REPAIRED THERAPEUTIC AND PROPHYLACTIC ANTIBODIES AGAINST SARS-CoV-2 VARIANTS

SEQUENCE LISTING

The instant application contains a Sequence Listing which has been submitted electronically in XML format and is hereby incorporated by reference in its entirety. Said XML copy, created Apr. 7, 2023, is named 093866-9303.xml and is 280,240 bytes in size.

TECHNICAL FIELD

The present disclosure relates generally to the fields of infectious disease and immunology. Specifically, the disclosure relates to engineered human antibodies against coronaviruses that cause Severe Acute Respiratory Syndrome (SARS), such as SARS-CoV-2 and its variants.

BACKGROUND

Coronaviruses are a family of viruses that can cause illnesses such as the common cold, pneumonia, severe acute respiratory syndrome (SARS) and Middle East respiratory syndrome (MERS). In 2019, a new coronavirus was identified as the causative agent for a severe acute respiratory syndrome (SARS) outbreak that originated in Wuhan, China. The virus is known as severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). The disease it causes is called coronavirus disease 2019 (COVID-19).

Vaccination against SARS-CoV-2 reduces the burden of COVID-19. However, vaccination is not safe for some persons, including immunocompromised persons and those who cannot be vaccinated. Accordingly, a potential option for COVID-19 immunoprophylaxis in those who cannot be vaccinated includes monoclonal antibodies, which protect against disease irrespective of immune system status and provide rapid protection. Such prophylactics can provide crucial immune protection, especially for immunocompromised individuals. Failure of such a prophylactic could result in illness or death of those who cannot be vaccinated.

Some combinations of monoclonal antibodies are already in use through the U.S. Food and Drug Administration (FDA)-approved Emergency or temporary Use Authorizations for preexposure (e.g., Ronapreve™ (casirivimab and imdevimab) 120 mg/mL solution for injection or infusion), postexposure (e.g., REGEN-COV™ (casirivimab and imdevimab)), prophylaxis against COVID-19, or treatment of mild-to-moderate disease (e.g., Bamlanivimab and etesevimab; sotrovimab; or Evusheld (AZD7442)). ZD7442 is a combination of two fully human, SARS-CoV-2-neutralizing monoclonal antibodies (tixagevimab and cilgavimab). The antibody COV2-2130 (Cilgavimab; AZD1061), which after further engineering, is one of two constituents of ZD7442. During the course of the COVID-19 pandemic, the SARS-CoV-2 virus, like most viruses, evolved gradually into new strains that evaded the protection offered by the human immune system and available medical countermeasures, including therapeutic or prophylactic monoclonal antibodies and aforementioned FDA-authorized countermeasures, and other candidates.

Accordingly, there is a need for monoclonal antibodies for use as immunoprophylaxis for preexposure, postexposure, prophylaxis, or treatment of mild-to-moderate pneumonia or severe acute respiratory syndrome caused by a coronavirus. This disclosure addresses these needs.

SUMMARY OF THE DISCLOSURE

One aspect of the present disclosure provides an isolated antibody or antibody fragment thereof that binds to a SARS-CoV-2 surface protein. In one aspect, the antibody or fragments thereof have enhanced binding and/or neutralization characteristics against all known SARS-CoV-2 strains when compared to antibodies previously described in the prior art, e.g., without limitation a monoclonal antibody selected from sotrovimab (S309), COV2-2381, cilgavimab (COV2-2130 or AZD1061), and/or tixagevimab (COV2-2196 or AZD8895). In some embodiments, the SARS-CoV-2 surface protein is a spike (S) glycoprotein. In some embodiments, the monoclonal antibody or antibody fragment binds to and/or neutralizes SARS-CoV-2 Omicron (B.1.1.529, BA.1, BA.1.1, BA.2, BA.3, BA.4 and BA.5 lineages).

In some embodiments, the antibody or antibody fragment comprises, or consists essentially of, or consists of: (a) a heavy chain complementary determining region 1 (CDRH1) comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 196, a CDRH2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 197, and a CDRH3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 202, and a light chain complementary determining region 1 (CDRL1) comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 226, a CDRL2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 242 or 243, and a CDRL3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 251; (b) a CDRH1 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 196, a CDRH2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 198, and a CDRH3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 203, and a CDRL1 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 227, a CDRL2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 242 or 243, and a CDRL3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 251; (c) a CDRH1 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 196, a CDRH2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 199, and a CDRH3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 204, and a CDRL1 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 227, a CDRL2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 242 or 243, and a CDRL3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 251; (d) a CDRH1 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 196, a CDRH2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 198, and a CDRH3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 204, and a CDRL1 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 228, a CDRL2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 242 or 243, and a CDRL3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 251; (e) a CDRH1 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 196, a CDRH2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 198, and a CDRH3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 204, and a CDRL1 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 229, a CDRL2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 242 or 243, and a CDRL3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 251; (f) a CDRH1 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 196, a CDRH2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 198, and a CDRH3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 204, and a CDRL1 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 230, a CDRL2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 242 or 244, and a CDRL3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 251; (g) a CDRH1 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 196, a CDRH2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 198, and a CDRH3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 204, and a CDRL1 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 231, a CDRL2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 242 or 245, and a CDRL3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 251; (h) a CDRH1 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 196, a CDRH2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 200, and a CDRH3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 204, and a CDRL1 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 232, a CDRL2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 242 or 245, and a CDRL3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 251; (i) a CDRH1 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 196, a CDRH2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 198, and a CDRH3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 205, and a CDRL1 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 233, a CDRL2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 242 or 243, and a CDRL3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 251; (j) a CDRH1 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 196, a CDRH2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 198, and a CDRH3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 205, and a CDRL1 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 234, a CDRL2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 242 or 245, and a CDRL3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 251; (k) a CDRH1 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 196, a CDRH2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 198, and a CDRH3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 206, and a CDRL1 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 235, a CDRL2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 242 or 246, and a CDRL3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 251; (l) a CDRH1 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 196, a CDRH2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 197, and a CDRH3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 207, and a CDRL1 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 236, a CDRL2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 242 or 243, and a CDRL3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 251; (m) a CDRH1 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 196, a CDRH2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 201, and a CDRH3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 208, and a CDRL1 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 228, a CDRL2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 242 or 247, and a CDRL3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 251; or (n) a CDRH1 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 196, a CDRH2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 198, and a CDRH3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 209, and a CDRL1 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 234, a CDRL2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 242 or 245, and a CDRL3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 251.

In some embodiments, the antibody or antibody fragment comprises, or consists essentially of, or consists of: (a) a heavy chain variable sequence having at least 70%, 80%, 90%, 95%, 96%, 97%, 98%, or 99% identity to the amino acid sequence of SEQ ID NO: 100, 101, 102, 103, 104, 105, 106, 107, 108, or 109; and/or (b) a light chain variable sequences having at least 70%, 80%, 90%, 95%, 96%, 97%, 98%, or 99% identity to the amino acid sequence of SEQ ID NO: 155, 156, 157, 158, 159, 160, 161, 162, 163, 164, 165, or 166.

In some embodiments, the antibody or antibody fragment comprises, or consists essentially of, or consists of a heavy chain variable sequence and a light chain variable sequence having at least 96% identity to an amino acid sequence comprising, or consisting essentially of, or consisting of an amino acid sequence selected from: (a) the amino acid sequence of SEQ ID NO: 100 and the amino acid sequence of SEQ ID NO: 155; (b) the amino acid sequence of SEQ ID NO: 101 and the amino acid sequence of SEQ ID NO: 156; (c) the amino acid sequence of SEQ ID NO: 102 and the amino acid sequence of SEQ ID NO: 156; (d) the amino acid sequence of SEQ ID NO: 103 and the amino acid sequence of SEQ ID NO: 157; (e) the amino acid sequence of SEQ ID NO: 103 and the amino acid sequence of SEQ ID NO: 158; (f) the amino acid sequence of SEQ ID NO: 103 and the amino acid sequence of SEQ ID NO: 159; (g) the amino acid sequence of SEQ ID NO: 103 and the amino acid sequence of SEQ ID NO: 160; (h) the amino acid sequence of SEQ ID NO: 104 and the amino acid sequence of SEQ ID NO: 161; (i) the amino acid sequence of SEQ ID NO: 105 and the amino acid sequence of SEQ ID NO: 162; (j) the amino acid sequence of SEQ ID NO: 105 and the amino acid sequence of SEQ ID NO: 163; (k) the amino acid sequence of SEQ ID NO: 106 and the amino acid sequence of SEQ ID NO: 164; (l) the amino acid sequence of SEQ ID NO: 107 and the amino acid sequence of SEQ ID NO: 165; (m) the amino acid sequence of SEQ ID NO: 108 and the amino acid sequence of SEQ ID NO: 166; or (n) the amino acid sequence of SEQ ID NO: 109 and the amino acid sequence of SEQ ID NO: 163. In one aspect the sequence that is at least 96% identical has a CDR sequence that is 100% identical to the CDR of the reference polypeptide. The reference CDRs are provided above, and further within this document. Stated another way, the amino acid substitutions and modifications are in regions outside of the CDR regions.

In some embodiments, the antibody fragment is a recombinant antibody variable domain fragment (Fv fragment) and/or the antibody fragment comprises, or consists essentially of, or consists of the amino acid sequence of SEQ ID NOs: 1-14 or an equivalent of each thereof. In one aspect the equivalent has at least 96% identity to any one of SEQ ID NOS: 1-14 and further in one embodiment has a CDR sequence that is 100% identical to the CDR of the reference polypeptide. The reference CDRs are provided above, and further within this document. Stated another way, the amino acid substitutions and modifications are in regions outside of the CDR regions.

In some embodiments, the antibody or antibody fragment comprises a heavy chain complementary determining region 1 (CDRH1) comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 196, a CDRH2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 198, and a CDRH3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 205, and a CDRL1 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 227, a CDRL2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 242 or 243, and a CDRL3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 251.

In some embodiments, the antibody or antibody fragment comprises a CDRH1 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 196, a CDRH2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 198, and a CDRH3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 204, and a CDRL1 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 237, a CDRL2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 242 or 243, and a CDRL3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 251.

In some embodiments, the antibody or antibody fragment comprises, or consisting essentially of, or consisting of a CDRH1 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 196, a CDRH2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 198, and a CDRH3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 204, and a CDRL1 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 238, a CDRL2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 242 or 243, and a CDRL3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 251.

In some embodiments, the antibody or antibody fragment comprises a CDRH1 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 196, a CDRH2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 198, and a CDRH3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 204, and a CDRL1 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 227, a CDRL2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 242 or 245, and a CDRL3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 251.

In some embodiments, the antibody or antibody fragment comprises a CDRH1 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 196, a CDRH2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 198, and a CDRH3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 204, and a CDRL1 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 227, a CDRL2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 242 or 244, and a CDRL3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 251.

In some embodiments, the antibody or antibody fragment comprises a CDRH1 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 196, a CDRH2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 197, and a CDRH3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 204, and a CDRL1 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 227, a CDRL2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 242 or 243, and a CDRL3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 251.

In some embodiments, the antibody or antibody fragment comprises a CDRH1 comprising or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 196, a CDRH2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 199, and a CDRH3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 209, and a CDRL1 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 234, a CDRL2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 242 or 245, and a CDRL3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 251.

In some embodiments, the antibody or antibody fragment comprises a CDRH1 comprising or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 196, a CDRH2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 197, and a CDRH3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 209, and a CDRL1 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 234, a CDRL2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 242 or 245, and a CDRL3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 251.

In some embodiments, the antibody or antibody fragment comprises a CDRH1 comprising or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 196, a CDRH2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 198, and a CDRH3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 210, and a CDRL1 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 234, a CDRL2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 242 or 245, and a CDRL3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 251.

In some embodiments, the antibody or antibody fragment comprises a CDRH1 comprising or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 196, a CDRH2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 198, and a CDRH3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 209, and a CDRL1 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 239, a CDRL2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 242 or 245, and a CDRL3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 251.

In some embodiments, the antibody or antibody fragment comprises a CDRH1 comprising or consisting essentially of, or consisting of g the amino acid sequence of SEQ ID NO: 196, a CDRH2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 199, and a CDRH3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 210, and a CDRL1 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 234, a CDRL2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 242 or 245, and a CDRL3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 251.

In some embodiments, the antibody or antibody fragment comprises a CDRH1 comprising or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 196, a CDRH2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 197, and a CDRH3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 210, and a CDRL1 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 234, a CDRL2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 242 or 245, and a CDRL3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 251.

In some embodiments, the antibody or antibody fragment comprises a CDRH1 comprising or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 196, a CDRH2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 199, and a CDRH3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 205, and a CDRL1 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 234, a CDRL2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 242 or 245, and a CDRL3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 251.

In some embodiments, the antibody or antibody fragment comprises a CDRH1 comprising or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 196, a CDRH2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 197, and a CDRH3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 205, and a CDRL1 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 234, a CDRL2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 242 or 245, and a CDRL3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 251.

In some embodiments, the antibody or antibody fragment comprises a CDRH1 comprising or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 196, a CDRH2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 198, and a CDRH3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 211, and a CDRL1 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 234, a CDRL2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 242 or 245, and a CDRL3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 251.

In some embodiments, the antibody or antibody fragment comprises a CDRH1 comprising or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 196, a CDRH2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 198, and a CDRH3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 205, and a CDRL1 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 239, a CDRL2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 242 or 245, and a CDRL3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 251.

In some embodiments, the antibody or antibody fragment comprises a CDRH1 comprising or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 196, a CDRH2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 199, and a CDRH3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 206, and a CDRL1 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 235, a CDRL2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 242 or 246, and a CDRL3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 251.

In some embodiments, the antibody or antibody fragment comprises a CDRH1 comprising or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 196, a CDRH2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 197 and a CDRH3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 206, and a CDRL1 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 235, a CDRL2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 242 or 246, and a CDRL3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 251.

In some embodiments, the antibody or antibody fragment comprises a CDRH1 comprising or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 196, a CDRH2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 198, and a CDRH3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 212, and a CDRL1 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 235, a CDRL2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 242 or 246, and a CDRL3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 251.

In some embodiments, the antibody or antibody fragment comprises a CDRH1 comprising or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 196, a CDRH2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 198, and a CDRH3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 213, and a CDRL1 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 235, a CDRL2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 242 or 246, and a CDRL3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 251.

In some embodiments, the antibody or antibody fragment comprises a CDRH1 comprising or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 196, a CDRH2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 198, and a CDRH3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 214, and a CDRL1 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 235, a CDRL2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 242 or 246, and a CDRL3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 251.

In some embodiments, the antibody or antibody fragment comprises a CDRH1 comprising or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 196, a CDRH2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 198, and a CDRH3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 206, and a CDRL1 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 235, a CDRL2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 242 or 248, and a CDRL3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 251.

In some embodiments, the antibody or antibody fragment comprises a CDRH1 comprising or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 196, a CDRH2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 198, and a CDRH3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 206, and a CDRL1 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 235, a CDRL2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 242 or 249, and a CDRL3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 251.

