NO:131, SEQ ID NO:132, SEQ ID NO:133, and SEQ ID NO:133. In some embodiments, the molecule comprises SEQ ID NO. 139, SEQ ID NO. 140, SEQ ID NO. 141 and SEQ ID NO. 142. In some embodiments, the molecule comprises SEQ ID NO:143, SEQ ID NO:144, SEQ ID NO:145, and SEQ ID NO:146. In some embodiments, the molecule comprises SEQ ID NO:147, SEQ ID NO:148, SEQ ID NO:149, and SEQ ID NO:150. In some embodiments, the molecule comprises SEQ ID NO:151, SEQ ID NO:152, SEQ ID NO:153, and SEQ ID NO:154. In some embodiments, the molecule comprises SEQ ID NO:155, SEQ ID NO:156, SEQ ID NO:157, and SEQ ID NO:158. In some embodiments, the molecule comprises SEQ ID NO 159, SEQ ID NO 160, SEQ ID NO 161 and SEQ ID NO 162. In some embodiments, the molecule comprises SEQ ID NO. 163, SEQ ID NO. 164, SEQ ID NO. 165 and SEQ ID NO. 166. In some embodiments, the molecule comprises SEQ ID NO 167, SEQ ID NO 168, SEQ ID NO 169, and SEQ ID NO 170. In some embodiments, the molecule comprises SEQ ID NO:171, SEQ ID NO:172, SEQ ID NO:173, and SEQ ID NO:174. In some embodiments, the molecule comprises SEQ ID NO:175, SEQ ID NO:176, SEQ ID NO:177, and SEQ ID NO:178. In some embodiments, the molecule comprises SEQ ID NO:179, SEQ ID NO:180, SEQ ID NO:181 and SEQ ID NO:182. In some embodiments, the molecule comprises SEQ ID NO. 183, SEQ ID NO. 184, SEQ ID NO. 185 and SEQ ID NO. 186.

In some embodiments, the heavy chain variable domain comprises SEQ ID NO. 5 and the light chain variable domain comprises SEQ ID NO. 6. In some embodiments, the heavy chain variable domain comprises SEQ ID NO. 41 and the light chain variable domain comprises SEQ ID NO. 42. In some embodiments, the heavy chain variable domain comprises SEQ ID NO:35 and the light chain variable domain comprises SEQ ID NO:36. In some embodiments, the heavy chain variable domain comprises SEQ ID NO. 1 and the light chain variable domain comprises SEQ ID NO. 2. In some embodiments, the heavy chain variable domain comprises SEQ ID NO. 23 and the light chain variable domain comprises SEQ ID NO. 24. In some embodiments, the heavy chain variable domain comprises SEQ ID NO. 13 and the light chain variable domain comprises SEQ ID NO. 14. In some embodiments, the heavy chain variable domain comprises SEQ ID NO. 27 and the light chain variable domain comprises SEQ ID NO. 28. In some embodiments, the heavy chain variable domain comprises the CDR1, CDR2 and CDR3 regions of SEQ ID NO. 5. In some embodiments, the heavy chain variable domain comprises the CDR1, CDR2 and CDR3 regions of SEQ ID NO 41. In some embodiments, the heavy chain variable domain comprises the CDR1, CDR2 and CDR3 regions of SEQ ID NO. 1.

In some embodiments, the variable domain of the first heavy chain comprises CDR1, CDR2 and CDR3 of SEQ ID NO 59; or CDR1, CDR2 and CDR3 of SEQ ID NO. 63. In some embodiments, the variable domain of the first light chain comprises CDR1, CDR2 and CDR3 of SEQ ID NO. 60; or CDR1, CDR2 and CDR3 of SEQ ID NO. 64. In some embodiments, the variable domain of the second heavy chain comprises CDR1, CDR2 and CDR3 of SEQ ID NO 61; or CDR1, CDR2 and CDR3 of SEQ ID NO. 65. In some embodiments, the variable domain of the second light chain comprises CDR1, CDR2 and CDR3 of SEQ ID NO. 62; or CDR1, CDR2 and CDR3 of SEQ ID NO 66.

In some embodiments, the first polypeptide chain and the second polypeptide chain are covalently bonded to each other. In some embodiments, the first heavy chain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID NO 59, or at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID NO 63. In some embodiments, the first light chain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 60, or at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 64. In some embodiments, the second heavy chain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 61, or at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 65. In some embodiments, the second light chain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 62, or at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 66.

In some embodiments, the first heavy chain comprises SEQ ID NO 59 or SEQ ID NO 63. In some embodiments, the first light chain comprises SEQ ID NO. 60 or SEQ ID NO. 64. In some embodiments, the second heavy chain comprises SEQ ID NO. 61 or SEQ ID NO. 65. In some embodiments, the second light chain comprises SEQ ID NO. 62 or SEQ ID NO. 66. In some embodiments, the variable domain of the first heavy chain comprises CDR1, CDR2 and CDR3 of SEQ ID NO 59; or CDR1, CDR2 and CDR3 of SEQ ID NO. 63. In some embodiments, the variable domain of the first light chain comprises CDR1, CDR2 and CDR3 of SEQ ID NO. 60; or CDR1, CDR2 and CDR3 of SEQ ID NO. 64. In some embodiments, the variable domain of the second heavy chain comprises CDR1, CDR2 and CDR3 of SEQ ID NO 61; or CDR1, CDR2 and CDR3 of SEQ ID NO. 65. In some embodiments, the variable domain of the second light chain comprises CDR1, CDR2 and CDR3 of SEQ ID NO. 62; or CDR1, CDR2 and CDR3 of SEQ ID NO 66.

In some embodiments, the heavy chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 15. In some embodiments, the light chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 16. In some embodiments, the heavy chain variable domain comprises SEQ ID NO. 15. In some embodiments, and the light chain variable domain comprises SEQ ID NO. 16.

In some embodiments, the antibody comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 15, and at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 16. In some embodiments, the antibody comprises SEQ ID NO. 15 and SEQ ID NO. 16.

Antibody structure

Antibodies are glycoproteins having immunoglobulin (Ig) monomer domains. The secreted antibody may have multiple Ig units. Ig monomers are "Y" shaped molecules with four polypeptide chains: two identical heavy chains and two identical light chains. These chains may be linked by disulfide bonds.

Mammalian Ig heavy chains are of five types, called: alpha, delta, epsilon, gamma and mu define the class of antibodies. Different heavy chains vary in size and composition. Each heavy chain comprises two regions, a constant region and a variable region. The constant regions are identical in all antibodies of the same isotype, but different in antibodies of different isotypes. The variable region of the heavy chain varies among antibodies produced by different B cells, but is the same among all antibodies produced by a single B cell or B cell clone. The variable region of each heavy chain is approximately 110 amino acids long.

There are two light chains in mammals-lambda (lambda) and kappa (kappa). The light chain comprises two domains, a constant domain and a variable domain. The approximate length of the light chain is 211 to 217 amino acids. Each antibody comprises two identical light chains.

The two arms of the Y-type antibody molecule include antibody binding sites that can bind to both antigens. The two antigens may be the same or different, as in bispecific antibodies. This region of the antibody is called the Fab (fragment, antigen binding) region, which confers antibody specificity. Thus, fab includes one constant domain and one variable domain from each heavy and each light chain of an antibody. The variable domain, also known as the Fv region, is the most important region for recognizing and binding to an antigen (e.g., a viral protein) or a receptor on the surface of a host cell. Antibodies have 3 variable loops on the light chain (VL) and 3 variable loops on the heavy chain (VH), which are responsible for binding to antigen. These loops are known as Complementarity Determining Regions (CDRs).

The base of the Y-shaped molecule is called the Fc (fragment, crystallizable) region, which is important for modulating immune cell activity. Fc comprises two heavy chains that can specifically bind to ensure that each antibody produces an appropriate immune response to a particular antigen. It can also mediate different physiological effects including opsonization, cytolysis and degranulation of mast cells, basophils and eosinophils. Furthermore, the two N-terminal fragments of an antibody are referred to as Fab regions and the C-terminal fragment is referred to as Fc region.

In one embodiment, the invention described herein relates to monoclonal and bispecific antibodies, variants, fusion proteins comprising an antibody portion having an antigen recognition site of a desired specificity. In some embodiments, the antigen binding fragment of the antibody is part of an antibody having at least one antigen recognition site. Fragments include, for example, but are not limited to, fab ', F (ab') 2 Fv) and single chain (scFv).

Antibodies may be recombinantly produced by any means known in the art. In one embodiment, such antibodies are sequenced and the polynucleotide sequence is then cloned into a vector for expression or propagation. In one embodiment, the antibody is synthetic. The sequences encoding the antibodies of interest may be maintained in vectors in host cells, which may then be expanded and frozen for future use. The polynucleotide sequences of such antibodies can be used in genetic manipulation to generate bispecific molecules described herein.

Antibody therapy

Antibody therapy uses monoclonal or bispecific antibodies to bind to predetermined antigens. Such therapies may stimulate the immune system of the patient to attack those antigens, such as viruses. In the clinical setting, the most common route of administration of therapeutic antibodies is Intravenous (IV) infusion. Subcutaneous administration may be followed, but intramuscular injection may also be performed. Oral routes of administration of antibodies are also under development. The monoclonal antibodies and bispecific antibodies described herein can be administered to a subject by any useful method known in the art. Monoclonal and bispecific antibodies described herein can be administered daily. Monoclonal antibodies and bispecific antibodies described herein can be administered twice, three times, four times, five times, or several times a day. The monoclonal antibodies and bispecific antibodies described herein can be administered for 1, 2, 3, 4, 5, 6 days, or 1 or 2 weeks. In some embodiments, the monoclonal antibodies and bispecific antibodies described herein can be administered for a period of more than two weeks. The monoclonal antibodies and bispecific antibodies described herein can be administered in combination with any other therapeutically effective agent or combination of agents.

Covd-19 therapy-mAb

The only therapy currently approved for the treatment of patients with covd-19 is adefovir (remdesired). Adefovir shows some benefit in shortening recovery time of severe covd-19. Currently, efforts are underway in this field to identify biological agents or small molecules that have a more efficient and widely affecting the transmission of the SARS-CoV-2 virus. FIG. 1A shows a schematic of SARS-CoV-2 virus particles. FIG. 1B shows a spike protein trimer that is located on the surface of a virus and binds to a cell receptor on a target cell. RBD is a key component of the viral spike glycoprotein found on coronaviruses, including SARS-CoV-2, and plays a major role in viral attachment, fusion and entry into host cells. The Receptor Binding Domain (RBD) is shown in green on top. The spike protein also has an N-terminal domain (NTD). RBD and NTD are targeted by antibodies against SARS-CoV-2 and variants. The spike protein trimer may be used to isolate B cells from convalescent patient samples. Fig. 1C shows an electron micrograph of coronavirus particles. Arrows indicate viral spike proteins on the viral membrane.

In some embodiments, the invention described herein relates to the isolation, characterization and provision of sequences of potent monoclonal antibodies (mAbs) against SARS-CoV-2 virus (resulting in COVID-19). In some embodiments, the invention described herein describes the optimal COVID-19 patient from which to isolate an effective mAb against SARS-CoV-2. The mAb may be isolated from a blood sample of a patient diagnosed with COVID-19 or a patient presenting with symptoms associated with COVID-19. Once the antibody sequences are determined from these samples, mabs can be synthesized in vitro. In some embodiments, the mAb sequences may be modified and optimized prior to synthesis. These mabs can be used in subsequent characterization experiments. In some embodiments, the invention described herein relates to identifying a mAb that is more potent than any other SARS-CoV-2mAb heretofore known in the art. In some embodiments, the mAb reduces IC50 and/or IC90 values.

The SARS-CoV-2 neutralizing mAbs described herein can be used to treat a subject infected with SARS-CoV-2. In some embodiments, the mabs described herein reduce SARS-CoV-2 viral load in a subject in need thereof. In some embodiments, the mabs described herein reduce the severity of disease in a subject in need thereof. In some embodiments, the mabs described herein improve the clinical outcome of covd-19 in a subject in need thereof.

In some embodiments, the mabs described herein can be used as a prophylaxis to prevent infection of SARS-CoV-2 in high risk subjects and individuals. In some embodiments, the high risk subject may be a healthcare worker in intimate contact with the covd-19 patient or a family member of such healthcare worker. In some embodiments, the mabs described herein can be used as prophylaxis in the general population.

Identification procedure for effective COVID-19mAb

In one embodiment, the invention disclosed herein relates to the identification, isolation and characterization of an effective mAb against SARS-CoV-2, a variant thereof, or any coronavirus from a patient. One embodiment of the methods described herein for isolating such antibodies is described below. In one embodiment, the invention disclosed herein relates to the administration of an antibody described herein to a subject in need thereof to treat or prevent a disease. In some embodiments, the treatment comprises administering a type of monoclonal antibody. In some embodiments, the treatment comprises administering a mixture of two or more monoclonal antibodies.

Plasma samples of the covd-19 patient may be isolated and evaluated for the following capabilities of potential antibodies contained in these plasma samples: a) Viral envelope binding to SARS-CoV-2 or variants (which can be analyzed by ELISA); b) In vitro neutralization of SARS-CoV-2 or variants. Plasma samples from patients with severe disease or symptoms of covd-19 have greater antiviral activity than plasma samples from patients with non-severe covd-19. The plasma sample determined to have the strongest activity can be used to select patients for further analysis of monoclonal antibodies.

Peripheral blood mononuclear cells (PMBC) can be isolated from patients with COVID-19, whose plasma samples exhibit the strongest activity in order to isolate individual B cells. These B cells contain mAb sequences that can lead to the strong plasma antiviral activity identified above. Antibody sequences may be recovered from these single B cells by any high throughput sequencing method known in the art. Genes encoding these mAb sequences from a single B cell can be synthesized and cloned into expression vectors. The encoded monoclonal antibody can then be expressed in vitro and purified for subsequent analysis and characterization.

Monoclonal antibodies produced and purified in vitro can then be tested for the following capacity: a) Viral envelope binding to SARS-CoV-2 or variants (which can be analyzed by ELISA); and B) neutralizing SARS-CoV-2 or the variant, thereby preventing or preventing infection by coronavirus. Neutralization assays can be performed by pseudovirus detection and/or live virus neutralization detection. Those mabs that perform best may then be subjected to additional characterization studies including, but not limited to, ELISA competition assays, surface Plasmon Resonance (SPR) binding affinity assays, and size exclusion chromatography. Antibody performance can be indicated by potency, IC50 concentration, and IC90 concentration. IC50 is a quantitative measure used to indicate the concentration of antibody required to inhibit a virus or viral population by 50%. IC90 is a quantitative measure used to indicate the concentration of antibody required to inhibit a virus or viral population by 90%. These are measures of antibody efficacy.

In one embodiment of the invention described herein, over 1 million (100 million) cells have been screened from 8 patients by the method described, 276 mAb sequences have been synthesized, and 66 mabs have been tested and characterized. In one embodiment, 6 mabs have been the subject of good antiviral activity against SARS-CoV-2. In one embodiment, a mAb appears that has excellent antiviral activity and potency against SARS-CoV-2. In one embodiment, the invention described herein relates to monoclonal antibodies that are significantly more potent than any other SARS-CoV-2mAb heretofore known in the art. In one embodiment, the invention described herein relates to monoclonal antibodies having IC50 and IC90 concentration values significantly lower than any other mAb against SARS-CoV-2 or variants heretofore known in the art.

In one embodiment, if additional plasma samples from a patient with covd-19 are screened for their ability to neutralize SARS-CoV-2, additional monoclonal antibodies are identified from the samples in the screening tube.

The mAbs produced by the methods described herein are more potent than other antibodies known in the art against SARS-CoV-2 or variants. In some embodiments, the invention described herein relates to a mAb against SARS-CoV-2 that has high antiviral activity in a subject. In some embodiments, the high antiviral activity is greater than the activity of SARS-CoV-2 known in the art. In some embodiments, the subject is a human. In some embodiments, the subject is a human patient.

In some embodiments, a more effective antibody requires administration of a smaller amount (lower concentration) to a subject in need thereof to achieve a desired covd-19 related clinical effect. This can be achieved by administering a lower dose to the subject. Thus, fewer potential side effects will be observed with a less frequent dosing regimen or a smaller amount of antibody. This may reduce the cost of antibody production. In some embodiments, the mAb against SARS-CoV-2 results in faster recovery of subjects with COVID-19 and greater overall efficacy.

In some embodiments, the mAbs described herein target multiple epitopes on SARS-CoV-2 virus or variants. In some embodiments, the invention disclosed herein relates to an antibody combination mixture to be administered to a subject. In some embodiments, the antibodies in the mixture target multiple epitopes on SARS-CoV-2 virus or variants. In some embodiments, the antibody combination mixture provides even higher efficacy and lower likelihood of viral resistance than each individual antibody in the mixture.

In some embodiments, the mAbs described herein can be used to treat and/or prevent a COVID-19 infection. In some embodiments, the mabs described herein can be administered to a subject as a prophylactic or prophylactic treatment. In some embodiments, the mAb may be used to diagnose exposure of a COVID-19 subject. In some embodiments, the mabs described herein can be used in laboratory research and development activities.

mAb sequences described herein against SARS-CoV-2 or variants

Underlined and italicized amino acids represent the respective Complementarity Determining Regions (CDRs). Each sequence has three CDRs, which occur in sequence as CDR1, CDR2 and CDR 3.

In some embodiments, monoclonal antibodies 2-4 comprise SEQ ID NO. 1 and SEQ ID NO. 2.

SEQ ID NO. 1 is the heavy chain variable region of antibody No. 2-4.

SEQ ID NO. 2 is the light chain variable region of antibody No. 2-4.

In some embodiments, monoclonal antibodies 4-8 comprise SEQ ID NO. 3 and SEQ ID NO. 4.

SEQ ID NO. 3 is the heavy chain variable region of antibody No. 4-8.

SEQ ID NO. 4 is the light chain variable region of antibody No. 4-8

In some embodiments, monoclonal antibodies 2-15 comprise SEQ ID NO. 5 and SEQ ID NO. 6.

SEQ ID NO. 5 is the heavy chain variable region of antibody Nos. 2-15.

SEQ ID NO. 6 is the light chain variable region of antibody Nos. 2-15.

In some embodiments, monoclonal antibodies 2-38 comprise SEQ ID NO. 7 and SEQ ID NO. 8.

SEQ ID NO. 7 is the heavy chain variable region of antibody Nos. 2-38.

SEQ ID NO. 8 is the light chain variable region of antibody Nos. 2-38.

In some embodiments, monoclonal antibodies 2-51 comprise SEQ ID NO 9 and SEQ ID NO 10.

SEQ ID NO. 9 is the heavy chain variable region of antibody No. 2-51.

SEQ ID NO. 10 is the light chain variable region of antibody Nos. 2-51.

In some embodiments, monoclonal antibodies 4-32 comprise SEQ ID NO. 11 and SEQ ID NO. 12.

SEQ ID NO. 11 is the heavy chain variable region of antibody No. 4-32.

SEQ ID NO. 12 is the light chain variable region of antibody No. 4-32.

In some embodiments, monoclonal antibodies 2-7 comprise SEQ ID NO. 13 and SEQ ID NO. 14.

SEQ ID NO. 13 is the heavy chain variable region of antibody Nos. 2-7, which specifically recognizes the Receptor Binding Domain (RBD).

SEQ ID NO. 14 is the light chain variable region of antibody Nos. 2-7, which specifically recognizes RBD.

In some embodiments, monoclonal antibodies 1-57 comprise SEQ ID NO. 15 and SEQ ID NO. 16.

SEQ ID NO. 15 is the heavy chain variable region of antibody Nos. 1-57, which specifically recognizes RBD.

SEQ ID NO. 16 is the light chain variable region of antibody Nos. 1-57, which specifically recognizes RBD.

In some embodiments, monoclonal antibodies 1-20 comprise SEQ ID NO. 17 and SEQ ID NO. 18.

SEQ ID NO. 17 is the heavy chain variable region of antibody Nos. 1-20, which specifically recognizes RBD.

SEQ ID NO. 18 is the light chain variable region of antibody Nos. 1-20, which specifically recognizes RBD.

In some embodiments, monoclonal antibodies 2-30 comprise SEQ ID NO. 19 and SEQ ID NO. 20.

SEQ ID NO. 19 is the heavy chain variable region of antibody Nos. 2-30, which specifically recognizes RBD.

SEQ ID NO. 20 is the light chain variable region of antibody Nos. 2-30, which specifically recognizes RBD.

In some embodiments, monoclonal antibodies 4-20 comprise SEQ ID NO. 21 and SEQ ID NO. 22.

SEQ ID NO. 21 is the heavy chain variable region of antibody Nos. 4-20, which specifically recognizes RBD.

SEQ ID NO. 22 is the light chain variable region of antibody Nos. 4-20, which specifically recognizes RBD.

In some embodiments, monoclonal antibodies 4-18 comprise SEQ ID NO. 23 and SEQ ID NO. 24.

SEQ ID NO. 23 is the heavy chain variable region of antibody Nos. 4-18, which specifically recognizes the N-terminal domain (NTD).

SEQ ID NO. 24 is the light chain variable region of antibody Nos. 4-18, which specifically recognizes NTD.

In some embodiments, monoclonal antibodies 5-24 comprise SEQ ID NO. 25 and SEQ ID NO. 26.

SEQ ID NO. 25 is the heavy chain variable region of antibody Nos. 5-24, which specifically recognizes NTD.

SEQ ID NO. 26 is the light chain variable region of antibody Nos. 5-24, which specifically recognizes NTD.

In some embodiments, monoclonal antibodies 5-7 comprise SEQ ID NO 27 and SEQ ID NO 28.

SEQ ID NO. 27 is the heavy chain variable region of antibody No. 5-7, which specifically recognizes NTD.

SEQ ID NO. 28 is the light chain variable region of antibody No. 5-7, which specifically recognizes NTD.

In some embodiments, monoclonal antibodies 1-68 comprise SEQ ID NO. 29 and SEQ ID NO. 30.

SEQ ID NO. 29 is the heavy chain variable region of antibody Nos. 1-68, which specifically recognizes NTD.

SEQ ID NO. 30 is the light chain variable region of antibody Nos. 1-68, which specifically recognizes NTD.

In some embodiments, monoclonal antibodies 2-43 comprise SEQ ID NO. 31 and SEQ ID NO. 32.

SEQ ID NO. 31 is the heavy chain variable region of antibody Nos. 2-43, which recognizes an epitope other than RBD or NTD.

SEQ ID NO. 32 is the light chain variable region of antibody Nos. 2-43, which recognizes an epitope other than RBD or NTD.

In some embodiments, monoclonal antibodies 4-19 comprise SEQ ID NO. 33 and SEQ ID NO. 34.

SEQ ID NO. 33 is the heavy chain variable region of antibody Nos. 4-19, which recognizes an epitope other than RBD or NTD.

SEQ ID NO. 34 is the light chain variable region of antibody Nos. 4-19, which recognizes an epitope other than RBD or NTD.

In some embodiments, monoclonal antibodies 2-17 comprise SEQ ID NO. 35 and SEQ ID NO. 36.

SEQ ID NO. 35 is the heavy chain variable region of antibody No. 2-17, which recognizes an epitope other than RBD or NTD.

SEQ ID NO. 36 is the light chain variable region of antibody Nos. 2-17, which recognizes an epitope other than RBD or NTD.

RBD binding agent

In some embodiments, monoclonal antibodies 2-36 comprise SEQ ID NO 37 and SEQ ID NO 38.

SEQ ID NO. 37 is the heavy chain variable region of antibody Nos. 2-36, which specifically recognizes RBD.

SEQ ID NO. 38 is the light chain variable region of antibody Nos. 2-36, which specifically recognizes RBD.

In some embodiments, monoclonal antibodies 1-87 comprise SEQ ID NO 39 and SEQ ID NO 40.

SEQ ID NO. 39 is the heavy chain variable region of antibody Nos. 1-87, which specifically recognizes NTD.

SEQ ID NO. 40 is the light chain variable region of antibody Nos. 1-87, which specifically recognizes NTD.

In some embodiments, monoclonal antibodies 2-15 comprise SEQ ID NO. 41 and SEQ ID NO. 42.

SEQ ID NO. 41 is the heavy chain variable region of 2-15mut mAb.

SEQ ID NO. 42 is the light chain variable region of 2-15mut mAb.

In one embodiment, the monoclonal antibody4-8(39/51)Comprising SEQ ID NO. 43 and SEQ ID NO. 44.

SEQ ID NO. 43 is the heavy chain variable region of 4-8 (39/51) mAb.

SEQ ID NO. 44 is the light chain variable region of 4-8 (39/51) mAb.

In one embodiment, monoclonal antibodies 4-8 (39/51/57) comprise SEQ ID NO. 45 and SEQ ID NO. 46.

SEQ ID NO. 45 is the heavy chain variable region of 4-8 (39/51/57) mAb.

SEQ ID NO. 46 is the light chain variable region of 4-8 (39/51/57) mAb.

Nucleic acid sequences of monoclonal antibodies directed against SARS-CoV-2 or variants described herein

In some embodiments, the invention disclosed herein relates to DNA sequences encoding any of the monoclonal antibodies disclosed herein. In some embodiments, the invention disclosed herein relates to mRNA sequences encoding any of the antibodies disclosed herein. As used herein, the terms "nucleic acid" and "nucleotide sequence" refer to DNA and RNA molecules, including base-, sugar-, or backbone-modified DNA or RNA molecules.

In some embodiments, the invention disclosed herein relates to an isolated nucleic acid having a nucleotide sequence encoding any of the antibodies of the disclosure. In some embodiments, the nucleotide sequence encoding any of the antibodies of the present disclosure is codon optimized for efficient expression in a host system. In some embodiments, the nucleotide sequence encoding any of the antibodies of the present disclosure is codon optimized to increase the translational efficiency of the nucleotide sequence. In some embodiments, the nucleotide sequence encoding any of the antibodies of the present disclosure is codon optimized to accommodate the codon bias of the host system. In some embodiments, the nucleotide sequence encoding any of the antibodies of the present disclosure is codon optimized to remove redundancy and/or evolutionary limitations, including, but not limited to, availability of tRNA isoreceptors (tRNAs), TATA boxes, and the Hien-Darcy sequences (Shine-Dalgarno sequence).

In some embodiments, the invention disclosed herein relates to a codon optimized isolated nucleic acid comprising a nucleic acid sequence encoding a monoclonal antibody comprising SEQ ID NO. 1. In some embodiments, the invention disclosed herein relates to a codon optimized isolated nucleic acid comprising a nucleic acid sequence encoding a monoclonal antibody comprising SEQ ID NO. 2. In some embodiments, the invention disclosed herein relates to a codon optimized isolated nucleic acid comprising a nucleic acid sequence encoding a monoclonal antibody comprising SEQ ID NO. 3. In some embodiments, the invention disclosed herein relates to a codon optimized isolated nucleic acid comprising a nucleic acid sequence encoding a monoclonal antibody comprising SEQ ID NO. 4. In some embodiments, the invention disclosed herein relates to a codon optimized isolated nucleic acid comprising a nucleic acid sequence encoding a monoclonal antibody comprising SEQ ID NO. 5. In some embodiments, the invention disclosed herein relates to a codon optimized isolated nucleic acid comprising a nucleic acid sequence encoding a monoclonal antibody comprising SEQ ID NO. 6. In some embodiments, the invention disclosed herein relates to a codon optimized isolated nucleic acid comprising a nucleic acid sequence encoding a monoclonal antibody comprising SEQ ID NO. 7. In some embodiments, the invention disclosed herein relates to a codon optimized isolated nucleic acid comprising a nucleic acid sequence encoding a monoclonal antibody comprising SEQ ID NO. 8. In some embodiments, the invention disclosed herein relates to a codon optimized isolated nucleic acid comprising a nucleic acid sequence encoding a monoclonal antibody comprising SEQ ID NO. 9. In some embodiments, the invention disclosed herein relates to a codon optimized isolated nucleic acid comprising a nucleic acid sequence encoding a monoclonal antibody comprising SEQ ID NO. 10. In some embodiments, the invention disclosed herein relates to a codon optimized isolated nucleic acid comprising a nucleic acid sequence encoding a monoclonal antibody comprising SEQ ID NO. 11. In some embodiments, the invention disclosed herein relates to a codon optimized isolated nucleic acid comprising a nucleic acid sequence encoding a monoclonal antibody comprising SEQ ID NO. 12. In some embodiments, the invention disclosed herein relates to a codon optimized isolated nucleic acid comprising a nucleic acid sequence encoding a monoclonal antibody comprising SEQ ID NO. 13. In some embodiments, the invention disclosed herein relates to a codon optimized isolated nucleic acid comprising a nucleic acid sequence encoding a monoclonal antibody comprising SEQ ID NO. 14. In some embodiments, the invention disclosed herein relates to a codon optimized isolated nucleic acid comprising a nucleic acid sequence encoding a monoclonal antibody comprising SEQ ID NO. 15. In some embodiments, the invention disclosed herein relates to a codon optimized isolated nucleic acid comprising a nucleic acid sequence encoding a monoclonal antibody comprising SEQ ID NO. 16. In some embodiments, the invention disclosed herein relates to a codon optimized isolated nucleic acid comprising a nucleic acid sequence encoding a monoclonal antibody comprising SEQ ID NO. 17. In some embodiments, the invention disclosed herein relates to a codon optimized isolated nucleic acid comprising a nucleic acid sequence encoding a monoclonal antibody comprising SEQ ID NO. 18. In some embodiments, the invention disclosed herein relates to a codon optimized isolated nucleic acid comprising a nucleic acid sequence encoding a monoclonal antibody comprising SEQ ID NO. 19. In some embodiments, the invention disclosed herein relates to a codon optimized isolated nucleic acid comprising a nucleic acid sequence encoding a monoclonal antibody comprising SEQ ID NO. 20. In some embodiments, the invention disclosed herein relates to a codon optimized isolated nucleic acid comprising a nucleic acid sequence encoding a monoclonal antibody comprising SEQ ID NO. 21. In some embodiments, the invention disclosed herein relates to a codon optimized isolated nucleic acid comprising a nucleic acid sequence encoding a monoclonal antibody comprising SEQ ID NO. 22. In some embodiments, the invention disclosed herein relates to a codon optimized isolated nucleic acid comprising a nucleic acid sequence encoding a monoclonal antibody comprising SEQ ID NO. 23. In some embodiments, the invention disclosed herein relates to a codon optimized isolated nucleic acid comprising a nucleic acid sequence encoding a monoclonal antibody comprising SEQ ID NO. 24. In some embodiments, the invention disclosed herein relates to a codon optimized isolated nucleic acid comprising a nucleic acid sequence encoding a monoclonal antibody comprising SEQ ID NO. 25. In some embodiments, the invention disclosed herein relates to a codon optimized isolated nucleic acid comprising a nucleic acid sequence encoding a monoclonal antibody comprising SEQ ID NO. 26. In some embodiments, the invention disclosed herein relates to a codon optimized isolated nucleic acid comprising a nucleic acid sequence encoding a monoclonal antibody comprising SEQ ID NO. 27. In some embodiments, the invention disclosed herein relates to a codon optimized isolated nucleic acid comprising a nucleic acid sequence encoding a monoclonal antibody comprising SEQ ID NO. 28. In some embodiments, the invention disclosed herein relates to a codon optimized isolated nucleic acid comprising a nucleic acid sequence encoding a monoclonal antibody comprising SEQ ID NO. 29. In some embodiments, the invention disclosed herein relates to a codon optimized isolated nucleic acid comprising a nucleic acid sequence encoding a monoclonal antibody comprising SEQ ID NO. 30. In some embodiments, the invention disclosed herein relates to a codon optimized isolated nucleic acid comprising a nucleic acid sequence encoding a monoclonal antibody comprising SEQ ID NO. 31. In some embodiments, the invention disclosed herein relates to a codon optimized isolated nucleic acid comprising a nucleic acid sequence encoding a monoclonal antibody comprising SEQ ID NO. 32. In some embodiments, the invention disclosed herein relates to a codon optimized isolated nucleic acid comprising a nucleic acid sequence encoding a monoclonal antibody comprising SEQ ID NO. 33. In some embodiments, the invention disclosed herein relates to a codon optimized isolated nucleic acid comprising a nucleic acid sequence encoding a monoclonal antibody comprising SEQ ID NO 34. In some embodiments, the invention disclosed herein relates to a codon optimized isolated nucleic acid comprising a nucleic acid sequence encoding a monoclonal antibody comprising SEQ ID NO. 35. In some embodiments, the invention disclosed herein relates to a codon optimized isolated nucleic acid comprising a nucleic acid sequence encoding a monoclonal antibody comprising SEQ ID NO. 36. In some embodiments, the invention disclosed herein relates to a codon optimized isolated nucleic acid comprising a nucleic acid sequence encoding a monoclonal antibody comprising SEQ ID NO. 37. In some embodiments, the invention disclosed herein relates to a codon optimized isolated nucleic acid comprising a nucleic acid sequence encoding a monoclonal antibody comprising SEQ ID NO. 38. In some embodiments, the invention disclosed herein relates to a codon optimized isolated nucleic acid comprising a nucleic acid sequence encoding a monoclonal antibody comprising SEQ ID NO 39. In some embodiments, the invention disclosed herein relates to a codon optimized isolated nucleic acid comprising a nucleic acid sequence encoding a monoclonal antibody comprising SEQ ID NO. 40. In some embodiments, the invention disclosed herein relates to a codon optimized isolated nucleic acid comprising a nucleic acid sequence encoding a monoclonal antibody comprising SEQ ID NO. 41. In some embodiments, the invention disclosed herein relates to a codon optimized isolated nucleic acid comprising a nucleic acid sequence encoding a monoclonal antibody comprising SEQ ID NO. 42. In some embodiments, the invention disclosed herein relates to a codon optimized isolated nucleic acid comprising a nucleic acid sequence encoding a monoclonal antibody comprising SEQ ID NO. 43. In some embodiments, the invention disclosed herein relates to a codon optimized isolated nucleic acid comprising a nucleic acid sequence encoding a monoclonal antibody comprising SEQ ID NO. 44. In some embodiments, the invention disclosed herein relates to a codon optimized isolated nucleic acid comprising a nucleic acid sequence encoding a monoclonal antibody comprising SEQ ID NO. 45. In some embodiments, the invention disclosed herein relates to a codon optimized isolated nucleic acid comprising a nucleic acid sequence encoding a monoclonal antibody comprising SEQ ID NO. 46.