In some embodiments, the antibody or antibody fragment comprises a CDRH1 comprising or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 196, a CDRH2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 197, and a CDRH3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 208, and a CDRL1 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 228, a CDRL2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 242 or 247, and a CDRL3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 251.

In some embodiments, the antibody or antibody fragment comprises a CDRH1 comprising or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 196, a CDRH2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 199, and a CDRH3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 208, and a CDRL1 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 228, a CDRL2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 242 or 247, and a CDRL3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 251.

In some embodiments, the antibody or antibody fragment comprises a CDRH1 comprising or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 196, a CDRH2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 201, and a CDRH3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 215, and a CDRL1 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 228, a CDRL2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 242 or 247, and a CDRL3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 251.

In some embodiments, the antibody or antibody fragment comprises a CDRH1 comprising or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 196, a CDRH2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 201, and a CDRH3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 216, and a CDRL1 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 228, a CDRL2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 242 or 247, and a CDRL3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 251.

In some embodiments, the antibody or antibody fragment comprises a CDRH1 comprising or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 196, a CDRH2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 201, and a CDRH3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 217, and a CDRL1 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 228, a CDRL2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 242 or 247, and a CDRL3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 251.

In some embodiments, the antibody or antibody fragment comprises a CDRH1 comprising or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 196, a CDRH2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 201, and a CDRH3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 208, and a CDRL1 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 228, a CDRL2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 242 or 250, and a CDRL3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 251.

In some embodiments, the antibody or antibody fragment comprises a CDRH1 comprising or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 196, a CDRH2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 197, and a CDRH3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 218, and a CDRL1 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 236, a CDRL2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 242 or 243, and a CDRL3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 251.

In some embodiments, the antibody or antibody fragment comprises a CDRH1 comprising or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 196, a CDRH2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 197, and a CDRH3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 219, and a CDRL1 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 236, a CDRL2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 242 or 243, and a CDRL3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 251.

In some embodiments, the antibody or antibody fragment comprises a CDRH1 comprising or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 196, a CDRH2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 197, and a CDRH3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 220, and a CDRL1 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 236, a CDRL2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 242 or 243, and a CDRL3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 251.

In some embodiments, the antibody or antibody fragment comprises a CDRH1 comprising or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 196, a CDRH2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 197, and a CDRH3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 218, and a CDRL1 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 236, a CDRL2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 242 or 245, and a CDRL3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 251.

In some embodiments, the antibody or antibody fragment comprises a CDRH1 comprising or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 196, a CDRH2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 197, and a CDRH3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 207, and a CDRL1 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 236, a CDRL2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 242 or 245, and a CDRL3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 251.

In some embodiments, the antibody or antibody fragment comprises a CDRH1 comprising or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 196, a CDRH2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 199, and a CDRH3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 207, and a CDRL1 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 236, a CDRL2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 242 or 243, and a CDRL3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 251.

In some embodiments, the antibody or antibody fragment comprises a CDRH1 comprising or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 196, a CDRH2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 199, and a CDRH3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 204, and a CDRL1 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 232, a CDRL2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 242 or 245, and a CDRL3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 251.

In some embodiments, the antibody or antibody fragment comprises a CDRH1 comprising or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 196, a CDRH2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 197, and a CDRH3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 204, and a CDRL1 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 232, a CDRL2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 242 or 245, and a CDRL3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 251.

In some embodiments, the antibody or antibody fragment comprises a CDRH1 comprising or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 196, a CDRH2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 200, and a CDRH3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 203, and a CDRL1 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 232, a CDRL2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 242 or 245, and a CDRL3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 251.

In some embodiments, the antibody or antibody fragment comprises a CDRH1 comprising or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 196, a CDRH2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 200, and a CDRH3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 205, and a CDRL1 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 232, a CDRL2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 242 or 245, and a CDRL3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 251.

In some embodiments, the antibody or antibody fragment comprises a CDRH1 comprising or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 196, a CDRH2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 200, and a CDRH3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 221, and a CDRL1 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 232, a CDRL2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 242 or 245, and a CDRL3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 251.

In some embodiments, the antibody or antibody fragment comprises a CDRH1 comprising or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 196, a CDRH2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 200, and a CDRH3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 209, and a CDRL1 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 232, a CDRL2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 242 or 245, and a CDRL3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 251.

In some embodiments, the antibody or antibody fragment comprises a CDRH1 comprising or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 196, a CDRH2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 200, and a CDRH3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 204, and a CDRL1 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 238, a CDRL2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 242 or 245, and a CDRL3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 251.

In some embodiments, the antibody or antibody fragment comprises a CDRH1 comprising or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 196, a CDRH2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 200, and a CDRH3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 204, and a CDRL1 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 229, a CDRL2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 242 or 245, and a CDRL3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 251.

In some embodiments, the antibody or antibody fragment comprises a CDRH1 comprising or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 196, a CDRH2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 200, and a CDRH3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 204, and a CDRL1 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 228, a CDRL2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 242 or 245, and a CDRL3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 251.

In some embodiments, the antibody or antibody fragment comprises a CDRH1 comprising or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 196, a CDRH2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 199, and a CDRH3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 202, and a CDRL1 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 226, a CDRL2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 242 or 243, and a CDRL3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 251.

In some embodiments, the antibody or antibody fragment comprises a CDRH1 comprising or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 196, a CDRH2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 197, and a CDRH3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 222, and a CDRL1 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 226, a CDRL2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 242 or 243, and a CDRL3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 251.

In some embodiments, the antibody or antibody fragment comprises a CDRH1 comprising or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 196, a CDRH2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 197, and a CDRH3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 223, and a CDRL1 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 226, a CDRL2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 242 or 243, and a CDRL3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 251.

In some embodiments, the antibody or antibody fragment comprises a CDRH1 comprising or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 196, a CDRH2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 197, and a CDRH3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 224, and a CDRL1 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 226, a CDRL2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 242 or 243, and a CDRL3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 251.

In some embodiments, the antibody or antibody fragment comprises a CDRH1 comprising or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 196, a CDRH2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 197, and a CDRH3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 225, and a CDRL1 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 226, a CDRL2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 242 or 243, and a CDRL3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 251.

In some embodiments, the antibody or antibody fragment comprises a CDRH1 comprising or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 196, a CDRH2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 197, and a CDRH3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 202, and a CDRL1 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 226, a CDRL2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 242 or 245, and a CDRL3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 251.

In some embodiments, the antibody or antibody fragment comprises a CDRH1 comprising or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 196, a CDRH2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 199, and a CDRH3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 204, and a CDRL1 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 231, a CDRL2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 242 or 245, and a CDRL3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 251.

In some embodiments, the antibody or antibody fragment comprises a CDRH1 comprising or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 196, a CDRH2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 197, and a CDRH3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 204, and a CDRL1 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 231, a CDRL2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 242 or 245, and a CDRL3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 251.

In some embodiments, the antibody or antibody fragment comprises a CDRH1 comprising or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 196, a CDRH2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 198, and a CDRH3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 203, and a CDRL1 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 231, a CDRL2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 242 or 245, and a CDRL3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 251.

In some embodiments, the antibody or antibody fragment comprises a CDRH1 comprising or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 196, a CDRH2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 198, and a CDRH3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 205, and a CDRL1 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 231, a CDRL2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 242 or 245, and a CDRL3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 251.

In some embodiments, the antibody or antibody fragment comprises a CDRH1 comprising or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 196, a CDRH2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 198, and a CDRH3 comprising the amino acid sequence of SEQ ID NO: 221, and a CDRL1 comprising the amino acid sequence of SEQ ID NO: 231, a CDRL2 comprising the amino acid sequence of SEQ ID NO: 242 or 245, and a CDRL3 comprising the amino acid sequence of SEQ ID NO: 251.

In some embodiments, the antibody or antibody fragment comprises a CDRH1 comprising or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 196, a CDRH2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 199, and a CDRH3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 203, and a CDRL1 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 231, a CDRL2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 242 or 245, and a CDRL3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 251.

In some embodiments, the antibody or antibody fragment comprises a CDRH1 comprising or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 196, a CDRH2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 199, and a CDRH3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 205, and a CDRL1 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 231, a CDRL2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 242 or 245, and a CDRL3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 251.

In some embodiments, the antibody or antibody fragment comprises a CDRH1 comprising or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 196, a CDRH2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 197, and a CDRH3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 203, and a CDRL1 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 231, a CDRL2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 242 or 245, and a CDRL3 comprising the amino acid sequence of SEQ ID NO: 251.

In some embodiments, the antibody or antibody fragment comprises a CDRH1 comprising or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 196, a CDRH2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 197, and a CDRH3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 205, and a CDRL1 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 231, a CDRL2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 242 or 245, and a CDRL3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 251.

In some embodiments, the antibody or antibody fragment comprises a CDRH1 comprising or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 196, a CDRH2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 198, and a CDRH3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 204, and a CDRL1 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 226, a CDRL2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 242 or 245, and a CDRL3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 251.

In some embodiments, the antibody or antibody fragment comprises a CDRH1 comprising or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 196, a CDRH2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 198, and a CDRH3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 209, and a CDRL1 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 231, a CDRL2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 242 or 245, and a CDRL3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 251.

In some embodiments, the antibody or antibody fragment comprises a CDRH1 comprising or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 196, a CDRH2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 199, and a CDRH3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 209, and a CDRL1 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 231, a CDRL2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 242 or 245, and a CDRL3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 251.

In some embodiments, the antibody or antibody fragment comprises a CDRH1 comprising or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 196, a CDRH2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 197, and a CDRH3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 209, and a CDRL1 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 231, a CDRL2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 242 or 245, and a CDRL3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 251.

In some embodiments, the antibody or antibody fragment comprises a CDRH1 comprising or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 196, a CDRH2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 199, and a CDRH3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 205, and a CDRL1 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 233, a CDRL2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 242 or 243, and a CDRL3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 251.

In some embodiments, the antibody or antibody fragment comprises a CDRH1 comprising or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 196, a CDRH2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 197, and a CDRH3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 205, and a CDRL1 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 233, a CDRL2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 242 or 243, and a CDRL3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 251.

In some embodiments, the antibody or antibody fragment comprises a CDRH1 comprising or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 196, a CDRH2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 198, and a CDRH3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 209, and a CDRL1 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 233, a CDRL2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 242 or 243, and a CDRL3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 251.

In some embodiments, the antibody or antibody fragment comprises a CDRH1 comprising or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 196, a CDRH2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 198, and a CDRH3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 211, and a CDRL1 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 233, a CDRL2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 242 or 243, and a CDRL3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 251.

In some embodiments, the antibody or antibody fragment comprises a CDRH1 comprising or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 196, a CDRH2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 198, and a CDRH3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 205, and a CDRL1 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 240, a CDRL2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 242 or 243, and a CDRL3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 251.

In some embodiments, the antibody or antibody fragment comprises a CDRH1 comprising or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 196, a CDRH2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 198, and a CDRH3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 205, and a CDRL1 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 241, a CDRL2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 242 or 243, and a CDRL3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 251.

In some embodiments, the antibody or antibody fragment comprises a CDRH1 comprising or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 196, a CDRH2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 199, and a CDRH3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 209, and a CDRL1 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 233, a CDRL2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 242 or 243, and a CDRL3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 251.

In some embodiments, the antibody or antibody fragment comprises a CDRH1 comprising or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 196, a CDRH2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 199, and a CDRH3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 209, and a CDRL1 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 233, a CDRL2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 242 or 245, and a CDRL3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 251.

In some embodiments, the antibody or antibody fragment comprises a CDRH1 comprising or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 196, a CDRH2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 199, and a CDRH3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 204, and a CDRL1 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 230, a CDRL2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 242 or 244, and a CDRL3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 251.

In some embodiments, the antibody or antibody fragment comprises a CDRH1 comprising or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 196, a CDRH2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 198, and a CDRH3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 203, and a CDRL1 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 230, a CDRL2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 242 or 244, and a CDRL3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 251.

In some embodiments, the antibody or antibody fragment comprises a CDRH1 comprising or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 196, a CDRH2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 198, and a CDRH3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 205, and a CDRL1 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 230, a CDRL2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 242 or 244, and a CDRL3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 251.

In some embodiments, the antibody or antibody fragment comprises a CDRH1 comprising or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 196, a CDRH2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 198, and a CDRH3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 221, and a CDRL1 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 230, a CDRL2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 242 or 244, and a CDRL3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 251.

In some embodiments, the antibody or antibody fragment comprises a CDRH1 comprising or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 196, a CDRH2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 197, and a CDRH3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 203, and a CDRL1 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 230, a CDRL2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 242 or 244, and a CDRL3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 251.

In some embodiments, the antibody or antibody fragment comprises a CDRH1 comprising or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 196, a CDRH2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 199, and a CDRH3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 203, and a CDRL1 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 230, a CDRL2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 242 or 244, and a CDRL3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 251.

In some embodiments, the antibody or antibody fragment comprises a CDRH1 comprising or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 196, a CDRH2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 197, and a CDRH3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 205, and a CDRL1 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 230, a CDRL2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 242 or 244, and a CDRL3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 251.

In some embodiments, the antibody or antibody fragment comprises a CDRH1 comprising or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 196, a CDRH2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 199, and a CDRH3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 205, and a CDRL1 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 230, a CDRL2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 242 or 244, and a CDRL3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 251.

In some embodiments, the antibody or antibody fragment comprises a CDRH1 comprising or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 196, a CDRH2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 197, and a CDRH3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 209, and a CDRL1 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 230, a CDRL2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 242 or 244, and a CDRL3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 251.

In some embodiments, the antibody or antibody fragment comprises a CDRH1 comprising or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 196, a CDRH2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 199, and a CDRH3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 209, and a CDRL1 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 230, a CDRL2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 242 or 244, and a CDRL3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 251.

In some embodiments, the antibody or antibody fragment comprises a CDRH1 comprising or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 196, a CDRH2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 199, and a CDRH3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 209, and a CDRL1 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 230, a CDRL2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 242 or 245, and a CDRL3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 251.

In some embodiments, the antibody or antibody fragment comprises a CDRH1 comprising or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 196, a CDRH2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 199, and a CDRH3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 209, and a CDRL1 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 229, a CDRL2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 242 or 243, and a CDRL3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 251.

In some embodiments, the antibody or antibody fragment comprises a CDRH1 comprising or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 196, a CDRH2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 199, and a CDRH3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 209, and a CDRL1 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 228, a CDRL2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 242 or 243, and a CDRL3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 251.

In some embodiments, the antibody or antibody fragment comprises a CDRH1 comprising or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 196, a CDRH2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 199, and a CDRH3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 209, and a CDRL1 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 238, a CDRL2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 242 or 243, and a CDRL3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 251.