In some embodiments, the invention disclosed herein relates to a pharmaceutical composition comprising an isolated nucleic acid having a nucleotide sequence encoding any of the antibodies of the present disclosure, and a pharmaceutically acceptable carrier. In some embodiments, the invention disclosed herein relates to a pharmaceutical composition comprising a codon-optimized isolated nucleic acid having a nucleotide sequence encoding any of the antibodies of the disclosure, and a pharmaceutically acceptable carrier.

Covd-19 therapy using bispecific antibodies

Monoclonal antibodies are homogeneous in the nature of their interaction with the ligand/antigen, thus giving absolute monospecificity to their target. However, sometimes this absolute specificity for a single antigen target can lead to certain limitations of the therapeutic agent, for example viruses that are known to evolve their target epitopes and thus escape the targeting of monoclonal antibodies. To overcome these limitations, scientists have generated antibodies that can be engineered to target multiple epitopes using a single antibody-like molecule. These antibody-like molecules can bind two different antigens simultaneously, known as "bispecific" antibodies. Their framework consists of fragments of two monoclonal antibodies, the regions of which have binding affinity for two different antigens. Such a framework may be used to bind non-conventional or rare antigens. In some embodiments, the invention described herein relates to the generation of bispecific antibodies capable of neutralizing SARS-CoV-2 virus and/or variants.

The SARS-CoV-2 bispecific antibodies described herein are useful for treating a subject infected with SARS-CoV-2 or a variant. In some embodiments, the bispecific antibodies described herein reduce viral load in a subject in need thereof. In some embodiments, the bispecific antibodies described herein reduce the severity of a disease in a subject in need thereof. In some embodiments, the bispecific antibodies described herein improve the clinical outcome of covd-19 in a subject in need thereof.

In some embodiments, the bispecific antibodies described herein can be used as a prophylaxis to prevent infection of a high risk subject and individual with SARS-CoV-2 or variant. In some embodiments, the high risk subject may be a healthcare worker in intimate contact with the covd-19 patient or a family member of such healthcare worker. In some embodiments, the bispecific antibodies described herein can be used as a prophylaxis for the general population.

In some embodiments, a more effective antibody requires administration of a smaller amount to the subject to achieve the desired covd-19 associated recovery effect. This can reduce potential side effects, reduce dosing regimen frequency, or reduce antibody production costs by administering lower doses to the subject. In some embodiments, potency is determined by IC50 and IC90 concentration values.

In some embodiments, the bispecific antibodies described herein target multiple epitopes on SARS-CoV-2 virus or variants. In some embodiments, the invention disclosed herein relates to bispecific antibody combination mixtures. In some embodiments, the antibodies in the mixture target multiple epitopes on SARS-CoV-2 virus or variants. In some embodiments, the bispecific antibody combination cocktail provides even higher efficacy and lower likelihood of viral resistance than each individual antibody in the cocktail. In some embodiments, the bispecific antibodies described herein can be used in laboratory research and development activities. In one embodiment, the bispecific antibodies described herein include variable regions of a first antibody that binds an epitope on the coronavirus Receptor Binding Domain (RBD) and a second antibody that binds an epitope on the coronavirus N-terminal domain (NTD).

Pairing anti-RBD and anti-NTD domain antibodies in this bispecific format will primarily provide a genetic barrier to increase the virus to achieve resistance to the antibody. It also allows the clinician to administer only a single formulation of one bispecific antibody to the patient, simplifying treatment. In some embodiments, the bispecific antibody is administered as a mixture of two or more antibodies. With regard to marketing and supply chain benefits, bispecific antibodies are an economical alternative to production in single molecule form, and combinations of these monoclonal antibodies are known to increase the threshold resistance of SARS-CoV-2.

In one embodiment, the invention disclosed herein relates to bispecific antibodies engineered for the treatment and prevention of a covd-19 infection. An engineered bispecific antibody can specifically bind the binding of two antibodies in one molecule. Bispecific antibodies can recognize two antigens on different cells or on the same cell. Thus, by specifically binding to two antigens, an engineered bispecific antibody can bring two cells close to or activate two receptors simultaneously. Furthermore, blocking two target proteins with one antibody instead of two antibodies may be more effective. In some embodiments, the bispecific antibody can target two viral epitopes that provide a higher barrier to viral escape. In some embodiments, bispecific antibodies may have a synergistic effect on potency against a viral target.

In one embodiment, the engineered antibody achieves dual specificity using the CrossMab format (Schaefer, 2011). The CrossMab engineering format allows bispecific antibodies to adopt structures that more closely resemble those of the natural antibodies. This may facilitate binding of bispecific antibodies to weakly expressed antigens while avoiding overstimulation of the immune response. In some embodiments, the bispecific antibody in the CrossMab format can be anchored to the host cell membrane. This allows for increasing local antibody concentrations, targeting continuous and/or interdependent virus entry steps, and compensating for monovalent binding.

The CrossMab format used to engineer bispecific antibodies as described herein can be applied to engineer bispecific antibodies against SARS-CoV-2 virus or variants. As shown in fig. 41, creating a "knob" in one heavy chain and a "hole" in the other heavy chain of a bispecific antibody favors heavy chain heterodimer formation, while the "crossover" of CL and CH1 sequences (constant domain, heavy chain, and light chain) in one arm of the antibody favors correct heavy-light chain pairing in both arms. The CrossMab format allows the correct assembly of two heavy and two light chains from different parent antibodies into one bispecific antibody molecule that is similar in quality and structure to a typical monoclonal antibody and does not require artificial linkers. Each antibody arm may be selected from a different effective neutralizing antibody against SARS-CoV-2 or a variant. Other bispecific forms combining the specific SARS-CoV-2 antibodies described herein can produce similar neutralizing activity. As shown in fig. 41, human IgG isotype 1Fc can be engineered to reduce Fc effector function by L234F, L235E and P331S mutations and to achieve half-life by M428L and N434S mutations.

In one embodiment, the bispecific antibody comprises two polypeptide chains, one from each parent monoclonal antibody. In one embodiment, the first parent polypeptide arm comprises the light chain and heavy chain polypeptides of the first parent antibody. The light chain may comprise a light chain variable domain (VL) comprising a light chain binding region and a light chain constant domain (CL) of the first parent antibody. The heavy chain may comprise a heavy chain variable domain (VH) comprising a heavy chain binding region and a heavy chain constant domain (CH) of the first parent antibody. In one embodiment, the first parent polypeptide arm further comprises a domain that promotes heterodimerization with a second polypeptide. In one embodiment, the heterodimerization domain can covalently bond a first parent polypeptide to a second parent polypeptide of a bispecific antibody.

In one embodiment, the second parent polypeptide arm comprises light chain and heavy chain polypeptides of the second parent antibody. The light chain may comprise a light chain variable domain (VL) comprising a light chain binding region and a light chain constant domain (CL) of the second parent antibody. The heavy chain may comprise a heavy chain variable domain (VH) comprising a heavy chain binding region and a heavy chain constant domain (CH) of the second parent antibody. In one embodiment, the second parent polypeptide arm further comprises a domain that promotes heterodimerization with a second polypeptide. In one embodiment, the heterodimerization domain can covalently bond a first parent polypeptide to a second parent polypeptide of a bispecific antibody. In one embodiment, the bispecific antibody further comprises a linker and disulfide bond linking the two parent polypeptides of the antibody. The linker may be any suitable amino acid sequence. In one embodiment of the present invention, in one embodiment,

Bispecific antibodies described herein include the variable regions of a first and a second antibody, wherein the first antibody binds to an epitope on the coronavirus Receptor Binding Domain (RBD) selected from 8 monoclonal Abs (1-20, 1-57, 2-7, 2-15, 2-30, 2-36, 2-43, and 4-20), and the second antibody binds to an epitope on the coronavirus N-terminal domain (NTD) also selected from 8 monoclonal Abs (1-68, 1-87, 2-17, 2-51, 4-8, 4-18, 4-19, and 5-24).

Cross Mabs described herein utilize the human IgG isotype (isotype) 1Fc backbone. The backbone may have additional mutations engineered through the L234F, L235E and P331S mutations to reduce Fc effector function and through the M428L and N434S mutations to achieve half-life.

To generate bispecific antibodies meeting these objectives, 58 candidate bispecific abs were screened using in vitro assays for their efficacy against the SARS-CoV-2 live infectious virus. Briefly, purified candidate bispecific abs were mixed with infectious live virus for 60 minutes and then added to monolayer Vero E6 cells to determine the degree of neutralization. The antibodies were diluted 5-fold from 10 μg/mL while the virus was incubated with the cells at a multiplicity of infection (MOI) =0.1. Three days after infection, the cytopathic effect of cells induced by live viral replication was measured. The difference in infection levels of control wells without antibody was converted to a percentage and values for inhibition of 50% virus (IC 50) and 90% virus (IC 90) were calculated using nonlinear regression analysis.

Using efficacy values based on the IC achieved by comparison with the parent antibody ₉₀ Fold enhancement of concentration was used to assign the bispecific antibody with advantage. With IC ₅₀ IC is less than or equal to 10ng/mL ₉₀ Bispecific antibodies with efficacy of 50ng/mL or less were selected as the antibodies that were able to achieve the largest barrier to SARS-CoV-2 resistance. Ab X and Ab Y are independent single arm Fab controls. The bispecific antibody candidate selected achieves efficacy similar to that of co-administration of the two parent mabs.

SARS-CoV-2 variant

In some embodiments, the bispecific antibody exhibits neutralizing activity against wild type SARS-CoV-2 virus (WA 1). In some embodiments, the bispecific antibody exhibits neutralizing activity against a SARS-CoV-2 variant. In some embodiments, the bispecific antibodies disclosed herein exhibit potent neutralizing activity against SARS-CoV-2 wild-type and various variant strains. In some embodiments, the SARS-CoV-2 variant is b.1.1.7 (uk variant), b.1.351 (south africa variant), b.1.526 (new york variant), or p.1 (brazil variant).

In one embodiment, each arm of the constructed bispecific antibody is selected from the group consisting of 19 potent neutralizing antibodies against SARS-CoV-2. Other bispecific antibodies 2-7/5-7, 2-7/1-57 and cross mab forms with inverted knuckles and pore arms, i.e., 5-7/2-7 or 1-57/2-7, show high efficacy when tested against wild type virus. Since 1-57 activity can be affected by SARS-CoV-2 variants (e.g., california variants) containing the L452R mutation, which can disrupt mAb 1-57 heavy chain binding as shown by structural modeling, 2-7/5-7 was chosen as a development candidate, further featuring neutralization of SARS-CoV-2 variants, and exhibiting effective neutralization of newly occurring variants, B.1.1.7 (British variants), B.1.351 (south Africa variants), B.1.526 (New York variants), and P.1 (Brazil variants), among others. For many bispecific antibodies tested herein, both arms of the bispecific antibody contribute to the neutralizing activity of the bispecific molecule. These bispecific antibodies with potent neutralizing activity against SARS-CoV-2 and its variants provide new therapies for the treatment and prevention of COVID-19, can provide a higher genetic barrier to viral escape as a single antibody molecule, and have lower costs than antibody combinations. The observed enhancement of activity was expected to extend to other engineered bispecific antibody formats.

Antibody sequences of bispecific antibodies

The 2-17/2-7 bispecific antibody sequences may comprise:

SEQ ID NO. 47 is the amino acid sequence 2-17HC-Hole-Cross defining the 2-17 derived heavy chain of the 2-17/2-7 antibody

SEQ ID NO. 48 is the amino acid sequence 2-17VLCH1 defining the 2-17 derived light chain of the 2-17/2-7 antibody

SEQ ID NO. 49 is the amino acid sequence 2-7HC-Knob defining the 2-7 derived heavy chain of the 2-17/2-7 antibody

SEQ ID NO. 50 is the amino acid sequence 2-7LC of the 2-7 derived light chain of the defined 2-17/2-7 antibody

PTECS

The 1-57/5-7 bispecific antibody sequences may comprise:

SEQ ID NO. 51 is an amino acid sequence 1-57HC-Hole-Cross defining the 1-57 derived heavy chain of the 1-57/5-7 antibody

SEQ ID NO. 52 is the amino acid sequence 1-57VLCH1 defining the 1-57 derived light chain of the 1-57/5-7 antibody

SEQ ID NO. 53 is the amino acid sequence 5-7HC-Knob defining the 5-7 derived heavy chain of the 1-57/5-7 antibody

SEQ ID NO. 54 is the amino acid sequence 5-7LC of the 5-7 derived light chain of the defined 1-57/5-7 antibody

The 5-7/1-57 bispecific antibody sequences may comprise:

SEQ ID NO. 55 is an amino acid sequence 5-7HC-Hole-Cross defining the 5-7 derived heavy chain of the 5-7/1-57 antibody

SEQ ID NO. 56 is the amino acid sequence 5-7VLCH1 defining the 5-7 derived light chain of the 5-7/1-57 antibody

SEQ ID NO. 57 is a 1-57HC-Knob amino acid sequence defining the 1-57 derived heavy chain of the 5-7/1-57 antibody

SEQ ID NO. 58 is the amino acid sequence 1-57LC of the 1-57 derived light chain of the 5-7/1-57 antibody

The 2-7/5-7 bispecific antibody sequences may comprise:

SEQ ID NO. 59 is the amino acid sequence 2-7HC-Hole-Cross defining the 2-7 derived heavy chain of the 2-7/5-7 antibody

SEQ ID NO. 60 is the amino acid sequence 2-7VLCH1 defining the 2-7 derived light chain of the 2-7/5-7 antibody

SEQ ID NO. 61 is the amino acid sequence 5-7HC-Knob defining the 5-7 derived heavy chain of the 2-7/5-7 antibody

SEQ ID NO. 62 is the amino acid sequence 5-7LC of the 5-7 derived light chain of the defined 2-7/5-7 antibody

The 5-7/2-7 bispecific antibody sequences may comprise:

SEQ ID NO. 63 is an amino acid sequence 5-7HC-Hole-Cross defining the 5-7 derived heavy chain of a 5-7/2-7 antibody

SEQ ID NO. 64 is the amino acid sequence 5-7VLCH1 defining the 5-7 derived light chain of the 5-7/2-7 antibody

SEQ ID NO. 65 is the amino acid sequence 2-7HC-Knob of the 2-7 derived heavy chain of the 5-7/2-7 antibody

SEQ ID NO. 66 is the amino acid sequence 2-7LC of the 2-7 derived light chain of the 5-7/2-7 antibody

The 2-17/1-57 bispecific antibody sequences may comprise:

SEQ ID NO. 67 is an amino acid sequence 2-17HC-Hole-Cross defining the 2-17 derived heavy chain of the 2-17/1-57 antibody

SEQ ID NO. 68 is the amino acid sequence 2-17VLCH1 defining the 2-17 derived light chain of the 2-17/1-57 antibody

SEQ ID NO. 69 shows the amino acid sequence 1-57HC-Knob of the 1-57 derived heavy chain of the 2-17/1-57 antibody

SEQ ID NO. 70 is an amino acid sequence defining the 1-57 derived light chain of the 2-17/1-57 antibody

The 2-17/2-15 bispecific antibody sequences may comprise:

SEQ ID NO. 71 is the amino acid sequence 2-17HC-Hole-Cross defining the 2-17 derived heavy chain of the 2-17/2-15 antibody

SEQ ID NO. 72 is the amino acid sequence 2-17VLCH1 defining the 2-17 derived light chain of the 2-17/2-15 antibody

SEQ ID NO. 73 is the amino acid sequence 2-15HC-Knob of the 2-15 derived heavy chain of the defined 2-17/2-15 antibody

SEQ ID NO. 74 is the amino acid sequence 2-15LC of the 2-15 derived light chain of the defined 2-17/2-15 antibody

The 2-17/2-30 bispecific antibody sequences may comprise:

SEQ ID NO. 75 is the amino acid sequence 2-17HC-Hole-Cross defining the 2-17 derived heavy chain of the 2-17/2-30 antibody

SEQ ID NO. 76 is the amino acid sequence 2-17VLCH1 defining the 2-17 derived light chain of the 2-17/2-30 antibody

SEQ ID NO. 77 is the amino acid sequence 2-30HC-Knob defining the 2-30 derived heavy chain of the 2-17/2-30 antibody

SEQ ID NO. 78 is an amino acid sequence 2-30 LC defining the 2-30 derived light chain of the 2-17/2-30 antibody

The 2-17/4-20 bispecific antibody sequences may comprise:

SEQ ID NO. 79 is the amino acid sequence 2-17HC-Hole-Cross defining the 2-17 derived heavy chain of the 2-17/4-20 antibody

SEQ ID NO. 80 is the amino acid sequence 2-17VLCH1 defining the 2-17 derived light chain of the 2-17/4-20 antibody

SEQ ID NO. 81 is the amino acid sequence 4-20HC-Knob defining the 4-20 derived heavy chain of the 2-17/4-20 antibody

SEQ ID NO. 82 is the amino acid sequence 4-20LC of the 4-20 derived light chain of the defined 2-17/4-20 antibody

The 5-24/1-57 bispecific antibody sequences may comprise:

SEQ ID NO. 83 is the amino acid sequence 5-24HC-Hole-Cross defining the 5-24 derived heavy chain of the 5-24/1-57 antibody

SEQ ID NO. 84 is the amino acid sequence 5-24VLCH1 defining the 5-24 derived light chain of the 5-24/1-57 antibody

SEQ ID NO. 85 is the amino acid sequence 1-57HC-Knob defining the 1-57 derived heavy chain of the 5-24/1-57 antibody

DSDGSFFLYSKLTVDKSRWQQGNVFSCSVLHEALHSHYTQKSLSLSPGK

SEQ ID NO. 86 is the amino acid sequence 1-57LC of the 1-57 derived light chain of the defined 5-24/1-57 antibody

The 5-24/2-15 bispecific antibody sequences may comprise:

SEQ ID NO. 87 is an amino acid sequence 5-24HC-Hole-Cross defining the 5-24 derived heavy chain of the 5-24/2-15 antibody

SEQ ID NO. 88 is the amino acid sequence 5-24VLCH1 defining the 5-24 derived light chain of the 5-24/2-15 antibody

SEQ ID NO. 89 is the amino acid sequence 2-15HC-Knob defining the 2-15 derived heavy chain of the 5-24/2-15 antibody

SEQ ID NO. 90 is the amino acid sequence 2-15LC of the 2-15 derived light chain of the 5-24/2-15 antibody

The 5-24/4-20 bispecific antibody sequences may comprise:

SEQ ID NO. 91 is an amino acid sequence 5-24HC-Hole-Cross defining the 5-24 derived heavy chain of the 5-24/4-20 antibody

SEQ ID NO. 92 is the amino acid sequence 5-24VLCH1 defining the 5-24 derived light chain of the 5-24/4-20 antibody

SEQ ID NO. 93 is the amino acid sequence 4-20HC-Knob of the 4-20 derived heavy chain of the 5-24/4-20 antibody

SEQ ID NO. 94 is the amino acid sequence 4-20LC of the 4-20 derived light chain of the 5-24/4-20 antibody

The 1-20/1-68 bispecific antibody sequences may comprise:

SEQ ID NO. 95 is the amino acid sequence 1-20HC-Hole-Cross defining the 1-20 derived heavy chain of the 1-20/1-68 antibody

SEQ ID NO. 96 is the amino acid sequence 1-20VLCH1 of the 1-20 derived light chain of the 1-20/1-68 antibody

SEQ ID NO. 97 is the amino acid sequence 1-68HC-Knob defining the 1-68 derived heavy chain of the 1-20/1-68 antibody

SEQ ID NO. 98 is the amino acid sequence 1-68LC of the 1-68 derived light chain of the 1-20/1-68 antibody

The 1-20/2-17 bispecific antibody sequences may comprise:

SEQ ID NO. 99 is the amino acid sequence 1-20HC-Hole-Cross defining the 1-20 derived heavy chain of the 1-20/2-17 antibody

SEQ ID NO. 100 is the amino acid sequence 1-20VLCH1 of the 1-20 derived light chain of the 1-20/2-17 antibody

SEQ ID NO. 101 is the amino acid sequence 2-17HC-Knob defining the 2-17 derived heavy chain of the 1-20/2-17 antibody

SEQ ID NO. 102 is the amino acid sequence 2-17LC of the 2-17 derived light chain of the 1-20/2-17 antibody

The 1-20/4-18 bispecific antibody sequences may comprise:

SEQ ID NO. 103 is the amino acid sequence 1-20HC-Hole-Cross defining the 1-20 derived heavy chain of the 1-20/4-18 antibody

SEQ ID NO. 104 is the amino acid sequence 1-20VLCH1 of the 1-20 derived light chain of the 1-20/4-18 antibody

SEQ ID NO. 105 is the amino acid sequence 4-18HC-Knob defining the 4-18 derived heavy chain of the 1-20/4-18 antibody

SEQ ID NO. 106 is the amino acid sequence 4-18LC of the 4-18 derived light chain of the 1-20/4-18 antibody

The 1-20/5-24 bispecific antibody sequences may comprise:

SEQ ID NO. 107 is an amino acid sequence 1-20HC-Hole-Cross defining the 1-20 derived heavy chain of the 1-20/5-24 antibody

SEQ ID NO. 108 is the amino acid sequence 1-20VLCH1 of the 1-20 derived light chain of the 1-20/5-24 antibody

SEQ ID NO. 109 is the amino acid sequence 5-24HC-Knob defining the 5-24 derived heavy chain of the 1-20/5-24 antibody

SEQ ID NO. 110 is the amino acid sequence 5-24LC of the 5-24 derived light chain of the defined 1-20/5-24 antibody

Bispecific antibodies with both LALA (TM) and LS mutations:

the 2-7/4-8 (39/51/57) bispecific antibody sequences may comprise:

SEQ ID NO. 111 is the amino acid sequence 2-7HC-Hole-Cross defining the 2-7 derived heavy chain of a 2-7/4-8 (39/51/57) antibody

SEQ ID NO. 112 is the amino acid sequence 2-7VLCH1 defining the 2-7 derived light chain of the 2-7/4-8 (39/51/57) antibody

SEQ ID NO. 113 is the amino acid sequence 4-8 (39/51/57) HC-Knob of the 4-8 (39/51/57) derived heavy chain of the 4-8 (39/51/57) antibody defining 2-7/4-8 (39/51/57)

SEQ ID NO. 114 is the amino acid sequence 4-8LC of the 4-8 (39/51/57) -derived light chain of the 2-7/4-8 (39/51/57) antibody

The 2-17/2-36 bispecific antibody sequences may comprise:

SEQ ID NO. 115 is the amino acid sequence 2-17HC-Hole-Cross defining the 2-17 derived heavy chain of the 2-17/2-36 antibody

SEQ ID NO. 116 is the amino acid sequence 2-17VLCH1 defining the 2-17 derived light chain of the 2-17/2-36 antibody

SEQ ID NO. 117 is the amino acid sequence 2-36HC-Knob of the 2-36 derived heavy chain of the defined 2-17/2-36 antibody

SEQ ID NO. 118 is the amino acid sequence 2-36LC of the 2-36 derived light chain of the defined 2-17/2-36 antibody

The 2-36/1-68 bispecific antibody sequence may comprise:

SEQ ID NO. 119 is an amino acid sequence 2-36HC-Hole-Cross defining the 2-36 derived heavy chain of the 2-36/1-68 antibody

SEQ ID NO. 120 is the amino acid sequence 2-36VLCH1 defining the 2-36 derived light chain of the 2-36/1-68 antibody

SEQ ID NO. 121 is the amino acid sequence 1-68HC-Knob defining the 1-68 derived heavy chain of the 2-36/1-68 antibody

SEQ ID NO. 122 is the amino acid sequence 1-68LC of the 1-68 derived light chain of the defined 2-36/1-68 antibody

The 2-36/4-18 bispecific antibody sequences may comprise:

SEQ ID NO. 123 is an amino acid sequence 2-36HC-Hole-Cross defining the 2-36 derived heavy chain of the 2-36/4-18 antibody

SEQ ID NO. 124 is the amino acid sequence 2-36VLCH1 defining the 2-36 derived light chain of the 2-36/4-18 antibody

SEQ ID NO. 125 is the amino acid sequence 4-18HC-Knob defining the 4-18 derived heavy chain of the 2-36/4-18 antibody

SEQ ID NO. 126 is the amino acid sequence 4-18LC of the 4-18 derived light chain of the defined 2-36/4-18 antibody

The 2-30/1-68 bispecific antibody sequences may comprise:

127 of the SEQ ID NO. 2-30HC-Hole-Cross, which defines the amino acid sequence of the 2-30 derived heavy chain of the 2-30/1-68 antibody

SEQ ID NO. 128 is the amino acid sequence 2-30VLCH1 defining the 2-30 derived light chain of the 2-30/1-68 antibody

SEQ ID NO. 129 is the amino acid sequence 1-68HC-Knob defining the 1-68 derived heavy chain of the 2-30/1-68 antibody

SEQ ID NO. 130 is the amino acid sequence 1-68LC of the 1-68 derived light chain of the defined 2-30/1-68 antibody

The 2-30/4-18 bispecific antibody may comprise:

SEQ ID NO. 131 is an amino acid sequence 2-30HC-Hole-Cross defining the 2-30 derived heavy chain of the 2-30/4-18 antibody

SEQ ID NO. 132 is the amino acid sequence 2-30VLCH1 defining the 2-30 derived light chain of the 2-30/4-18 antibody

SEQ ID NO 133 is the amino acid sequence 4-18HC-Knob defining the 4-18 derived heavy chain of the 2-30/4-18 antibody

SEQ ID NO. 134 is the amino acid sequence 4-18LC of the 4-18 derived light chain of the defined 2-30/4-18 antibody

The 2-7/4-8 (39/51) bispecific antibody sequences may comprise:

SEQ ID NO. 135 is the amino acid sequence 2-7HC-Hole-Cross defining the 2-7 derived heavy chain of the 2-7/4-8 (39/51) antibody

SEQ ID NO. 136 is the amino acid sequence 2-7VLCH1 defining the 2-7 derived light chain of the 2-7/4-8 (39/51) antibody

SEQ ID NO. 137 is the amino acid sequence 4-8 (39/51) HC-Knob of the 4-8 (39/51) derived heavy chain of the definition 2-7/4-8 (39/51) antibody

SEQ ID NO. 138 is the amino acid sequence 4-8LC of the 4-8 (39/51) -derived light chain of the 2-7/4-8 (39/51) antibody

The 1-57/1-87 bispecific antibody sequences may comprise:

SEQ ID NO. 139 is the amino acid sequence 1-57HC-Hole-Cross defining the 1-57 derived heavy chain of the 1-57/1-87 antibody

SEQ ID NO. 140 is the amino acid sequence 1-57VLCH1 defining the 1-57 derived light chain of the 1-57/1-87 antibody

SEQ ID NO. 141 is the amino acid sequence 1-87HC-Knob of the 1-87 derived heavy chain of the 1-57/1-87 antibody

SEQ ID NO. 142 is the amino acid sequence 1-87LC of the 1-87 derived light chain of the 1-57/1-87 antibody

1-57/4-8 (39/51) bispecific antibody may comprise:

SEQ ID NO. 143 is the amino acid sequence 1-57HC-Hole-Cross defining the 1-57 derived heavy chain of the 1-57/4-8 (39/51) antibody

SEQ ID NO. 144 is the amino acid sequence 1-57VLCH1 defining the 1-57 derived light chain of the 1-57/4-8 (39/51) antibody

SEQ ID NO. 145 is the amino acid sequence 4-8 (39/51) HC-Knob of the 4-8 (39/51) derived heavy chain of the 4-8 (39/51) antibody defining 1-57/4-8 (39/51)

SEQ ID NO. 146 is the amino acid sequence 4-8 (39/51) LC of the 4-8 (39/51) derived light chain of the 1-57/4-8 (39/51) antibody

2-7/2-51 bispecific antibody:

SEQ ID NO 147 is the amino acid sequence 2-7HC-Hole-Cross defining the 2-7 derived heavy chain of the 2-7/2-51 antibody

SEQ ID NO. 148 is the amino acid sequence 2-7VLCH1 defining the 2-7 derived light chain of the 2-7/2-51 antibody

149 is the amino acid sequence 2-51HC-Knob defining the 2-51 derived heavy chain of the 2-7/2-51 antibody

SEQ ID NO. 150 is the amino acid sequence 2-51LC of the 2-51 derived light chain of the defined 2-7/2-51 antibody

The 2-15/4-8 (39/51) bispecific antibody sequence may comprise:

SEQ ID NO. 151 is the amino acid sequence 2-15HC-Hole-Cross defining the 2-15 derived heavy chain of a 2-15/4-8 (39/51) antibody

SEQ ID NO. 152 is the amino acid sequence 2-15VLCH1 defining the 2-15 derived light chain of the 2-15/4-8 (39/51) antibody

153 is the amino acid sequence 4-8 (39/51) HC-Knob of the 4-8 (39/51) derived heavy chain of the 4-8 (39/51) antibody defining 2-15/4-8 (39/51)

SEQ ID NO. 154 is the amino acid sequence 4-8 (39/51) LC of the 4-8 (39/51) derived light chain of the defining 2-15/4-8 (39/51) antibody

The 1-57/4-18 bispecific antibody sequences may comprise:

SEQ ID NO. 155 is the amino acid sequence 1-57HC-Hole-Cross defining the 1-57 derived heavy chain of the 1-57/4-18 antibody

SEQ ID NO. 156 is the amino acid sequence 1-57VLCH1 defining the 1-57 derived light chain of the 1-57/4-18 antibody

SEQ ID NO. 157 is the amino acid sequence 4-18HC-Knob of the 4-18 derived heavy chain of the 1-57/4-18 antibody

SEQ ID NO. 158 is the amino acid sequence 4-18LC of the 4-18 derived light chain of the 1-57/4-18 antibody

The 2-7/1-68 bispecific antibody may comprise:

SEQ ID NO. 159 is the amino acid sequence 2-7HC-Hole-Cross defining the 2-7 derived heavy chain of the 2-7/1-68 antibody

160 is the amino acid sequence 2-7VLCH1 defining the 2-7 derived light chain of the 2-7/1-68 antibody

SEQ ID NO. 161 is the amino acid sequence 1-68HC-Knob defining the 1-68 derived heavy chain of the 2-7/1-68 antibody

162 is the amino acid sequence 1-68LC of the 1-68 derived light chain of the 1-68 defining 2-7/1-68 antibody

The 4-20/4-18 bispecific antibody sequences may comprise:

163 is the amino acid sequence 4-20HC-Hole-Cross defining the 4-20 derived heavy chain of the 4-20/4-18 antibody

SEQ ID NO. 164 is the amino acid sequence 4-20VLCH1 defining the 4-20 derived light chain of the 4-20/4-18 antibody

SEQ ID NO. 165 is the amino acid sequence 4-18HC-Knob of the 4-18 derived heavy chain of the 4-20/4-18 antibody

SEQ ID NO 166 is the amino acid sequence 4-18LC of the 4-18 derived light chain of the 4-20/4-18 antibody

The 2-15mut/2-51 bispecific antibody sequence may comprise:

167 is the amino acid sequence 2-15HC-Hole-Cross defining the 2-15mut derived heavy chain of the 2-15mut/2-51 antibody

SEQ ID NO. 168 is the amino acid sequence 2-15mutVLCH1 defining the 2-15mut derived light chain of the 2-15mut/2-51 antibody

SEQ ID NO. 169 is the amino acid sequence 2-51HC-Knob defining the 2-51 derived heavy chain of a 2-15mut/2-51 antibody

SEQ ID NO. 170 is the amino acid sequence 2-51LC of the 2-51 derived light chain of the 2-15mut/2-51 antibody

4-20/4-8(39/51)Bispecific antibody sequences may comprise:

SEQ ID NO. 171 is the amino acid sequence 4-20HC-Hole-Cross defining the 4-20 derived heavy chain of the 4-20/4-8 (39/51) antibody

SEQ ID NO. 172 is the amino acid sequence 4-20VLCH1 defining the 4-20 derived light chain of the 4-20/4-8 (39/51) antibody

SEQ ID NO. 173 is the amino acid sequence 4-8 (39/51) HC-Knob of the 4-8 (39/51) derived heavy chain of the 4-20/4-8 (39/51) antibody

SEQ ID NO. 174 is the amino acid sequence 4-8 (39/51) LC of the 4-8 (39/51) derived light chain of the 4-20/4-8 (39/51) antibody

The 1-57/2-51 bispecific antibody sequence may comprise:

SEQ ID NO. 175 is an amino acid sequence 1-57HC-Hole-Cross defining the 1-57 derived heavy chain of the 1-57/2-51 antibody

SEQ ID NO. 176 is the amino acid sequence 1-57VLCH1 of the 1-57 derived light chain of the 1-57/2-51 antibody

177 is the amino acid sequence 2-51HC-Knob defining the 2-51 derived heavy chain of the 1-57/2-51 antibody

SEQ ID NO. 178 is the amino acid sequence 2-51LC of the 2-51 derived light chain of the defined 1-57/2-51 antibody

The 4-20/1-87 bispecific antibody sequences may comprise:

179 is the amino acid sequence 4-20HC-Hole-Cross defining the 4-20 derived heavy chain of the 4-20/1-87 antibody

SEQ ID NO. 180 is the amino acid sequence 4-20VLCH1 defining the 4-20 derived light chain of the 4-20/1-87 antibody

181 of SEQ ID NO. 1-87HC-Knob is the amino acid sequence defining the 1-87 derived heavy chain of the 4-20/1-87 antibody

SEQ ID NO. 182 is the amino acid sequence 1-87LC of the 1-87 derived light chain of the defined 4-20/1-87 antibody

The 1-57/1-68 bispecific antibody sequences may comprise:

SEQ ID NO. 183 is an amino acid sequence 1-57HC-Hole-Cross defining the 1-57 derived heavy chain of the 1-57/1-68 antibody

SEQ ID NO. 184 is the amino acid sequence 1-57VLCH1 of the 1-57 derived light chain of the 1-57/1-68 antibody

SEQ ID NO. 185 is the amino acid sequence 1-68HC-Knob defining the 1-68 derived heavy chain of the 1-57/1-68 antibody

186 is the amino acid sequence 1-68LC of the 1-68 derived light chain of the 1-57/1-68 antibody

Bispecific antibodies with only LS mutations

The 2-7/4-8 (39/51/57) bispecific antibody sequences may comprise:

SEQ ID NO. 187 is an amino acid sequence 2-7HC-Hole-Cross defining the 2-7 derived heavy chain of a 2-7/4-8 (39/51/57) antibody

SEQ ID NO. 188 is the amino acid sequence 2-7VLCH1 defining the 2-7 derived light chain of the 2-7/4-8 (39/51/57) antibody

SEQ ID NO. 189 is the amino acid sequence 4-8 (39/51/57) HC-Knob of the 4-8 (39/51/57) derived heavy chain of the 4-8 (39/51/57) antibody defining 2-7/4-8 (39/51/57)

SEQ ID NO. 190 is an amino acid sequence 4-8LC of the 4-8 (39/51/57) -derived light chain of a 2-7/4-8 (39/51/57) antibody

The 2-17/2-36 bispecific antibody sequences may comprise:

SEQ ID NO. 191 is the amino acid sequence 2-17HC-Hole-Cross defining the 2-17 derived heavy chain of the 2-17/2-36 antibody

SEQ ID NO. 192 is the amino acid sequence 2-17VLCH1 defining the 2-17 derived light chain of the 2-17/2-36 antibody

SEQ ID NO 193 defines the amino acid sequence 2-36HC-Knob of the 2-36 derived heavy chain of the 2-17/2-36 antibody

SEQ ID NO. 194 is the amino acid sequence 2-36LC of the 2-36 derived light chain of the defined 2-17/2-36 antibody

2-36/1-68Bispecific antibody sequences:

195 is the amino acid sequence 2-36HC-Hole-Cross defining the 2-36 derived heavy chain of the 2-36/1-68 antibody

SEQ ID NO. 196 is the amino acid sequence 2-36VLCH1 defining the 2-36 derived light chain of the 2-36/1-68 antibody

SEQ ID NO 197 shows the amino acid sequence 1-68HC-Knob of the 1-68 derived heavy chain of the 2-36/1-68 antibody

SEQ ID NO. 198 is the amino acid sequence 1-68LC of the 1-68 derived light chain of the defined 2-36/1-68 antibody

The 2-36/4-18 bispecific antibody sequences may comprise:

199 is the amino acid sequence 2-36HC-Hole-Cross defining the 2-36 derived heavy chain of the 2-36/4-18 antibody

SEQ ID NO. 200 is the amino acid sequence 2-36VLCH1 defining the 2-36 derived light chain of the 2-36/4-18 antibody

SEQ ID NO. 201 is the amino acid sequence 4-18HC-Knob defining the 4-18 derived heavy chain of the 2-36/4-18 antibody

SEQ ID NO 202 is the amino acid sequence 4-18LC of the 4-18 derived light chain of the defined 2-36/4-18 antibody

The 2-30/1-68 bispecific antibody sequences may comprise:

SEQ ID NO. 203 is an amino acid sequence 2-30HC-Hole-Cross defining the 2-30 derived heavy chain of the 2-30/1-68 antibody

SEQ ID NO. 204 is the amino acid sequence 2-30VLCH1 defining the 2-30 derived light chain of the 2-30/1-68 antibody

SEQ ID NO. 205 is the amino acid sequence 1-68HC-Knob defining the 1-68 derived heavy chain of the 2-30/1-68 antibody

SEQ ID NO. 206 is the amino acid sequence 1-68LC of the 1-68 derived light chain of the defined 2-30/1-68 antibody

The 2-30/4-18 bispecific antibody sequences may comprise:

SEQ ID NO. 207 is the amino acid sequence 2-30HC-Hole-Cross defining the 2-30 derived heavy chain of the 2-30/4-18 antibody

SEQ ID NO. 208 is the amino acid sequence 2-30VLCH1 defining the 2-30 derived light chain of the 2-30/4-18 antibody

SEQ ID NO. 209 is the amino acid sequence 4-18HC-Knob defining the 4-18 derived heavy chain of the 2-30/4-18 antibody

SEQ ID NO. 210 is the amino acid sequence 4-18LC of the 4-18 derived light chain of the defined 2-30/4-18 antibody

The 2-7/4-8 (39/51) bispecific antibody sequences may comprise:

SEQ ID NO. 211 is the amino acid sequence 2-7HC-Hole-Cross defining the 2-7 derived heavy chain of a 2-7/4-8 (39/51) antibody

SEQ ID NO. 212 is the amino acid sequence 2-7VLCH1 defining the 2-7 derived light chain of the 2-7/4-8 (39/51) antibody

SEQ ID NO. 213 is the amino acid sequence 4-8 (39/51) HC-Knob of the 4-8 (39/51) derived heavy chain of the 4-8 (39/51) antibody defining 2-7/4-8 (39/51)

SEQ ID NO. 214 is the amino acid sequence 4-8LC of the 4-8 (39/51) -derived light chain of the 2-7/4-8 (39/51) antibody

The 1-57/1-87 bispecific antibody sequences may comprise:

SEQ ID NO. 215 is the amino acid sequence 1-57HC-Hole-Cross defining the 1-57 derived heavy chain of the 1-57/1-87 antibody

SEQ ID NO. 216 is the amino acid sequence 1-57VLCH1 of the 1-57 derived light chain of the 1-57/1-87 antibody

SEQ ID NO. 217 is the amino acid sequence 1-87HC-Knob of the 1-87 derived heavy chain of the 1-57/1-87 antibody

SEQ ID NO. 218 is the amino acid sequence 1-87LC of the 1-87 derived light chain of the 1-57/1-87 antibody

1-57/4-8 (39/51) bispecific antibody sequences may comprise:

SEQ ID NO. 219 is the amino acid sequence 1-57HC-Hole-Cross defining the 1-57 derived heavy chain of the 1-57/4-8 (39/51) antibody

SEQ ID NO. 220 is the amino acid sequence 1-57VLCH1 defining the 1-57 derived light chain of the 1-57/4-8 (39/51) antibody

SEQ ID NO. 221 is the amino acid sequence 4-8 (39/51) HC-Knob of the 4-8 (39/51) derived heavy chain of the 1-57/4-8 (39/51) antibody

SEQ ID NO. 222 is the amino acid sequence 4-8LC of the 4-8 (39/51) -derived light chain of the 1-57/4-8 (39/51) antibody

The 2-7/2-51 bispecific antibody sequences may comprise:

SEQ ID NO. 223 is the amino acid sequence 2-7HC-Hole-Cross defining the 2-7 derived heavy chain of the 2-7/2-51 antibody

224 is the amino acid sequence 2-7VLCH1 defining the 2-7 derived light chain of the 2-7/2-51 antibody

SEQ ID NO. 225 is the amino acid sequence 2-51HC-Knob defining the 2-51 derived heavy chain of the 2-7/2-51 antibody

SEQ ID NO. 226 is the amino acid sequence 2-51LC of the 2-51 derived light chain of the defined 2-7/2-51 antibody

The 2-15/4-8 (39/51) bispecific antibody sequence may comprise:

SEQ ID NO. 227 is the amino acid sequence 2-15HC-Hole-Cross defining the 2-15 derived heavy chain of a 2-15/4-8 (39/51) antibody

SEQ ID NO. 228 is the amino acid sequence 2-15VLCH1 defining the 2-15 derived light chain of a 2-15/4-8 (39/51) antibody

SEQ ID NO. 229 is the amino acid sequence 4-8 (39/51) HC-Knob of the 4-8 (39/51) derived heavy chain of the 4-8 (39/51) antibody defining 2-15/4-8 (39/51)

SEQ ID NO. 230 is the amino acid sequence 4-8LC of the 4-8 (39/51) -derived light chain of the defined 2-15/4-8 (39/51) antibody

The 1-57/4-18 bispecific antibody sequences may comprise:

231 is the amino acid sequence 1-57HC-Hole-Cross defining the 1-57 derived heavy chain of the 1-57/4-18 antibody

SEQ ID NO. 232 is the amino acid sequence 1-57VLCH1 defining the 1-57 derived light chain of the 1-57/4-18 antibody

SEQ ID NO. 233 is the amino acid sequence 4-18HC-Knob defining the 4-18 derived heavy chain of a 1-57/4-18 antibody

SEQ ID NO. 234 is the amino acid sequence 4-18LC of the 4-18 derived light chain of the 1-57/4-18 antibody

The 2-7/1-68 bispecific antibody sequences may comprise:

SEQ ID NO. 235 is an amino acid sequence 2-7HC-Hole-Cross defining the 2-7 derived heavy chain of the 2-7/1-68 antibody

SEQ ID NO. 236 is the amino acid sequence 2-7VLCH1 defining the 2-7 derived light chain of the 2-7/1-68 antibody

SEQ ID NO. 237 is the amino acid sequence 1-68HC-Knob defining the 1-68 derived heavy chain of the 2-7/1-68 antibody

SEQ ID NO. 238 is the amino acid sequence 1-68LC of the 1-68 derived light chain of the defined 2-7/1-68 antibody

The 4-20/4-18 bispecific antibody sequences may comprise:

239 is the amino acid sequence 4-20HC-Hole-Cross defining the 4-20 derived heavy chain of the 4-20/4-18 antibody

SEQ ID NO. 240 is the amino acid sequence 4-20VLCH1 defining the 4-20 derived light chain of the 4-20/4-18 antibody

SEQ ID NO. 241 is the amino acid sequence 4-18HC-Knob of the 4-18 derived heavy chain of the 4-20/4-18 antibody

SEQ ID NO. 242 is the amino acid sequence 4-18LC of the 4-18 derived light chain of the 4-20/4-18 antibody

The 2-15mut/2-51 bispecific antibody sequence may comprise:

243 is the amino acid sequence 2-15HC-Hole-Cross defining the 2-15mut derived heavy chain of the 2-15mut/2-51 antibody

SEQ ID NO. 244 is the amino acid sequence 2-15mutVLCH1 defining the 2-15mut derived light chain of the 2-15mut/2-51 antibody

SEQ ID NO. 245 is the amino acid sequence 2-51HC-Knob defining the 2-51 derived heavy chain of a 2-15mut/2-51 antibody

SEQ ID NO. 246 is the amino acid sequence 2-51LC of the 2-51 derived light chain of the 2-15mut/2-51 antibody

The 4-20/4-8 (39/51) bispecific antibody sequence may comprise:

SEQ ID NO. 247 is the amino acid sequence 4-20HC-Hole-Cross defining the 4-20 derived heavy chain of the 4-20/4-8 (39/51) antibody

SEQ ID NO. 248 is the amino acid sequence 4-20VLCH1 defining the 4-20 derived light chain of the 4-20/4-8 (39/51) antibody

SEQ ID NO. 249 is the amino acid sequence 4-8 (39/51) HC-Knob of the 4-8 (39/51) derived heavy chain of the 4-20/4-8 (39/51) antibody

SEQ ID NO. 250 is the amino acid sequence 4-8LC of the 4-8 (39/51) -derived light chain of the 4-20/4-8 (39/51) antibody

The 1-57/2-51 bispecific antibody sequence may comprise:

SEQ ID NO. 251 is an amino acid sequence 1-57HC-Hole-Cross defining the 1-57 derived heavy chain of the 1-57/2-51 antibody

SEQ ID NO. 252 is the amino acid sequence 1-57VLCH1 of the 1-57 derived light chain of the 1-57/2-51 antibody

SEQ ID NO 253 shows the amino acid sequence 2-51HC-Knob of the 2-51 derived heavy chain of the 1-57/2-51 antibody

254 is the amino acid sequence 2-51LC defining the 2-51 derived light chain of the 1-57/2-51 antibody

The 4-20/1-87 bispecific antibody sequences may comprise:

255 is the amino acid sequence 4-20HC-Hole-Cross defining the 4-20 derived heavy chain of the 4-20/1-87 antibody

SEQ ID NO. 256 is the amino acid sequence 4-20VLCH1 defining the 4-20 derived light chain of the 4-20/1-87 antibody

SEQ ID NO 257 is the amino acid sequence 1-87HC-Knob of the 1-87 derived heavy chain of the antibody defining 4-20/1-87

258 SEQ ID NO. 1-87LC, the amino acid sequence of the 1-87 derived light chain of the 4-20/1-87 antibody

The 1-57/1-68 bispecific antibody sequences may comprise:

SEQ ID NO. 259 defines the amino acid sequence 1-57HC-Hole-Cross of the 1-57 derived heavy chain of the 1-57/1-68 antibody

SEQ ID NO. 260 is the amino acid sequence 1-57VLCH1 of the 1-57 derived light chain of the 1-57/1-68 antibody

SEQ ID NO 261 is the amino acid sequence 1-68HC-Knob defining the 1-68 derived heavy chain of the 1-57/1-68 antibody

SEQ ID NO. 262 is the amino acid sequence 1-68LC of the 1-68 derived light chain of the 1-57/1-68 antibody

In some embodiments, bispecific antibodies can be generated from any combination of the two monoclonal antibodies described herein. In some embodiments, the invention disclosed herein relates to DNA sequences encoding bispecific antibodies comprising any sequence from SEQ ID NOs1-262 or a combination of SEQ ID NOs1 to 262. In some embodiments, the invention disclosed herein relates to RNA sequences encoding bispecific antibodies encoding a polypeptide comprising any sequence from SEQ ID NOs1-262 or a combination of SEQ ID NOs1 to 262.

Examples

The following examples are provided to facilitate a more complete understanding of the present invention. The following examples illustrate exemplary modes of making and implementing the present invention. However, the scope of the invention is not limited to the specific embodiments disclosed in these examples, which are for illustrative purposes only, as alternative methods may be used to achieve similar results.

Examples of monoclonal antibodies

Example 1 patient antibody response

By carefully studying 40 cases of covd-19 patients, it was found that more potent coronavirus antibodies were produced in the blood of the heavier patients. These antibodies include antibodies specific for the spike protein trimer. For a patient of covd-19 classified as critical, mechanical ventilation is required in the critical care unit and is treated with hydroxychloroquine, adefovir and/or tolizumab. For a patient of covd-19 who is classified as non-critical, intensive care is not required. FIGS. 2A-G show ELISA binding data in which various plasma samples of patients with COVID-19 (represented by lines of different colors) were tested for their ability to bind to SARS-CoV-2 nucleocapsid antigen (FIGS. 2A and D), SARS-CoV-2 envelope trimer antigen (FIGS. 2B and E), or SARS-CoV envelope trimer antigen (FIGS. 2C and F). This binding assay is used to determine whether a plasma sample contains elements (most likely antibodies) that can interact with SARS-CoV-2 and SARS-CoV antigens, indicating an immune response to the antigen. Those plasma samples that interact with SARS-CoV-2 and SARS-CoV antigens demonstrate the potential to use these samples to neutralize antigens and virus-carrying antigens. FIG. 2G shows a data plot from FIGS. 2A-F showing the presence of a higher SARS-CoV-2 antibody response in a plasma sample from a patient with severe COVID-19 than a patient with non-severe COVID-19.

Example 2-efficacy related to neutralizing antibody response to SARS-CoV-2

FIGS. 3A-J show that a stronger binding antibody response is closely related to a stronger efficacy in neutralizing SARS-CoV-2. Neutralization assay wherein plasma samples of various covd-19 patients, represented by lines of different colors, were tested for their ability to protect target cells from SARS-CoV-2 infection in a single cycle pseudovirus infectivity assay (fig. 3A and fig. D), their ability to protect target cells from SARS-CoV infection in a single cycle pseudovirus infectivity assay (fig. 3B and fig. E), and their ability to protect target cells from SARS-CoV-2 infection in a live virus infectivity assay with replication ability (fig. 3C and fig. F). This test was used to determine if the plasma samples contained elements (most likely antibodies) that could effectively block SARS-CoV-2 or SARS-CoV infection. FIG. 3G shows the dilution (ID) plotted according to FIGS. 3A-F ₅₀ ) Wherein the plasma inhibits 50% of the viral infection. The presence of higher dilution of SARS-CoV-2 inhibition in plasma samples of severe patients with COVID-19 suggests that the antibody responses of these patients may be more effective. FIGS. 3H-J show ICs obtained by a pseudovirus neutralization assay ₅₀ Value and IC obtained by live virus neutralization assay (FIG. 3H) ₅₀ Values, reaching the OD by ELISA test in FIG. 2 (FIG. 3I) ₄₅₀ Half of the value readings were of plasma dilution, and plasma OD bound to S trimer when diluted 400-fold in figure 2 (figure 3J) ₄₅₀ The values are closely related. The data herein show that the binding capacity of the antibody to SARS-CoV-2S trimer in a COVID-19 plasma sample is closely related to the neutralizing capacity, indicating that individual B cells from the separation of SARS-CoV-2S trimer from COIVD-19 patients should have neutralizing capacity in their secreted antibodies。

Example 3-schematic representation of identification of potent neutralizing antibodies against COVID-19

From the cases of the patient with covd-19, where the most potent antibody response was shown in the experiments of fig. 2 and 3, it can be seen that blood samples were obtained and the protocols described in fig. 4A-C were performed to identify monoclonal antibodies that were able to effectively neutralize the SARS-CoV-2 virus. From each blood sample, antibody-producing cells called cd27+ memory B cells were isolated. In particular, a subpopulation of cells is isolated that can bind to the virus spike protein trimer.

The protocol may include the following steps. PBMC from infected individuals were collected and treated with LIVE/DEAD at room temperature ^TM Yellow dead cell staining kit (Ingium (Invitrogen), cat# L34959) can be fixed for 20 minutes to exclude dead cells. Subsequently, cells were first incubated with 10 μg/ml spike trimer, followed by incubation of cells with antibody mixtures to identify SARS-CoV-2S trimer-specific B cells. The mixture consisted of CD3-PE-CF594 (BD Biosciences, cat# 562406), CD19-PE-cy7 (bioleged, cat# 302216), CD20-APC-cy7 (bioleged, cat# 302314), igM-V450 (BD Biosciences, cat# 561286), CD27-PerCP-Cy5.5 (BD Biosciences, cat# 560612) and Anti-His-PE (bioleged, cat# 362603). Spike trimer specific single B cell gating (gated) to CD3-CD19 CD27+ spike trimer+, and sorting to LoBind microcentrifuge tube (Sigma, cat# Z666505). After enrichment of antigen specific cd27+ memory B cells, 1310B cells were loaded onto a 10X chromasum chip. All experimental procedures were performed prior to library preparation in the BSL 3-grade laboratory. According to the manufacturer's protocol, 10X Chromium Single Cell 5'Library is used&Gel head Kit performs single cell lysis and RNA first strand synthesis. The following RNA and VDJ library preparations were performed in the BSL-2 laboratory according to the manufacturer's protocol (Chromium Single Cell V (D) J Reagent kit, 10X Genomics). Sequencing was performed on a 26X91 run of Rapid SBS Kit V2 2X100bp Kit (Illumina)Dual read mode Hiseq 2500 platform. With this information, each monoclonal antibody was reconstituted, and more than 250 monoclonal antibodies were now obtained by this method.

As shown in fig. 9, a total of 254 monoclonal antibodies (mabs) were currently synthesized and tested. These include binding antibodies, pseudovirus neutralizing mabs, and live virus neutralizing antibodies. Binding antibodies included 121 targeted spike protein trimer mabs, 38 RBD-targeted mabs, and 83 non-RBD-targeted mabs. Pseudovirus neutralizing mabs included 52 mabs targeting spike protein trimers, 28 mabs targeting RBDs, and 24 mabs not targeting RBDs. The live virus neutralizing antibodies included 32 spike protein-targeted mabs, 13 RBD-targeted mabs, and 19 non-RBD-targeted mabs. Live virus neutralization tests were performed on 217 mabs.

Example 4

FIGS. 5A-D show characteristics of six synthetic and purified monoclonal antibodies initially identified, including high binding capacity to SARS-CoV-2S trimer (FIG. 5A) and viral receptor binding domain (FIG. 5B), and neutralization capacity of viruses with high potency in pseudovirus (FIG. 5C) and live virus with replication capacity (FIG. 5D) assays. Each color line represents a different monoclonal antibody. Half maximal effective concentration (EC ₅₀ ) And half maximal Inhibitory Concentration (IC) tested by neutralization assay ₅₀ ) Provided alongside each antibody. SARS-CoV antibody CR3022 is a control that binds but does not neutralize SARS-CoV-2. These six antibodies were specific for SARS-CoV-2S trimer, three of which were RBD binders and three of which were non-RBD binders. All six antibodies have neutralizing activity against SARS-CoV-2 pseudovirus and live virus, of which 2-15 is the most effective one. Binding data suggests that these antibodies can be divided into at least two groups, which have different mechanisms in neutralizing viruses.

To determine the binding sites of these monoclonal antibodies on the SAR-CoV-2 envelope, competitive ELISA experiments were performed to map the epitopes of these novel monoclonal antibodies. As shown in fig. 6A-F, each antibody was biotinylated, mixed with other serially diluted antibodies, and then incubated with spike protein (S) trimer. Binding of biotinylated antibodies to S trimer in the presence of other monoclonal antibodies was detected by ELISA. Each color line represents a different monoclonal antibody. Six characterized monoclonal antibodies are divided into two classes: RBD binding (fig. 6A-C) and non-RBD binding (fig. 6D-F). There is no competition between the two classes of antibodies, but within the two classes of antibodies, the combination of RBD and non-RBD binding agents offers the possibility to combat covd-19, as they have different mechanisms for neutralizing viruses.

Example 5

To further characterize the binding affinity of these novel monoclonal antibodies, surface plasmon resonance experiments were performed to determine the binding affinity of the most promising monoclonal antibodies to SARS-CoV-2 spike trimer. As shown in FIGS. 7A-E, each figure represents the binding affinity and binding kinetics of the indicated monoclonal antibodies (i.e., 2-4, 2-15, 2-38, etc.) to SARS-CoV-2 envelope trimer. Lower KD1 values indicate higher antibody binding affinities. The color bars represent the different dilutions of the corresponding antibodies tested.

Example 6

FIGS. 8A-D show a preliminary screen of mAbs that bind to and neutralize SARS-CoV-2 virus. Figure 8A shows mabs binding to spike trimer proteins. Fig. 8B shows mabs binding to the Receptor Binding Domain (RBD). FIG. 8C shows an IC infected with a pseudovirus ₅₀ Values. Fig. 8D shows the% inhibition values of live virus infection. Fig. 9 shows a summary of synthetic antibodies.

Example 7

As shown in fig. 10A-I, the effective neutralizing mabs disclosed herein can bind to the receptor binding domain of a spike protein trimer, the N-terminal domain of a spike protein trimer, or different epitopes on a spike protein trimer. Each color line shown in fig. 10A-I represents a different monoclonal antibody used in the ELISA binding assay. Fig. 10A, D and G show binding to a spike protein trimer. Fig. 10B, E and H show the combination with RBD. Fig. 10C, F, and I show the combination with NTD. Figures 10A-C show binding of mabs that bind efficiently to RBD. Figures 10D-F show binding of mabs that bind efficiently to NTD. Fig. 10G-I show binding of mabs that bind to epitopes other than RBD or NTD. Monoclonal antibodies 2-15, 2-7, 1-57, 2-4, 2-38, 1-20, 2-30 and 4-20 specifically target the RBD of the spike protein trimer. They have no binding affinity for the NTD of the spike protein trimer as shown in fig. 10C. Monoclonal antibodies 4-18, 5-24, 5-7 and 1-68 specifically target the NTD of the spike protein trimer. These antibodies have no binding affinity for the RBD of the spike protein trimer, as shown in fig. 10E. Monoclonal antibodies 2-43, 4-8, 4-19, 2-51 and 2-17 target at least one other epitope on the spike protein trimer. These antibodies did not show binding affinity to RBD or NTD, as shown in fig. 10H and fig. I, respectively.

Example 8

FIGS. 11A-F show SARS-CoV-2 neutralization activity of selected mAbs. Fig. 11A shows the neutralizing activity of mabs targeting RBD to neutralize pseudoviral infection. IC of these antibodies ₅₀ The concentration ranges from 0.005 μg/ml to 0.512 μg/ml. Fig. 11B shows the neutralizing activity of mabs targeting NTD to neutralize pseudovirus infection. IC of these antibodies ₅₀ The concentration ranges from 0.013 μg/ml to 0.767 μg/ml. Fig. 11C shows neutralizing activity of mabs targeting epitopes other than RBD or NTD to neutralize pseudovirus infection. IC of these antibodies ₅₀ The concentration ranges from 0.070. Mu.g/ml to 0.652. Mu.g/ml. Fig. 11D shows the neutralizing activity of mabs targeting RBD to neutralize live virus infection. IC of these antibodies ₅₀ The concentration ranges from 0.001. Mu.g/ml to 0.103. Mu.g/ml. Fig. 11E shows neutralizing activity of mabs targeting NTD to neutralize live virus infection. IC of these antibodies ₅₀ The concentration ranges from 0.011 μg/ml to 0.034 μg/ml. Fig. 11F shows neutralizing activity of mabs targeting epitopes other than RBD or NTD to neutralize live virus infection. IC of these antibodies ₅₀ The concentration ranges from 0.003. Mu.g/ml to 0.079. Mu.g/ml.

Example 9

Fig. 12A-B show the CryoEM structure of RBD-targeting monoclonal antibodies. Fig. 12A shows a side view of the structure, and 12B shows a top view of the structure. The mAb is shown in the blue band, immediately adjacent to the RBD, which is shown in the green band. The CryoEM structure indicates that RBD-targeting mabs do not directly interact with NTD, which is shown as a brown band.

Fig. 13A-B show the CryoEM structure of monoclonal antibodies targeting NTD. Fig. 13A shows a side view of the structure, and fig. 13B shows a top view of the structure. The mAb is shown in blue bands, immediately adjacent to NTD, which is shown in brown bands. The CryoEM structure indicates that the NTD-targeted mabs do not interact directly with RBDs, which are shown as green sheets.

FIG. 14 shows three SARS-CoV-2 neutralization epitope clusters identified on spike protein trimers. NTD clusters are shown in blue bands. RBD is shown in green bands. Other colors represent potential neutralizing epitopes on spike proteins that are different from the NTD and RBD clusters.

Example 10 efficient neutralizing monoclonal antibodies against multiple epitopes on SARS-CoV-2 spike

Abstract

SARS-CoV-2 is a pandemic with damaging consequences for human life and global economy. The discovery and development of virus-neutralizing monoclonal antibodies may be a method of treating or preventing such novel coronavirus infections. Here we report the isolation of 61 SARS-CoV-2 neutralizing monoclonal antibodies from 5 critically ill hospitalized infected patients. Of these, 19 antibodies effectively neutralized true SARS-CoV-2 in vitro, 9 exhibited excellent efficacy, with 50% viral inhibition concentrations ranging from 1 to 9ng/mL. Epitope mapping showed that the collection of 19 antibodies was approximately equally divided between the antibody directed against the Receptor Binding Domain (RBD) and the antibody directed against the N-terminal domain (NTD), indicating that both regions at the top of the viral spike were strongly immunogenic. In addition, two other powerful neutralizing antibodies can recognize quaternary epitopes overlapping the spike-top domain. The cryo-electron microscope structure of RBD-targeting antibodies, second NTD-targeting antibodies, and third RBD-and NTD-bridging antibodies reveals the blocking of spikes, recognition of the "all RBD down" conformation. Several of these monoclonal antibodies are promising candidates for clinical development as potential therapeutic and/or prophylactic agents against SARS-CoV-2.

Background

Novel coronavirus-now called SARS-CoV-2 ^1,2 Nearly 800 thousands of diagnosed infections have been caused worldwide, resulting in about 45 thousands of deaths. The pandemic also places many areas of the world in a standstill, causing unprecedented damage to life and unprecedented damage to economy. Restoring to a certain normal state will rely on science to provide an effective solution, while the scientific community's response is compelling. Drug development is proceeding smoothly and candidate vaccines are entering clinical trials. Another promising approach is to isolate SARS-CoV-2 neutralizing monoclonal antibodies (mAbs) that can be used as therapeutic or prophylactic agents. The main target of this antibody is the virus spike, a trimeric protein ^3,4 Which is responsible for binding to the ACE2 receptor on host cells ^1,3,5,6 . The spike protein consists of two subunits. The S1 subunit has two major structural elements: RBD and NTD; after RBD binds ACE2, the S2 subunit mediates virus-cell membrane fusion. Report of finding neutralizing mAbs targeting RBD has recently been published ^7-11 . We now describe our efforts in isolating and characterizing a set of mAbs that are not only directed against multiple epitopes on the spike of the virus, but also show excellent efficacy in neutralizing SARS-CoV-2.

Patient selection

40 patients who were confirmed to be infected with SARS-CoV-2 by PCR participated in an observational cohort study with respect to virus neutralizing antibodies. All subjects were first tested for their neutralizing activity against SARS-CoV-2 pseudovirus (spike pseudovesicular stomatitis virus). Widely different neutralization titers were observed, with IC50 ranging from a reverse plasma dilution (Reciprocal plasma dilutions) <100 to about 13,000 (fig. 15A). Five patients were selected for mAb isolation because their plasma virus neutralization titers were highest. Fig. 19 summarizes the clinical characteristics of these 5 cases. Briefly, all patients suffer from severe acute respiratory distress syndrome requiring mechanical ventilation. Their age is between 50 and 71 years old. Two are spanish women, two are white men and one is black men. One patient died and the other patient recovered. Importantly, the blood used to isolate SARS-CoV-2mAb was obtained from day 18 to 32 after symptoms had occurred.

Monoclonal antibody isolation and construction

Peripheral blood mononuclear cells from each patient were subjected to an experimental protocol (fig. 22A) starting with cell sorting by flow cytometry. Sorting strategies focused on CD 3-negative, CD 19-positive and CD 27-positive live memory B lymphocytes (fig. 22B). The last step focused on those cells that bound SARS-CoV-2 spike trimer (S-trimer) ⁴ . S trimer positive memory B cells were enriched (0.4% to 1.4%) in 5 patients compared to normal healthy donors (0.2%) (fig. 22C). A total of 602, 325, 14, 147 and 145 such B cells from patient 1, patient 2, patient 3, patient 4 and patient 5, respectively, were labeled with unique tags. The cells were then placed in 10X chromoum (10X Genomics) for single cell 5' mRNA and V (D) J sequencing to obtain paired heavy (H) and light (L) chain sequences. Careful bioinformatics analysis was performed on 1,145 paired sequences to screen for "high confidence" antigen-specific mabs. A total of 331 mAb sequences were recovered, representing 252 individual clones. Only 6 mabs were from patient 3, while 44 to 100 mabs were identified from each other patient (figure 20). The VH and VL sequences of 252 antibodies (one for each clone) were codon optimized and synthesized, then each VH and VL gene was cloned into an expression plasmid with the H and L chain constant regions of the corresponding human IgG1, and mabs were expressed by co-transfection of pairs of full length H and L chain genes into Expi293 cells. Supernatants from each transfection were collected for screening assays and antibody purification.

Monoclonal antibody screening

All 252 parts of the transfection supernatant were screened by enzyme-linked immunosorbent assay (ELISA) to bind S trimer and RBD, and to neutralize SARS-CoV-2 pseudovirus and live virus. These results are graphically represented in fig. 15B-E and made into the table of fig. 20. It is evident that a significant proportion of the mAb in the supernatant bound to the S trimer and some of it bound to the RBD. Specifically, 121 supernatants were rated positive for S trimer binding, yielding a 48% overall hit rate (over hit rate). Of these, 38 RBD binding was positive, while the remaining 83 were negative. Interestingly, none of the 13 trimer-specific mabs from patient 5 recognized RBD. In the pseudo-virus neutralization screen, 61 supernatants were rated positive, indicating half of the trimer-specific mabs had virus neutralization. In particular, even when diluted 1,000-fold or more, 15 parts of the supernatant retained the neutralization activity. In the neutralization screening against SARS-CoV-2 (strain USA-WA 1/2020), 41 parts of the supernatant liquid WAs evaluated as positive, of which 10 parts are completely neutral and virus (+++). Overall, this screening strategy was very effective in picking neutralizing mabs (vertical lines and bottom labeled antibodies of fig. 15B) that were later identified as effective.

Sequence analysis of S trimer-specific monoclonal antibodies

Of the 121 mabs that bound the S trimer, 88% were of IgG isotypes with IgG1 predominating (fig. 23A-D). Antibodies of small amounts of IgM and IgA isotypes were also found. Compared with the IgG library of three healthy human donors ¹² Statistically significant overexpression of IGHV3-30, IGKV3-20 and IGHJ6 genes was observed in this group of SARS-CoV-2 mAbs (FIG. 23E). Longer CDRH3 lengths were also noted (fig. 23F). Interestingly, the average percentages of somatic hypermutations in VH and VL were 2.1 and 2.5, respectively, significantly lower than levels in healthy individuals (fig. 23G) and very close to germline sequences.

Antigen binding and virus neutralization

Since the screening for pseudovirus neutralization was performed by serial dilution quantification of the transfection supernatants, we plotted in FIG. 24 a best fit neutralization curve of 130 samples positive in at least one of the screens shown in FIG. 15B. Most were non-neutralized or weakly neutralized, but 18 samples showed significantly better efficacy. One additional supernatant was originally omitted in the pseudovirus screen (patient 1 in fig. 24), but was later found to be an effective neutralizing mAb. These 19 mabs were purified together from the transfection supernatant and further characterized for their binding and neutralization properties. As shown in fig. 16A, D and G, all mabs except one (2-43) bound the S trimer by quantitative ELISA. Two additional quantitative ELISA were used, of which nine antibodies clearly bound RBD (fig. 16C, fig. E and fig. H), with little or no binding to NTD (fig. 16C, fig. F and fig. I). Eight antibodies bound NTD to varying degrees, but did not bind RBD. Two mabs bound neither RBD nor NTD and were therefore classified as "other.

The pseudo-virus neutralization spectra of these purified 19 mabs are shown at the top of fig. 16J-L. Antibodies to RBD with IC of 0.005 to 0.512 μg/mL ₅₀ Neutralizing the pseudovirus; antibodies to NTD are slightly less potent, IC ₅₀ Ranging from 0.013 to 0.767 μg/mL. One common feature of NTD mabs is that virus neutralization water stabilizes at a level below 100%. Two antibodies classified as "other" were tested at IC's of 0.071 and 0.652. Mu.g/mL ₅₀ Neutralization, mAb 2-51 exhibited the typical stabilization of NTD antibodies. The real or live SARS-CoV-2 (strain USA-WA 1/2020) WAs antibody-neutralized using Vero cells seeded with a multiplicity of infection of 0.1. As shown in the bottom of FIG. 16M-O, antibodies to RBD again neutralize the virus, but IC ₅₀ 0.0007 to 0.209 μg/mL; antibodies against NTD showed similar efficacy, IC ₅₀ Ranging from 0.007 to 0.109 μg/mL. Here, the stabilization seen in the pseudovirus neutralization test is less pronounced. Antibodies 2-43 and 2-51 were tested at IC's of 0.003 and 0.007 μg/mL, respectively ₅₀ Neutralizing live virus. Overall, nine mabs were tested in vitro at IC of 0.009 μg/mL or less ₅₀ Excellent efficacy was demonstrated in neutralizing real SARS-CoV-2, including four for RBD (2-15, 2-7, 1-57 and 1-20), three for NTD (2-17, 5-24 and 4-8), and two for undefined regions on S trimer (2-43 and 2-51). Notably, patient 2 alone contributed to 5 of the first 9 SARS-CoV-2 neutralizing mAbs.

Epitope mapping

All 19 potent neutralizing mabs (fig. 16) were further studied in an antibody competition experiment to gain insight into their epitopes. In addition, 12 mAbs that bind strongly to S trimer during initial supernatant screening were also selected, including 5 that bind RBD (1-97, 2-26, 4-13, 4-24 and 4-29) and 7 that do not bind RBD (1-21, 2-29, 4-15, 4-32, 4-33, 4-41 and 5-45). Of these mAbs, 4 were very weak in neutralizing SARS-CoV-2 pseudovirus, and the remaining 8 were non-neutralizing (FIG. 25). First, the competition of binding of a total of 16 non-RBD mabs to S trimer was assessed by ELISA in a "checkerboard" experiment. The extent of antibody competition is reflected by the intensity of the heat map shown in fig. 17A. There is a large cluster of mabs (a), where these mabs compete with each other, which partially overlap with a small cluster (B). The third cluster (C) does not overlap at all. Note that all antibodies in cluster a except one recognize NTD. Antibody 2-51 is apparently directed against the NTD region even though it cannot bind NTD. In addition, each mAb from clusters B and C also recognized NTD, indicating that all three clusters are within or near NTD. A mAb,1-21, appears to have a unique non-overlapping epitope (epitope region D).

Second, we were tested for 14 mAbs against RBD and CR3022 by ELISA ^13,14 A similar "checkerboard" competition experiment was performed. Here, the heat map (hotmap) shows four consecutively overlapping epitope clusters (fig. 17B). In fig. 17C-D, there is one large cluster (E) comprising mabs that are primarily capable of blocking ACE2 binding. Furthermore, the 4 antibodies in this cluster lost binding to mutant RBD (L455R, A475R, G502R) which could no longer bind ACE2 (our unpublished findings). Taken together, these results indicate that most mabs in the E cluster target the ACE2 binding site of RBD. The next cluster (F) is connected to cluster E and cluster G, the position of which is defined by its member CR3022 ¹⁵ And (3) limiting. Finally, cluster G overlaps another cluster (H), which includes 1-97 that highly inhibits 2-30 binding to S trimer. This finding suggests that cluster H may be near one edge of cluster E.

An effective neutralizing mAb 2-43 did not bind to S trimer and was determined by ELISA (fig. 16) and therefore could not be tested in the competition experiments described above. However, 2-43 did bind strongly to S-trimer upon cell surface expression as determined by flow cytometry, and this binding was competitive by itself, not by RBD, NTD, ACE2 or soluble S-trimer 4 (fig. 26A-C). Mabs against NTD had only a modest effect on their binding to cell surface S trimers, but many mabs against RBD in cluster E blocked effectively the binding of 2-43, suggesting that this antibody might target a quaternary epitope on top of RBD, including a portion of the binding site to ACE 2.

The results in FIGS. 17A-B and 26A-C can be represented by two sets of Ween (Venn) plots as shown in FIGS. 17C-D. In the non-RBD area, is effectiveThe neutralizing mAb is only located in cluster a and binds to patches on NTD. Weaker neutralizing mabs recognize the region at the junction between clusters a and B. In the RBD region, the most potent neutralizing mabs also cluster in one cluster (E). Given that all of these block ACE2 binding, they are likely to recognize the top of RBD and neutralize the virus by competitively inhibiting receptor binding. Cluster G contains CR3022, a technique known to be in the "up" position at RBD ¹⁵ A monoclonal antibody directed against an epitope on one of the cryptic sites on the RBD side. Thus, cluster F may be located between the top and this "hidden" site. The wien (Venn) plot also shows that cluster H may occupy a different side near the top of the RBD, possibly in the area identified in S3098.

Freezing electron microscope

To further understand antibody recognition of viral spikes and to help explain the mapping studies, we determined the cryo-electron microscope structure of Fab of three mabs complexed with S trimer 4. First, single particle analysis of complexes with 2-4Fab (directed against RBD) resulted in high quality spectra (FIG. 18a; FIG. 21; FIGS. 27A-F), where the most abundant particle class represented the 3-Fab-per-oligomer complex, refined to Is used for the overall resolution of (a). Although the density of Fab constant portions is visible, only variable domains are included in the molecular model, as they are obscured by molecular motion. 2-4 binds to spike protein near the apex, all RBDs are in a "downward" orientation, and the structure of the antibody-bound spike protein is highly similar to the ligand-free spike structure of the previously published "all-downward" conformation 3, 4. The 2-4 epitope on RBD has 751->Is shared 284 +.>Detailed interactions between 2-4 and RBD are discussed and comparative analysis is shown in fig. 28. In summary, FIG. 18A shows that mAb 2-4 neutralizes SARS-CoV-2 probably due toLocking the RBD in the downward conformation also blocks ACE2 passage.

Next, we also performed 3D cryo-electron microscopy reconstruction of Fab of 4-8 (for NTD) complexed with S trimer (fig. 21; fig. 29). Two main particle classes were observed-one is a 3-Fab bound complex, where all RBDs are inAt resolution "down" (FIG. 18B), and the other is a 3-Fab-bound complex, where one RBD is at +.>Resolution "up" (fig. 30). However, molecular motion prevents visualization of interactions at high resolution. Nevertheless, the densities in figures 4-8 reveal the overall position of antibody chains targeting NTD and help determine the outcome of antibody competition experiments. It is not clear how this binding to the NTD tip results in SARS-CoV-2 neutralization.

Third, 2-43 Fab complexed with S trimerResolution reconstruction (FIG. 21; FIG. 31) reveals three bound Fab's, each targeting a quaternary epitope at the top of the spike, including the RBD of one promoter (promoter) and the NTD of the other promoter (FIG. 4C), consistent with epitope mapping results (FIGS. 26 and 17B). In view of these findings, the inability of 2-43 to bind to S trimer by ELISA may be an artifact of the assay format, as the mAb did bind to cell surface and spikes expressed in the cryo-electron microscopy study. FIG. 18C shows that 2-43 can block SARS-CoV-2 infection by blocking the site necessary for ACE2 binding.

Using these three frozen EM reconstructions, we mapped epitopes of our various SARS-CoV-2 neutralizing mAbs onto the spike surface using the Ween (Venn) diagram of FIG. 17B (FIG. 18D). This is clearly a rough approximation, as the antibody footprint is much larger than the area occupied by the mAb number. However, the spatial relationship of antibody epitopes should be reasonably represented by fig. 18D.

Discussion of the invention

We have found a group of SARS-CoV-2 neutralizing mAbs that are not only potent but also diverse. Nine of these antibodies could neutralize real virus in vitro at concentrations below 9ng/mL (fig. 16B), including 4 for RBD, 3 for NTD, and 2 for nearby quaternary epitopes. Surprisingly, many of these mabs had V (D) J sequences close to germline sequences without extensive somatic hypermutation (fig. 23E), a finding that suggests vaccine development. The RBD specific mAbs (e.g., 2-15, 2-7, 1-57, and 1-20) that were most effective were comparable to the recently reported antibodies ^7,8,10,16-20 Including those antibodies with high potency ^9,11,21,22 . From epitope mapping studies, the mAb against the top of RBD competed strongly with ACE2 binding and effectively neutralized the virus, while the mAb against the side surface of RBD does not compete with ACE2 and is less neutralizing (fig. 3B and 4D). non-RBD neutralizing mabs we collect were unprecedented because such antibodies were only sporadically reported and were much less potent ^22-24 . The most effective of these mabs are those directed against patches on NTD (e.g., 2-17, 5-24, and 4-8) or those that overlap patches on NTD (2-51) (fig. 3B and 4D). It is not clear how mabs against NTD block SARS-CoV-2 infection and why their neutralization spectra differ from antibodies against RBD (fig. 16B). However, vaccine strategies that do not include NTD will not induce an important class of virus neutralizing antibodies.

The separation of the two mabs (2-43 and 2-51) against epitopes that do not map to RBD and NTD was also unprecedented. The 2-43 cryo-electron microscope binding to S trimer has demonstrated that its epitope is quaternary in nature, spanning from the top of RBD to the top of the adjacent NTD (fig. 18C). It is also useful to know the epitope of 2-51. In this study, we also demonstrated the first evidence of neutralizing mAb (4-8) binding to the NTD of the viral spike by cryo-electron microscopy (FIG. 18B), and another high resolution structure of mAb (2-4) binding to RBD (FIG. 18A). Overall, these findings will help to understand how antibodies bind and neutralize SARS-CoV-2.

The efficacy and diversity of our SARS-CoV-2 neutralizing mAb may be attributed toIn the selection of the patient. Previously, we observed that severe infected individuals generated a more robust virus neutralizing antibody response ²⁵ . If patient 2 is not included, the five most neutralizing mabs will be lost. The diversity of our antibodies is also partly due to the selection of S trimers for sorting from memory B cells, while most groups focus on the use of RBDs ^{7,9-11,16-19,21} . The nature of this diverse collection of mabs enabled us to observe that all of the effective SARS-CoV-2 neutralizing antibodies described so far were directed against the top of the viral spike. Undoubtedly, RBD and NTD are very immunogenic. Neutralizing antibodies against the S trimer dry region remain to be discovered. In summary, we believe that our several monoclonal antibodies with excellent virus neutralization activity are candidates for development as a method of treating or preventing SARS-CoV-2 infection.

Method

Expression and purification of SARS-CoV-2 protein

Jason McLellan-friendly provides a mammalian expression vector encoding the extracellular domain of SARS-CoV-2S trimer and a vector encoding the RBD fused to SD1 at the N-terminus and followed by an mFc tag and an 8xHis tag fusion to the HRV-3C protease cleavage site at the C-terminus ⁴ . SARS-CoV-2NTD (aa 1-290) with HRV-3C protease cleavage site, mFc tag and 8XHis tag at C-terminus was also cloned into mammalian expression vector pCAGGS. Each expression vector was transiently transfected into Expi293 cells using 1mg/mL polyethylenimine (Polysciences). Five days after transfection, S trimer was purified using Strep-Tactin XT resin (Zymo Research) and RBD-mFc and NTD-mFc were purified using protein A agarose (ThermoFisher Scientific). To obtain RBD-SD1 and NTD, the C-terminal mFc and 8XHis tags were removed by HRV-3C protease (Millipore-Sigma), then purified using Ni-NTA resin (Invitrogen), then protein A agarose was used.

S trimer specific B cell sorting and single cell BCR sequencing

Peripheral blood mononuclear cells from five patients and one healthy donor were used at ambient temperature with LIVE/DEAD ^TM Yellow dead cell staining kit (Invitrogen) can be fixed for 20 minutes and then stained with RPMI-1640 was washed with complete medium and incubated with 10. Mu.g/mL S trimer for 45 min at 4 ℃. The cells were then washed again and incubated with a mixture of flow cytometry and labeled antibodies at 4℃for 1 hour containing CD3 PE-CF594 (Bidi Biosciences), CD19 PE-Cy7 (Biolegend), CD20 APC-Cy7 (Biolegend), igM V450 (BD Biosciences), CD27 PerCP-Cy5.5 (BD Biosciences), anti-His PE (Biosciences) and human Hashtag 3 (Biolegend). The stained cells were then washed, resuspended in RPMI-1640 complete medium, and S trimer specific memory B cells (CD 3-CD19+CD27+S trimer+live single lymphocytes) were sorted. The sorted cells were mixed with monocytes from the same donor, labeled with tag 1, and loaded into a 10X Chromium chip of the 5' single cell immunospectral analysis (10X Genomics) of the university of Columbia human immunomonitoring center (HIMC; RRID: SCR_ 016740). Library preparation and quality control were performed according to the manufacturer's protocol and sequenced on a NextSeq 500 sequencer (Illumina).

Identification of S trimer-specific antibody transcripts

For each sample, full-length antibody transcripts were assembled using VDJ modules in Cell range (version 3.1.0, 10X Genomics) with default parameters and GRCh38 genome as references. To identify cells from antigen classification, we first calculated copies of all tags (hashatag) in each Cell from Illumina NGS raw reads using a counting module in Cell range. High confidence antigen-specific cells were identified as follows. Briefly, depending on the number of copies of the tag observed, the cell must contain more than 100 copies of the antigen class-specific tag to qualify as an antigen-specific cell. Since the tag can be detached from the cells and bind to different populations of cells in the sample mixture, each cell typically has a specific tag that is classified and incorporated. To enrich for truly antigen-specific cells, the copy number of a specific tag must be at least 1.5 times higher than the copy number of an unspecific tag. Low quality cells were identified and removed using the Cell calling algorithm in Cell range. Cells that do not have productive H and L chain pairs are excluded. If the cell contains more than two H or/and L chain transcripts, transcripts with less than 3 unique molecular identifiers are removed. Cells with identical H and L chain sequences, which can be caused by mRNA leakage, are pooled into one cell. Additional filters are applied during the antibody gene annotation process to remove low quality cells and/or transcripts.

Antibody transcript annotation and selection criteria

Treatment of antigen-specific antibody transcripts for quality control and annotation using our bioinformatics pipeline SONAR ²⁶ . Briefly, BLAST is used ²⁷ According to the information System (IMGT) database from International ImmunoGenetics ^26,28 The obtained germline gene database customization parameters assign V (D) J genes to each transcript. CDR3 was identified using a conserved second cysteine in the V region and the WGXG (H chain) or FGXG (L chain) motif (X represents any amino acid) in the J region based on BLAST alignment of the V and J regions. For H chain transcripts, constant domain 1 (CH 1) sequences were used to assign isotypes using BLAST according to the default parameters of the human CH1 gene database obtained from IMGT. The BLAST E value threshold of 1E-6 was used to find important isotype assignments and the CH1 allele with the lowest E value was used. Sequences outside the V (D) J region are removed, excluding transcripts containing incomplete V (D) J or/and frame shift. We then used CLUSTALO ²⁹ Each remaining transcript was aligned with the assigned germline V gene and somatic hypermutation levels were calculated.

To select representative antibodies for functional characterization, we first used userch ³⁰ All antibodies were clustered as follows: the heavy chain V and J genes are equally distributed, CDRH3 is identical in length, and CDRH3 identity is higher than 0.9. For each cluster, cells with the same light chain V and J gene assignments were grouped into one clone. All clone assignments were checked manually. Then we calculated the clone size for each clone, each clone selecting a pair of H and L chains for antibody synthesis. For clones with multiple members, the member with the highest level of somatic hypermutation was selected for synthesis. For cells with multiple high quality H or L chains (possibly from a doublet), we synthesized all H and L chain combinations.

S trimer specific antibody library analysis

Since 88% of the S trimer-specific antibodies are of the IgG isotype, we characterized the pool with that from three healthy donors ³¹ Is compared (17,243H chains, 27,575 kappa L chains, 20,889 lambda L chains). Library data from three healthy donors were pooled and annotated using sonor, the procedure being the same as described above.

Antibody expression and purification

For each antibody, the variable genes were optimized for human cell expression and synthesized by GenScript. VH and VL were inserted into plasmids encoding the H chain and L chain constant regions, respectively (gWiz or pcdna 3.4). Monoclonal antibodies were expressed in an Expi293 (thermo fisher, a 14527), by co-transfection of H-and L-chain expression plasmids using polyethylenimine, and in a shaker at 37 ℃ at 125RPM and 8% co ₂ And (5) culturing. On day 3 post-transfection, 400. Mu.L of supernatant was collected to screen for binding to S trimer and RBD by ELISA and to neutralize SARS-CoV-2 pseudovirus and real virus. Supernatants were also collected on day 5 to purify the antibodies by affinity chromatography by rProtein ASepharose (GE, 17-1279-01).

Pseudovirus production

Recombinant Indiana VSV (rVSV) expressing SARS-CoV-2 spike was generated as described previously ³² _, ³³ . HEK293T cells were grown to 80% confluence prior to transfection with pCMV3-SARS-CoV-2-spike (Sino Biological) using fugenee 6 (Promega). Cells at 37℃and 5% CO ₂ The culture was carried out overnight under the conditions. The next day, the medium was removed and cells in DMEM were infected with VSV-G pseudotyped Δg-luciferase (g×Δg-luciferase, kerafast) at a MOI of 3 for 1 hour, and then washed 3 times with 1X DPBS. DMEM supplemented with 2% fetal bovine serum and 100i.u./mL penicillin and 100 μg/mL streptomycin was added to the inoculated cells and cultured overnight as described above. The next day the supernatant was collected and clarified by centrifugation at 300g for 10 minutes, then sub-packaged and stored at-80 ℃.

Pseudovirus neutralization

By mixing pseudoviruses with serial dilutions of heat-inactivated plasma and plasma The clear or purified antibodies were incubated together for neutralization assays and scored by a decrease in luciferase gene expression. Briefly, vero E6 cells (ATCC) were used at 2X 10 per well ⁴ The concentration of individual cells was seeded in 96-well plates. The following day, pseudoviruses were incubated with serial dilutions of test samples in duplicate or triplicate for 30 minutes at 37 ℃. The mixture was added to the cultured cells and incubated for an additional 24 hours. Luminescence was measured by the briitelite plus reporter gene detection system (PerkinElmer). IC (integrated circuit) ₅₀ Defined as the dilution of 50% decrease in relative light units compared to the virus control wells (virus + cells) after subtraction of the background of the cell-only control group. IC50 values were calculated using nonlinear regression in GraphPad Prism 8.0.

True SARS-CoV-2 neutralization

Supernatants containing expressed mAbs were diluted 1:10 and 1:50 in EMEM containing 7.5% inactivated fetal bovine serum and incubated with authentic SARS-CoV-2 (strain USA-WA1/2020; MOI 0.1) for 1 hour at 37 ℃. After incubation, the mixture was transferred to monolayer Vero E6 cells cultured overnight. After incubating the cells with the mixture at 37 ℃ for 70 hours, each well was scored for cytopathic effect (CPE) caused by infection, ranging from 0 to 4, to indicate the extent of viral inhibition. Semi-quantitative representations of inhibition of each antibody-containing supernatant at a dilution of 1:50 are shown in the bottom panel of figure 15B, the neutralization level ranged from (-) (none to (+++) and (3) completely neutralizing).