In some embodiments, the antibody or antibody fragment disclosed herein comprises, or consists essentially of, or consists of a heavy chain variable sequence and a light chain variable sequence having at least 96%, at least 97%, at least 98%, at least 99% or 100% identity to an amino acid sequence selected from: the amino acid sequence of SEQ ID NO: 110 and the amino acid sequence of SEQ ID NO: 167; the amino acid sequence of SEQ ID NO: 111 and the amino acid sequence of SEQ ID NO: 168; the amino acid sequence of SEQ ID NO: 111 and the amino acid sequence of SEQ ID NO: 169; the amino acid sequence of SEQ ID NO: 111 and the amino acid sequence of SEQ ID NO: 170; the amino acid sequence of SEQ ID NO: 111 and the amino acid sequence of SEQ ID NO: 171; the amino acid sequence of SEQ ID NO: 112 and the amino acid sequence of SEQ ID NO: 173; the amino acid sequence of SEQ ID NO: 113 and the amino acid sequence of SEQ ID NO: 172; the amino acid sequence of SEQ ID NO: 114 and the amino acid sequence of SEQ ID NO: 172; the amino acid sequence of SEQ ID NO: 115 and the amino acid sequence of SEQ ID NO: 172; the amino acid sequence of SEQ ID NO: 116 and the amino acid sequence of SEQ ID NO: 173; the amino acid sequence of SEQ ID NO: 117 and the amino acid sequence of SEQ ID NO: 172; the amino acid sequence of SEQ ID NO: 118 and the amino acid sequence of SEQ ID NO: 172; the amino acid sequence of SEQ ID NO: 119 and the amino acid sequence of SEQ ID NO: 172; the amino acid sequence of SEQ ID NO: 120 and the amino acid sequence of SEQ ID NO: 172; the amino acid sequence of SEQ ID NO: 121 and the amino acid sequence of SEQ ID NO: 172; the amino acid sequence of SEQ ID NO: 110 and the amino acid sequence of SEQ ID NO: 173; the amino acid sequence of SEQ ID NO: 122 and the amino acid sequence of SEQ ID NO: 174; the amino acid sequence of SEQ ID NO: 123 and the amino acid sequence of SEQ ID NO: 174; the amino acid sequence of SEQ ID NO: 124 and the amino acid sequence of SEQ ID NO: 174; the amino acid sequence of SEQ ID NO: 125 and the amino acid sequence of SEQ ID NO: 174; the amino acid sequence of SEQ ID NO: 126 and the amino acid sequence of SEQ ID NO: 174; the amino acid sequence of SEQ ID NO: 127 and the amino acid sequence of SEQ ID NO: 175; the amino acid sequence of SEQ ID NO: 127 and the amino acid sequence of SEQ ID NO: 176; the amino acid sequence of SEQ ID NO: 128 and the amino acid sequence of SEQ ID NO: 177; the amino acid sequence of SEQ ID NO: 129 and the amino acid sequence of SEQ ID NO: 177; the amino acid sequence of SEQ ID NO: 130 and the amino acid sequence of SEQ ID NO: 177; the amino acid sequence of SEQ ID NO: 131 and the amino acid sequence of SEQ ID NO: 177; the amino acid sequence of SEQ ID NO: 132 and the amino acid sequence of SEQ ID NO: 177; the amino acid sequence of SEQ ID NO: 133 and the amino acid sequence of SEQ ID NO: 178; the amino acid sequence of SEQ ID NO: 134 and the amino acid sequence of SEQ ID NO: 179; the amino acid sequence of SEQ ID NO: 135 and the amino acid sequence of SEQ ID NO: 179; the amino acid sequence of SEQ ID NO: 136 and the amino acid sequence of SEQ ID NO: 179; the amino acid sequence of SEQ ID NO: 134 and the amino acid sequence of SEQ ID NO: 180; the amino acid sequence of SEQ ID NO: 137 and the amino acid sequence of SEQ ID NO: 180; the amino acid sequence of SEQ ID NO: 138 and the amino acid sequence of SEQ ID NO: 179; the amino acid sequence of SEQ ID NO: 139 and the amino acid sequence of SEQ ID NO: 181; the amino acid sequence of SEQ ID NO: 112 and the amino acid sequence of SEQ ID NO: 181; the amino acid sequence of SEQ ID NO: 140 and the amino acid sequence of SEQ ID NO: 181; the amino acid sequence of SEQ ID NO: 141 and the amino acid sequence of SEQ ID NO: 181; the amino acid sequence of SEQ ID NO: 142 and the amino acid sequence of SEQ ID NO: 181; the amino acid sequence of SEQ ID NO: 143 and the amino acid sequence of SEQ ID NO: 181; the amino acid sequence of SEQ ID NO: 144 and the amino acid sequence of SEQ ID NO: 182; the amino acid sequence of SEQ ID NO: 144 and the amino acid sequence of SEQ ID NO: 183; the amino acid sequence of SEQ ID NO: 144 and the amino acid sequence of SEQ ID NO: 184; the amino acid sequence of SEQ ID NO: 145 and the amino acid sequence of SEQ ID NO: 185; the amino acid sequence of SEQ ID NO: 146 and the amino acid sequence of SEQ ID NO: 185; the amino acid sequence of SEQ ID NO: 147 and the amino acid sequence of SEQ ID NO: 185; the amino acid sequence of SEQ ID NO: 148 and the amino acid sequence of SEQ ID NO: 185; the amino acid sequence of SEQ ID NO: 149 and the amino acid sequence of SEQ ID NO: 185; the amino acid sequence of SEQ ID NO: 150 and the amino acid sequence of SEQ ID NO: 186; the amino acid sequence of SEQ ID NO: 139 and the amino acid sequence of SEQ ID NO: 187; the amino acid sequence of SEQ ID NO: 112 and the amino acid sequence of SEQ ID NO: 187; the amino acid sequence of SEQ ID NO: 151 and the amino acid sequence of SEQ ID NO: 187; the amino acid sequence of SEQ ID NO: 110 and the amino acid sequence of SEQ ID NO: 187; the amino acid sequence of SEQ ID NO: 152 and the amino acid sequence of SEQ ID NO: 187; the amino acid sequence of SEQ ID NO: 153 and the amino acid sequence of SEQ ID NO: 187; the amino acid sequence of SEQ ID NO: 119 and the amino acid sequence of SEQ ID NO: 187; the amino acid sequence of SEQ ID NO: 154 and the amino acid sequence of SEQ ID NO: 187; the amino acid sequence of SEQ ID NO: 120 and the amino acid sequence of SEQ ID NO: 187; the amino acid sequence of SEQ ID NO: 111 and the amino acid sequence of SEQ ID NO: 186; the amino acid sequence of SEQ ID NO: 116 and the amino acid sequence of SEQ ID NO: 187; the amino acid sequence of SEQ ID NO: 113 and the amino acid sequence of SEQ ID NO: 187; the amino acid sequence of SEQ ID NO: 114 and the amino acid sequence of SEQ ID NO: 187; the amino acid sequence of SEQ ID NO: 119 and the amino acid sequence of SEQ ID NO: 188; the amino acid sequence of SEQ ID NO: 120 and the amino acid sequence of SEQ ID NO: 188; the amino acid sequence of SEQ ID NO: 116 and the amino acid sequence of SEQ ID NO: 188; the amino acid sequence of SEQ ID NO: 121 and the amino acid sequence of SEQ ID NO: 188; the amino acid sequence of SEQ ID NO: 110 and the amino acid sequence of SEQ ID NO: 189; the amino acid sequence of SEQ ID NO: 110 and the amino acid sequence of SEQ ID NO: 190; the amino acid sequence of SEQ ID NO: 113 and the amino acid sequence of SEQ ID NO: 188; the amino acid sequence of SEQ ID NO: 113 and the amino acid sequence of SEQ ID NO: 191; the amino acid sequence of SEQ ID NO: 139 and the amino acid sequence of SEQ ID NO: 192; the amino acid sequence of SEQ ID NO: 151 and the amino acid sequence of SEQ ID NO: 192; the amino acid sequence of SEQ ID NO: 110 and the amino acid sequence of SEQ ID NO: 192; the amino acid sequence of SEQ ID NO: 152 and the amino acid sequence of SEQ ID NO: 192; the amino acid sequence of SEQ ID NO: 154 and the amino acid sequence of SEQ ID NO: 192; the amino acid sequence of SEQ ID NO: 153 and the amino acid sequence of SEQ ID NO: 192; the amino acid sequence of SEQ ID NO: 120 and the amino acid sequence of SEQ ID NO: 192; the amino acid sequence of SEQ ID NO: 119 and the amino acid sequence of SEQ ID NO: 192; the amino acid sequence of SEQ ID NO: 114 and the amino acid sequence of SEQ ID NO: 192; the amino acid sequence of SEQ ID NO: 113 and the amino acid sequence of SEQ ID NO: 192; the amino acid sequence of SEQ ID NO: 113 and the amino acid sequence of SEQ ID NO: 193; the amino acid sequence of SEQ ID NO: 113 and the amino acid sequence of SEQ ID NO: 194; the amino acid sequence of SEQ ID NO: 113 and the amino acid sequence of SEQ ID NO: 195; or the amino acid sequence of SEQ ID NO: 113 and the amino acid sequence of SEQ ID NO: 169.

In some embodiments, the antibody or antibody fragment disclosed herein comprises, or consists essentially of, or consists of a heavy chain variable sequence having at least 96%, at least 97%, at least 98% or at least 99% identity to a heavy chain variable amino acid sequence in Table 14 and/or a light chain variable sequence having at least 96%, at least 97%, at least 98% or at least 99% identity to a light chain variable amino acid sequence in Table 14, with the proviso that the amino acid mutations are not altered in the equivalent from those mutations identified in Table 13.

In another aspect, the present disclosure provides an isolated nucleic acid molecule comprising, or consisting essentially of, or consisting of a nucleotide sequence encoding the CDR, heavy chain, light chain, scFV, antibody or antibody fragment and equivalents as disclosed herein that are optionally detectably labeled. In some embodiments, the nucleic acid comprises a nucleotide sequence encoding an Fv fragment comprising an amino acid sequence selected from any one of SEQ ID NO: 15-99; or an amino acid sequence disclosed in Table 13 or an equivalent thereof with the proviso that the amino acid mutations are not altered in the equivalent from those mutations identified in Table 13.

Another aspect of the present disclosure provides an isolated nucleic acid comprising, or consisting essentially of, or consisting of a nucleotide sequence encoding the antibody or antibody fragment disclosed herein. In some embodiments, the nucleic acid comprises, or consists essentially of, or consists of the nucleotide sequence encoding: (a) an antibody fragment comprising an amino acid sequence selected from SEQ ID NO: 1-14 or an equivalent thereof; or (b) a heavy chain variable sequence comprising an amino acid sequence selected from SEQ ID NO: 100, 101, 102, 103, 104, 105, 106, 107, 108, or 109; or an equivalent thereof and/or (b) a light chain variable sequence comprising an amino acid sequence selected from SEQ ID NO: 155, 156, 157, 158, 159, 160, 161, 162, 163, 164, 165, or 166 or an equivalent thereof. In one aspect the equivalent nucleic acid encodes a polypeptide that has at least 96% identity to any one of the reference polypeptides and further in one embodiment has a CDR sequence that is 100% identical to the CDR of the reference polypeptide. The reference CDRs are provided above, and further within this document. Stated another way, the amino acid substitutions and modifications are in regions outside of the CDR regions.

In some embodiments, the nucleic acid comprises a nucleotide sequence encoding a heavy chain variable sequence comprising an amino acid sequence selected from any one of SEQ ID NOs: 110-154. In some embodiments, the nucleic acid comprises a nucleotide sequence encoding a heavy chain variable sequence comprising a heavy chain variable amino acid sequence disclosed in Table 14. In some embodiments, the nucleic acid comprises a nucleotide sequence encoding a heavy chain variable sequence comprising an amino acid sequence having at least 70%, 80%, 90%, 95%, 96%, 97%, 98%, or 99% identity a heavy chain variable amino acid sequence disclosed in Table 14, with the proviso that the amino acid mutations are not altered in the equivalent from those mutations identified in Table 14.

In some embodiments, the nucleic acid comprises a nucleotide sequence encoding a light chain variable sequence comprising an amino acid sequence selected from SEQ ID NO: 167, 168, 169, 170, 171, 172, 173, 174, 175, 176, 177, 178, 179, 180, 181, 182, 183, 184, 185, 186, 187, 188, 189, 190, 191, 192, 193, 194, or 195. In some embodiments, the nucleic acid comprises a nucleotide sequence encoding a light chain variable amino acid sequence comprising a light chain variable amino acid sequence disclosed in Table 14. In some embodiments, the nucleic acid comprises a nucleotide sequence encoding a light chain variable sequence comprising a light chain variable amino acid sequence having at least 70%, 80%, 90%, 95%, 96%, 97%, 98%, or 99% identity to the amino acid sequence disclosed in Table 14, with the proviso that the amino acid mutations are not altered in the equivalent from those mutations identified in Table 13.

Another aspect of the present disclosure provides a vector comprising, or consisting essentially of, or consisting of an isolated nucleic acid disclosed herein.

Another aspect of the present disclosure provides an engineered cell comprising, or consisting essentially of, or consisting of the isolated nucleic acid disclosed herein; or the vector disclosed herein.

Another aspect of the present disclosure provides a composition comprising, or consisting essentially of, or consisting of one or more antibodies or antibody fragments disclosed herein, the isolated nucleic acid disclosed herein, or the vector disclosed herein and a carrier. In some embodiments, the composition is a pharmaceutically acceptable composition.

Another aspect of the present disclosure provides a method of one or more of: treating a subject infected with SARS-CoV-2; reducing the likelihood of infection with SARS-CoV-2 in a subject at risk of infection; reducing the likelihood of symptomatic infection with SARS-CoV-2, treating a subject at risk of contracting SARS-CoV-2, the method comprising, or consisting essentially of, or consisting of administering to the subject the antibody or antibody fragment disclosed herein.

Another aspect of the present disclosure provides a method of one or more of: treating a subject infected with SARS-CoV-2; reducing the likelihood of infection of a subject at risk of infection with SARS-CoV-2; reducing the likelihood of symptomatic infection with SARS-CoV-2; treating a subject at risk of contracting SARS-CoV-2, the method comprising, or consisting essentially of, or consisting of administering to the subject a first antibody or antibody fragment and a second antibody or antibody fragment. In some embodiments, the first and second antibodies or antibody fragments synergistically neutralize the SARS-CoV-2 virus. In some embodiments, the first antibody or antibody fragment and the second antibody or antibody fragment bind to non-overlapping sites on the S glycoprotein. In some embodiments, the first antibody or antibody fragment and the second antibody or antibody fragment have a synergy score of about 10, about 12, about 14, about 15, about 17, about 18, about 20 or more. In some embodiments, the synergy score is based on experimental combination response in a focus reduction neutralization test (FRNT) and/or by synergy-scoring models.

Another aspect of the present disclosure provides a vaccine formulation comprising, or consisting essentially of, or consisting of one or more antibodies or antibody fragments disclosed herein. In some embodiments, the vaccine formulation further comprises, or consists essentially of, or consists of a second antibody or antibody fragment that binds to a SARS-CoV-2 surface spike protein. In some embodiments, the second antibody or antibody fragment is an antibody or antibody fragment disclosed herein. In some embodiments, the second antibody or antibody fragment is an antibody or antibody fragment selected from sotrovimab (S309), COV2-2381, cilgavimab (COV2-2130 or AZD1061), or tixagevimab (COV2-2196 or AZD8895). The vaccine compositions can optionally comprise adjuvants, stabilizers and/or preservatives.

Another aspect of the present disclosure provides a vaccine formulation comprising, or consisting essentially of, or consisting of one or more expression vectors. In some embodiments, the one or more expression vectors comprise at least one nucleic acid encoding an antibody or antibody fragment disclosed herein. In some embodiments, the nucleic acid is DNA, RNA, or mRNA. The vaccine compositions can optionally comprise adjuvants, stabilizers and/or preservatives.

Another aspect of the present disclosure provides a method of detecting infection with SARS-CoV-2 in a subject the method comprising, or consisting essentially of, or consisting of (a) contacting a sample isolated from the subject with the antibody or antibody fragment disclosed herein; and (b) detecting SARS-CoV-2 in the sample by detecting the binding of the antibody or antibody fragment to a SARS-CoV-2 antigen in the sample.

Another aspect of the present disclosure provides a method of determining a SARS-CoV-2 virus variant antigenic profile the method comprising, or consisting essentially of, or consisting of: (a) contacting a sample comprising the SARS-CoV-2 virus variant with the antibody or antibody fragment disclosed herein; and (b) determining the binding of the antibody or antibody fragment to the SARS-CoV-2 virus variant, wherein absence of binding indicates a conformational change and/or a mutation in the SARS-CoV-2 surface spike protein.

Another aspect of the present disclosure provides a biosensor for detecting the presence of a SARS-CoV-2 antigen in a sample, the biosensor comprising, or consisting essentially of, or consisting of (a) a transducer component comprising an electrode operatively connected to a microprocessor, the microprocessor being adapted to receive, process and transmit a signal; (b) a receptor component having a sensing element capable of detecting and binding to the SARS-CoV-2 antigen; and (c) the transducer component and the receptor component are capable of being brought into direct contact with the sample in situ. In some embodiments, the sensing element comprises the antibody or antibody fragment disclosed herein.

Another aspect of the present disclosure provides a kit comprising, or consisting essentially of, or consisting of the antibody or antibody fragment disclosed herein and instructions for using the first antibody or antibody fragment.

Another aspect of the present disclosure provides a kit comprising, or consisting essentially of, or consisting of a first antibody or antibody fragment that binds to a SARS-CoV-2 surface spike protein and a second antibody or antibody fragment that binds to a SAR-CoV-2 surface spike protein. In some embodiments, the first and/or second antibody and antibody fragment is antibody or antibody fragment disclosed herein. In some embodiments, at least one of the first and/or second antibody and antibody fragment is antibody or antibody fragment disclosed herein. In some embodiments, the kit further comprises, consists essentially of, or consists of instructions for using the first antibody or antibody fragment and the second antibody or antibody fragment for treating a subject infected with SARS-CoV-2 or for reducing the likelihood of infection of a subject at risk of contracting SARS-CoV-2.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1: shows a table demonstrating the improved binding affinity (EC₅₀) of the isolated monoclonal antibodies disclosed herein against the SARS-CoV-2 Wuhan and Omicron strains when compared to monoclonal antibodies known in the art.

FIG. 2: shows a table demonstrating the estimated binding affinity (K_D) of the isolated monoclonal antibodies of the present disclosure.

FIG. 3: shows a table demonstrating the enhanced neutralization effects (IC₅₀) of the isolated monoclonal antibodies disclosed herein against SARS-CoV-2 Omicron strains when compared to the COV2-2130 monoclonal antibody

FIG. 4: discloses the amino acid sequences of the variable domain fragments (Fv fragments) of the novel class of anti-SARS-CoV-2 monoclonal antibodies disclosed herein.

FIGS. 5A-5E: show binding and thermostability characteristics of the disclosed IgG antibodies. FIG. 5A shows the parental COV2-2130 (pink circles) and antibodies (2130-1-0114-112 (also referred to herein as LLNL199) highlighted in orange asterisks; remainder in black) were assayed for thermal shift (n=3, technical replicates). Bars indicate the mean and error bars indicate standard deviation. FIGS. 5B-5E show the parental COV2-2130 antibody and the disclosed antibodies (represented by the same symbols as in FIG. 5A) and cross-reactive positive control antibody s309 (blue squares), which were analyzed for relative binding against four SARS-CoV-2 Spike-RBD variants in Gyrolab immunoassay: Wild-type (FIG. 5B), Delta (FIG. 5C), Omicron BA.1 (FIG. 5D) and Omicron BA.1.1 (FIG. 5E). Lines represent 4-parameter logistic regression model fit using GraphPad Prism to each titration, executed without technical replicates.

FIGS. 6A-6E: show pseudoviral neutralization. The parental COV2-2130 antibody (circles), cross-reactive positive control antibody 5309 (squares), and down-selected by the disclosed antibodies (2130-1-0114-112 (also referred to herein as LLNL199) highlighted in orange asterisks; others in black) were assayed for neutralization of lentivirus pseudotyped with Spike variants D614G (FIG. 6A), Omicron BA.1 (FIG. 6B), BA.1.1 (FIG. 6C), BA.2 (FIG. 6D), and BA.4 (FIG. 6E). Symbols indicate the mean and standard deviation of two technical replicates; curves are 4-parameter logistic regression models fit using GraphPad Prism.

FIGS. 7A-7G: show authentic virus neutralization by focus reduction neutralization test in Vero-TMPRSS2 cells. 2130-1-0114-112 (also referred to herein as LNLL199) potently neutralizes (FIG. 7A) WA1/2020 D614G (FIG. 7B) Omicron BA.1, (FIG. 7C) Omicron BA.1.1, (FIG. 7D) Omicron BA.2, (FIG. 7E) Omicron BA.2.12.1, (FIG. 7F) Omicron BA.4, and (FIG. 7G) Omicron BA.5 authentic viruses in focus reduction neutralization assays in Vero-TMPRSS2 cells. All experiments were performed in technical duplicate, symbols represent the mean of the duplicates, and fits are four-parameter logistic curves. All analysis performed in GraphPad Prism.

FIG. 8: show in vivo prophylaxis by 2130-1-0114-112 (also referred to herein as LLNL199) against SARS-CoV-2 variants. Eight-week-old female K18-hACE2 mice were administered 100 μg (about 5 mg/kg) of the indicated mAb treatment by intraperitoneal injection one day before intranasal inoculation with 104 FFU of WA1/2020 D614G (Left), BA.1.1 (Center) or BA.5 (Right). Tissues were collected four days after inoculation and viral RNA levels in the lungs (Top), nasal turbinates (Center), and nasal washes (Bottom) were determined by RT-qPCR (lines indicate median±SEM.; n=9-10 mice per group, two experiments; Kruskal-Wallis test with Dunn's multiple comparisons test; ns, not significant; *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001). All analysis conducted in GraphPad Prism.

FIGS. 9A-9E: show Cryo-EM structure of neutralizing antibodies 2130-1-0114-112 (also referred to herein as LLNL199) in complex with Cov2 BA.2 RBD. FIG. 9A shows Cryo-EM map and model of the RBD-Fab complex. The map is transparent and colored by chain with RBD red, 2130-1-0114-112 (also referred to herein as LNLL199) HC yellow and 2130-1-0114-112 (also referred to herein as LNLL199) LC green. FIG. 9B shows atomic model of the RBD-Fab complex, which were labeled as in FIG. 9A, with hydrogen bond shown in dashed line; BA.2 RBD mutation in orange, and The heavy chain (HC) and light chain variable regions of the 2130-1-0114-112 (also referred to herein as also referred to herein as LLNL199) mutation shown in cyan and blue respectively. FIG. 9C shows detail illustrating the interaction between the heavy chain complementary-determining region (HCDR) and Light chain CDR (LCDR) of 2130-1-0114-112 (also referred to herein as LLNL199) modified residues and the Cov2 BA.2 RBD. Left, HCDR3 Glu112. Middle, LCDR1 Ala32 and Ala33 hydrophobic network. Right, LCDR2 Glu59 salt bridge with Arg498. Orange and green dashed lines indicate H-bonds and hydrophobic interactions, respectively; yellow dashed lines are labeled with distances. FIG. 9D shows the HCDR3 shown as in (FIG. 9C) with surface color by electrostatic potential, showing the positive and negative charges of Lys444 and Glu112; and A32 and A33 in the LCDR1 with the nearby RBD surface colored by hydrophobicity (orange to cyan hydrophobic to hydrophilic). FIG. 9E shows a 2D diagram of Fab 2130-1-0114-112 (also referred to herein as LLNL199) paratope and epitope residues involved in hydrogen bonding (dashed lines) and hydrophobic interactions. Residues involved in hydrophobic interactions are shown as curved lines with rays, where atoms are shown as circles, with oxygen red, carbon black, and nitrogen blue, and interacting residues that belong to the CDR loops are colored in different shade. Asterisks correspond to mutated residues. Image created with Ligplot+[Laskowski 2011].

FIGS. 10A-10D: show optimized, single-concentration screening data in Gyrolab immunoassays, which allowed for the selection of candidates from Set 1 (n=230) for down-stream characterization. Parental mAb COV2-2130 (magenta circles) and positive control mAb 5309 (blue squares, [Pinto 2020]) served as references for disclosed mAbs (gray diamonds) in single-concentration immunoassays. Target antigens were Wild Type (FIG. 10A), Omicron BA.1 (FIG. 10B), Delta (FIG. 10C), and Omicron BA.1.1 (FIG. 10D). Each screened antibody and antigen combination was evaluated in two replicate assays; each of the controls was replicated in two replicate assays for each of three groups of antibodies, resulting in six replicate assays for each point on the control curves. All error bars indicate standard deviation.

FIGS. 11A-11D: show line graphs demonstrating results of ELISA screening, which allowed for down selection of candidates from Set 2 (n=204), for down-stream characterization. Parental mAb COV-22130 (red circles; 3 technical replicates) and positive control mAb 5309 (blue squares; 3 technical replicates [pinto2020]) serve as references for disclosed mAbs (black curves). Orange squares are 2130-1-0114-112 (also referred to herein as also referred to herein as LLNL199). Each designed antibody had a single measurement (n=1) at each concentration. All curves were 4-parameter logistic fits, produced in GraphPad Prism. Target antigens are Wild Type (FIG. 11A), Omicron (Acro) (FIG. 11B), Omicron BA.1 (FIG. 11C), and Omicron BA.1.1 (FIG. 11D).

FIGS. 12A-12G: show normalized pseudoviral neutralization of recently emerged VOCs by 2130-1-0114-112 (also referred to herein as LLNL199). FIG. 12A provides a legend for antibodies tested. FIG. 12B shows that COV2-2130 and 2130-1-0114-112 (also referred to herein as LLNL199) potently neutralize D614G. FIG. 12C shows that only 2130-1-0114-112 (also referred to herein as LLNL199) potently neutralized BA.1, consistent with other pseudoviral neutralization assays. FIG. 12D shows 2130-1-0114-112 (also referred to herein as LLNL199) potently neutralizes BA.2.75, outperforming COV2-2130 by 90-fold—12E-12F show that 2130-1-0114-112 (also referred to herein as LLNL199) loses substantial potency in the context of BA.4.6 and artificially-produced BA.2.75+R346T (here listed as “BA.2.76”) but retains measurable neutralization, demonstrating mitigation of this important weakness of COV2-2130. FIGS. 12B-12F show the mean of two technical replicates and curves show a fitted four-parameter logistic curve with fixed minimum and maximum values. FIG. 12G shows neutralization IC50 values in ng/ml. “>” indicates IC50 greater than 10,000 ng/mL. All analysis conducted in GraphPad Prism.

FIGS. 13A-13F: show authentic virus neutralization by focus reduction neutralization test in Vero-TMPRSS2 cells. 2130-1-0114-112 (also referred to herein as LLNL199) potently neutralizes (FIG. 13A) Delta (B.1.617.2), (FIG. 13B) Omicron BA.1, (FIG. 13C) Omicron BA.1.1, (FIG. 13D) Omicron BA.4, and (FIG. 13E) Omicron BA.5.5 authentic viruses in focus reduction neutralization assays in Vero-TMPRSS2 cells. All experiments were performed in technical duplicate, symbols represent the mean of the duplicates, and fits are four-parameter logistic curves. FIG. 13F shows IC₅₀values in μg/ml. >indicates greater than 10 μg/ml. All analysis performed in GraphPad Prism.

FIGS. 14A-14D: show authentic virus neutralization in plaque assays using Vero E6-TMPRSS2-T2A-ACE2 (VAT) cells. All SARS-CoV-2 viral stocks as well as VAT cells were obtained through BEI Resources, NIAID, NIH. FIG. 14A shows Delta variant isolate hCoV-19/USA/MD-HP05647/2021, lineage B.1.617.2; NR-55672. FIG. 14B shows BA.1 isolate hCoV-19/USA/GA-EHC-2811C/2021, lineage B.1.1.529; NR-56481). FIG. 14C shows BA1.1 (isolate hCoV-19/USA/HI-CDC-4359259-001/2021, lineage B.1.1.529; NR-56475). Viral stocks were amplified in Vero E6 cells (Delta variant) or VAT cells (Omicron variants). Serial dilutions of mAbs were incubated with virus at a concentration of 400 PFU/mL at 37° C. with 5% CO2 for 30 min. Antibody-virus complexes were then added to VAT cells in 12-well plates and incubated for 30 min, then overlaid with 2 mL per well of 0.6% microcrystalline cellulose (Sigma) in minimal essential media (ThermoFisher) supplemented with 0.3% bovine serum albumin (Sigma), and 1% penicillin/streptomycin (ThermoFisher). After 72 hours incubation, plaques were visualized by incubation in 0.25% crystal violet in 100% methanol for 10 minutes. Data are represented as the normalized infection of mAb-treated virus to virus treated with control human IgG (Invitrogen). For S309, each point shows the mean of four technical replicates; all other points are means of two technical replicates; error bars show standard error. Curves are two-parameter (IC50, hill-slope) logistic fits to normalized response. FIG. 14D shows IC₅₀values (μg/ml) illustrating that 2130-1-0104-024 (also referred to herein as LLNL193), while having only two mutations from COV2-2130, remained strongly potent against BA.1 and suffered a 20-times loss in potency against BA.1.1. 2130-1-0114-112 (LLNL199) was strongly potent against all three tested variants. All analysis performed in GraphPad Prism.