An end point dilution assay in the form of a 96-well plate was performed to measure the neutralizing activity of the selected purified mAb. Briefly, each antibody was serially diluted (5-fold dilution) starting from 20. Mu.g/mL. Each mAb dilution was incubated with SARS-CoV-2 in triplicate at an MOI of 0.1 in EMEM containing 7.5% inactivated fetal bovine serum at 37℃for 1 hour. After incubation, the virus-antibody mixture was transferred to monolayer Vero-E6 cells for overnight growth. Cells were incubated with the mixture for 70 hours. CPE for each well was scored visually blindly by two independent observers. The results were then converted to percent neutralization at the given mAb concentration and the mean ± SEM were plotted using a five-parameter dose response curve in GraphPad Prism 8.0.

Epitope mapping using ELISA

50 ng/well of S trimer, 50 ng/well of RBD and 100 ng/well of NTD were plated on ELISA plates overnight at 4 ℃. The ELISA plates were then blocked with 300. Mu.L of blocking buffer (1% BSA and 10% calf serum (BCS) (Sigma)) in PBS for 2 hours at 37 ℃. Thereafter, the supernatant of the antibody transfected or purified antibody was serially diluted with a dilution buffer (containing 1% BSA and 20% BCS in PBS) and incubated at 37℃for 1 hour. Next, 100 μl of 10,000-fold diluted peroxidase AffiniPure goat anti-human IgG (h+l) antibody (Jackson ImmunoResearch) was added to each well and incubated for 1 hour at 37 ℃. Between each step, the plates were washed with PBST (0.5% Tween-20 in PBS). Finally, TMB substrate (Sigma) was added and incubated, and then the reaction was stopped using 1M sulfuric acid. Absorbance was measured at 450 nm.

For the competition ELISA, purified mabs were biotinylated using a one-step antibody biotinylation kit (meitian gentle biotechnology (Miltenyi Biotec)) as suggested by the manufacturer and purified using a 40K MWCO desalting column (sameifeishi technology (ThermoFisher Scientific)). mu.L of serially diluted competitor antibody was added to the S trimer pre-coated ELISA plate, followed by 50. Mu.L of biotinylated antibody at a concentration that reached an OD450 reading of 1.5 without competitor antibody. Plates were incubated at 37℃for 1 hour, 100. Mu.L of 500-fold diluted Avidin-HRP (ThermoFisher Scientific) was added to each well, and incubated at 37℃for an additional 1 hour. Between each of the preceding steps, the plates were washed with PBST. The plate was then developed with TMB and the absorbance read at 450nm after the reaction stopped.

For the ACE2 competition ELISA, 100ng of the ACE2 protein (Abcam) was immobilized on a plate overnight at 4 ℃. Unbound ACE2 was washed out with PBST and the plates were then blocked. After washing, 100ng of S trimer in 50. Mu.L of dilution buffer was added to each well, followed by an additional 50. Mu.L of serial dilutions of competing antibody, and the plates were incubated for 1 hour at 37 ℃. ELISA plates were washed 4 times with PBST, then 100. Mu.L of 2000-fold diluted anti-Streptococcus HRP (Millipore Sigma) was added to each well and left at 37℃for an additional 1 hour. The plate was then washed, developed with TMB, and the absorbance read at 450nm after the reaction was stopped.

For all competitive ELISA experiments, the relative binding of biotinylated antibody or ACE2 to S trimer in the presence of competitor was normalized by comparison to a control without competitor. The relative binding curve and area under the curve (AUC) were generated by fitting a nonlinear five-parameter dose response curve in GraphPad Prism 8.0.

Cell surface competitive binding assay

Expi293 cells were co-transfected with vectors encoding pRRL-cPPT-PGK-GFP (Addgene) and pCMV3-SARS-CoV-2 (2019-nCoV) Spike (Sino Biological) at a 1:1 ratio. Two days after transfection, the cells were incubated with a mixture of biotinylated mAb 2-43 (0.25. Mu.g/mL) and serial dilutions of competing antibodies for 1 hour at 4 ℃. mu.L of diluted APC-streptavidin (Biolegend) was then added to the cells and incubated at 4℃for 45 minutes. Prior to each step, cells were washed 3 times with FACS buffer. Finally, the cells were resuspended and the 2-43 binding to cell surface S trimer was quantified on an LSRII flow cytometer (BD Biosciences). The average fluorescence intensity of APCs in GFP positive cells was analyzed using FlowJo and the relative binding of 2-43 to S trimer in the presence of competitors was calculated as a percentage of the average fluorescence intensity compared to a competitor-free control.

Data collection and processing for cryo-electron microscope

A final concentration of 2mg/ml SARS-CoV-2S trimer was incubated with a 6-fold molar excess of each spike monomer of the antibody Fab fragment in 10mM Tris-HCl, 150mM NaCl and 0.005% n-dodecyl- β -D-maltoside (DDM) for 30 minutes. mu.L of the sample was incubated on a C-flat 1.2/1.3 carbon grid for 30 seconds and then vitrified using a Leica EM GP plunger freezer. The data were collected on a Titan Krios electron microscope, operating at 300kV, equipped with a Gatan K3 direct detector and an energy filter using the Leginon software package 34.Is not less than a total electron of (a)Fluence was graded over 40 frames with a total exposure time of 2 seconds. Magnification of 81,000 times makes the pixel size +.>And a defocus range of-0.4 to-3.5 μm is used. All treatments were done using crysparc v 2.14.235. The original film is aligned and dose weighted using patch motion correction and CTF is estimated using patch CTF estimation. A small portion of approximately 200 micrographs was selected using a blob picker (blob picker), then the particle pick was 2D classified and manually curated (manual curation) and used to train Topaz neural networks ³⁶ . The network was then used to pick particles from the remaining micrographs, which were extracted with a frame size of 384 pixels. For the Fab 2-4 dataset, 2D classification, de novo modeling (ab initio modelling) and 3D heterogeneous refinement showed 83,927 particles with three 2-4Fab binding, with one RBD per 2-4Fab binding. We used non-uniform refinement with forced C3 symmetry to construct these particles, resulting in +. >The graph is determined by the gold standard FSC. In view of the relatively low resolution of the RBD-Fab junction, a significantly improved density is achieved using masking local refinement> A drawing. Masking local refinement of the S trimer remainder results in +.>And (5) reconstructing. Using UCSF Chimera ³⁷ The vop maximum function of (c) aligns and combines the two local refinements. This operation is repeated for the half-graph used and the refinement mask from the global non-uniform refinement to calculate 3DFSC38 and obtain +.>Is used for the resolution estimation of (a). All figures have been submitted to the EMDB with an ID of EMD-22156. For the Fab 4-8 dataset, image pre-processing and particle pickup were performed as described above. The 2D classification, de novo modeling and 3D heterogeneous classification showed 47,555 particles with 3 Fab binding, with each Fab binding one NTD and all 3 RBDs in a downward conformation. Although this particle stack is refined to +.>But significant molecular movement prevented the visualization of Fab epitopes at high resolution (EMD-22159). Furthermore, 105,278 particles showed binding to 3 Fabs, but 1 RBD was in the up-conformation. These particles were refined to +. >And has the same conformational flexibility as the full RBD-down particle. This flexibility is visualized using 3D variability analysis in crysparc. For the following purposesImage preprocessing and particle pickup were performed as described above, but motion correction was performed using a MotionCor 239. 2D classification, de novo modeling and 3D heterogeneous classification showed 18,884 particles with 3 Fab binding, which were refined to +.>Resolution 601 (EMD-22157).

Model fitting for cryoelectron microscope

The initial homology model for 2-4Fab was performed using Schrodinger Release 2020-2:bioluminate ⁴⁰ Constructed by the method. The RBD is initially modeled using coordinates in PDB ID 6W 41. The remainder of the S trimer was modeled using coordinates in PDB ID 6 VSB. These models were docked into the consensus diagram using Chimera. Then using ISOLDE 1.0b541 and COOT 0.8.9.242And fits the model interactively using real-space refinements in Phenix 1.1843. In the case where the side chains are not visible in the experimental data, they are truncated to alanine. Using molprobit ⁴⁴ And EMringer ⁴⁵ And (5) performing verification. The model has been submitted to PDB ID 6 XEY. The graph is made using ChimeraX ⁴⁶ The preparation is carried out.

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EXAMPLE 11 effective neutralizing monoclonal antibodies

FIGS. 32A-B show monoclonal antibodies 2-15mut. FIG. 32A shows virus neutralization using 2-15mut antibodies. FIG. 32B shows the efficacy of 2-15mut antibodies. The 2-15mut antibody showed an IC of less than 0.00064. Mu.g/ml ₅₀ IC with concentration of 0.011 μg/ml ₉₀ Concentration.

FIGS. 33A-B show selection mutants of the 4-8 monoclonal antibodies. FIG. 33A shows neutralization of antibodies 4-8 (39/51) and 4-8 (39/51/57). FIG. 33B shows the efficacy of antibodies 4-8 (39/51) and 4-8 (39/51/57). The 4-8 (39/51) antibody showed an IC of 0.002. Mu.g/ml ₅₀ IC with concentration and 0.108. Mu.g/ml ₉₀ Concentration. The 4-8 (39/51/57) antibody showed an IC of 0.003. Mu.g/ml ₅₀ IC at a concentration of 0.014. Mu.g/ml ₉₀ Concentration.

EXAMPLE 12 identification and characterization of Cross-neutralizing monoclonal antibodies 2-36

SARS-CoV-2 is phylogenetically closely related to SARS-CoV, which caused human epidemics in 2002-2003. SARS-CoV and SARS-CoV-2 have 76% amino acid identity in their spike proteins, which increases the likelihood of a conserved immunogenic surface on these antigens. Many human monoclonal antibodies have been shown to target SARS-CoV-2 spike protein, but cross-neutralizing antibodies are relatively unusual in patients with COVID-19. There are also some SARS-like viruses, including sand Bei Bingdu (sarbecoviruses) common to some humans and animals in bats or pangolins, which can infect human cells and thus potentially cause the next pandemic.

Our invention requires isolation and characterization of monoclonal antibodies that can neutralize not only SARS-CoV-2 and SARS-CoV, but also sand Bei Bingdu in bats and pangolins. Such cross-neutralizing antibodies are of great value to understand how to provide broader protection against human SARS-like viruses, including human and animal co-morbid sand Bei Bingdu that is widely found within bats, pangolins, and the like.

To find antibodies with cross-neutralizing activity, we screened anti-SARS-CoV antibodies from patients with convalescence of COVID-19. SARS-CoV-2 spike-specific antibodies were isolated by single B cell sorting and 10X genomics sequencing, then antibodies were synthesized and the transfected supernatants were used to screen anti-SARS-CoV antibodies using SARS-CoV pseudovirus. While most antibodies did not or had very weak neutralizing activity, antibody 2-36 was found to be an effective anti-SARS-CoV neutralizer (FIG. 35).

Next, we first characterized the antibody by ELISA testing for binding to SARS-CoV-2 and SARS-CoV spike using CR3022 (an antibody previously reported to have cross-activity as a control). We demonstrated that antibody 2-36 binds to SARS-CoV-2 and SARS-CoV spike (FIGS. 36A-B). We then measured the binding affinity of antibody 2-36 to these spikes by SPR, and we further demonstrated that antibody 2-36 could bind to SARS-CoV-2 (FIG. 37A) and SARS-CoV (FIG. 37B) spikes, but had a higher affinity for SARS-CoV-2 spikes. As a control, SARS-CoV-2 specific antibody 2-4 bound only to SARS-CoV-2 spike and not to SARS-CoV spike (FIGS. 37C-D).

We then tested the neutralizing activity of antibody 2-36 against SARS-CoV-2 and SARS-CoV Sex, the result shows that antibody 2-36 can effectively neutralize SARS-CoV-2, IC ₅₀ 0.004. Mu.g/ml (FIG. 38A). Antibody 2-36 also neutralized SARS-CoV, but was less potent, IC ₅₀ 0.386. Mu.g/ml (FIG. 38B). Control antibody CR3022 neutralized SARS-CoV, but had very little effect on SARS-CoV-2; antibody 2-4 only neutralized SARS-CoV-2 (FIGS. 38A-B).

We also tested antibodies 2-36 against a set of other coronaviruses, including MERS-CoV, human common cold coronavirus 229E, and some bat and pangolin coronaviruses (fig. 39A), including 6 of the SARS-CoV related lineages (e.g., WIV1, SHC014, LYRa11, rs7327, rs4231, rs 4084) and 3 of the SARS-CoV-2 related lineages (e.g., raTG13, GDPPangolin, GXPangolin). Pseudoviruses with these coronavirus spikes were prepared and tested. As shown in FIG. 39B, antibodies 2-36 neutralized SARS-CoV and SARS-CoV-2 and its related lineage viruses, but antibodies 2-36 were unable to neutralize MERS-CoV and 229E, indicating its breadth.

We also solved the cryo-electron microscope structure of the complex of antibody 2-36 and SARS-CoV-2 spike trimer (fig. 40A), which shows in detail how the antibody interacts with the spike (fig. 40B). Antibodies 2-36 bind to one side of RBD and can compete with ACE2 for binding to RBD, although their epitopes do not overlap with the ACE2 binding site (fig. 40C). The 2-36 epitope on spike is highly conserved among SARS-CoV-2, SARS-CoV and other SARS-associated coronaviruses (FIG. 40D), which can explain why 2-36 exhibits such broad neutralization properties.

Thus, antibody 2-36 may serve as a starting point for antibodies engineered to be more effective against a variety of bat coronaviruses with human pandemic potential. Such antibodies may be further developed and placed in national reserves for pandemic. Furthermore, we define a conserved site in the spike that can be used to develop a broad protective vaccine against bat coronaviruses with pandemic potential.

EXAMPLE 13 monoclonal antibody nos. 2-36

Antibodies 2-36 are monoclonal antibodies that neutralize SARS-CoV-2 variants, SARS-CoV and other beta coronaviruses. To screen monoclonal antibodies with cross-neutralizing activity against other beta coronaviruses200 mAbs isolated from patients with convalescence of COVID-19 (Liu et al, nature 2020) were tested on SARS-CoV pseudovirus. Monoclonal antibody 2-36 not only uses IC ₅₀ About 0.04. Mu.g/mL neutralizes SARS-CoV-2 pseudovirus and the actual virus at an IC50 of about 0.1. Mu.g/mL, and also shows neutralizing activity against SARS-CoV, although less potent (IC 50 for pseudovirus about 0.2. Mu.g/mL, IC50 for actual virus about 7.5. Mu.g/mL). Monoclonal antibody 2-36 blocks the binding of SARS-CoV-2 spike trimer to ACE 2. Conservation analysis of SARS-CoV-2, SARS-CoV and part of the inter-strain Receptor Binding Domain (RBD) of bats and pangolins shows that the 2-36 binding site is highly conserved. Antibody 2-36 was tested on different SARS-CoV-2 variants of interest/interest (including b.1.1.7, b.1.351, p.1, b.1.526, r.1, b.1.429) and various single mutations at high cycling frequencies. The results indicate that antibody 2-36 can neutralize all of these variants/mutations without any significant loss of activity. In addition, in addition to SARS-CoV-2 and SARS-CoV, antibody 2-36 can neutralize some bat and pangolin coronaviruses that can enter host cells using human ACE2 as a receptor. These coronaviruses include, but are not limited to, raTG13, WIV1, SHC014, gdpanglin and GXPangolin viruses. Antibody 2-36 escape mutations were selected by in vitro passaging of SARS-CoV-2 in Vero E6 cells in the presence of 2-36 and demonstrated that K378T in spike RBD (very low frequency in nature) resulted in viral resistance. Together, these results, 2-36 represent a promising candidate for strategic reserves for the prevention and treatment of possible diseases caused by pre-emergent SARS-associated coronavirus.

FIGS. 96A-D show the neutralization of SARS-like coronavirus with monoclonal antibodies 2-36 using hACE 2. FIG. 96A shows virus neutralization with 2-36 antibodies. Fig. 96B shows virus neutralization with S309 antibody. FIG. 96C shows virus neutralization with COVA1-16 antibodies. Figure 96D shows virus neutralization with CR3022 antibody.

Examples of bispecific antibodies

Bispecific antibodies with 1400 arms or a32 arms as described herein indicate that those arms of the bispecific antibody are not specific for nor bind to spike proteins of SARS-CoV-2 virus. These antibodies provide an "unrelated" control to demonstrate the need for avidity by controlling the "two" arms of the bispecific antibody. The construct with these unrelated controls was called the single arm IgG control. 1400 (PGDM 1400) is a neutralizing monoclonal antibody against Human Immunodeficiency Virus (HIV). A32 is a non-neutralizing monoclonal antibody against HIV. The same control antibodies were used throughout the study. AbX and AbY are controls, x=pgdm 1400 (HIV neutralizing antibody). Y=a32 (HIV neutralizing antibody). Since they do not neutralize SARS-CoV-2 or any variant, they are used herein to construct bispecific antibodies, one of which is resistant to SARS-CoV-2 and the other of which does not have any activity against SARS-CoV-2. Thus, these control antibodies enable one to accurately attribute the contribution of each arm to the activity of the bispecific antibody.

Example 14

FIG. 42 shows that the 2-17/1-57 bispecific antibody against SARS-CoV-2 does not enhance efficacy compared to the corresponding single arm counterpart. Ab X and Ab Y are single arm independent controls. X=pgdm 1400 (HIV neutralization Ab). Y=a32 (HIV neutralization Ab). IC of 2-17/1-57 bispecific antibody ₅₀ Is 0.023 μg/mL. IC of 2-17/1-57 bispecific antibody ₉₀ 0.167. Mu.g/mL. Ab X/1-57 control IC ₅₀ 0.014 μg/mL. Ab X/1-57 control IC ₉₀ Is 0.424. Mu.g/mL. IC for 2-17/Ab Y control ₅₀ 1.451. Mu.g/mL. IC for 2-17/Ab Y control ₉₀ Is 3.572 mug/mL.

FIG. 43 shows the effective neutralization of bispecific antibodies 2-17/2-7 against SARS-CoV-2 as compared to the corresponding single arm counterparts. Ab X and Ab Y are single arm independent controls. X=pgdm 1400 (HIV neutralization Ab). Y=a32 (HIV neutralization Ab). IC of 2-17/2-7 bispecific antibody ₅₀ 0.008 μg/mL. IC of 2-17/2-7 bispecific antibody ₉₀ Is 0.025 μg/mL. Ab X/2-7 control IC ₅₀ Is 0.028 mug/mL. Ab X/2-7 control IC ₉₀ 0.162. Mu.g/mL. IC for 2-17/Ab Y control ₅₀ 1.451. Mu.g/mL. IC of 2-17/Ab Y bispecific antibody ₉₀ Is 3.572 mug/mL.

FIG. 44 shows the effective neutralization of bispecific antibodies 2-17/2-15 against SARS-CoV-2 as compared to the corresponding single arm counterparts. Ab X and Ab Y are single arm independent controls. X=pgdm 1400 (HIV neutralization Ab). Y=a32 (HIV neutralization Ab). IC of 2-17/2-15 bispecific antibody ₅₀ Is 0.006 μg/mL. IC of 2-17/2-15 bispecific antibody ₉₀ 0.085 μg/mL. Ab X/2-15 control IC ₅₀ 0.043. Mu.g/mL. Ab X/2-15 bispecific antibody IC ₉₀ 0.232 μg/mL. IC for 2-17/Ab Y control ₅₀ 1.451. Mu.g/mL. IC for 2-17/Ab Y control ₉₀ Is 3.572 mug/mL.

FIG. 45 shows the effective neutralization of bispecific antibody 2-17/2-30 against SARS-CoV-2 as compared to the corresponding single arm counterpart. Ab X and Ab Y are single arm independent controls. X=pgdm 1400 (HIV neutralization Ab). Y=a32 (HIV neutralization Ab). IC of 2-17/2-30 bispecific antibody ₅₀ Is 0.065 mug/mL. IC of 2-17/2-30 bispecific antibody ₉₀ Is 0.598 mug/mL. Ab X/2-30 control IC ₅₀ 0.349 μg/mL. Ab X/2-30 control IC ₉₀ Is 4.939 mug/mL. IC for 2-17/Ab Y control ₅₀ 1.451. Mu.g/mL. IC for 2-17/Ab Y control ₉₀ Is 3.572 mug/mL.

FIG. 46 shows the effective neutralization of bispecific antibody 2-17/4-20 against SARS-CoV-2 as compared to the corresponding single arm counterpart. Ab X and Ab Y are single arm independent controls. X=pgdm 1400 (HIV neutralization Ab). Y=a32 (HIV neutralization Ab). IC of 2-17/4-20 bispecific antibody ₅₀ 0.030 μg/mL. IC of 2-17/4-20 bispecific antibody ₉₀ 0.164. Mu.g/mL. Ab X/4-20 control IC ₅₀ 0.084. Mu.g/mL. Ab X/4-20 control IC ₉₀ Is 3.066 mug/mL. IC for 2-17/Ab Y control ₅₀ 1.451. Mu.g/mL. IC for 2-17/Ab Y control ₉₀ Is 3.572 mug/mL.

FIG. 47 shows the effective neutralization of bispecific antibody 5-24/1-57 against SARS-CoV-2 as compared to the corresponding single arm counterpart. Ab X and Ab Y are single arm absent And (5) closing a control. X=pgdm 1400 (HIV neutralization Ab). Y=a32 (HIV neutralization Ab). IC of 5-24/1-57 bispecific antibody ₅₀ 0.004. Mu.g/mL. IC of 5-24/1-57 bispecific antibody ₉₀ 0.046. Mu.g/mL. Ab X/1-57 control IC ₅₀ 0.014 μg/mL. Ab X/1-57 control IC ₉₀ Is 0.424. Mu.g/mL. IC for 5-24/Ab Y control ₅₀ 0.137. Mu.g/mL. IC for 5-24/Ab Y control ₉₀ 2.320. Mu.g/mL.

FIG. 48 shows the effective neutralization of bispecific antibodies 5-24/2-15 against SARS-CoV-2 as compared to the corresponding single arm counterparts. Ab X and Ab Y are single arm independent controls. X=pgdm 1400 (HIV neutralization Ab). Y=a32 (HIV neutralization Ab). IC of 5-24/2-15 bispecific antibody ₅₀ 0.004. Mu.g/mL. IC of 5-24/2-15 bispecific antibody ₉₀ 0.033 μg/mL. Ab X/2-15 control IC ₅₀ 0.043. Mu.g/mL. Ab X/2-15 control IC ₉₀ 0.232 μg/mL. IC for 5-24/Ab Y control ₅₀ 0.137. Mu.g/mL. IC for 5-24/Ab Y control ₉₀ 2.320. Mu.g/mL.

FIG. 49 shows that bispecific antibody 5-24/4-20 against SARS-CoV-2 does not enhance efficacy (IC 50) as compared to the corresponding single arm counterpart. Ab X and Ab Y are single arm independent controls. X=pgdm 1400 (HIV neutralization Ab). Y=a32 (HIV neutralization Ab). IC of 5-24/4-20 bispecific antibody ₅₀ Is 0.063 mug/mL. IC of 5-24/4-20 bispecific antibody ₉₀ Is 0.222 mug/mL. Ab X/4-20 control IC ₅₀ 0.084. Mu.g/mL. Ab X/4-20 control IC ₉₀ Is 3.066 mug/mL. IC for 5-24/Ab Y control ₅₀ 0.137. Mu.g/mL. IC for 5-24/Ab Y control ₉₀ 2.320. Mu.g/mL.

FIG. 50 shows the effective neutralization of bispecific antibody 1-20/1-68 against SARS-CoV-2 as compared to the corresponding single arm counterpart. Ab X and Ab Y are single arm independent controls. X=pgdm 1400 (HIV neutralization Ab). Y=a32 (HIV neutralization Ab). IC of 1-20/1-68 bispecific antibody ₅₀ 0.010. Mu.g/mL. IC of 1-20/1-68 bispecific antibody ₉₀ 0.157 μg/mL.1-20/Ab Y control IC ₅₀ 0.078 μg/mL.1-20/Ab Y control IC ₉₀ 1.241. Mu.g/mL. Ab X/1-68 control IC ₅₀ Is 0.035. Mu.g/mL. Ab X/1-68 control IC ₉₀ At 0.332. Mu.g/mL。

FIG. 51 shows the effective neutralization of bispecific antibodies 1-20/2-17 against SARS-CoV-2 as compared to the corresponding single arm counterparts. Ab X and Ab Y are single arm independent controls. X=pgdm 1400 (HIV neutralization Ab). Y=a32 (HIV neutralization Ab). IC of 1-20/2-17 bispecific antibody ₅₀ Is 0.025 mug/mL. IC of 1-20/2-17 bispecific antibody ₉₀ 0.213. Mu.g/mL. 1-20/Ab Y control IC ₅₀ 0.078 μg/mL.1-20/Ab Y control IC ₉₀ 1.241. Mu.g/mL. Ab X/2-17 control IC ₅₀ Is 0.380 mug/mL. Ab X/2-17 control IC ₉₀ Is 3.912 mug/mL.

FIG. 52 shows the effective neutralization of bispecific antibody 1-20/4-18 against SARS-CoV-2 as compared to the corresponding single arm counterpart. Ab X and Ab Y are single arm independent controls. X=pgdm 1400 (HIV neutralization Ab). Y=a32 (HIV neutralization Ab). IC of 1-20/4-18 bispecific antibody ₅₀ 0.009. Mu.g/mL. IC of 1-20/4-18 bispecific antibody ₉₀ 0.043. Mu.g/mL. 1-20/Ab Y control IC ₅₀ 0.078 μg/mL.1-20/Ab Y control IC ₉₀ 1.241. Mu.g/mL. Ab X/4-18 control IC ₅₀ 0.049. Mu.g/mL. Ab X/4-18 control IC ₉₀ Is 3.463 mug/mL.

FIG. 53 shows the effective neutralization of bispecific antibodies 1-20/5-24 against SARS-CoV-2 as compared to the corresponding single arm counterparts. Ab X and Ab Y are single arm independent controls. X=pgdm 1400 (HIV neutralization Ab). Y=a32 (HIV neutralization Ab). IC of 1-20/5-24 bispecific antibody ₅₀ 0.009. Mu.g/mL. IC of 1-20/5-24 bispecific antibody ₉₀ Is 0.035. Mu.g/mL. 1-20/Ab Y control IC ₅₀ 0.078 μg/mL.1-20/Ab Y control IC ₉₀ 1.241. Mu.g/mL. Ab X/5-24 control IC ₅₀ Is 0.500 mug/mL. Ab X/5-24 control IC ₉₀ 4.120. Mu.g/mL.

The constructed bispecific antibodies showed effective neutralization of SARS-CoV-2 at levels similar to or better than the parent antibody. Both arms of the bispecific antibody contribute to the neutralizing activity of the bispecific molecule. Bispecific antibodies are single molecules with high viral escape barriers and/or synergistic efficacy for the treatment and prevention of covd-19. The observed enhancement of activity can be extended to any engineered bispecific antibody format.

FIG. 54 shows effective neutralization against SARS-CoV-2. Bispecific antibodies with 2-17 as a component were more effective than the single arm control (2-17/a 32, a32 being a control antibody that was inactive against SARS-CoV-2), suggesting that the other arm targeting a different viral epitope together with the 2-17 arm contributed to the overall neutralization activity of the bispecific molecule. FIG. 54 shows an IC containing bispecific antibodies 2-17 ₅₀ And IC ₉₀ Values.

FIG. 55 shows the effective neutralization of SARS-CoV-2 with a bispecific antibody containing 5-24 as a component. FIG. 55 shows an IC containing bispecific antibody 5-24 ₅₀ And IC ₉₀ Values.

FIG. 56 shows the effective neutralization of SARS-CoV-2 with bispecific antibodies containing 1-20 as components. FIG. 56 shows an IC containing bispecific antibodies 1-20 ₅₀ And IC ₉₀ Values.

Example 15

FIGS. 57A-E show bispecific antibodies with 1-20 (RBD) as parent antibodies. FIG. 57A shows virus neutralization with bispecific antibodies 1-20/5-24, 1-20/4-18, 1-20/2-17 and 1-20/1-68. FIG. 57B shows virus neutralization of bispecific antibodies 1-20/5-24 as compared to control antibodies. FIG. 57C shows virus neutralization of bispecific antibodies 1-20/4-18 as compared to control antibodies. FIG. 57D shows virus neutralization of bispecific antibodies 1-20/2-17 as compared to control antibodies. FIG. 57E shows virus neutralization of bispecific antibody 1-20/1-68 as compared to control antibody. The control antibody is generated using an unrelated antibody such as PGDM1400 (Ab X) or a32 (Ab Y) instead of one arm of the bispecific antibody.

FIG. 58 shows the efficacy of bispecific antibodies with 1-20 (RBD) as parent antibodies. IC of bispecific antibody 1-20/5-24 ₅₀ 0.009. Mu.g/ml. IC of bispecific antibody 1-20/5-24 ₉₀ 0.035. Mu.g/ml. IC of bispecific antibody 1-20/4-18 ₅₀ 0.009. Mu.g/ml. IC of bispecific antibody 1-20/4-18 ₉₀ 0.043. Mu.g/ml. IC of bispecific antibody 1-20/2-17 ₅₀ 0.025. Mu.g/ml. IC of bispecific antibody 1-20/2-17 ₉₀ 0.212. Mu.g/ml. Bispecific antibodies1-20/1-68 IC ₅₀ 0.009. Mu.g/ml. IC of bispecific antibody 1-20/1-68 ₉₀ 0.028. Mu.g/ml.

FIG. 60 shows the efficacy of bispecific antibodies with 2-17 (NTD) as the parent antibody. IC of bispecific antibody 2-17/1-57 ₅₀ 0.075 μg/ml. IC of bispecific antibody 2-17/1-57 ₉₀ 0.144. Mu.g/ml. IC of bispecific antibody 2-17/2-7 ₅₀ 0.009. Mu.g/ml. IC of bispecific antibody 2-17/2-7 ₉₀ 0.027. Mu.g/ml. IC of bispecific antibody 2-17/2-15 ₅₀ 0.005 μg/ml. IC of bispecific antibody 2-17/2-15 ₉₀ 0.087. Mu.g/ml. IC of bispecific antibody 2-17/2-30 ₅₀ 0.065 μg/ml. IC of bispecific antibody 2-17/2-30 ₉₀ Is 0.598 mug/ml. IC of bispecific antibody 2-17/4-20 ₅₀ 0.029. Mu.g/ml. IC of bispecific antibody 2-17/4-20 ₉₀ 0.164. Mu.g/ml. IC of bispecific antibody 2-36/2-17 ₅₀ 0.063. Mu.g/ml. IC of bispecific antibody 2-36/2-17 ₉₀ 4.55. Mu.g/ml.

FIGS. 61A-D show bispecific antibodies with 5-24 (NTD) as the parent antibody. FIG. 61A shows virus neutralization of bispecific antibodies 5-24/4-20, 5-24/1-57, 5-24/2-15 and 1-20/5-24. FIG. 61B shows virus neutralization of bispecific antibody 5-24/2-15 as compared to control antibody. FIG. 61C shows virus neutralization of bispecific antibody 5-24/1-57 as compared to control antibody. FIG. 61D shows virus neutralization of bispecific antibody 5-24/4-20 as compared to control antibody.

FIG. 62 shows the efficacy of bispecific antibodies with 5-24 (NTD) as the parent antibody. IC of bispecific antibody 1-20/5-24 ₅₀ 0.009. Mu.g/ml. IC of bispecific antibody 1-20/5-24 ₉₀ 0.035. Mu.g/ml. IC of bispecific antibody 5-24/2-15 ₅₀ 0.003. Mu.g/ml. IC of bispecific antibody 5-24/2-15 ₉₀ 0.033. Mu.g/ml. IC of bispecific antibody 5-24/1-57 ₅₀ 0.004. Mu.g/ml. IC of bispecific antibody 5-24/1-57 ₉₀ 0.045. Mu.g/ml. IC of bispecific antibody 5-24/4-20 ₅₀ 0.062. Mu.g/ml. IC of bispecific antibody 5-24/4-20 ₉₀ 0.222. Mu.g/ml.

FIGS. 63A-D show bispecific antibodies with 2-36 (RBD) as parent antibodies. FIG. 63A shows virus neutralization of bispecific antibodies 2-36/1-68, 2-36/4-18 and 2-36/2-17. FIG. 63B shows virus neutralization of bispecific antibody 2-36/2-17 as compared to control antibody. FIG. 63C shows virus neutralization of bispecific antibody 2-36/1-68 as compared to control antibody. FIG. 63D shows virus neutralization of bispecific antibody 2-36/4-18 as compared to control antibody.

FIG. 64 shows the efficacy of bispecific antibodies with 2-36 (RBD) as parent antibody. IC of bispecific antibody 2-36/1-69 ₅₀ 0.096 μg/ml. IC of bispecific antibody 2-36/1-68 ₉₀ 9.948. Mu.g/ml. IC of bispecific antibody 2-36/4-18 ₅₀ 0.03. Mu.g/ml. IC of bispecific antibody 2-36/4-18 ₉₀ 0.982. Mu.g/ml. IC of bispecific antibody 2-36/2-17 ₅₀ 0.063. Mu.g/ml. IC of bispecific antibody 2-36/2-17 ₉₀ 4.55. Mu.g/ml.

FIG. 66 shows the efficacy of bispecific antibodies with 2-30 (RBD) as parent antibody. Bispecific antibodyIC of sex antibody 2-30/4-18 ₅₀ 0.026. Mu.g/ml. IC of bispecific antibody 2-30/4-18 ₉₀ 0.096 μg/ml. IC of bispecific antibody 2-30/1-68 ₅₀ 0.008. Mu.g/ml. IC of bispecific antibody 2-30/1-68 ₉₀ 0.036. Mu.g/ml.

FIGS. 67A-D show comparative evaluation of bispecific antibody 2-17/2-7. FIG. 67A shows the efficacy of a 2-17/2-7 bispecific antibody compared to an antibody in which one arm is similar to the single parent component of the bispecific antibody and the other arm is an unrelated control. It also compares the efficacy of bispecific antibodies to a combination of two single arm IgG controls. FIG. 67B shows the combination of parent antibodies and virus neutralization of each parent antibody. FIG. 67C shows virus neutralization of bispecific antibody 2-17/2-7 as compared to the combination of parent antibodies. Figure 67D shows the efficacy of the combined parent monoclonal antibodies each tested in half the amount of the corresponding bispecific antibody. IC of bispecific antibody 2-17/2-7 ₅₀ 0.005 μg/ml. IC of bispecific antibody 2-17/2-7 ₉₀ 0.02 μg/ml. Parent antibody combined IC ₅₀ 0.014. Mu.g/ml. Parent antibody combined IC ₉₀ 0.07. Mu.g/ml.

FIGS. 68A-D show comparative evaluations of bispecific antibodies 1-20/4-18. FIG. 68A shows the efficacy of 1-20/4-18 bispecific antibodies compared to an antibody in which one arm is similar to the single parent component of the bispecific antibody and the other arm is an unrelated control. It also compares the efficacy of bispecific antibodies to a combination of two single arm IgG controls. FIG. 68B shows the combination of parent antibodies and virus neutralization of each parent antibody. FIG. 68C shows virus neutralization of bispecific antibodies 1-20/4-18 as compared to the combination of parent antibodies. Figure 68D shows the efficacy of the combined parent monoclonal antibodies each tested in half the amount of the corresponding bispecific antibody. IC of bispecific antibody 1-20/4-18 ₅₀ Is 0.016. Mu.g/ml. IC of bispecific antibody 1-20/4-18 ₉₀ 0.076. Mu.g/ml. Parent antibody combined IC ₅₀ 0.014. Mu.g/ml. Parent antibody combined IC ₉₀ 0.057. Mu.g/ml.

FIGS. 69A-D show comparative evaluations of bispecific antibodies 1-20/5-24. FIG. 69A shows a view similar to one of the armsEfficacy of 1-20/5-24 bispecific antibodies compared to the single parent component of bispecific antibody, while the other arm is the antibody of the unrelated control. It also compares the efficacy of bispecific antibodies to a combination of two single arm IgG controls. FIG. 69B shows virus neutralization with a combination of parent antibodies and each parent antibody. FIG. 69C shows virus neutralization of bispecific antibodies 1-20/5-24 as compared to the combination of parent antibodies. Figure 69D shows the efficacy of the combined parent monoclonal antibodies each tested in half the amount of the corresponding bispecific antibody. IC of bispecific antibody 1-20/5-24 ₅₀ 0.012. Mu.g/ml. IC of bispecific antibody 1-20/5-24 ₉₀ 0.023. Mu.g/ml. Parent antibody combined IC ₅₀ 0.004. Mu.g/ml. Parent antibody combined IC ₉₀ Is 0.016. Mu.g/ml.

FIGS. 70A-D show comparative evaluations of bispecific antibody 2-17/4-20. FIG. 70A shows the efficacy of 2-17/4-20 bispecific antibodies compared to an antibody in which one arm is similar to the single parent component of the bispecific antibody and the other arm is an unrelated control. It also compares the efficacy of bispecific antibodies to a combination of two single arm IgG controls. FIG. 70B shows virus neutralization with a combination of parent antibodies and each parent antibody. FIG. 70C shows virus neutralization of bispecific antibody 2-17/4-20 as compared to the combination of parent antibodies. Figure 70D shows the efficacy of the combined parent monoclonal antibodies each tested in half the amount of the corresponding bispecific antibody. IC of bispecific antibody 2-17/4-20 ₅₀ 0.042. Mu.g/ml. IC of bispecific antibody 2-17/4-20 ₉₀ 0.097 μg/ml. Parent antibody combined IC ₅₀ 0.021. Mu.g/ml. Parent antibody combined IC ₉₀ 0.135. Mu.g/ml.

FIGS. 71A-B show the in vitro efficacy of bispecific antibodies against SARS-CoV-2 real virus strain USA/WA 1. Fig. 71A shows an IC ₉₀ Neutralization of ten bispecific antibodies at a concentration between 0.003 μg/ml and 0.023 μg/ml. Fig. 71B shows an IC ₉₀ Nine bispecific antibodies were neutralised at concentrations between 0.025. Mu.g/ml and 0.046. Mu.g/ml.

FIG. 72 shows the process performed by each IC ₅₀ And IC ₉₀ The concentration shows the in vitro efficacy of bispecific antibodies.

FIGS. 73A-B show a display IC ₉₀ And IC ₅₀ In vitro potency of bispecific antibody at concentration. FIG. 73A shows an IC showing a threshold value relative to 0.05 μg/ml ₉₀ And IC ₅₀ In vitro potency of bispecific antibody at concentration. FIG. 73B shows an IC showing a threshold value relative to 0.01 μg/ml ₉₀ And IC ₅₀ In vitro potency of bispecific antibody at concentration. Fig. 73A and 73B are two separate "sort" data. In a, data were sorted for the first 20 bispecific antibodies with highest to lowest inhibition of 90% of viruses (IC ₉₀ ). In B, the data were sorted by the first 20 bispecific antibodies with highest to lowest inhibition of 50% of the viruses (IC ₅₀ )。

FIGS. 74A-D show comparative evaluations of bispecific antibodies 1-57/1-68. FIG. 74A shows virus neutralization of bispecific antibodies 1-57/1-68 compared to two control bispecific antibodies (each antibody comprising a parent antibody arm and a control antibody arm), and compared to a mixture of two control bispecific antibodies. AbX is an irrelevant control antibody arm, where AbX is PGDM1400. FIG. 74B shows virus neutralization with a combination of parent antibodies and each parent antibody. FIG. 74C shows virus neutralization of bispecific antibody 1-57/1-68 as compared to the combination of parent antibodies. FIG. 74D shows the efficacy of a mixture of bispecific antibodies 1-57/1-68 and each parent monoclonal antibody, each tested in half the amount of the corresponding bispecific antibody. Concentration IC of bispecific antibody 1-57/1-68 ₅₀ 0.002. Mu.g/ml. Concentration IC of bispecific antibody 1-57/1-68 ₉₀ 0.005 μg/ml. Parent antibody combined IC ₅₀ Is 0.006 mug/ml. Parent antibody combined IC ₉₀ 0.02 μg/ml. The comparison data is from a single experiment, while the data in the summary table is the average of multiple replicates.

FIGS. 75A-D show comparative evaluation of bispecific antibody 2-17/2-7. FIG. 75A shows virus neutralization of bispecific antibodies 2-17/2-7 compared to two control bispecific antibodies (each antibody comprising a parent antibody arm and a control antibody arm), and compared to a mixture of two control bispecific antibodies. AbX and AbY are independent control antibody arms, abX of whichPGDM1400, abY a32. FIG. 75B shows virus neutralization with a combination of parent antibodies and each parent antibody. FIG. 75C shows virus neutralization of bispecific antibody 2-17/2-7 as compared to the combination of parent antibodies. FIG. 75D shows the efficacy of a mixture of bispecific antibodies 2-17/2-7 and each parent monoclonal antibody, each tested in half the amount of the corresponding bispecific antibody. IC of bispecific antibody 2-17/2-7 ₅₀ 0.005 μg/ml. IC of bispecific antibody 2-17/2-7 ₉₀ 0.02 μg/ml. Parent antibody combined IC ₅₀ 0.014. Mu.g/ml. Parent antibody combined IC ₉₀ 0.07. Mu.g/ml. The comparison data is from a single experiment, while the data in the summary table is the average of multiple replicates.

FIGS. 76A-D show comparative evaluations of bispecific antibodies 1-20/4-18. FIG. 76A shows virus neutralization of bispecific antibodies 1-20/4-18 as compared to two control bispecific antibodies (each antibody comprising a parent antibody arm and a control antibody arm) and as compared to a mixture of two control bispecific antibodies. AbX and AbY are unrelated control antibody arms, where AbX is PGDM1400 and AbY is a32. FIG. 76B shows the combination of parent antibodies and virus neutralization of each parent antibody. FIG. 76C shows virus neutralization of bispecific antibodies 1-20/4-18 as compared to the combination of parent antibodies. FIG. 76D shows the efficacy of a mixture of bispecific antibodies 1-20/4-18 and each parent monoclonal antibody, each tested in half the amount of the corresponding bispecific antibody. IC of bispecific antibody 1-20/4-18 ₅₀ Is 0.016. Mu.g/ml. IC of bispecific antibody 1-20/4-18 ₉₀ 0.077. Mu.g/ml. Parent antibody combined IC ₅₀ 0.014. Mu.g/ml. Parent antibody combined IC ₉₀ 0.057. Mu.g/ml. The comparison data is from a single experiment, while the data in the summary table is the average of multiple replicates.

FIGS. 77A-D show comparative evaluations of bispecific antibodies 1-20/5-24. FIG. 77A shows virus neutralization of bispecific antibodies 1-20/5-24 compared to two control bispecific antibodies (each antibody comprising a parent antibody arm and a control antibody arm) and compared to a mixture of two control bispecific antibodies. AbX and AbY are unrelated controlsAntibody arm, wherein AbX is PGDM1400 and AbY is a32. FIG. 77B shows the combination of parent antibodies and virus neutralization of each parent antibody. FIG. 77C shows virus neutralization of bispecific antibodies 1-20/5-24 as compared to the combination of parent antibodies. FIG. 77D shows the efficacy of a mixture of bispecific antibodies 1-20/5-24 and each parent monoclonal antibody, each tested in half the amount of the corresponding bispecific antibody. IC of bispecific antibody 1-20/5-24 ₅₀ 0.012. Mu.g/ml. IC of bispecific antibody 1-20/5-24 ₉₀ 0.023. Mu.g/ml. Parent antibody combined IC ₅₀ 0.004. Mu.g/ml. Parent antibody combined IC ₉₀ Is 0.016. Mu.g/ml. The comparison data is from a single experiment, while the data in the summary table is the average of multiple replicates.

FIGS. 78A-D show comparative evaluations of bispecific antibodies 2-30/1-68. FIG. 78A shows virus neutralization of bispecific antibodies 2-30/1-68 compared to two control bispecific antibodies (each antibody comprising a parent antibody arm and a control antibody arm) and compared to a mixture of two control bispecific antibodies. AbX is an irrelevant control antibody arm, where AbX is PGDM1400. FIG. 78B shows virus neutralization with a combination of parent antibodies and each parent antibody. FIG. 78C shows virus neutralization of bispecific antibody 2-30/1-68 as compared to the combination of parent antibodies. FIG. 78D shows the efficacy of a mixture of bispecific antibodies 2-30/1-68 and each parent monoclonal antibody, each tested in half the amount of the corresponding bispecific antibody. IC of bispecific antibody 2-30/1-68 ₅₀ 0.009. Mu.g/ml. IC of bispecific antibody 2-30/1-68 ₉₀ 0.034. Mu.g/ml. Parent antibody combined IC ₅₀ 0.009. Mu.g/ml. Parent antibody combined IC ₉₀ 0.086. Mu.g/ml. The comparison data is from a single experiment, while the data in the summary table is the average of multiple replicates.

FIGS. 79A-D show comparative evaluation of bispecific antibodies 2-7/4-8 (39/51). FIG. 79A shows virus neutralization of bispecific antibodies 2-7/4-8 (39/51) compared to two control bispecific antibodies, each comprising a parent antibody arm and a control antibody arm, and compared to a mixture of two control bispecific antibodies. AbXIs an unrelated control antibody arm, wherein AbX is PGDM1400. FIG. 79B shows the combination of parent antibodies and virus neutralization of each parent antibody. FIG. 79C shows virus neutralization of bispecific antibodies 2-7/4-8 (39/51) as compared to the combination of parent antibodies. FIG. 79D shows the efficacy of a mixture of bispecific antibodies 2-7/4-8 (39/51) and each parent monoclonal antibody, each tested in half the amount of the corresponding bispecific antibody. IC of bispecific antibody 2-7/4-8 (39/51) ₅₀ 0.0008. Mu.g/ml. IC of bispecific antibody 2-7/4-8 (39/51) ₉₀ 0.002. Mu.g/ml. The comparison data is from a single experiment, while the data in the summary table is the average of multiple replicates.

FIGS. 80A-D show comparative evaluations of bispecific antibodies 1-57/1-87. FIG. 80A shows virus neutralization of bispecific antibodies 1-57/1-87 as compared to two control bispecific antibodies, each comprising a parent antibody arm and a control antibody arm, and as compared to a mixture of two control bispecific antibodies. AbX is an irrelevant control antibody arm, where AbX is PGDM1400. FIG. 80B shows virus neutralization with a combination of parent antibodies and each parent antibody. FIG. 80C shows virus neutralization of bispecific antibodies 1-57/1-87 as compared to the combination of parent antibodies. FIG. 80D shows the efficacy of a mixture of bispecific antibodies 1-57/1-87 and each parent monoclonal antibody, each tested in half the amount of the corresponding bispecific antibody. IC of bispecific antibody 1-57/1-87 ₅₀ 0.0012. Mu.g/ml. IC of bispecific antibody 1-57/1-87 ₉₀ 0.002. Mu.g/ml. Parent antibody combined IC ₅₀ 0.0012. Mu.g/ml. Parent antibody combined IC ₉₀ 0.010. Mu.g/ml. The comparison data is from a single experiment, while the data in the summary table is the average of multiple replicates.

Figure 81 shows the efficacy of other bispecific antibodies.

EXAMPLE 16 bispecific anti-SARS-CoV-2 variant

FIGS. 82A-B show the neutralizing activity of antibodies specific for worker Cheng Huashuang. FIG. 82A shows the neutralizing activity of bispecific antibodies 2-17/2-7, 1-57/5-7, 5-7/1-57, 2-7/5-7, 5-7/2-7 against wild type virus (WA 1) at various antibody concentrations. FIG. 82B shows the antibody potency (IC) of bispecific antibodies 2-17/2-7 (control), 1-57/5-7, 5-7/1-57, 2-7/5-7, 5-7/2-7 against wild type virus (WA 1) ₅₀ And IC ₉₀ ). IC of bispecific antibody 2-17/2-7 ₅₀ 0.005-0.008 mug/ml. IC of bispecific antibody 2-17/2-7 ₉₀ 0.025-0.055 mug/ml. IC of bispecific antibody 1-57/5-7 ₅₀ 0.005 μg/ml. IC of bispecific antibody 1-57/5-7 ₉₀ 0.037. Mu.g/ml. IC of bispecific antibody 5-7/1-57 ₅₀ Is 0.006 mug/ml. IC of bispecific antibody 5-7/1-57 ₉₀ 0.054. Mu.g/ml. IC of bispecific antibody 2-7/5-7 ₅₀ 0.005 μg/ml. IC of bispecific antibody 2-7/5-7 ₉₀ 0.020. Mu.g/ml. IC of bispecific antibody 5-7/2-7 ₅₀ 0.007. Mu.g/ml. IC of bispecific antibody 5-7/2-7 ₉₀ 0.049. Mu.g/ml.

FIGS. 83A-B show 2-7/5-7 bispecific and affinity antibody activity. FIG. 83A shows the neutralizing activity of bispecific antibody 2-7/5-7, parent antibodies 2-7, 5-7, and combinations of 2-7 and 5-7 parent antibodies against wild type virus (WA 1) at various antibody concentrations. FIG. 83B shows the IC50 concentrations of bispecific antibody 2-7/5-7, parent antibodies 2-7, 5-7, and combinations of parent antibodies 2-7 and 5-7. IC of parent antibody 2-7 ₅₀ 0.009. Mu.g/ml. IC of parent antibody 5-7 ₅₀ 0.044. Mu.g/ml. IC of bispecific antibody 2-7/5-7 ₅₀ 0.002. Mu.g/ml. IC of the combination of parent antibodies 2-7 and 5-7 ₅₀ 0.020. Mu.g/ml.

FIGS. 84A-B show the activity of bispecific antibodies 2-7/5-7 against SARS-CoV-2 wild-type (WA 1) and variants B1.1.7, B1.526, B1351 and P.1. FIG. 84A shows the neutralizing activity of bispecific antibody 2-7/5-7 against various SARS-CoV-2 strains at different antibody concentrations. SARS-CoV-2 strains WA1, B1.1.7, B1.351, B1.526 and P.1. FIG. 84B shows the IC of bispecific antibody 2-7/5-7 against multiple SARS-CoV-2 strains ₅₀ And IC ₉₀ Concentration. IC of bispecific antibody 2-7/5-7 against WA1 (wild type strain) ₅₀ 0.002. Mu.g/ml. IC of bispecific antibody 2-7/5-7 against WA1 (wild type strain) ₉₀ 0.019. Mu.g/ml. IC of bispecific antibody 2-7/5-7 against B1.1.7 ₅₀ Is 0.002 μg/ml. IC of bispecific antibody 2-7/5-7 against B1.1.7 ₉₀ 0.011 μg/ml. IC of bispecific antibody 2-7/5-7 against B1.526 ₅₀ 0.002. Mu.g/ml. IC of bispecific antibody 2-7/5-7 against B1.526 ₉₀ 0.01. Mu.g/ml. IC of bispecific antibody 2-7/5-7 against B1.351 ₅₀ 0.025. Mu.g/ml. IC of bispecific antibody 2-7/5-7 against B1.351 ₉₀ 0.156. Mu.g/ml. IC of bispecific antibody 2-7/5-7 against P.1 ₅₀ 0.013. Mu.g/ml. IC of bispecific antibody 2-7/5-7 against P.1 ₉₀ 0.188 μg/ml.

EXAMPLE 17 bispecific 2-7/5-7 antibodies

FIGS. 85A-D show neutralization of a pseudovirus having mutations associated with SARS-CoV-2 variant by a bispecific 2-7/5-7 antibody. FIG. 85A shows the antibody neutralization profile against a circulating SARS-CoV-2 virus variant. FIG. 85B shows antibody efficacy against circulating SARS-CoV-2 virus variants. FIG. 85C shows an antibody neutralization profile for SARS-CoV-2 variant with high frequency mutations. FIG. 85D shows antibody efficacy against SARS-CoV-2 variant with high frequency mutation.

FIGS. 86A-F show the use of bispecific antibodies 2-7/5-7 to neutralize pseudoviruses with SARS-CoV-2 mutation corresponding to the "related variant". Fig. 86A shows the antibody neutralization profile against b.1.1.7 (uk virus variant) with NTD mutations. Figure 86B shows antibody efficacy against b.1.1.7 (uk virus variant) with NTD mutations. Fig. 86C shows an antibody neutralization profile for b.1.352 (SA) virus variants with NTD mutations. Figure 86D shows antibody efficacy against b.1.352 (SA) virus variants with NTD mutations. Figure 86E shows an antibody neutralization profile for p.1 (BZ) virus variants with NTD mutations. Figure 86F shows antibody efficacy against p.1 (BZ) virus variants with NTD mutations.

Claims

1. A synthetic monoclonal antibody, or a functional fragment thereof, comprising a heavy chain having a variable domain and a constant domain and a light chain having a variable domain and a constant domain, wherein the antibody binds an antigen on a coronavirus particle.

2. The antibody of claim 1, wherein the heavy chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 3, a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 7, a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 11, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 17, 62%, 67%, 70%, 90%, 92%, 80%, 98%, 99% identical to SEQ ID No. 9.

3. The antibody of claim 1, wherein the heavy chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 19, a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 21, a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 29, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 31, 62%, 67%, 70%, 90%, 80%, 98%, 99% identical to SEQ ID No. 25.

4. The antibody of claim 1, wherein the heavy chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID NO 33, a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID NO 37, a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID NO 43, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID NO, or a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 77%, 80%, 98%, 99% identical to SEQ ID NO.

5. The antibody of claim 1, wherein the light chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 4, a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 8, a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 12, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 18, 62%, 67%, 70%, 72%, 80%, 98%, 99% identical to SEQ ID No. 10, at least 60%, 62%, 67%, 72%, 77%, 80%, 98%, 99% and the polypeptide sequence.

6. The antibody of claim 1, wherein the light chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID NO, 32%, 62%, 67%, 70%, 72%, 75%, 80%, 98%, 99% identical to SEQ ID NO.

7. The antibody of claim 1, wherein the light chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID NO 34, a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID NO 38, a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID NO 44, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID NO 46, at least 60%, 62%, 65%, 67%, 70%, 72%, 77%, 80%, 98%, 99% identical to SEQ ID NO.

8. The antibody of claim 1, wherein the heavy chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID NO 3 and the light chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID NO 4.

9. The antibody of claim 1, wherein the heavy chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 7 and the light chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 8.

10. The antibody of claim 1, wherein the heavy chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID NO 9 and the light chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID NO 10.

11. The antibody of claim 1, wherein the heavy chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 11 and the light chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 12.

12. The antibody of claim 1, wherein the heavy chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 17 and the light chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 18.

13. The antibody of claim 1, wherein the heavy chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 19 and the light chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 20.

14. The antibody of claim 1, wherein the heavy chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 21 and the light chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 22.

15. The antibody of claim 1, wherein the heavy chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 25 and the light chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 26.

16. The antibody of claim 1, wherein the heavy chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 29 and the light chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 30.

17. The antibody of claim 1, wherein the heavy chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 31 and the light chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 32.

18. The antibody of claim 1, wherein the heavy chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID NO 33 and the light chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID NO 34.

19. The antibody of claim 1, wherein the heavy chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 37 and the light chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 38.

20. The antibody of claim 1, wherein the heavy chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID NO 39 and the light chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID NO 40.

21. The antibody of claim 1, wherein the heavy chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 43 and the light chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 44.

22. The antibody of claim 1, wherein the heavy chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 45 and the light chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 46.

23. The antibody of claim 1, wherein the heavy chain variable domain comprises SEQ ID No. 3 and the light chain variable domain comprises SEQ ID No. 4.

24. The antibody of claim 1, wherein the heavy chain variable domain comprises SEQ ID No. 7 and the light chain variable domain comprises SEQ ID No. 8.

25. The antibody of claim 1, wherein the heavy chain variable domain comprises SEQ ID No. 9 and the light chain variable domain comprises SEQ ID No. 10.

26. The antibody of claim 1, wherein the heavy chain variable domain comprises SEQ ID No. 11 and the light chain variable domain comprises SEQ ID No. 12.

27. The antibody of claim 1, wherein the heavy chain variable domain comprises SEQ ID No. 17 and the light chain variable domain comprises SEQ ID No. 18.

28. The antibody of claim 1, wherein the heavy chain variable domain comprises SEQ ID No. 19 and the light chain variable domain comprises SEQ ID No. 20.

29. The antibody of claim 1, wherein the heavy chain variable domain comprises SEQ ID No. 21 and the light chain variable domain comprises SEQ ID No. 22.

30. The antibody of claim 1, wherein the heavy chain variable domain comprises SEQ ID No. 25 and the light chain variable domain comprises SEQ ID No. 26.

31. The antibody of claim 1, wherein the heavy chain variable domain comprises SEQ ID No. 29 and the light chain variable domain comprises SEQ ID No. 30.

32. The antibody of claim 1, wherein the heavy chain variable domain comprises SEQ ID No. 31 and the light chain variable domain comprises SEQ ID No. 32.

33. The antibody of claim 1, wherein the heavy chain variable domain comprises SEQ ID No. 33 and the light chain variable domain comprises SEQ ID No. 34.

34. The antibody of claim 1, wherein the heavy chain variable domain comprises SEQ ID No. 37 and the light chain variable domain comprises SEQ ID No. 38.

35. The antibody of claim 1, wherein the heavy chain variable domain comprises SEQ ID No. 39 and the light chain variable domain comprises SEQ ID No. 40.

36. The antibody of claim 1, wherein the heavy chain variable domain comprises SEQ ID No. 43 and the light chain variable domain comprises SEQ ID No. 44.

37. The antibody of claim 1, wherein the heavy chain variable domain comprises SEQ ID No. 45 and the light chain variable domain comprises SEQ ID No. 46.

38. The antibody of claim 1, wherein the heavy chain variable domain comprises CDR1, CDR2, and CDR3 regions of SEQ ID No. 3; CDR1, CDR2 and CDR3 regions of SEQ ID No. 7; CDR1, CDR2 and CDR3 regions of SEQ ID No. 9: CDR1, CDR2 and CDR3 regions of SEQ ID No. 11; or CDR1, CDR2 and CDR3 regions of SEQ ID NO 17.

39. The antibody of claim 1, wherein the heavy chain variable domain comprises CDR1, CDR2, and CDR3 regions of SEQ ID No. 19; CDR1, CDR2 and CDR3 regions of SEQ ID No. 21; CDR1, CDR2 and CDR3 regions of SEQ ID No. 25: CDR1, CDR2 and CDR3 regions of SEQ ID No. 29; or CDR1, CDR2 and CDR3 regions of SEQ ID NO. 31.

40. The antibody of claim 1, wherein the heavy chain variable domain comprises CDR1, CDR2, and CDR3 regions of SEQ ID No. 33; CDR1, CDR2 and CDR3 regions of SEQ ID No. 37; CDR1, CDR2 and CDR3 regions of SEQ ID NO: 39: CDR1, CDR2 and CDR3 regions of SEQ ID No. 43; or CDR1, CDR2 and CDR3 regions of SEQ ID NO 45.

41. The antibody of claim 1, wherein the light chain variable domain comprises CDR1, CDR2, and CDR3 regions of SEQ ID No. 4; CDR1, CDR2 and CDR3 regions of SEQ ID No. 8; CDR1, CDR2 and CDR3 regions of SEQ ID No. 10: CDR1, CDR2 and CDR3 regions of SEQ ID No. 12; or CDR1, CDR2 and CDR3 regions of SEQ ID NO. 18.

42. The antibody of claim 1, wherein the light chain variable domain comprises CDR1, CDR2, and CDR3 regions of SEQ ID No. 20; CDR1, CDR2 and CDR3 regions of SEQ ID No. 22; CDR1, CDR2 and CDR3 regions of SEQ ID No. 26: CDR1, CDR2 and CDR3 regions of SEQ ID No. 30; or CDR1, CDR2 and CDR3 regions of SEQ ID NO. 32.

43. The antibody of claim 1, wherein the light chain variable domain comprises CDR1, CDR2, and CDR3 regions of SEQ ID No. 34; CDR1, CDR2 and CDR3 regions of SEQ ID No. 38; CDR1, CDR2, and CDR3 regions of SEQ ID No. 40: CDR1, CDR2 and CDR3 regions of SEQ ID No. 44; or CDR1, CDR2 and CDR3 regions of SEQ ID NO 46.