FIGS. 15A-15E: show Cryo-EM structure of neutralizing antibodies 2130-1-0114-112 (also referred to herein as LLNL199) in complex with Cov2 BA.2 RBD. FIG. 15A shows RBD residues in 7 Å distance form 2130-1-0114-112 (also referred to herein as LLNL199). RBD in red, with BA.2 mutated residues in orange, and 2130-1-0114-112 (also referred to herein as LLNL199) in yellow/green. FIG. 15B shows Left, 3D representation of the interaction plot between RBD and 2130-1-0114-112 (also referred to herein as LLNL199) HC. 2130-1-0114-112 (also referred to herein as LLNL199) is shown as stick (yellow) and RBD as gray sphere with the contact residues in red. Contact residues labelled and numbered. Right, 3D representation of the interaction plot between RBD and 2130-1-0114-112 (also referred to herein as LLNL199) LC. 2130-1-0114-112 (also referred to herein as LLNL199) is shown as stick (green) and RBD as gray sphere with the contact residues in red. Contact residues are labelled and numbered. FIG. 15C shows Glu112 and Lys440 with the EM map and distance between the sidechains. FIG. 15D shows Fab COV-2130 paratope and epitope residues involved in hydrogen bonding (dashed lines) and hydrophobic interactions. Hydrophobic interactions residues are shown as curved lines with rays. Atoms shown as circles, with oxygen red, carbon black, and nitrogen blue. Image was created with Ligplot+. FIG. 15E Representative micrograph.

FIG. 16: shows CryoEM workflow.

FIGS. 17A-17D: show full-curve Gyros binding of strain (FIG. 17A) Wuhan; (FIG. 17B) Delta; (FIG. 17C) Omicron BA.1; (FIG. 17D) Omicron BA.1.1.

FIGS. 18A-18D: show the neutralization of strain (FIG. 18A) BA1; (FIG. 18B) BA1.1; (FIG. 18C) BA1.1 (bottom); (FIG. 18D) BA2.

FIGS. 19A-19E: show authentic virus neutralization by focus reduction neutralization test in Vero-TMPRSS2 cells. 2130-1-0114-112 (also referred to herein as LLNL199) and 2130-1-0121-354 (also referred to herein as B-133) potently neutralizes (FIG. 19A) B.1.617.2; (FIG. 19B) BA.1; (FIG. 19C) BA1.1; (FIG. 19D) BA.4; (FIG. 19E) BA.5.5 authentic viruses in focus reduction neutralization assays in Vero-TMPRSS2 cells. All experiments were performed in technical duplicate, symbols represent the mean of the duplicates, and fits are four-parameter logistic curves.

FIGS. 20A-20F: Show designed antibodies improve pseudoviral neutralization over COV2-2130. The parental COV2-2130 antibody (orange circles), cross-reactive positive control antibody rS2K146 (magenta squares), and down-selected computationally designed antibodies were assayed by neutralization with lentiviruses pseudotyped with spike variants of Wuhan-1 D614G (FIG. 20A), Omicron BA.1 (FIG. 20B), BA.1.1 (FIG. 20C), BA.2 (FIG. 20D), and BA.4 (FIG. 20E). FIG. 20F shows a key for the symbols of FIGS. 20A-20E. Symbols indicate the mean and standard deviation of two technical replicates; curves are 4-parameter logistic regression models fit using GraphPad Prism.

DETAILED DESCRIPTION

The COVID-19 pandemic has highlighted how viral variants that escape monoclonal antibody drugs can undermine drug development efforts. The substantially mutated SARS-CoV-2 variants Omicron BA.1 and BA.1.1 escaped many clinical antibody drug products, including AZD1061 [VanBlargan 2022]. Rapidly modifying existing antibody-based drugs products to restore efficacy to emerging variants is a potential mitigation strategy. To that end, the design of a novel antibody derived from COV2-2130, the progenitor of AZD1061, that could potently neutralize BA.1 and BA.1.1 while maintaining efficacy against the Delta variant were sought. Accordingly, the present disclosure provides disclosed derivatives of COV2-2130 that achieve this goal. Notably, the antibody designs described herein achieved potent neutralization of Delta, BA.1, BA.1.1, and BA.2, and also potently neutralized more recent BA.4 and BA.5.5 Omicron strains, where the parental COV2-2130 again suffers significant losses in potency. This work provides a novel computational system for rapid antibody adaptation addressing escape variants. The present disclosure also provides a platform that combines high-performance computing-enabled simulation, machine learning, and structural bioinformatics to optimize multiple antibody properties simultaneously. Combined with knowledge of potential escape liabilities, the approach described herein could support pre-emptive design of broadly protective monoclonal antibodies that are robust across a wide range of possible antigenic variants.

Viruses evolve gradually or suddenly, which can result in evasion of the protection offered by medical countermeasures, including therapeutic or prophylactic monoclonal antibodies. During the course of the COVID-19 pandemic, new variants of the SARS-CoV-2 virus have emerged that evade FDA-approved countermeasures, candidates, and the human immune system, posing new dangers to the public.

AZD7442 (Evusheld), an FDA Emergency Authorized monoclonal antibody, is a combination of two fully human, SARS-CoV-2-neutralizing monoclonal antibodies (tixagevimab and cilgavimab) that are derived from antibodies isolated from B cells obtained from persons infected with SARS-CoV-2. Cilgavimab (AZD1061) is a derivative of the monoclonal antibody COV2-2130. Another derivative of COV2-2130 is also used in the antibody product being produced by Ology Bioservices (e.g., COV2-2130 and COV2-2381) under contract with the Department of Defense (DOD). While the combination of Tixagevimab and cilgavimab is fully active against the Delta variant of SARS-CoV-2 (B.1.617.2 and AY lineages), the combination of Tixagevimab and cilgavimab has decreased neutralizing activity in vitro against the Omicron variants. See e.g., JAMA, 327(4):384-385 (2022). Specifically, the antibody COV-2130 fails to neutralize the BA.1 and BA.1.1 (“Omicron”) strains of SARS-CoV-2 in vitro. Given the continuous evolution of the virus that leads to SARS-CoV-2 variants and the constant developments in our understanding of the impacts of these variants, there is a need for novel prophylactic monoclonal antibodies that can bind to and neutralize old and emerging strains of SARS-CoV-2.

One aspect of the present disclosure provides a novel class of anti-SARS-CoV-2 antibodies that exhibited dramatically improved in vitro binding and/or neutralization effects against all known SARS-CoV-2 variants, including but not limited to the “Omicron” variants BA.1 and BA.1.1 (e.g, B.1.1.529, BA.1, BA.1.1, BA.2, BA.3, BA.4 and BA.5 lineages). The novel class anti-SARS-CoV-2 antibodies showed enhanced binding and/or neutralization of older SARS-CoV strains (e.g., Alpha (B.1.1.7 and Q lineages); Beta (B.1.351 and descendent lineages); Gamma (P.1 and descendent lineages); Delta (B.1.617.2 and AY lineages); Epsilon (B.1.427 and B.1.429); Eta (B.1.525); Iota (B.1.526); Kappa (B.1.617.1); 1.617.3; Mu (B.1.621, B.1.621.1); Zeta (P.2)). In particular, the novel class anti-SARS-CoV-2 antibodies showed enhanced binding and/or neutralization of the “Omicron” variants such as B.1.1.529, BA.1, BA.1.1, BA.2, BA.3, BA.4 and BA.5 lineages. This novel class antibodies disclosed herein were derived from the COV-2130 monoclonal antibody and were re-engineered specifically for improved and enhanced binding and neutralization characteristics against several SARS-CoV-2 strains (i.e., old and emerging strains).

The novel antibodies of the present disclosure can be used as part of a therapeutic or prophylactic drug substance or drug product. They also can be used in diagnostics, detection and as personalized therapies. In some embodiments, the novel antibodies are used as: (1) a “drop-in” replacement of existing AZD1061/COV-2130 prophylactic, (2) a parallel prophylactic (delivered prior to infection) or therapeutic (delivered after infection) products, or (3) in combination with other anti-SARS-CoV-2 antibodies or therapeutics. In some embodiments, the novel antibodies of the present disclosure are used as part of a diagnostic or biosensor for identifying which strain or strains of coronavirus, severe acute respiratory syndrome coronavirus, or SARS-CoV-2 is or are present in a sample. In some embodiments, the novel antibodies are used to predict the efficacy of Evusheld, Cilgavimab, Resilience or related therapeutics, and/or prophylactics against a patient's individual COVID-19 case. In some embodiments, the novel antibodies are used as reagents in laboratory processes, or for similar functions.

The state of the art method for dealing with the emergence of novel SARS-CoV-2 strains is to abandon or substantially modify the drug product or drug substance, resulting in large delays and/or total loss of the product line. The antibodies of this disclosure and the methods of making them (1) can avoid this outcome for the deployed Evusheld/Cilgavimab product, and (2) are far less expensive and have a less uncertain path to minimal, robust repair of approved and deployed antibody or protein drug substances and products. In the context of the COVID-19 pandemic in particular, better protection against BA.1 and BA.1.1 and future related strains could save lives and substantial sums of money, depending on how the pandemic continues to evolve. Because the antibodies of the present disclosure introduced targeted new chemistries to the antibody/antigen interface, they showed excellent binding and neutralization characteristics against old SARS-CoV-2 strains, while also achieving the critical goal of improving/restoring efficacy against new emerging variants, such as BA.1 and BA.1.1 and other Omicron lineages and related strains (e.g, B.1.1.529, BA.1, BA.1.1, BA.2, BA.3, BA.4, and BA.5 lineages). Indeed, FIG. 1 and Table 15 shows the antibodies of the present disclosure show improved affinity (EC₅₀) against either the BA1 variant and the Wuhan variant compared to the parental antibody or the BA.1.1 variant (e.g., EC₅₀=1.353, 1.955 or 2.614) than known COV2-2130 variants (e.g., EC₅₀=3.542 or 4.132). Moreover, as shown in FIG. 3, the neutralization characteristics (IC₅₀) of the antibodies of the present disclosure are also superior to those of the classic COV2-2130 antibody. FIG. 3 shows that the IC₅₀of the antibodies of the present disclosure ranges from 0.1-1.1 ng/ml vs 6.1 ng/ml for COV2-2130; 17 ng/ml vs>3687 ng/ml for COV2-2130; or 7.5 ng/ml v. 1769 ng/ml for COV2-2130.

The monoclonal antibodies of the present disclosure are an improvement over previously described anti-SARS-CoV-2 monoclonal antibodies particularly because of their enhancements in binding and neutralization of known SARS-CoV-2 strains when compared to the COV-2130 antibody, even for some strains that COV-2130 binds and neutralizes well. The novel class of antibodies of the present disclosure have unique mutations in the complementary determining regions of the heavy (V_H) and light chain (V_L) variable domains. These mutations were surprising because they alter residues in the COV2-2130 monoclonal antibody that offered surprising, and across-the-board improvements for binding old and emerging strains of SARS-CoV-2, including the Omicron variants (e.g, B.1.1.529, BA.1, BA.1.1, BA.2, BA.3, BA.4, and BA.5 lineages). For example, in the CDR2 of the heavy chain variable domain, the isoleucine residue at position 55 of the COV2-2130 sequence was mutated to either histidine (I55H; SEQ ID NO: 108), glutamic acid (I55E; SEQ ID NO: 100 or 107), aspartate (I55D; SEQ ID NO: 104) or tryptophan (I55W; SEQ ID NO: 102). In the CDR3 of the heavy chain variable domain, the glycine residue at position 112 of the COV2-2130 sequence was mutated to glutamic acid (G112E), which altered the characteristics of the H3 loop. Other mutations were surprising because they delivered large empirical effects from chemically modest mutant residues. For example, in the CDR1 of the light chain variable domain of the COV2-2130 sequence, the serine residues at positions 32 and 33 of the COV2-2130 sequence were mutated to alanine (e.g., S32A, S33A; SEQ ID NO: 163). See Table 13 for the list of specific mutations in the disclosed antibodies.

In addition, the antibodies provided by the present disclosure repair a clinically deployed prophylactic product (e.g., COV2-2130; Cilgavimab (AZD1061)) rendered dramatically less effective by the emergence of novel SARS-CoV-2 strains, such as the current BA.1, BA.1.1, and related “Omicron” strains.

Accordingly, in one aspect, the present disclosure provides an isolated antibody or antibody fragment that binds to a SARS-CoV-2 surface protein (e.g., a spike (S) glycoprotein) having improved binding and/or neutralization characteristics against all known SARS-CoV-2 strains when compared to a monoclonal antibody selected from sotrovimab (S309), COV2-2381, cilgavimab (COV2-2130 or AZD1061), and/or tixagevimab (COV2-2196 or AZD8895). The SARS-CoV-2 variant is selected from Alpha (B.1.1.7 and Q lineages); Beta (B.1.351 and descendent lineages); Gamma (P.1 and descendent lineages); Delta (B.1.617.2 and AY lineages); Epsilon (B.1.427 and B.1.429); Eta (B.1.525); Iota (B.1.526); Kappa (B.1.617.1); 1.617.3; Mu (B.1.621, B.1.621.1); Zeta (P.2); or Omicron (B.1.1.529, BA.1, BA.1.1, BA.2, BA.3, BA.4 and BA.5 lineages). In particular, the SARS-CoV-2 strain is an “Omicron” strain or variants thereof, such as B.1.1.529, BA.1, BA.1.1, BA.2, BA.3, BA.4 and BA.5 lineages. In some embodiments, the isolated antibody or antibody fragment comprises, or consists essentially of, or consists of heavy chain complementary determining regions 1-3 (CDRH1-3) sequences; and a light chain complementary determining regions 1-3 (CDRL1-3) sequences disclosed in Tables 1˜4 or Table 13. In some embodiments, the isolated antibody or antibody fragment comprises a heavy chain variable sequence and a light chain variable sequence disclosed in Table 4. The antibody fragment is a recombinant antibody variable domain fragment (Fv fragment) and/or comprises the amino acid sequence of SEQ ID NOs: 1-14 or 15-99, or an amino acid sequence SEQ ID NO disclosed in Table 4, Table 13 or Table 14.

Another aspect of the present disclosure provides an isolated nucleic acid comprising, or consisting essentially of, or consisting of a nucleotide sequence encoding the antibody or antibody fragment disclosed herein. Another aspect of the present disclosure provides a vector comprising, or consisting essentially of, or consisting of an isolated nucleic acid comprising a nucleotide sequence encoding the antibody or antibody fragment disclosed herein. Another aspect of the present disclosure provides an engineered cell comprising, or consisting essentially of, or consisting of the isolated nucleic acid comprising a nucleotide sequence encoding the antibody or antibody fragment disclosed herein. Another aspect of the present disclosure provides a composition comprising, or consisting essentially of, or consisting of one or more of the antibodies or antibody fragments disclosed herein, the isolated nucleic acid, or the vector disclosed herein. Another aspect of the present disclosure provides a vaccine formulation comprising, or consisting essentially of, or consisting of one or more antibodies or antibody fragments disclosed herein.

Another aspect of the present disclosure provides a method of treating an infectious disease, e.g., viral infection, coronavirus infection, SARS-CoV-2 infection. In one aspect, the present disclosure provides a method of treating a subject infected with SARS-CoV-2 or reducing the likelihood of infection of a subject at risk of contracting SARS-CoV-2, comprising, or consisting essentially of, or consisting of delivering to the subject the antibody or antibody fragment disclosed herein. Another aspect provides a method of treating a subject infected with SARS-CoV-2 or reducing the likelihood of infection of a subject at risk of contracting SARS-CoV-2, comprising, or consisting essentially of, or consisting of delivering to the subject a first antibody or antibody fragment and a second antibody or antibody fragment. In some embodiments, the first and second antibodies or antibody fragments synergistically neutralize the SARS-CoV-2 virus.

Another aspect of the present disclosure provides a method of detecting an infectious agent in a subject suspected of being infected by a coronavirus. In one aspect, the disclosure provides a method of detecting infection with SARS-CoV-2 in a subject comprising, or consisting essentially of, or consisting of contacting a sample isolated from the subject with the antibody or antibody fragment disclosed herein; and detecting SARS-CoV-2 in the sample by assessing the binding of the antibody or antibody fragment disclosed herein to a SARS-CoV-2 antigen in the sample.

Another aspect of the present disclosure provides a method of determining a SARS-CoV-2 virus variant antigenic profile comprising, or consisting essentially of, or consisting of: contacting a sample comprising the SARS-CoV-2 virus variant with the antibody or antibody fragment disclosed herein; and determining the binding of the antibody or antibody fragment to the SARS-CoV-2 virus variant, wherein absence of binding indicates a conformational change and/or a mutation in the SARS-CoV-2 surface spike protein.

Another aspect of the present disclosure provides a biosensor for detecting the presence of a SARS-CoV-2 antigen in a sample, the biosensor comprising a transducer component comprising, or consisting essentially of, or consisting of an electrode operatively connected to a microprocessor, the microprocessor being adapted to receive, process and transmit a signal; a receptor component having a sensing element capable of detecting and binding to the SARS-CoV-2 antigen; and the transducer component and the receptor component are capable of being brought into direct contact with the sample in situ. For example, the sensing element comprises the antibody or antibody fragment disclosed herein.

It will be understood that monoclonal antibodies binding to SARS-CoV-2 will have several applications. These include the production of diagnostic kits for use in detecting and diagnosing SARS-CoV-2 infection, as well as for treating the same. In these contexts, one may link such antibodies to diagnostic or therapeutic agents, use them as capture agents or competitors in competitive assays, or use them individually without additional agents being attached thereto. The antibodies may be mutated or modified, as discussed further below. Methods for preparing and characterizing antibodies are well known in the art (see, e.g., Antibodies: A Laboratory Manual, Cold Spring Harbor Laboratory, 1988; U.S. Pat. No. 4,196,265).

Definitions

Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure belongs. Although methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present disclosure, suitable methods and materials are described herein.

It is also to be understood that the terminology used herein is for the purpose of describing particular embodiments only, and is not intended to be limiting.

The practice of the present disclosure will employ, unless otherwise indicated, conventional techniques of tissue culture, immunology, molecular biology, microbiology, cell biology and recombinant DNA, which are within the skill of the art. See e.g., Green and Sambrook eds. (2012) Molecular Cloning: A Laboratory Manual, 4th edition; the series Ausubel et al. eds. (2015) Current Protocols in Molecular Biology; the series Methods in Enzymology (Academic Press, Inc., N.Y.); MacPherson et al. (2015) PCR 1: A Practical Approach (IRL Press at Oxford University Press); MacPherson et al. (1995) PCR 2: A Practical Approach; McPherson et al. (2006) PCR: The Basics (Garland Science); Harlow and Lane eds. (1999) Antibodies, A Laboratory Manual; Greenfield ed. (2014) Antibodies, A Laboratory Manual; Freshney (2010) Culture of Animal Cells: A Manual of Basic Technique, 6th edition; Gait ed. (1984) Oligonucleotide Synthesis; Hames and Higgins eds. (1984) Nucleic Acid Hybridization; Anderson (1999) Nucleic Acid Hybridization; Herdewijn ed. (2005) Oligonucleotide Synthesis: Methods and Applications; Hames and Higgins eds. (1984) Transcription and Translation; Buzdin and Lukyanov ed. (2007) Nucleic Acids Hybridization: Modern Applications; Immobilized Cells and Enzymes (IRL Press (1986)); Grandi ed. (2007) In Vitro Transcription and Translation Protocols, 2nd edition; Guisan ed. (2006) Immobilization of Enzymes and Cells; Perbal (1988) A Practical Guide to Molecular Cloning, 2nd edition; Miller and Calos eds, (1987) Gene Transfer Vectors for Mammalian Cells (Cold Spring Harbor Laboratory); Makrides ed. (2003) Gene Transfer and Expression in Mammalian Cells; Mayer and Walker eds. (1987) Immunochemical Methods in Cell and Molecular Biology (Academic Press, London); Lundblad and Macdonald eds. (2010) Handbook of Biochemistry and Molecular Biology, 4th edition; and Herzenberg et al. eds (1996) Weir's Handbook of Experimental Immunology, 5th edition.

As used herein, the singular forms “a”, “an,” and “the” include plural referents unless the context clearly indicates otherwise. For example, the term “a cell” includes a plurality of cells, including mixtures thereof, and means one cell or more than one cell

As used herein, the term “about” is used to indicate that a value includes the standard deviation of error for the device or method being employed to determine the value. The term “about” when used before a numerical designation, e.g., temperature, time, amount, and concentration, including range, indicates approximations which may vary by (+) or (—) (±) 20%, 15%, 10%, 5%, 3%, 2%, or 1%. Preferably ±5%, more preferably ±1%, and still more preferably ±0.1% from the specified value, as such variations are appropriate to perform the disclosed methods.

Administration or treatment in “combination” refers to administering two agents such that their pharmacological effects are manifest at the same time. Combination does not require administration at the same time or substantially the same time, although combination can include such administrations.

The term “antigen” as used herein is defined as a molecule that provokes an immune response. This immune response may involve either antibody production, or the activation of specific immunologically-competent cells, or both. The skilled artisan will understand that any macromolecule, including virtually all proteins or peptides, can serve as an antigen. Furthermore, antigens can be derived from recombinant or genomic DNA. A skilled artisan will understand that any DNA, which comprises a nucleotide sequences or a partial nucleotide sequence encoding a protein that elicits an immune response therefore encodes an “Antigen” as that term is used herein. Furthermore, one skilled in the art will understand that an antigen need not be encoded solely by a full-length nucleotide sequence of a gene. It is readily apparent that the present invention includes, but is not limited to, the use of partial nucleotide sequences of more than one gene and that these nucleotide sequences are arranged in various combinations to elicit the desired immune response.

As used herein, the term “antibody” refers to an immunoglobulin molecule, which specifically binds with an antigen. Antibodies can be intact immunoglobulins derived from natural sources or from recombinant sources and can be immunoreactive portions of intact immunoglobulins. Antibodies are typically tetramers of immunoglobulin molecules. The antibodies in the present invention may exist in a variety of forms including, for example, polyclonal antibodies, monoclonal antibodies, variable domain fragment (Fv), Fab and F(ab)2, as well as single chain antibodies (scFv) and humanized antibodies In some embodiments, antibody refers to such assemblies (e.g., intact antibody molecules, immunoadhesins, or variants thereof) which have significant known specific immunoreactive activity to an antigen of interest (e.g., SARS-CoV-2 protein). Antibodies and immunoglobulins comprise light and heavy chains, with or without an interchain covalent linkage between them. Basic immunoglobulin structures in vertebrate systems are relatively well understood.

In some embodiments, an antibody is a protein, or polypeptide sequence derived from an immunoglobulin molecule which specifically binds with an antigen. Antibodies can be polyclonal or monoclonal, multiple or single chain, or intact immunoglobulins, and may be derived from natural sources or from recombinant sources. Antibodies can be tetramers of immunoglobulin molecules. In one embodiment, the antibody or antibody molecule comprises, e.g., consists of, an antibody fragment.

The term “antibody fragment” refers to a portion of an intact antibody and refers to the antigenic determining variable regions of an intact antibody. In some embodiments, the term “antibody fragment” refers to at least one portion of an intact antibody, or recombinant variants thereof, and refers to the antigen binding domain, e.g., an antigenic determining variable region of an intact antibody, that is sufficient to confer recognition and specific binding of the antibody fragment to a target, such as an antigen. Examples of antibody fragments include, but are not limited to, Fab, Fab′, F(ab′)2, Fv fragments, scFv antibody fragments, linear antibodies, single domain antibodies such as sdAb (either V_Lor V_H), camelid VHH domains, and multi-specific antibodies formed from antibody fragments such as a bivalent fragment comprising two Fab fragments linked by a disulfide bridge at the hinge region, and an isolated CDR or other epitope binding fragments of an antibody. An antigen binding fragment can also be incorporated into single domain antibodies, maxibodies, minibodies, nanobodies, intrabodies, diabodies, triabodies, tetrabodies, v-NAR and bis-scFv (see, e.g., Hollinger and Hudson, Nature Biotechnology 23:1126-1136, 2005). Antigen binding fragments can also be grafted into scaffolds based on polypeptides such as a fibronectin type III (Fn3)(see U.S. Pat. No. 6,703,199, which describes fibronectin polypeptide minibodies). “Fab” means a monovalent antigen-binding fragment of an immunoglobulin that is composed of the light chain and part of the heavy chain. F(ab′)2 means a bivalent antigen-binding fragment of an immunoglobulin that contains both light chains and part of both heavy chains.

As used herein, the term “Fv fragment” or “variable domain fragment” refers to a VH domain and a VL domain of an antibody specifically binding to an antigen, both domains forming together a Fv fragment. In some embodiment, Fv fragments means an antibody fragment comprising the V_Hand V_Ldomains of an antibody, wherein these domains are present in a single polypeptide chain as disclosed in Table 4 herein. Generally, the Fv fragment polypeptide further comprises a polypeptide linker between the V_Hand V_Ldomains polypeptide that enables the scFv to form.

The term “scFv” refers to a fusion protein comprising at least one antibody fragment comprising a variable region of a light chain and at least one antibody fragment comprising a variable region of a heavy chain, wherein the light and heavy chain variable regions are contiguously linked via a short flexible polypeptide linker, and capable of being expressed as a single chain polypeptide, and wherein the scFv retains the specificity of the intact antibody from which it is derived. Unless specified, as used herein an scFv may have the V_Land V_Hvariable regions in either order, e.g., with respect to the N-terminal and C-terminal ends of the polypeptide, the scFv may comprise V_L-linker-V_Hor may comprise V_H-linker-V_L.

As used herein, the term “antibody heavy chain” refers to the larger of the two types of polypeptide chains present in all antibody molecules in their naturally occurring conformations.

As used herein, the term “antibody light chain” refers to the smaller of the two types of polypeptide chains present in all antibody molecules in their naturally occurring conformations. Kappa (κ) and lambda (λ) light chains refer to the two major antibody light chain isotypes.

As used herein, the term “synthetic antibody” means an antibody, which is generated using recombinant DNA technology, such as, for example, an antibody expressed by a bacteriophage. The term should also be construed to mean an antibody, which has been generated by the synthesis of a DNA molecule encoding the antibody and which DNA molecule expresses an antibody protein, or an amino acid sequence specifying the antibody, wherein the DNA or amino acid sequence has been obtained using synthetic DNA or amino acid sequence technology which is available and well known in the art.

As used herein, the term “antibody variant” includes synthetic and engineered forms of antibodies which are altered such that they are not naturally occurring, e.g., antibodies that comprise at least two heavy chain portions but not two complete heavy chains (such as, domain deleted antibodies or minibodies); multi-specific forms of antibodies (e.g., bispecific, tri-specific, etc.) altered to bind to two or more different antigens or to different epitopes on a single antigen); heavy chain molecules joined to scFv molecules and the like. In addition, the term “antibody variant” includes multivalent forms of antibodies (e.g., trivalent, tetravalent, etc., antibodies that bind to three, four or more copies of the same antigen.

As used herein, the term “antigen” or “Ag” is defined as a molecule that provokes an immune response. This immune response may involve either antibody production, or the activation of specific immunologically-competent cells, or both. The skilled artisan will understand that any macromolecule, including virtually all proteins or peptides, can serve as an antigen. Furthermore, antigens can be derived from recombinant or genomic DNA. A skilled artisan will understand that any DNA, which comprises a nucleotide sequence or a partial nucleotide sequence encoding a protein that elicits an immune response therefore encodes an “antigen” as that term is used herein. Furthermore, one skilled in the art will understand that an antigen need not be encoded solely by a full-length nucleotide sequence of a gene. It is readily apparent that the present invention includes, but is not limited to, the use of partial nucleotide sequences of more than one gene and that these nucleotide sequences are arranged in various combinations to elicit a desired immune response. Moreover, the skilled artisan will understand that an antigen need not be encoded by a “gene” at all. It is readily apparent that an antigen can be generated synthesized or can be derived from a biological sample. Such a biological sample can include, but is not limited to a tissue sample, a tumor sample, a cell or a biological fluid.

As used herein, the term “complementary” as used in connection with nucleic acid, refers to the pairing of bases, A with T or U, and G with C. The term complementary refers to nucleic acid molecules that are completely complementary, that is, form A to T or U pairs and G to C pairs across the entire reference sequence, as well as molecules that are at least 80%, 85%, 90%, 95%, 99% complementary.

As used herein, the term “complementarity determining region” or “CDR” refers to the sequences of amino acids within antibody variable regions which confer antigen specificity and binding affinity. For example, in general, there are three CDRs in each heavy chain variable region (e.g., CDRH1, CDRH2, and CDRH3) and three CDRs in each light chain variable region (CDRL1, CDRL2, and CDRL3). The precise amino acid sequence boundaries of a given CDR can be determined using any of a number of well-known schemes, including those described by Kabat et al. (1991), “Sequences of Proteins of Immunological Interest,” 5th Ed. Public Health Service, National Institutes of Health, Bethesda, Md. (“Kabat” numbering scheme), Al-Lazikani et al., (1997) JMB 273, 927-948 (“Chothia” numbering scheme), or a combination thereof, North et al., J. Molecular Biology, 406 (2): 228-256 (2011) and; Lefranc et al. Nucl. Acids Res. 27:209-212 (1999) or Ruiz et al. Nucl. Acids Res. 28:219-221 (2000)) (IMGT numbering scheme).

Under the Kabat numbering scheme, in some embodiments, the CDR amino acid residues in the heavy chain variable domain (VH) are numbered 31-35 (HCDR1), 50-65 (HCDR2), and 95-102 (HCDR3); and the CDR amino acid residues in the light chain variable domain (VL) are numbered 24-34 (LCDR1), 50-56 (LCDR2), and 89-97 (LCDR3). Under the Chothia numbering scheme, in some embodiments, the CDR amino acids in the VH are numbered 26-32 (HCDR1), 52-56 (HCDR2), and 95-102 (HCDR3); and the CDR amino acid residues in the VL are numbered 26-32 (LCDR1), 50-52 (LCDR2), and 91-96 (LCDR3). Under the IMGT numbering scheme, in some embodiments, the CDR amino acids in the VH are numbered 27-38 (HCDR1), 56-65 (HCDR2), and 105-120 (HCDR3); and the CDR amino acid residues in the VL are numbered 27-38 (LCDR1), 56-65 (LCDR2), and 105-120 (LCDR3).