44. The antibody of claim 1, wherein the antibody neutralizes coronavirus.

45. The antibody of claim 44, wherein the coronavirus is SARS-CoV-2 virus.

46. The antibody of claim 45, wherein the coronavirus is a SARS-CoV-2 variant.

47. The antibody of claim 46, wherein the variant strain is selected from the group consisting of WA1, b.1.1.7, B1.526, B1.351, P1, B1.352, and b.1.427.

48. The antibody of claim 1, wherein the antibody specifically binds to an epitope on the surface of a coronavirus.

49. The antibody of claim 48, wherein the epitope comprises at least a portion of a coronavirus spike protein.

50. The antibody of claim 49, wherein at least a portion of the coronavirus spike protein comprises a Receptor Binding Domain (RBD) of a spike protein.

51. The antibody of claim 49, wherein at least a portion of the coronavirus spike protein comprises an N-terminal domain (NTD) of a spike protein.

52. The antibody of any one of claims 48-51, wherein the coronavirus is a SARS-CoV-2 coronavirus.

53. A method of treating or preventing a covd-19 infection in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of a synthetic monoclonal antibody, or a functional fragment thereof, comprising a heavy chain having a variable domain and a constant domain and a light chain having a variable domain and a constant domain, wherein the antibody binds an antigen on a coronavirus particle.

54. The method of claim 53, wherein the heavy chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 3, a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 7, a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 11, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 17, 62%, 67%, 70%, 90%, 92%, 80%, 98%, 99% identical to SEQ ID No. 9.

55. The method of claim 53, wherein the heavy chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 19, a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 21, a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 29, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 31, 62%, 67%, 70%, 90%, 92%, 80%, 98%, 99% identical to SEQ ID No. 25.

56. The method of claim 53, wherein the heavy chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID NO 33, a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID NO 37, a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID NO 43, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID NO, or a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 90%, 92%, 80%, 98%, 99% identical to SEQ ID NO.

57. The method of claim 53, wherein the light chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 4, a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 8, a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 12, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 18, 62%, 67%, 70%, 90%, 92%, 80%, 98%, 99% identical to SEQ ID No. 10, at least 60%, 62%, 67%, 75%, 77%, 80%, 98%, 99% and the polypeptide sequence.

58. The method of claim 53, wherein the light chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID NO:20, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID NO:22, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID NO:30, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID NO:32, at least 60%, 62%, 67%, 70%, 72%, 80%, 98%, 99% identical to SEQ ID NO.

59. The method of claim 53, wherein the light chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID NO:34, a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID NO:38, or a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID NO:44, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID NO:46, at least 60%, 62%, 67%, 70%, 72%, 80%, 98%, 99% identical to SEQ ID NO: 40.

60. The method of claim 53, wherein the heavy chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID NO 3 and the light chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID NO 4.

61. The method of claim 53, wherein the heavy chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 7 and the light chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 8.

62. The method of claim 53, wherein the heavy chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID NO 9 and the light chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID NO 10.

63. The method of claim 53, wherein the heavy chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 11 and the light chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 12.

64. The method of claim 53, wherein the heavy chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 17 and the light chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 18.

65. The method of claim 53, wherein the heavy chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 19 and the light chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 20.

66. The method of claim 53, wherein the heavy chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 21 and the light chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 22.

67. The method of claim 53, wherein the heavy chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 25 and the light chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 26.

68. The method of claim 53, wherein the heavy chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID NO 29 and the light chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID NO 30.

69. The method of claim 53, wherein the heavy chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 31 and the light chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 32.

70. The method of claim 53, wherein the heavy chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID NO 33 and the light chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID NO 34.

71. The method of claim 53, wherein the heavy chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 37 and the light chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 38.

72. The method of claim 53, wherein the heavy chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID NO 39 and the light chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID NO 40.

73. The method of claim 53, wherein the heavy chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 43 and the light chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 44.

74. The method of claim 53, wherein the heavy chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 45 and the light chain variable domain comprises a polypeptide sequence that is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 46.

75. The method of claim 53, wherein the heavy chain variable domain comprises SEQ ID NO. 3 and the light chain variable domain comprises SEQ ID NO. 4.

76. The method of claim 53, wherein the heavy chain variable domain comprises SEQ ID NO. 7 and the light chain variable domain comprises SEQ ID NO. 8.

77. The method of claim 53, wherein the heavy chain variable domain comprises SEQ ID NO 9 and the light chain variable domain comprises SEQ ID NO 10.

78. The method of claim 53, wherein the heavy chain variable domain comprises SEQ ID NO. 11 and the light chain variable domain comprises SEQ ID NO. 12.

79. The method of claim 53, wherein the heavy chain variable domain comprises SEQ ID NO. 17 and the light chain variable domain comprises SEQ ID NO. 18.

80. The method of claim 53, wherein the heavy chain variable domain comprises SEQ ID NO. 19 and the light chain variable domain comprises SEQ ID NO. 20.

81. The method of claim 53, wherein the heavy chain variable domain comprises SEQ ID NO. 21 and the light chain variable domain comprises SEQ ID NO. 22.

82. The method of claim 53, wherein the heavy chain variable domain comprises SEQ ID NO. 25 and the light chain variable domain comprises SEQ ID NO. 26.

83. The method of claim 53, wherein the heavy chain variable domain comprises SEQ ID NO. 29 and the light chain variable domain comprises SEQ ID NO. 30.

84. The method of claim 53, wherein the heavy chain variable domain comprises SEQ ID NO. 31 and the light chain variable domain comprises SEQ ID NO. 32.

85. The method of claim 53, wherein the heavy chain variable domain comprises SEQ ID NO. 33 and the light chain variable domain comprises SEQ ID NO. 34.

86. The method of claim 53, wherein the heavy chain variable domain comprises SEQ ID NO. 37 and the light chain variable domain comprises SEQ ID NO. 38.

87. The method of claim 53, wherein the heavy chain variable domain comprises SEQ ID NO 39 and the light chain variable domain comprises SEQ ID NO 40.

88. The method of claim 53, wherein the heavy chain variable domain comprises SEQ ID NO. 43 and the light chain variable domain comprises SEQ ID NO. 44.

89. The method of claim 53, wherein the heavy chain variable domain comprises SEQ ID NO. 45 and the light chain variable domain comprises SEQ ID NO. 46.

90. The method of claim 53, wherein the heavy chain variable domain comprises CDR1, CDR2 and CDR3 regions of SEQ ID NO 3; CDR1, CDR2 and CDR3 regions of SEQ ID No. 7; CDR1, CDR2 and CDR3 regions of SEQ ID No. 9: CDR1, CDR2 and CDR3 regions of SEQ ID No. 11; or CDR1, CDR2 and CDR3 regions of SEQ ID NO 17.

91. The method of claim 53, wherein the heavy chain variable domain comprises CDR1, CDR2 and CDR3 regions of SEQ ID NO 19; CDR1, CDR2 and CDR3 regions of SEQ ID No. 21; CDR1, CDR2 and CDR3 regions of SEQ ID No. 25: CDR1, CDR2 and CDR3 regions of SEQ ID No. 29; or CDR1, CDR2 and CDR3 regions of SEQ ID NO. 31.

92. The method of claim 53, wherein the heavy chain variable domain comprises the CDR1, CDR2 and CDR3 regions of SEQ ID NO 33; CDR1, CDR2 and CDR3 regions of SEQ ID No. 37; CDR1, CDR2 and CDR3 regions of SEQ ID NO: 39: CDR1, CDR2 and CDR3 regions of SEQ ID No. 43; or CDR1, CDR2 and CDR3 regions of SEQ ID NO 45.

93. The method of claim 53, wherein the light chain variable domain comprises the CDR1, CDR2 and CDR3 regions of SEQ ID NO. 4; CDR1, CDR2 and CDR3 regions of SEQ ID No. 8; CDR1, CDR2 and CDR3 regions of SEQ ID No. 10: CDR1, CDR2 and CDR3 regions of SEQ ID No. 12; or CDR1, CDR2 and CDR3 regions of SEQ ID NO. 18.

94. The method of claim 53, wherein the light chain variable domain comprises the CDR1, CDR2 and CDR3 regions of SEQ ID NO. 20; CDR1, CDR2 and CDR3 regions of SEQ ID No. 22; CDR1, CDR2 and CDR3 regions of SEQ ID No. 26: CDR1, CDR2 and CDR3 regions of SEQ ID No. 30; or CDR1, CDR2 and CDR3 regions of SEQ ID NO. 32.

95. The method of claim 53, wherein the light chain variable domain comprises the CDR1, CDR2 and CDR3 regions of SEQ ID NO 34; CDR1, CDR2 and CDR3 regions of SEQ ID No. 38; CDR1, CDR2, and CDR3 regions of SEQ ID No. 40: CDR1, CDR2 and CDR3 regions of SEQ ID No. 44; or CDR1, CDR2 and CDR3 regions of SEQ ID NO 46.

96. The method of claim 53, wherein the antibody neutralizes coronavirus.

97. The method of claim 96, wherein the coronavirus is a SARS-CoV-2 virus.

98. The method of claim 97, wherein the coronavirus is a SARS-CoV-2 variant.

99. The method of claim 98, wherein the variant plant is selected from the group consisting of WA1, b.1.1.7, B1.526, B1.351, P1, B1.352, and b.1.427.

100. The method of claim 53, wherein the antibody specifically binds to an epitope on the surface of a coronavirus.

101. The method of claim 100, wherein the epitope comprises at least a portion of a coronavirus spike protein.

102. The method of claim 101, wherein at least a portion of the coronavirus spike protein comprises a Receptor Binding Domain (RBD) of a spike protein.

103. The method of claim 101, wherein at least a portion of the coronavirus spike protein comprises an N-terminal domain (NTD) of a spike protein.

104. The method of any one of claims 100-103, wherein the coronavirus is a SARS-CoV-2 coronavirus.

105. The method of claim 104, wherein the composition is administered in combination with a second therapeutic agent.

106. A bispecific molecule comprising a first polypeptide chain and a second polypeptide chain, wherein:

the first polypeptide chain comprises:

a first light chain comprising a variable domain and a constant domain;

a first heavy chain comprising a variable domain and a constant domain;

the second polypeptide chain comprises:

a second light chain comprising a variable domain and a constant domain; and

a second heavy chain comprising a variable domain and a constant domain.

107. The molecule of claim 106, wherein the first polypeptide chain and the second polypeptide chain are covalently bonded to each other.

108. The molecule of claim 106, wherein the first heavy chain comprises at least one amino acid sequence that hybridizes to SEQ ID NO:51, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to a polypeptide sequence of SEQ ID NO:55, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to a polypeptide sequence of SEQ ID NO: 67%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical polypeptide sequences to at least 60%, 62%, 65%, 82%, 85%, 87%, 90%, 95%, 98%, 99% identical polypeptide sequences to at least 60%, 83%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98% of SEQ ID NO A polypeptide sequence 99% identical to SEQ ID NO:87, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to a polypeptide sequence of SEQ ID NO:91 at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to a polypeptide sequence of SEQ ID NO:95 at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical polypeptide sequences with at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical polypeptide sequences with at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 95%, 97%, 98%, 99% identical polypeptide sequences with at least 60%, 62%, 65%, 98%, 99%, 95%, 98%, 99%, 95%, 99%, 119% identical polypeptide sequences with at least 60% of SEQ ID NO 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical polypeptide sequences to those of SEQ ID NO:123, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to a polypeptide sequence of SEQ ID NO:127 at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to a polypeptide sequence of SEQ ID NO:131 at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID NO 139 at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID NO 143 at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID NO 147 at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID NO 151 at least 60%, 62%, 65%, 67%, 70%, 72%, 75% of SEQ ID NO, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical polypeptide sequences to SEQ ID NO:155, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to a polypeptide sequence of SEQ ID NO:159 at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to a polypeptide sequence of SEQ ID NO:163 is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to polypeptide sequence which is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90% identical to polypeptide sequence which is at least 60%, 67%, 70%, 72%, 75%, 67%, 70%, 72%, 75%, 80%, 85%, 87%, 90% of SEQ ID NO 92%, 95%, 97%, 98%, 99% identical or a polypeptide sequence at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 183.

109. The molecule of claim 106, wherein the first light chain comprises a sequence that hybridizes to SEQ ID NO:52, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to a polypeptide sequence of SEQ ID NO:56 at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to a polypeptide sequence of SEQ ID NO:68 at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical polypeptide sequence to at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 99% identical polypeptide sequence to at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 95%, 97%, 98%, 99% identical polypeptide sequence to at least 60%, 62%, 65%, 82%, 98%, 99% identical polypeptide sequence to SEQ ID NO 84 at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical polypeptide sequence to SEQ ID NO, and SEQ ID NO:88 at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to a polypeptide sequence of SEQ ID NO:92, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to a polypeptide sequence of SEQ ID NO:96 at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical polypeptide sequence to at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 99% identical polypeptide sequence to at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 95%, 97%, 98%, 99% identical polypeptide sequence to at least 60%, 62%, 65%, 82%, 98%, 99% identical polypeptide sequence to at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical polypeptide sequence to at least 60% of SEQ ID NO, 60%, 120% polypeptide sequence to at least 60% of SEQ ID NO 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical polypeptide sequences to those of SEQ ID NO:124 at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to a polypeptide sequence of SEQ ID NO:128 at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to a polypeptide sequence of SEQ ID NO:132, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID NO:140, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID NO:144, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID NO:148, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID NO:152, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 75% amino acid sequence identical to SEQ ID NO:152, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical polypeptide sequences to SEQ ID NO:156, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to a polypeptide sequence of SEQ ID NO:160, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to a polypeptide sequence of SEQ ID NO:164 at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical polypeptide sequence to at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical polypeptide sequence to at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 95%, 97%, 98%, 99% identical polypeptide sequence to at least 60%, 180%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92% identical polypeptide sequence to at least 60%, 180% to SEQ ID NO 95%, 97%, 98%, 99% identical or a polypeptide sequence which is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID NO 184.

110. The molecule of claim 106, wherein the second heavy chain comprises a sequence identical to SEQ ID NO:53 at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to a polypeptide sequence of SEQ ID NO:57, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to a polypeptide sequence of SEQ ID NO:69 at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical polypeptide sequence, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical polypeptide sequence, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 95%, 97%, 98%, 99% identical polypeptide sequence of SEQ ID NO, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical polypeptide sequence of SEQ ID NO, and SEQ ID NO:89, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to a polypeptide sequence of SEQ ID NO:93 at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to a polypeptide sequence of SEQ ID NO:97 at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical polypeptide sequence to at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical polypeptide sequence to at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 95%, 97%, 98%, 99% identical polypeptide sequence to at least 60%, 117 at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical polypeptide sequence to at least 60%, 62%, 121% of SEQ ID NO 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical polypeptide sequences to those of SEQ ID NO:125 at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to a polypeptide sequence of SEQ ID NO:129 at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical polypeptide sequence to at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical polypeptide sequence to at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 95%, 97%, 98%, 99% identical polypeptide sequence to at least 60%, 62%, 65%, 95%, 97%, 98%, 99% identical polypeptide sequence to at least 60%, 62%, 80%, 95%, 97%, 98%, 99% identical polypeptide sequence to at least 60%, 149%, 95%, 67%, 98%, 99% identical polypeptide sequence to at least 60%, 67%, 70%, 75%, 98%, 99% of SEQ ID NO 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical polypeptide sequences to SEQ ID NO:157 at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to the polypeptide sequence of SEQ ID NO:161, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to a polypeptide sequence of SEQ ID NO:165 at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical polypeptide sequence to SEQ ID NO 169 at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical polypeptide sequence to SEQ ID NO 173 at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical polypeptide sequence to SEQ ID NO 177 at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical polypeptide sequence to SEQ ID NO 181 at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92% identical polypeptide sequence to SEQ ID NO 181 95%, 97%, 98%, 99% identical polypeptide sequence, or a polypeptide sequence at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 185.

111. The molecule of claim 106, wherein the second light chain comprises a sequence that hybridizes to SEQ ID NO:54, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to a polypeptide sequence of SEQ ID NO:58, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to a polypeptide sequence of SEQ ID NO:70, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical polypeptide sequence, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical polypeptide sequence, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 95%, 97%, 98%, 99% identical polypeptide sequence to SEQ ID NO 74, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical polypeptide sequence to SEQ ID NO 86, and SEQ ID NO:90, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to a polypeptide sequence of SEQ ID NO:94, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to a polypeptide sequence of SEQ ID NO:98 at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical polypeptide sequence to at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 99% identical polypeptide sequence to at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 95%, 97%, 98%, 99% identical polypeptide sequence to at least 60%, 62%, 80%, 95%, 98%, 99% identical polypeptide sequence to at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical polypeptide sequence to at least 60%, 118%, 62%, 98%, 99% of SEQ ID sequence to that of SEQ ID NO, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical polypeptide sequences to those of SEQ ID NO:126 at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to a polypeptide sequence of SEQ ID NO:130, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical polypeptide sequence, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical polypeptide sequence, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 95%, 97%, 98%, 99% identical polypeptide sequence, at least 150%, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical polypeptide sequence, at least 60%, 62%, 65%, 67%, 70%, 75%, 98%, 99% identical polypeptide sequence to SEQ ID NO 146, at least 60%, 62%, 65%, 80%, 75%, 80%, 90%, 98%, 99% 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical polypeptide sequences to SEQ ID NO:158, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to a polypeptide sequence of SEQ ID NO:162, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to a polypeptide sequence of SEQ ID NO:166 at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to a polypeptide sequence at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 95%, 97%, 98%, 99% identical to a polypeptide sequence at least 60%, 182%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92% to a polypeptide sequence at least 60%, 182% to a polypeptide sequence of SEQ ID NO of at least 60%, 62%, 65%, 67%, 87%, 90%, 98%, 99% 95%, 97%, 98%, 99% identical or a polypeptide sequence at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 186.

112. The molecule of claim 106, wherein the variable domain of the first heavy chain comprises a sequence that hybridizes to SEQ ID NO:15, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to a polypeptide sequence of SEQ ID NO:27, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to a polypeptide sequence of SEQ ID NO:35 at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical polypeptide sequence to at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical polypeptide sequence to at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 95%, 97%, 98%, 99% identical polypeptide sequence to at least 60%, 62%, 65%, 67%, 70%, 72%, 90%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 95%, 97% identical polypeptide sequence to at least 60%, 75% to SEQ ID NO 19., 98%, 99% identical polypeptide sequence to at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical polypeptide sequence to at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 98%, 99% identical polypeptide sequence to at least 60%, 62%, 80%, 82%, 85%, 87%, 90%, 98%, 99% identical polypeptide sequence to at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical polypeptide sequence to at least 60%, 41.

113. The molecule of claim 106, wherein the variable domain of the first light chain comprises a sequence that hybridizes to SEQ ID NO:16, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to a polypeptide sequence of SEQ ID NO:28, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to a polypeptide sequence of SEQ ID NO:36 at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical polypeptide sequence to at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97% identical polypeptide sequence to at least 20% to SEQ ID NO, 98%, 99% identical polypeptide sequence to at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical polypeptide sequence to at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 98%, 99% identical polypeptide sequence to at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical polypeptide sequence to at least 60%, 42.

114. The molecule of claim 106, wherein the variable domain of the second heavy chain comprises a sequence that hybridizes to SEQ ID NO:27, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to a polypeptide sequence of SEQ ID NO:15, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to a polypeptide sequence of SEQ ID NO:5 at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical polypeptide sequence to at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 99% identical polypeptide sequence to at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical polypeptide sequence to at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97% identical polypeptide sequence to at least 60%, 29% to SEQ ID NO, 98%, 99% identical polypeptide sequence to SEQ ID NO:35, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to a polypeptide sequence of SEQ ID NO:23, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to a polypeptide sequence of SEQ ID NO:25, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID NO:37, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID NO:39, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID NO:43, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 95%, 97%, 98%, 99% identical to SEQ ID NO:9, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID NO.

115. The molecule of claim 106, wherein the variable domain of the second light chain comprises a sequence that hybridizes to SEQ ID NO:28, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to a polypeptide sequence of SEQ ID NO:16, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to a polypeptide sequence of SEQ ID NO:6 at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical polypeptide sequence to at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 99% identical polypeptide sequence to at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical polypeptide sequence to at least 60%, 62%, 65%, 82%, 98%, 99% identical polypeptide sequence to SEQ ID NO 30 at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97% identical to SEQ ID NO, 98%, 99% identical polypeptide sequence to SEQ ID NO:36, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to a polypeptide sequence of SEQ ID NO:24, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to a polypeptide sequence of SEQ ID NO:26, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical polypeptide sequence to SEQ ID NO, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 95%, 98%, 99% identical polypeptide sequence to SEQ ID NO, or at least 10, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 98%, 99% identical polypeptide sequence to SEQ ID NO.

116. The molecule of claim 106, wherein the variable domain of the first heavy chain comprises CDR1, CDR2, and CDR3 of SEQ ID No. 51; CDR1, CDR2 and CDR3 of SEQ ID NO. 55; CDR1, CDR2 and CDR3 of SEQ ID NO 67; CDR1, CDR2 and CDR3 of SEQ ID NO 71; CDR1, CDR2 and CDR3 of SEQ ID NO 75; CDR1, CDR2 and CDR3 of SEQ ID NO 79; CDR1, CDR2 and CDR3 of SEQ ID NO 83; CDR1, CDR2 and CDR3 of SEQ ID NO 87; CDR1, CDR2 and CDR3 of SEQ ID NO. 91; CDR1, CDR2 and CDR3 of SEQ ID NO 95; CDR1, CDR2 and CDR3 of SEQ ID No. 99; CDR1, CDR2 and CDR3 of SEQ ID NO 103; CDR1, CDR2 and CDR3 of SEQ ID NO 107; CDR1, CDR2 and CDR3 of SEQ ID NO 115; CDR1, CDR2 and CDR3 of SEQ ID No. 119; CDR1, CDR2 and CDR3 of SEQ ID NO. 123; CDR1, CDR2 and CDR3 of SEQ ID NO 127; CDR1, CDR2 and CDR3 of SEQ ID NO. 131; CDR1, CDR2 and CDR3 of SEQ ID NO 139; CDR1, CDR2 and CDR3 of SEQ ID NO 143; CDR1, CDR2 and CDR3 of SEQ ID No. 147; CDR1, CDR2 and CDR3 of SEQ ID NO 151; CDR1, CDR2 and CDR3 of SEQ ID NO 155; CDR1, CDR2 and CDR3 of SEQ ID NO 159; CDR1, CDR2 and CDR3 of SEQ ID NO 163; CDR1, CDR2 and CDR3 of SEQ ID NO 167; CDR1, CDR2 and CDR3 of SEQ ID NO. 171; CDR1, CDR2 and CDR3 of SEQ ID NO 175; CDR1, CDR2 and CDR3 of SEQ ID NO 179; or CDR1, CDR2 and CDR3 of SEQ ID NO 183.

117. The molecule of claim 106, wherein the variable domain of the first light chain comprises CDR1, CDR2, and CDR3 of SEQ ID No. 52; CDR1, CDR2 and CDR3 of SEQ ID NO. 56; CDR1, CDR2 and CDR3 of SEQ ID NO. 68; CDR1, CDR2 and CDR3 of SEQ ID NO 72; CDR1, CDR2 and CDR3 of SEQ ID NO 76; CDR1, CDR2 and CDR3 of SEQ ID NO 80; CDR1, CDR2 and CDR3 of SEQ ID NO 84; CDR1, CDR2 and CDR3 of SEQ ID NO 88; CDR1, CDR2 and CDR3 of SEQ ID NO. 92; CDR1, CDR2 and CDR3 of SEQ ID No. 96; CDR1, CDR2 and CDR3 of SEQ ID NO. 100; CDR1, CDR2 and CDR3 of SEQ ID No. 104; CDR1, CDR2 and CDR3 of SEQ ID NO 108; CDR1, CDR2 and CDR3 of SEQ ID NO. 116; CDR1, CDR2 and CDR3 of SEQ ID NO. 120; CDR1, CDR2 and CDR3 of SEQ ID NO 124; CDR1, CDR2 and CDR3 of SEQ ID NO 128; CDR1, CDR2 and CDR3 of SEQ ID NO. 132; CDR1, CDR2 and CDR3 of SEQ ID NO. 140; CDR1, CDR2 and CDR3 of SEQ ID NO. 144; CDR1, CDR2 and CDR3 of SEQ ID NO. 148; CDR1, CDR2 and CDR3 of SEQ ID NO 152; CDR1, CDR2 and CDR3 of SEQ ID NO 156; CDR1, CDR2 and CDR3 of SEQ ID NO. 160; CDR1, CDR2 and CDR3 of SEQ ID NO. 164; CDR1, CDR2 and CDR3 of SEQ ID No. 168; CDR1, CDR2 and CDR3 of SEQ ID NO 172; CDR1, CDR2 and CDR3 of SEQ ID NO. 176; CDR1, CDR2 and CDR3 of SEQ ID NO 180; or CDR1, CDR2 and CDR3 of SEQ ID NO 184.

118. The molecule of claim 106, wherein the variable domain of the second heavy chain comprises CDR1, CDR2, and CDR3 of SEQ ID No. 53; CDR1, CDR2 and CDR3 of SEQ ID NO 57; CDR1, CDR2 and CDR3 of SEQ ID NO 69; CDR1, CDR2 and CDR3 of SEQ ID NO 73; CDR1, CDR2 and CDR3 of SEQ ID NO 77; CDR1, CDR2 and CDR3 of SEQ ID NO 81; CDR1, CDR2 and CDR3 of SEQ ID NO 85; CDR1, CDR2 and CDR3 of SEQ ID NO 89; CDR1, CDR2 and CDR3 of SEQ ID NO. 93; CDR1, CDR2 and CDR3 of SEQ ID NO 97; CDR1, CDR2 and CDR3 of SEQ ID NO 101; CDR1, CDR2 and CDR3 of SEQ ID NO 105; CDR1, CDR2 and CDR3 of SEQ ID NO 109; CDR1, CDR2 and CDR3 of SEQ ID NO 117; CDR1, CDR2 and CDR3 of SEQ ID NO. 121; CDR1, CDR2 and CDR3 of SEQ ID No. 125; CDR1, CDR2 and CDR3 of SEQ ID No. 129; CDR1, CDR2 and CDR3 of SEQ ID NO 133; CDR1, CDR2 and CDR3 of SEQ ID NO 141; CDR1, CDR2 and CDR3 of SEQ ID NO. 145; CDR1, CDR2 and CDR3 of SEQ ID NO 149; CDR1, CDR2 and CDR3 of SEQ ID NO 153; CDR1, CDR2 and CDR3 of SEQ ID NO. 157; CDR1, CDR2 and CDR3 of SEQ ID NO 161; CDR1, CDR2 and CDR3 of SEQ ID NO. 165; CDR1, CDR2 and CDR3 of SEQ ID NO 169; CDR1, CDR2 and CDR3 of SEQ ID NO 173; CDR1, CDR2 and CDR3 of SEQ ID NO 177; CDR1, CDR2 and CDR3 of SEQ ID NO 181; or CDR1, CDR2 and CDR3 of SEQ ID NO 185.

119. The molecule of claim 106, wherein the variable domain of the second light chain comprises CDR1, CDR2, and CDR3 of SEQ ID No. 54; CDR1, CDR2 and CDR3 of SEQ ID NO 58; CDR1, CDR2 and CDR3 of SEQ ID NO 70; CDR1, CDR2 and CDR3 of SEQ ID NO 74; CDR1, CDR2 and CDR3 of SEQ ID NO. 78; CDR1, CDR2 and CDR3 of SEQ ID No. 82; CDR1, CDR2 and CDR3 of SEQ ID NO 86; CDR1, CDR2 and CDR3 of SEQ ID NO. 90; CDR1, CDR2 and CDR3 of SEQ ID No. 94; CDR1, CDR2 and CDR3 of SEQ ID NO 98; CDR1, CDR2 and CDR3 of SEQ ID No. 102; CDR1, CDR2 and CDR3 of SEQ ID NO 106; CDR1, CDR2 and CDR3 of SEQ ID NO. 110; CDR1, CDR2 and CDR3 of SEQ ID NO 118; CDR1, CDR2 and CDR3 of SEQ ID NO. 122; CDR1, CDR2 and CDR3 of SEQ ID NO. 126; CDR1, CDR2 and CDR3 of SEQ ID NO. 130; CDR1, CDR2 and CDR3 of SEQ ID No. 134; CDR1, CDR2 and CDR3 of SEQ ID NO. 142; CDR1, CDR2 and CDR3 of SEQ ID NO 146; CDR1, CDR2 and CDR3 of SEQ ID NO. 150; CDR1, CDR2 and CDR3 of SEQ ID NO 154; CDR1, CDR2 and CDR3 of SEQ ID NO 158; CDR1, CDR2 and CDR3 of SEQ ID NO. 162; CDR1, CDR2 and CDR3 of SEQ ID NO 166; CDR1, CDR2 and CDR3 of SEQ ID NO. 170; CDR1, CDR2 and CDR3 of SEQ ID NO 174; CDR1, CDR2 and CDR3 of SEQ ID NO 178; CDR1, CDR2 and CDR3 of SEQ ID NO 182; or CDR1, CDR2 and CDR3 of SEQ ID NO. 186.

120. The molecule of claim 106, wherein the molecule comprises SEQ ID No. 51, SEQ ID No. 52, SEQ ID No. 53, and SEQ ID No. 54.

121. The molecule of claim 106, wherein the molecule comprises SEQ ID No. 55, SEQ ID No. 56, SEQ ID No. 57, and SEQ ID No. 58.

122. The molecule of claim 106, wherein the molecule comprises SEQ ID No. 67, SEQ ID No. 68, SEQ ID No. 69, and SEQ ID No. 70.

123. The molecule of claim 106, wherein the molecule comprises SEQ ID No. 71, SEQ ID No. 72, SEQ ID No. 73, and SEQ ID No. 74.

124. The molecule of claim 106, wherein the molecule comprises SEQ ID No. 75, SEQ ID No. 76, SEQ ID No. 77, and SEQ ID No. 78.

125. The molecule of claim 106, wherein the molecule comprises SEQ ID No. 79, SEQ ID No. 80, SEQ ID No. 81, and SEQ ID No. 82.