In a combined Kabat and Chothia numbering scheme, in some embodiments, the CDRs correspond to the amino acid residues that are part of a Kabat CDR, a Chothia CDR, or both. For instance, in some embodiments, the CDRs correspond to amino acid residues 26-35 (HCDR1), 50-65 (HCDR2), and 95-102 (HCDR3) in a VH, e.g., a mammalian VH, e.g., a human VH; and amino acid residues 24-34 (LCDR1), 50-56 (LCDR2), and 89-97 (LCDR3) in a VL, e.g., a mammalian VL, e.g., a human VL.

As used herein, the term “conservative sequence modifications” is intended to refer to amino acid modifications that do not significantly affect or alter the binding characteristics of the antibody containing the amino acid sequence. Such conservative modifications include amino acid substitutions, additions and deletions. Modifications can be introduced into an antibody of the invention by standard techniques known in the art, such as site-directed mutagenesis and PCR-mediated mutagenesis. Conservative amino acid substitutions are ones in which the amino acid residue is replaced with an amino acid residue having a similar side chain. Families of amino acid residues having similar side chains have been defined in the art. These families include amino acids with basic side chains (e.g., lysine, arginine, histidine), acidic side chains (e.g., aspartic acid, glutamic acid), uncharged polar side chains (e.g., glycine, asparagine, glutamine, serine, threonine, tyrosine, cysteine, tryptophan), nonpolar side chains (e.g., alanine, valine, leucine, isoleucine, proline, phenylalanine, methionine), beta-branched side chains (e.g., threonine, valine, isoleucine) and aromatic side chains (e.g., tyrosine, phenylalanine, tryptophan, histidine). Thus, one or more amino acid residues within the CDR regions of an antibody can be replaced with other amino acid residues from the same side chain family and the altered antibody can be tested for the ability to bind antigens using the functional assays described herein.

As used herein, the term “disease” refers to a state of health of an animal wherein the animal cannot maintain homeostasis, and wherein if the disease is not ameliorated then the animal's health continues to deteriorate. In contrast, the term “disorder” in an animal refers to a state of health in which the animal is able to maintain homeostasis, but in which the animal's state of health is less favorable than it would be in the absence of the disorder. Left untreated, a disorder does not necessarily cause a further decrease in the animal's state of health.

A “composition” as used herein, refers to an active agent, such as a compound as disclosed herein and a carrier, inert or active. In some embodiments, a carrier can be, without limitation, solid such as a bead or resin, or liquid, such as phosphate buffered saline. In some embodiments, a carrier also includes pharmaceutical excipients and additives proteins, peptides, amino acids, lipids, and carbohydrates (e.g., sugars, including monosaccharides, di-, tri, tetra-oligosaccharides, and oligosaccharides; derivatized sugars such as alditols, aldonic acids, esterified sugars and the like; and polysaccharides or sugar polymers), which can be present singly or in combination, comprising alone or in combination 1-99.99% by weight or volume. Exemplary protein excipients include serum albumin such as human serum albumin (HSA), recombinant human albumin (rHA), gelatin, casein, and the like. Representative amino acid/antibody components, which can also function in a buffering capacity, include alanine, arginine, glycine, arginine, betaine, histidine, glutamic acid, aspartic acid, cysteine, lysine, leucine, isoleucine, valine, methionine, phenylalanine, aspartame, and the like. Carbohydrate excipients are also intended within the scope of this technology, examples of which include but are not limited to monosaccharides such as fructose, maltose, galactose, glucose, D-mannose, sorbose, and the like; disaccharides, such as lactose, sucrose, trehalose, cellobiose, and the like; polysaccharides, such as raffinose, melezitose, maltodextrins, dextrans, starches, and the like; and alditols, such as mannitol, xylitol, maltitol, lactitol, xylitol sorbitol (glucitol) and myoinositol.

As used herein, the term “encoding” refers to the inherent property of specific sequences of nucleotides in a polynucleotide (such as a gene, a cDNA, or an mRNA), to serve as templates for synthesis of other polymers and macromolecules in biological processes having either a defined sequence of nucleotides (e.g., rRNA, tRNA and mRNA) or a defined sequence of amino acids and the biological properties resulting therefrom. Thus, a gene, cDNA, or RNA, encodes a protein if transcription and translation of mRNA corresponding to that gene produces the protein in a cell or other biological system. Both the coding strand, the nucleotide sequence of which is identical to the mRNA sequence and is usually provided in sequence listings, and the non-coding strand, used as the template for transcription of a gene or cDNA, can be referred to as encoding the protein or other product of that gene or cDNA. Unless otherwise specified, a “nucleotide sequence encoding an amino acid sequence” includes all nucleotide sequences that are degenerate versions of each other and that encode the same amino acid sequence. The phrase nucleotide sequence that encodes a protein or a RNA may also include introns to the extent that the nucleotide sequence encoding the protein may in some version contain an intron(s).

As used herein, the term “recombinant host cell,” “recombinant cell,” “engineered host cell,” or “engineered cell,” means a cell into which a recombinant expression vector has been introduced. It should be understood that such terms are intended to refer not only to the particular subject cell but to the progeny of such a cell. Because certain modifications may occur in succeeding generations due to either mutation or environmental influences, such progeny may not, in fact, be identical to the parent cell, but are still included within the scope of the term “host cell” or “cell” as used herein. Host cells include progeny of a single host cell, and the progeny may not necessarily be completely identical (in morphology or in total DNA complement) to the original parent cell due to natural, accidental, or deliberate mutation and/or change. A host cell includes a cell transfected or infected in vivo or in vitro with a recombinant vector, an expression vector, or a nucleic acid encoding an antibody of the present disclosure. A host cell, which comprises a recombinant vector, expression vector, or a nucleic acid encoding an antibody disclosed herein, may also be referred to as a “recombinant host cell,” “engineered host cell,” or “engineered cell”.

As used herein, the term “host cell” refers to a cell, which may be used in a process for purifying an immunogenic protein or recombinant antibody in accordance with the present disclosure. Such host cell expresses the protein of interest (the antibody disclosed herein). A host cell may also be referred to as a protein-expressing cell. “Host cell” refers not only to the particular subject cell but to the progeny or potential progeny of such a cell. Because certain modifications may occur in succeeding generations due to either mutation or environmental influences, such progeny may not, in fact, be identical to the parent cell, but are still included within the scope of the term as used herein.

A host cell, according to the present disclosure, may be, but is not limited to, prokaryotic cells, eukaryotic cells, archeobacteria, bacterial cells, insect cells, yeast, mammal cells, and/or plant cells. Bacteria envisioned as host cells can be either gram-negative or gram-positive, e.g. Escherichia coli, Erwinia sp., Klebsellia sp., Lactobacillus sp. or Bacillus subtilis. In some embodiments, the host cell is a yeast cell. In that embodiment, the yeast host cell is selected from the group consisting of Saccharomyces cerevisiae, Hansenula polymorpha, and Pichia pastoris.

“Eukaryotic cells” comprise all of the life kingdoms except Monera. They can be easily distinguished through a membrane-bound nucleus. Animals, plants, fungi, and protists are eukaryotes or organisms whose cells are organized into complex structures by internal membranes and a cytoskeleton. The most characteristic membrane-bound structure is the nucleus. Unless specifically recited, the term “host” includes a eukaryotic host, including, for example, yeast, higher plant, insect and mammalian cells. Non-limiting examples of eukaryotic cells or hosts include simian, bovine, porcine, murine, rat, avian, reptilian and human.

“Prokaryotic cells” that usually lack a nucleus or any other membrane-bound organelles and are divided into two domains, bacteria and archaea. In addition to chromosomal DNA, these cells can also contain genetic information in a circular loop called an episome. Bacterial cells are very small, roughly the size of an animal mitochondrion (about 1-2 μm in diameter and 10 μm long). Prokaryotic cells feature three major shapes: rod shaped, spherical, and spiral. Instead of going through elaborate replication processes like eukaryotes, bacterial cells divide by binary fission. Examples include but are not limited to Bacillus bacteria, E. coli bacterium, and Salmonella bacterium.

“Effective amount” or “therapeutically effective amount” as used interchangeably herein, refer to an amount of a compound, formulation, material, pharmaceutical agent, or composition, as described herein effective to achieve a desired physiological, therapeutic, or prophylactic outcome in a subject in need thereof. Such results may include, but are not limited to an amount that when administered to a mammal, causes a detectable level of immune response compared to the immune response detected in the absence of the composition of the invention. The immune response can be readily assessed by a plethora of art-recognized methods. The skilled artisan would understand that the amount of the composition administered herein varies and can be readily determined based on a number of factors such as the disease or condition being treated, the age and health and physical condition of the mammal being treated, the severity of the disease, the particular compound being administered, and the like. The effective amount may vary among subjects depending on the health and physical condition of the subject to be treated, the taxonomic group of the subjects to be treated, the formulation of the composition, assessment of the subject's medical condition, and other relevant factors.

In some embodiments, an “effective amount” is an amount sufficient to effect beneficial or desired results. An effective amount can be administered in one or more administrations, applications or dosages. Such delivery is dependent on a number of variables including the time period for which the individual dosage unit is to be used, the bioavailability of the therapeutic agent, the route of administration, etc. It is understood, however, that specific dose levels of the therapeutic agents disclosed herein for any particular subject depends upon a variety of factors including the activity of the specific compound employed, bioavailability of the compound, the route of administration, the age of the animal and its body weight, general health, sex, the diet of the animal, the time of administration, the rate of excretion, the drug combination, and the severity of the particular disorder being treated and form of administration. Provided herein in the experimental examples are effective amounts determined in a murine or rat animal model, which can be converted to an effective dose by converting the reported μg of particle per g of the mouse. A typical adult mouse is from about 15 to about 35 g for a female mouse and about 20 g to about 30 g for a male mouse. In general, one will desire to administer an amount of the compound that is effective to achieve a serum level commensurate with the concentrations found to be effective in vivo. These considerations, as well as effective formulations and administration procedures are well known in the art and are described in standard textbooks.

As used herein, the term “expression vector” refers to a vector comprising a recombinant polynucleotide comprising expression control sequences operatively linked to a nucleotide sequence to be expressed. An expression vector comprises sufficient cis-acting elements for expression; other elements for expression can be supplied by the host cell or in an in vitro expression system. Expression vectors include all those known in the art, such as cosmids, plasmids (e.g., naked or contained in liposomes) and viruses (e.g., Sendai viruses, lentiviruses, retroviruses, adenoviruses, and adeno-associated viruses) that incorporate the recombinant polynucleotide.

As used herein, the term “ex vivo,” refers to cells that have been removed from a living organism, (e.g., a human) and propagated outside the organism (e.g., in a culture dish, test tube, or bioreactor).

As used herein, the term “humanized antibodies” refers to human forms of non-human (e.g., murine) antibodies, and are chimeric immunoglobulins, immunoglobulin chains or fragments thereof (such as Fv, Fab, Fab′, F(ab′)2 or other antigen-binding subsequences of antibodies), which contain minimal sequence derived from non-human immunoglobulin. For the most part, humanized antibodies are human immunoglobulins (recipient antibody) in which residues from a complementary-determining region (CDR) of the recipient are replaced by residues from a CDR of a non-human species (donor antibody) such as mouse, rat or rabbit having the desired specificity, affinity, and capacity. In some instances, Fv framework region (FR) residues of the human immunoglobulin are replaced by corresponding non-human residues.

Furthermore, humanized antibodies can comprise residues which are found neither in the recipient antibody nor in the imported CDR or framework sequences. These modifications are made to further refine and optimize antibody performance. In general, the humanized antibody will comprise substantially all of at least one, and typically two, variable domains, in which all or substantially all of the CDR regions correspond to those of a non-human immunoglobulin and all or substantially all of the FR regions are those of a human immunoglobulin sequence. The humanized antibody optimally also will comprise at least a portion of an immunoglobulin constant region (Fc), typically that of a human immunoglobulin. For further details, see Jones et al., Nature 321: 522-525 (1986); Reichmann et al., Nature 332: 323-329 (1988); Presta, Curr. Op. Struct. Biol. 2: 593-596 (1992).

As used herein, the term “fully human antibody” refers to an immunoglobulin, such as an antibody, where the whole molecule is of human origin or consists of an amino acid sequence identical to a human form of the antibody. In some embodiments, the antibodies disclosed herein are fully human antibodies. In some embodiments, the antibodies are humanized antibodies.

As used herein, the term “sequence identity” refers to the subunit sequence identity between two polymeric molecules particularly between two amino acid molecules, such as, between two polypeptide molecules. When two amino acid sequences have the same residues at the same positions, then they are identical at that position. For example, if a position in each of two polypeptide molecules is occupied by an Arginine, then the two polypeptides are identical. The identity or extent to which two amino acid sequences have the same residues at the same positions in an alignment is often expressed as a percentage. The identity between two amino acid sequences is a direct function of the number of matching or identical positions. For example, if half (e.g., five positions in a polymer ten amino acids in length) of the positions in two sequences are identical, the two sequences are 50% identical; if 90% of the positions (e.g., 9 of 10), are matched or identical, the two amino acids sequences are 90% identical.

As used herein, the term “immunoglobulin” or “Ig,” defines a class of proteins, which function as antibodies. Antibodies expressed by B cells are sometimes referred to as the BCR (B cell receptor) or antigen receptor. The five members included in this class of proteins are IgA, IgG, IgM, IgD, and IgE. IgA is the primary antibody that is present in body secretions, such as saliva, tears, breast milk, gastrointestinal secretions and mucus secretions of the respiratory and genitourinary tracts. IgG is the most common circulating antibody. IgM is the main immunoglobulin produced in the primary immune response in most subjects. It is the most efficient immunoglobulin in agglutination, complement fixation, and other antibody responses, and is important in defense against bacteria and viruses. IgD is the immunoglobulin that has no known antibody function, but may serve as an antigen receptor. IgE is the immunoglobulin that mediates immediate hypersensitivity by causing release of mediators from mast cells and basophils upon exposure to allergen.

As used herein, the term “immune response” as used herein is defined as a cellular response to an antigen that occurs when lymphocytes identify antigenic molecules as foreign and induce the formation of antibodies and/or activate lymphocytes to remove the antigen.

As used herein, the term “immune effector cell,” refers to a cell that is involved in an immune response, e.g., in the promotion of an immune effector response. Examples of immune effector cells include T cells (e.g., alpha/beta T cells and gamma/delta T cells), B cells, natural killer (NK) cells, natural killer T (NKT) cells, mast cells, and myeloid-derived phagocytes.

As used herein, the term “immune effector function or immune effector response,” refers to a function or response that enhances or promotes an immune attack of a target cell. In some embodiment, an immune effector function or response refers to a property of a T or NK cell that promotes the killing or the inhibition of growth or proliferation, of a target cell. In the case of a T cell, primary stimulation and co-stimulation are examples of immune effector function or response.