126. The molecule of claim 106, wherein the molecule comprises SEQ ID No. 83, SEQ ID No. 84, SEQ ID No. 85, and SEQ ID No. 86.

127. The molecule of claim 106, wherein the molecule comprises SEQ ID No. 87, SEQ ID No. 88, SEQ ID No. 89, and SEQ ID No. 90.

128. The molecule of claim 106, wherein the molecule comprises SEQ ID No. 91, SEQ ID No. 92, SEQ ID No. 93, and SEQ ID No. 94.

129. The molecule of claim 106, wherein the molecule comprises SEQ ID No. 95, SEQ ID No. 96, SEQ ID No. 97, and SEQ ID No. 98.

130. The molecule of claim 106, wherein the molecule comprises SEQ ID No. 99, SEQ ID No. 100, SEQ ID No. 101, and SEQ ID No. 102.

131. The molecule of claim 106, wherein the molecule comprises SEQ ID No. 103,SEQ ID NO:104,SEQ ID NO:105 and SEQ ID No. 106.

132. The molecule of claim 106, wherein the molecule comprises SEQ ID No. 107, SEQ ID No. 108, SEQ ID No. 109, and SEQ ID No. 110.

133. The molecule of claim 106, wherein the molecule comprises SEQ ID No. 115, SEQ ID No. 116, SEQ ID No. 117, and SEQ ID No. 118.

134. The molecule of claim 106, wherein the molecule comprises SEQ ID No. 119, SEQ ID No. 120, SEQ ID No. 121, and SEQ ID No. 122.

135. The molecule of claim 106, wherein the molecule comprises SEQ ID No. 123, SEQ ID No. 124, SEQ ID No. 125, and SEQ ID No. 126.

136. The molecule of claim 106, wherein the molecule comprises SEQ ID No. 127, SEQ ID No. 128, SEQ ID No. 129, and SEQ ID No. 130.

137. The molecule of claim 106, wherein the molecule comprises SEQ ID No. 131, SEQ ID No. 132, SEQ ID No. 133, and SEQ ID No. 133.

138. The molecule of claim 106, wherein the molecule comprises SEQ ID No. 139, SEQ ID No. 140, SEQ ID No. 141, and SEQ ID No. 142.

139. The molecule of claim 106, wherein the molecule comprises SEQ ID No. 143, SEQ ID No. 144, SEQ ID No. 145, and SEQ ID No. 146.

140. The molecule of claim 106, wherein the molecule comprises SEQ ID No. 147, SEQ ID No. 148, SEQ ID No. 149, and SEQ ID No. 150.

141. The molecule of claim 106, wherein the molecule comprises SEQ ID No. 151, SEQ ID No. 152, SEQ ID No. 153, and SEQ ID No. 154.

142. The molecule of claim 106, wherein the molecule comprises SEQ ID No. 155, SEQ ID No. 156, SEQ ID No. 157, and SEQ ID No. 158.

143. The molecule of claim 106, wherein the molecule comprises SEQ ID No. 159, SEQ ID No. 160, SEQ ID No. 161, and SEQ ID No. 162.

144. The molecule of claim 106, wherein the molecule comprises SEQ ID No. 163, SEQ ID No. 164, SEQ ID No. 165, and SEQ ID No. 166.

145. The molecule of claim 106, wherein the molecule comprises SEQ ID No. 167, SEQ ID No. 168, SEQ ID No. 169, and SEQ ID No. 170.

146. The molecule of claim 106, wherein the molecule comprises SEQ ID No. 171, SEQ ID No. 172, SEQ ID No. 173, and SEQ ID No. 174.

147. The molecule of claim 106, wherein the molecule comprises SEQ ID No. 175, SEQ ID No. 176, SEQ ID No. 177, and SEQ ID No. 178.

148. The molecule of claim 106, wherein the molecule comprises SEQ ID No. 179, SEQ ID No. 180, SEQ ID No. 181, and SEQ ID No. 182.

149. The molecule of claim 106, wherein the molecule comprises SEQ ID No. 183, SEQ ID No. 184, SEQ ID No. 185, and SEQ ID No. 186.

150. The molecule of claim 106, wherein the molecule further comprises one or more disulfide bonds.

151. The molecule of claim 106, wherein the molecule further comprises one or more linker sequences.

152. The molecule of claim 106, wherein the molecule is capable of neutralizing coronavirus.

153. The molecule of claim 152, wherein the coronavirus is a SARS-CoV-2 virus.

154. The molecule of claim 153, wherein the SARS-CoV-2 virus strain is selected from the group consisting of WA1, b.1.1.7, B1.526, B1.351, P1, B1.352, and b.1.427.

155. The molecule of claim 106, wherein the molecule specifically recognizes a first epitope and a second epitope on a SARS-CoV-2 particle.

156. The molecule of claim 155, wherein the first epitope comprises at least a portion of a Receptor Binding Domain (RBD) on a spike protein of a SARS-CoV-2 particle.

157. The molecule of claim 155, wherein the second epitope comprises at least a portion of an N-terminal domain (NTD) on a spike protein of a SARS-CoV-2 particle.

158. A method of treating or preventing a covd-19 infection in a subject in need thereof, the method comprising administering to the subject a composition comprising a bispecific molecule comprising a first polypeptide chain and a second polypeptide chain, wherein:

the first polypeptide chain comprises:

a first light chain comprising a variable domain and a constant domain;

a first heavy chain comprising a variable domain and a constant domain;

the second polypeptide chain comprises:

a second light chain comprising a variable domain and a constant domain; and

A second heavy chain comprising a variable domain and a constant domain.

159. The method of claim 158, wherein the first polypeptide chain and the second polypeptide chain are covalently bonded to each other.

160. The method of claim 158, wherein the first heavy chain comprises a sequence that hybridizes at least to SEQ ID NO:51, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to a polypeptide sequence of SEQ ID NO:55, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to a polypeptide sequence of SEQ ID NO: 67%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical polypeptide sequences to at least 60%, 62%, 65%, 82%, 85%, 87%, 90%, 95%, 98%, 99% identical polypeptide sequences to at least 60%, 83%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98% of SEQ ID NO A polypeptide sequence 99% identical to SEQ ID NO:87, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to a polypeptide sequence of SEQ ID NO:91 at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to a polypeptide sequence of SEQ ID NO:95 at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical polypeptide sequences with at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical polypeptide sequences with at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 95%, 97%, 98%, 99% identical polypeptide sequences with at least 60%, 62%, 65%, 98%, 99%, 95%, 98%, 99%, 95%, 99%, 119% identical polypeptide sequences with at least 60% of SEQ ID NO 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical polypeptide sequences to those of SEQ ID NO:123, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to a polypeptide sequence of SEQ ID NO:127 at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to a polypeptide sequence of SEQ ID NO:131 at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID NO 139 at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID NO 143 at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID NO 147 at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID NO 151 at least 60%, 62%, 65%, 67%, 70%, 72%, 75% of SEQ ID NO, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical polypeptide sequences to SEQ ID NO:155, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to a polypeptide sequence of SEQ ID NO:159 at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to a polypeptide sequence of SEQ ID NO:163 is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to polypeptide sequence which is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90% identical to polypeptide sequence which is at least 60%, 67%, 70%, 72%, 75%, 67%, 70%, 72%, 75%, 80%, 85%, 87%, 90% of SEQ ID NO 92%, 95%, 97%, 98%, 99% identical or a polypeptide sequence at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 183.

161. The method of claim 158, wherein the first light chain comprises a sequence that hybridizes to SEQ ID NO:52, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to a polypeptide sequence of SEQ ID NO:56 at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to a polypeptide sequence of SEQ ID NO:68 at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical polypeptide sequence to at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 99% identical polypeptide sequence to at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 95%, 97%, 98%, 99% identical polypeptide sequence to at least 60%, 62%, 65%, 82%, 98%, 99% identical polypeptide sequence to SEQ ID NO 84 at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical polypeptide sequence to SEQ ID NO, and SEQ ID NO:88 at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to a polypeptide sequence of SEQ ID NO:92, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to a polypeptide sequence of SEQ ID NO:96 at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical polypeptide sequence to at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 99% identical polypeptide sequence to at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 95%, 97%, 98%, 99% identical polypeptide sequence to at least 60%, 62%, 65%, 82%, 98%, 99% identical polypeptide sequence to at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical polypeptide sequence to at least 60% of SEQ ID NO, 60%, 120% polypeptide sequence to at least 60% of SEQ ID NO 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical polypeptide sequences to those of SEQ ID NO:124 at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to a polypeptide sequence of SEQ ID NO:128 at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to a polypeptide sequence of SEQ ID NO:132, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID NO:140, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID NO:144, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID NO:148, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID NO:152, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 75% amino acid sequence identical to SEQ ID NO:152, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical polypeptide sequences to SEQ ID NO:156, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to a polypeptide sequence of SEQ ID NO:160, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to a polypeptide sequence of SEQ ID NO:164 at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical polypeptide sequence to at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical polypeptide sequence to at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 95%, 97%, 98%, 99% identical polypeptide sequence to at least 60%, 180%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92% identical polypeptide sequence to at least 60%, 180% to SEQ ID NO 95%, 97%, 98%, 99% identical or a polypeptide sequence which is at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID NO 184.

162. The method of claim 158, wherein the second heavy chain comprises a sequence identical to SEQ ID NO:53 at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to a polypeptide sequence of SEQ ID NO:57, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to a polypeptide sequence of SEQ ID NO:69 at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical polypeptide sequence, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical polypeptide sequence, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 95%, 97%, 98%, 99% identical polypeptide sequence of SEQ ID NO, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical polypeptide sequence of SEQ ID NO, and SEQ ID NO:89, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to a polypeptide sequence of SEQ ID NO:93 at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to a polypeptide sequence of SEQ ID NO:97 at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical polypeptide sequence to at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical polypeptide sequence to at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 95%, 97%, 98%, 99% identical polypeptide sequence to at least 60%, 117 at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical polypeptide sequence to at least 60%, 62%, 121% of SEQ ID NO 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical polypeptide sequences to those of SEQ ID NO:125 at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to a polypeptide sequence of SEQ ID NO:129 at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical polypeptide sequence to at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical polypeptide sequence to at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 95%, 97%, 98%, 99% identical polypeptide sequence to at least 60%, 62%, 65%, 95%, 97%, 98%, 99% identical polypeptide sequence to at least 60%, 62%, 80%, 95%, 97%, 98%, 99% identical polypeptide sequence to at least 60%, 149%, 95%, 67%, 98%, 99% identical polypeptide sequence to at least 60%, 67%, 70%, 75%, 98%, 99% of SEQ ID NO 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical polypeptide sequences to SEQ ID NO:157 at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to the polypeptide sequence of SEQ ID NO:161, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to a polypeptide sequence of SEQ ID NO:165 at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical polypeptide sequence to SEQ ID NO 169 at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical polypeptide sequence to SEQ ID NO 173 at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical polypeptide sequence to SEQ ID NO 177 at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical polypeptide sequence to SEQ ID NO 181 at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92% identical polypeptide sequence to SEQ ID NO 181 95%, 97%, 98%, 99% identical polypeptide sequence, or a polypeptide sequence at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 185.

163. The method of claim 158, wherein the second light chain comprises a sequence that hybridizes to SEQ ID NO:54, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to a polypeptide sequence of SEQ ID NO:58, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to a polypeptide sequence of SEQ ID NO:70, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical polypeptide sequence, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical polypeptide sequence, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 95%, 97%, 98%, 99% identical polypeptide sequence to SEQ ID NO 74, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical polypeptide sequence to SEQ ID NO 86, and SEQ ID NO:90, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to a polypeptide sequence of SEQ ID NO:94, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to a polypeptide sequence of SEQ ID NO:98 at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical polypeptide sequence to at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 99% identical polypeptide sequence to at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 95%, 97%, 98%, 99% identical polypeptide sequence to at least 60%, 62%, 80%, 95%, 98%, 99% identical polypeptide sequence to at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical polypeptide sequence to at least 60%, 118%, 62%, 98%, 99% of SEQ ID sequence to that of SEQ ID NO, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical polypeptide sequences to those of SEQ ID NO:126 at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to a polypeptide sequence of SEQ ID NO:130, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical polypeptide sequence, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical polypeptide sequence, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 95%, 97%, 98%, 99% identical polypeptide sequence, at least 150%, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical polypeptide sequence, at least 60%, 62%, 65%, 67%, 70%, 75%, 98%, 99% identical polypeptide sequence to SEQ ID NO 146, at least 60%, 62%, 65%, 80%, 75%, 80%, 90%, 98%, 99% 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical polypeptide sequences to SEQ ID NO:158, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to a polypeptide sequence of SEQ ID NO:162, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to a polypeptide sequence of SEQ ID NO:166 at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to a polypeptide sequence at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 95%, 97%, 98%, 99% identical to a polypeptide sequence at least 60%, 182%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92% to a polypeptide sequence at least 60%, 182% to a polypeptide sequence of SEQ ID NO of at least 60%, 62%, 65%, 67%, 87%, 90%, 98%, 99% 95%, 97%, 98%, 99% identical or a polypeptide sequence at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID No. 186.

164. The method of claim 158, wherein the variable domain of the first heavy chain comprises a sequence that hybridizes to SEQ ID NO:15, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to a polypeptide sequence of SEQ ID NO:27, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to a polypeptide sequence of SEQ ID NO:35 at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical polypeptide sequence to at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical polypeptide sequence to at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 95%, 97%, 98%, 99% identical polypeptide sequence to at least 60%, 62%, 65%, 67%, 70%, 72%, 90%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 95%, 97% identical polypeptide sequence to at least 60%, 75% to SEQ ID NO 19., 98%, 99% identical polypeptide sequence to at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical polypeptide sequence to at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 98%, 99% identical polypeptide sequence to at least 60%, 62%, 80%, 82%, 85%, 87%, 90%, 98%, 99% identical polypeptide sequence to at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical polypeptide sequence to at least 60%, 41.

165. The method of claim 158, wherein the variable domain of the first light chain comprises a sequence that hybridizes to SEQ ID NO:16, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to a polypeptide sequence of SEQ ID NO:28, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to a polypeptide sequence of SEQ ID NO:36 at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical polypeptide sequence to at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97% identical polypeptide sequence to at least 20% to SEQ ID NO, 98%, 99% identical polypeptide sequence to at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical polypeptide sequence to at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 98%, 99% identical polypeptide sequence to at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical polypeptide sequence to at least 60%, 42.

166. The method of claim 158, wherein the variable domain of the second heavy chain comprises a sequence that hybridizes to SEQ ID NO:27, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to a polypeptide sequence of SEQ ID NO:15, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to a polypeptide sequence of SEQ ID NO:5 at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical polypeptide sequence to at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 99% identical polypeptide sequence to at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical polypeptide sequence to at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97% identical polypeptide sequence to at least 60%, 29% to SEQ ID NO, 98%, 99% identical polypeptide sequence to SEQ ID NO:35, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to a polypeptide sequence of SEQ ID NO:23, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to a polypeptide sequence of SEQ ID NO:25, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID NO:37, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID NO:39, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID NO:43, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 95%, 97%, 98%, 99% identical to SEQ ID NO:9, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to SEQ ID NO.

167. The method of claim 158, wherein the variable domain of the second light chain comprises a sequence that hybridizes to SEQ ID NO:28, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to a polypeptide sequence of SEQ ID NO:16, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to a polypeptide sequence of SEQ ID NO:6 at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical polypeptide sequence to at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 99% identical polypeptide sequence to at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical polypeptide sequence to at least 60%, 62%, 65%, 82%, 98%, 99% identical polypeptide sequence to SEQ ID NO 30 at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97% identical to SEQ ID NO, 98%, 99% identical polypeptide sequence to SEQ ID NO:36, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to a polypeptide sequence of SEQ ID NO:24, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical to a polypeptide sequence of SEQ ID NO:26, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 97%, 98%, 99% identical polypeptide sequence to SEQ ID NO, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 95%, 98%, 99% identical polypeptide sequence to SEQ ID NO, or at least 10, at least 60%, 62%, 65%, 67%, 70%, 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 92%, 95%, 98%, 99% identical polypeptide sequence to SEQ ID NO.

168. The method of claim 158, wherein the variable domain of the first heavy chain comprises CDR1, CDR2, and CDR3 of SEQ ID No. 51; CDR1, CDR2 and CDR3 of SEQ ID NO. 55; CDR1, CDR2 and CDR3 of SEQ ID NO 67; CDR1, CDR2 and CDR3 of SEQ ID NO 71; CDR1, CDR2 and CDR3 of SEQ ID NO 75; CDR1, CDR2 and CDR3 of SEQ ID NO 79; CDR1, CDR2 and CDR3 of SEQ ID NO 83; CDR1, CDR2 and CDR3 of SEQ ID NO 87; CDR1, CDR2 and CDR3 of SEQ ID NO. 91; CDR1, CDR2 and CDR3 of SEQ ID NO 95; CDR1, CDR2 and CDR3 of SEQ ID No. 99; CDR1, CDR2 and CDR3 of SEQ ID NO 103; CDR1, CDR2 and CDR3 of SEQ ID NO 107; CDR1, CDR2 and CDR3 of SEQ ID NO 115; CDR1, CDR2 and CDR3 of SEQ ID No. 119; CDR1, CDR2 and CDR3 of SEQ ID NO. 123; CDR1, CDR2 and CDR3 of SEQ ID NO 127; CDR1, CDR2 and CDR3 of SEQ ID NO. 131; CDR1, CDR2 and CDR3 of SEQ ID NO 139; CDR1, CDR2 and CDR3 of SEQ ID NO 143; CDR1, CDR2 and CDR3 of SEQ ID No. 147; CDR1, CDR2 and CDR3 of SEQ ID NO 151; CDR1, CDR2 and CDR3 of SEQ ID NO 155; CDR1, CDR2 and CDR3 of SEQ ID NO 159; CDR1, CDR2 and CDR3 of SEQ ID NO 163; CDR1, CDR2 and CDR3 of SEQ ID NO 167; CDR1, CDR2 and CDR3 of SEQ ID NO. 171; CDR1, CDR2 and CDR3 of SEQ ID NO 175; CDR1, CDR2 and CDR3 of SEQ ID NO 179; or CDR1, CDR2 and CDR3 of SEQ ID NO 183.

169. The method of claim 158, wherein the variable domain of the first light chain comprises CDR1, CDR2, and CDR3 of SEQ ID No. 52; CDR1, CDR2 and CDR3 of SEQ ID NO. 56; CDR1, CDR2 and CDR3 of SEQ ID NO. 68; CDR1, CDR2 and CDR3 of SEQ ID NO 72; CDR1, CDR2 and CDR3 of SEQ ID NO 76; CDR1, CDR2 and CDR3 of SEQ ID NO 80; CDR1, CDR2 and CDR3 of SEQ ID NO 84; CDR1, CDR2 and CDR3 of SEQ ID NO 88; CDR1, CDR2 and CDR3 of SEQ ID NO. 92; CDR1, CDR2 and CDR3 of SEQ ID No. 96; CDR1, CDR2 and CDR3 of SEQ ID NO. 100; CDR1, CDR2 and CDR3 of SEQ ID No. 104; CDR1, CDR2 and CDR3 of SEQ ID NO 108; CDR1, CDR2 and CDR3 of SEQ ID NO. 116; CDR1, CDR2 and CDR3 of SEQ ID NO. 120; CDR1, CDR2 and CDR3 of SEQ ID NO 124; CDR1, CDR2 and CDR3 of SEQ ID NO 128; CDR1, CDR2 and CDR3 of SEQ ID NO. 132; CDR1, CDR2 and CDR3 of SEQ ID NO. 140; CDR1, CDR2 and CDR3 of SEQ ID NO. 144; CDR1, CDR2 and CDR3 of SEQ ID NO. 148; CDR1, CDR2 and CDR3 of SEQ ID NO 152; CDR1, CDR2 and CDR3 of SEQ ID NO 156; CDR1, CDR2 and CDR3 of SEQ ID NO. 160; CDR1, CDR2 and CDR3 of SEQ ID NO. 164; CDR1, CDR2 and CDR3 of SEQ ID No. 168; CDR1, CDR2 and CDR3 of SEQ ID NO 172; CDR1, CDR2 and CDR3 of SEQ ID NO. 176; CDR1, CDR2 and CDR3 of SEQ ID NO 180; or CDR1, CDR2 and CDR3 of SEQ ID NO 184.

170. The method of claim 158, wherein the variable domain of the second heavy chain comprises CDR1, CDR2, and CDR3 of SEQ ID No. 53; CDR1, CDR2 and CDR3 of SEQ ID NO 57; CDR1, CDR2 and CDR3 of SEQ ID NO 69; CDR1, CDR2 and CDR3 of SEQ ID NO 73; CDR1, CDR2 and CDR3 of SEQ ID NO 77; CDR1, CDR2 and CDR3 of SEQ ID NO 81; CDR1, CDR2 and CDR3 of SEQ ID NO 85; CDR1, CDR2 and CDR3 of SEQ ID NO 89; CDR1, CDR2 and CDR3 of SEQ ID NO. 93; CDR1, CDR2 and CDR3 of SEQ ID NO 97; CDR1, CDR2 and CDR3 of SEQ ID NO 101; CDR1, CDR2 and CDR3 of SEQ ID NO 105; CDR1, CDR2 and CDR3 of SEQ ID NO 109; CDR1, CDR2 and CDR3 of SEQ ID NO 117; CDR1, CDR2 and CDR3 of SEQ ID NO. 121; CDR1, CDR2 and CDR3 of SEQ ID No. 125; CDR1, CDR2 and CDR3 of SEQ ID No. 129; CDR1, CDR2 and CDR3 of SEQ ID NO 133; CDR1, CDR2 and CDR3 of SEQ ID NO 141; CDR1, CDR2 and CDR3 of SEQ ID NO. 145; CDR1, CDR2 and CDR3 of SEQ ID NO 149; CDR1, CDR2 and CDR3 of SEQ ID NO 153; CDR1, CDR2 and CDR3 of SEQ ID NO. 157; CDR1, CDR2 and CDR3 of SEQ ID NO 161; CDR1, CDR2 and CDR3 of SEQ ID NO. 165; CDR1, CDR2 and CDR3 of SEQ ID NO 169; CDR1, CDR2 and CDR3 of SEQ ID NO 173; CDR1, CDR2 and CDR3 of SEQ ID NO 177; CDR1, CDR2 and CDR3 of SEQ ID NO 181; or CDR1, CDR2 and CDR3 of SEQ ID NO 185.

171. The method of claim 158, wherein the variable domain of the second light chain comprises CDR1, CDR2, and CDR3 of SEQ ID No. 54; CDR1, CDR2 and CDR3 of SEQ ID NO 58; CDR1, CDR2 and CDR3 of SEQ ID NO 70; CDR1, CDR2 and CDR3 of SEQ ID NO 74; CDR1, CDR2 and CDR3 of SEQ ID NO. 78; CDR1, CDR2 and CDR3 of SEQ ID No. 82; CDR1, CDR2 and CDR3 of SEQ ID NO 86; CDR1, CDR2 and CDR3 of SEQ ID NO. 90; CDR1, CDR2 and CDR3 of SEQ ID No. 94; CDR1, CDR2 and CDR3 of SEQ ID NO 98; CDR1, CDR2 and CDR3 of SEQ ID No. 102; CDR1, CDR2 and CDR3 of SEQ ID NO 106; CDR1, CDR2 and CDR3 of SEQ ID NO. 110; CDR1, CDR2 and CDR3 of SEQ ID NO 118; CDR1, CDR2 and CDR3 of SEQ ID NO. 122; CDR1, CDR2 and CDR3 of SEQ ID NO. 126; CDR1, CDR2 and CDR3 of SEQ ID NO. 130; CDR1, CDR2 and CDR3 of SEQ ID No. 134; CDR1, CDR2 and CDR3 of SEQ ID NO. 142; CDR1, CDR2 and CDR3 of SEQ ID NO 146; CDR1, CDR2 and CDR3 of SEQ ID NO. 150; CDR1, CDR2 and CDR3 of SEQ ID NO 154; CDR1, CDR2 and CDR3 of SEQ ID NO 158; CDR1, CDR2 and CDR3 of SEQ ID NO. 162; CDR1, CDR2 and CDR3 of SEQ ID NO 166; CDR1, CDR2 and CDR3 of SEQ ID NO. 170; CDR1, CDR2 and CDR3 of SEQ ID NO 174; CDR1, CDR2 and CDR3 of SEQ ID NO 178; CDR1, CDR2 and CDR3 of SEQ ID NO 182; or CDR1, CDR2 and CDR3 of SEQ ID NO. 186.

172. The method of claim 158, wherein the molecule comprises SEQ ID No. 51, SEQ ID No. 52, SEQ ID No. 53, and SEQ ID No. 54.

173. The method of claim 158, wherein the molecule comprises SEQ ID No. 55, SEQ ID No. 56, SEQ ID No. 57, and SEQ ID No. 58.

174. The method of claim 158, wherein the molecule comprises SEQ ID No. 67, SEQ ID No. 68, SEQ ID No. 69, and SEQ ID No. 70.

175. The method of claim 158, wherein the molecule comprises SEQ ID No. 71, SEQ ID No. 72, SEQ ID No. 73, and SEQ ID No. 74.

176. The method of claim 158, wherein the molecule comprises SEQ ID No. 75, SEQ ID No. 76, SEQ ID No. 77, and SEQ ID No. 78.

177. The method of claim 158, wherein the molecule comprises SEQ ID No. 79, SEQ ID No. 80, SEQ ID No. 81, and SEQ ID No. 82.

178. The method of claim 158, wherein the molecule comprises SEQ ID No. 83, SEQ ID No. 84, SEQ ID No. 85, and SEQ ID No. 86.

179. The method of claim 158, wherein the molecule comprises SEQ ID No. 87, SEQ ID No. 88, SEQ ID No. 89, and SEQ ID No. 90.

180. The method of claim 158, wherein the molecule comprises SEQ ID No. 91, SEQ ID No. 92, SEQ ID No. 93, and SEQ ID No. 94.

181. The method of claim 158, wherein the molecule comprises SEQ ID No. 95, SEQ ID No. 96, SEQ ID No. 97, and SEQ ID No. 98.

182. The method of claim 158, wherein the molecule comprises SEQ ID No. 99, SEQ ID No. 100, SEQ ID No. 101, and SEQ ID No. 102.

183. The method of claim 158, wherein the molecule comprises SEQ ID No. 103,SEQ ID NO:104,SEQ ID NO:105 and SEQ ID No. 106.

184. The method of claim 158, wherein the molecule comprises SEQ ID No. 107, SEQ ID No. 108, SEQ ID No. 109, and SEQ ID No. 110.

185. The method of claim 158, wherein the molecule comprises SEQ ID No. 115, SEQ ID No. 116, SEQ ID No. 117, and SEQ ID No. 118.

186. The method of claim 158, wherein the molecule comprises SEQ ID No. 119, SEQ ID No. 120, SEQ ID No. 121, and SEQ ID No. 122.

187. The method of claim 158, wherein the molecule comprises SEQ ID No. 123, SEQ ID No. 124, SEQ ID No. 125, and SEQ ID No. 126.

188. The method of claim 158, wherein the molecule comprises SEQ ID No. 127, SEQ ID No. 128, SEQ ID No. 129, and SEQ ID No. 130.

189. The method of claim 158, wherein the molecule comprises SEQ ID No. 131, SEQ ID No. 132, SEQ ID No. 133, and SEQ ID No. 133.

190. The method of claim 158, wherein the molecule comprises SEQ ID No. 139, SEQ ID No. 140, SEQ ID No. 141, and SEQ ID No. 142.

191. The method of claim 158, wherein the molecule comprises SEQ ID No. 143, SEQ ID No. 144, SEQ ID No. 145, and SEQ ID No. 146.

192. The method of claim 158, wherein the molecule comprises SEQ ID No. 147, SEQ ID No. 148, SEQ ID No. 149 and SEQ ID No. 150.

193. The method of claim 158, wherein the molecule comprises SEQ ID No. 151, SEQ ID No. 152, SEQ ID No. 153, and SEQ ID No. 154.

194. The method of claim 158, wherein the molecule comprises SEQ ID No. 155, SEQ ID No. 156, SEQ ID No. 157, and SEQ ID No. 158.

195. The method of claim 158, wherein the molecule comprises SEQ ID No. 159, SEQ ID No. 160, SEQ ID No. 161, and SEQ ID No. 162.

196. The method of claim 158, wherein the molecule comprises SEQ ID No. 163, SEQ ID No. 164, SEQ ID No. 165, and SEQ ID No. 166.

197. The method of claim 158, wherein the molecule comprises SEQ ID No. 167, SEQ ID No. 168, SEQ ID No. 169, and SEQ ID No. 170.

198. The method of claim 158, wherein the molecule comprises SEQ ID No. 171, SEQ ID No. 172, SEQ ID No. 173, and SEQ ID No. 174.

199. The method of claim 158, wherein the molecule comprises SEQ ID No. 175, SEQ ID No. 176, SEQ ID No. 177, and SEQ ID No. 178.

200. The method of claim 158, wherein the molecule comprises SEQ ID No. 179, SEQ ID No. 180, SEQ ID No. 181, and SEQ ID No. 182.

201. The method of claim 158, wherein the molecule comprises SEQ ID No. 183, SEQ ID No. 184, SEQ ID No. 185, and SEQ ID No. 186.

202. The method of claim 158, wherein the molecule further comprises one or more disulfide bonds.

203. The method of claim 158, wherein the molecule further comprises one or more linker sequences.

204. The method of claim 158, wherein the molecule is capable of neutralizing a coronavirus.

205. The method of claim 204, wherein the coronavirus is a SARS-CoV-2 virus.

206. The method of claim 205, wherein the SARS-CoV-2 virus strain is selected from the group consisting of WA1, b.1.1.7, B1.526, B1.351, P1, B1.352, and b.1.427.

207. The method of claim 158, wherein the molecule specifically recognizes a first epitope and a second epitope on a SARS-CoV-2 particle.

208. The method of claim 207, wherein the first epitope comprises at least a portion of a Receptor Binding Domain (RBD) on a spike protein of a SARS-CoV-2 particle.

209. The method of claim 207, wherein the second epitope comprises at least a portion of an N-terminal domain (NTD) on a spike protein of a SARS-CoV-2 particle.

210. The method of claim 158, wherein the composition is administered in combination with a second therapeutic agent.