As used herein, the term “instructional material” includes a publication, a recording, a diagram, or any other medium of expression which can be used to communicate the usefulness of the compositions and methods of the disclosure. The instructional material of the kit of the invention may, for example, be affixed to a container which contains the nucleic acid, peptide, and/or composition of the disclosure or be shipped together with a container which contains the nucleic acid, peptide, and/or composition. Alternatively, the instructional material may be shipped separately from the container with the intention that the instructional material and the compound be used cooperatively by the recipient.

As used herein, the term “isolated” means altered or removed from the natural state. For example, a nucleic acid or a peptide naturally present in a living animal is not “isolated,” but the same nucleic acid or peptide partially or completely separated from the coexisting materials of its natural state is “isolated.” An isolated nucleic acid or protein can exist in substantially purified form, or can exist in a non-native environment such as, for example, a host cell.

As used herein, the term “lentivirus” refers to a genus of the retroviridae family. Lentiviruses are unique among the retroviruses in being able to infect non-dividing cells; they can deliver a significant amount of genetic information into the DNA of the host cell, so they are one of the most efficient methods of a gene delivery vector. HIV, SIV, and FIV are all examples of lentiviruses. Vectors derived from lentiviruses offer the means to achieve significant levels of gene transfer in vivo.

As used herein, the term “flexible polypeptide linker” or “linker” as used in the context of a scFv refers to a peptide linker that consists of amino acids such as glycine and/or serine residues used alone or in combination, to link variable heavy and variable light chain regions together. In one embodiment, the flexible polypeptide linker is a Gly/Ser linker and comprises the amino acid sequence (Gly-Gly-Gly-Ser)n, where n is a positive integer equal to or greater than 1. For example, n=1, n=2, n=3. n=4, n=5 and n=6, n=7, n=8, n=9 and n=10. In one embodiment, the flexible polypeptide linkers include, but are not limited to, (Gly4 Ser)4 or (Gly4 Ser)3. In another embodiment, the linkers include multiple repeats of (Gly2Ser), (GlySer) or (Gly3Ser).

As used herein, the term “modified” means a changed state or structure of a molecule or cell of the invention. Molecules may be modified in many ways, including chemically, structurally, and functionally. Cells may be modified through the introduction of nucleic acids.

As used herein the term, “monoclonal antibody” refers to an antibody obtained from a population of substantially homogeneous antibodies, i.e., the individual antibodies comprising the population are identical except for possible naturally occurring mutations that may be present in minor amounts. Monoclonal antibodies are highly specific, being directed against a single antigenic site. Furthermore, in contrast to polyclonal antibody preparations that include different antibodies directed against different determinants (epitopes), each monoclonal antibody is directed against a single determinant on the antigen. In addition to their specificity, the monoclonal antibodies are advantageous in that they may be synthesized uncontaminated by other antibodies. The modifier “monoclonal” is not to be construed as requiring production of the antibody by any particular method. For example, the monoclonal antibodies may be prepared by the hybridoma methodology first described by Kohler et al., Nature, 256:495 (1975), or may be made using recombinant DNA methods in bacterial, eukaryotic animal or plant cells (see, e.g., U.S. Pat. No. 4,816,567) after single cell sorting of an antigen specific B cell, an antigen specific plasmablast responding to an infection or immunization, or capture of linked heavy and light chains from single cells in a bulk sorted antigen specific collection. The “monoclonal antibodies” may also be isolated from phage antibody libraries using the techniques described in Clackson et al., Nature, 352:624-628 (1991) and Marks et al., J. Mol. Biol., 222:581-597 (1991), for example.

In the context of the present disclosure, the following abbreviations for the commonly occurring nucleic acid bases are used. “A” refers to adenosine, “C” refers to cytosine, “G” refers to guanosine, “T” refers to thymidine, and “U” refers to uridine.

As used herein, the term “neutralization” refers to the ability of an antibody by itself to inhibit infection of susceptible cells or, in the case of some extracellular organisms, to inhibit an initial pathogenic step. In some embodiments, neutralization of infectivity in vitro means that antibodies are capable of blocking the infectivity or pathogenesis of viruses, bacteria, parasites, and fungi. Neutralization generally occurs as a result of interfering with an organism's attachment to host tissues. Several mechanisms account for the neutralization of a given organism. In addition, a single antibody or antibodies with different specificities can neutralize a given organism, at least in vitro, through multiple mechanisms.

“Neutralization” refers to the function of the antibody disclosed herein. An antibody function refers to the biological effect that the antibody has on a pathogen or its toxin. Additional examples of antibody functions include phagocytosis, antibody-dependent cellular cytotoxicity (ADCC), and complement-mediated lysis of pathogens or of infected cells. See e.g., Forthal, Microbiol Spectr. 2(4): 1-17 (2014).

Unless otherwise specified, a “nucleotide sequence encoding an amino acid sequence” includes all nucleotide sequences that are degenerate versions of each other and that encode the same amino acid sequence. The phrase nucleotide sequence that encodes a protein or an RNA may also include introns to the extent that the nucleotide sequence encoding the protein may in some version contain an intron(s).

As used herein, the term “operably linked” refers to functional linkage between a regulatory sequence and a heterologous nucleic acid sequence resulting in expression of the latter. For example, a first nucleic acid sequence is operably linked with a second nucleic acid sequence when the first nucleic acid sequence is placed in a functional relationship with the second nucleic acid sequence. For instance, a promoter is operably linked to a coding sequence if the promoter affects the transcription or expression of the coding sequence. Generally, operably linked DNA sequences are contiguous and, where necessary to join two protein coding regions, in the same reading frame.

As used herein, the term “parenteral administration” of an immunogenic composition includes, e.g., subcutaneous (s.c.), intravenous (i.v.), intramuscular (i.m.), or intrasternal injection, or infusion techniques.

A “pharmaceutical composition” is intended to include the combination of an active agent with a carrier, inert or active, making the composition suitable for diagnostic or therapeutic use in vitro, in vivo or ex vivo.

“Pharmaceutically acceptable carriers” refers to any diluents, excipients, or carriers that may be used in the compositions disclosed herein. Pharmaceutically acceptable carriers include ion exchangers, alumina, aluminum stearate, lecithin, serum proteins, such as human serum albumin, buffer substances, such as phosphates, glycine, sorbic acid, potassium sorbate, partial glyceride mixtures of saturated vegetable fatty acids, water, salts or electrolytes, such as protamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salts, colloidal silica, magnesium trisilicate, polyvinyl pyrrolidone, cellulose-based substances, polyethylene glycol, sodium carboxymethylcellulose, polyacrylates, waxes, polyethylene-polyoxypropylene-block polymers, polyethylene glycol and wool fat. Suitable pharmaceutical carriers are described in Remington's Pharmaceutical Sciences, Mack Publishing Company, a standard reference text in this field. They may be selected with respect to the intended form of administration, that is, oral tablets, capsules, elixirs, syrups and the like, and consistent with conventional pharmaceutical practices.

The compositions used in accordance with the disclosure can be packaged in dosage unit form for ease of administration and uniformity of dosage. The term “unit dose” or “dosage” refers to physically discrete units suitable for use in a subject, each unit containing a predetermined quantity of the composition calculated to produce the desired responses in association with its administration, i.e., the appropriate route and regimen. The quantity to be administered, both according to number of treatments and unit dose, depends on the result and/or protection desired. Precise amounts of the composition also depend on the judgment of the practitioner and are peculiar to each individual. Factors affecting dose include physical and clinical state of the subject, route of administration, intended goal of treatment (alleviation of symptoms versus cure), and potency, stability, and toxicity of the particular composition. Upon formulation, solutions are administered in a manner compatible with the dosage formulation and in such amount as is therapeutically or prophylactically effective. The formulations are easily administered in a variety of dosage forms, such as the type of injectable solutions described herein.

As used herein, the term “polynucleotide” as used herein is defined as a chain of nucleotides. Furthermore, nucleic acids are polymers of nucleotides. Thus, nucleic acids and polynucleotides as used herein are interchangeable. One skilled in the art has the general knowledge that nucleic acids are polynucleotides, which can be hydrolyzed into the monomeric “nucleotides.” The monomeric nucleotides can be hydrolyzed into nucleosides. As used herein polynucleotides include, but are not limited to, all nucleic acid sequences which are obtained by any means available in the art, including, without limitation, recombinant means, i.e., the cloning of nucleic acid sequences from a recombinant library or a cell genome, using ordinary cloning technology and PCR™, and the like, and by synthetic means.

As used herein, the terms “peptide,” “polypeptide,” and “protein” are used interchangeably, and refer to a compound comprised of amino acid residues covalently linked by peptide bonds. A protein or peptide must contain at least two amino acids, and no limitation is placed on the maximum number of amino acids that can comprise a protein's or peptide's sequence. Polypeptides include any peptide or protein comprising two or more amino acids joined to each other by peptide bonds. As used herein, the term refers to both short chains, which also commonly are referred to in the art as peptides, oligopeptides and oligomers, for example, and to longer chains, which generally are referred to in the art as proteins, of which there are many types. “Polypeptides” include, for example, biologically active fragments, substantially homologous polypeptides, oligopeptides, homodimers, heterodimers, variants of polypeptides, modified polypeptides, derivatives, analogs, fusion proteins, among others. The polypeptides include natural peptides, recombinant peptides, synthetic peptides, or a combination thereof.

As used herein, the term “promoter” is defined as a DNA sequence recognized by the synthetic machinery of the cell, or introduced synthetic machinery, required to initiate the specific transcription of a polynucleotide sequence.

As used herein, the term “promoter/regulatory sequence” means a nucleic acid sequence which is required for expression of a gene product operably linked to the promoter/regulatory sequence. In some instances, this sequence may be the core promoter sequence and in other instances, this sequence may also include an enhancer sequence and other regulatory elements which are required for expression of the gene product. The promoter/regulatory sequence may, for example, be one which expresses the gene product in a tissue specific manner.

As used herein, the term “sample” and “biological sample” and “agricultural sample” are used interchangeably, referring to sample material derived from a subject. Biological samples may include tissues, cells, protein or membrane extracts of cells, and biological fluids (e.g., ascites fluid or cerebrospinal fluid (CSF)) isolated from a subject, as well as tissues, cells and fluids present within a subject. Biological samples may include, but are not limited to, samples taken from breast tissue, renal tissue, the uterine cervix, the endometrium, the head or neck, the gallbladder, parotid tissue, the prostate, the brain, the pituitary gland, kidney tissue, muscle, the esophagus, the stomach, the small intestine, the colon, the liver, the spleen, the pancreas, thyroid tissue, heart tissue, lung tissue, the bladder, adipose tissue, lymph node tissue, the uterus, ovarian tissue, adrenal tissue, testis tissue, the tonsils, thymus, blood, hair, buccal, skin, serum, plasma, CSF, semen, prostate fluid, seminal fluid, urine, feces, sweat, saliva, sputum, mucus, bone marrow, lymph, and tears. Agricultural samples include soil, foliage or any plant tissue or surface or other sample suspected of harboring virus. In addition, the sample can include industrial samples, such as those isolated from surfaces and the environment.

In some embodiments, the sample may be an upper respiratory specimen, such as a nasopharyngeal (NP) specimen, an oropharyngeal (OP) specimen, a nasal mid-turbinate swab, an anterior nares (nasal swab) specimen, or nasopharyngeal wash/aspirate or nasal wash/aspirate (NW) specimen.

In some embodiments, the samples include fluid from a subject, including, without limitation, blood or a blood product (e.g., serum, plasma, or the like), umbilical cord blood, amniotic fluid, cerebrospinal fluid, spinal fluid, lavage fluid (e.g., bronchoalveolar, gastric, peritoneal, ductal, ear, arthroscopic), washings of female reproductive tract, urine, feces, sputum, saliva, nasal mucous, prostate fluid, lavage, semen, lymphatic fluid, bile, tears, sweat, breast milk, breast fluid, the like or combinations thereof. In some embodiments, a liquid biological sample is a blood plasma or serum sample. The term “blood” as used herein refers to a blood sample or preparation from a subject. The term encompasses whole blood, blood product or any fraction of blood, such as serum, plasma, buffy coat, or the like as conventionally defined. In some embodiments, the term “blood” refers to peripheral blood. Blood plasma refers to the fraction of whole blood resulting from centrifugation of blood treated with anticoagulants. Blood serum refers to the watery portion of fluid remaining after a blood sample has coagulated. Fluid samples often are collected in accordance with standard protocols hospitals or clinics generally follow. For blood, an appropriate amount of peripheral blood (e.g., between 3-40 milliliters) often is collected and can be stored according to standard procedures prior to or after preparation.

As used herein, the term “Sendai virus” refers to a genus of the Paramyxoviridae family. Sendai viruses are negative, single stranded RNA viruses that do not integrate into the host genome or alter the genetic information of the host cell. Sendai viruses have an exceptionally broad host range and are not pathogenic to humans. Used as a recombinant viral vector, Sendai viruses are capable of transient but strong gene expression.

As used herein, the term “single chain antibodies” refer to antibodies formed by recombinant DNA techniques in which immunoglobulin heavy and light chain fragments are linked to the Fv region via an engineered span of amino acids. Various methods of generating single chain antibodies are known, including those described in U.S. Pat. No. 4,694,778; Bird, Science 242:423-442 (1988); Huston et al., Proc. Natl. Acad. Sci. USA 85:5879-5883 (1988); Ward et al., Nature 334:54454 (1989); Skerra et al., Science 242:1038-1041 (1988).

As used herein, the term “specificity” refers to the ability to specifically bind (e.g., immunoreact with) a given target antigen (e.g., a human target antigen). A chimeric antigen receptor may be monospecific and contain one or more binding sites, which specifically bind a target or a chimeric antigen receptor may be multi-specific and contain two or more binding sites which specifically bind the same or different targets. In certain embodiments, a chimeric antigen receptor is specific for two different (e.g., non-overlapping) portions of the same target. In certain embodiments, a chimeric antigen receptor is specific for more than one target.

As used herein, the term “specifically binds,” with respect to an antibody, means an antibody or binding fragment thereof (e.g., Fv fragment or scFv) which recognizes a specific antigen, but does not substantially recognize or bind other molecules in a sample. For example, an antibody that specifically binds to an antigen from one species may also bind to that antigen from one or more species. But, such cross-species reactivity does not itself alter the classification of an antibody as specific. In another example, an antibody that specifically binds to an antigen may also bind to different allelic forms of the antigen. However, such cross reactivity does not itself alter the classification of an antibody as specific. In some instances, the terms “specific binding” or “specifically binding,” can be used in reference to the interaction of an antibody, a protein, a chimeric antigen receptor, or a peptide with a second chemical species, to mean that the interaction is dependent upon the presence of a particular structure (e.g., an antigenic determinant or epitope) on the chemical species; for example, a chimeric antigen receptor recognizes and binds to a specific protein structure rather than to proteins generally. If an antibody is specific for epitope “A,” the presence of a molecule containing epitope A (or free, unlabeled A), in a reaction containing labeled “A” and the antibody, will reduce the amount of labeled A bound to the antibody.

As used herein, the terms “subject” and “patient” are used interchangeably. As used herein, a subject is can be a mammal, such as a non-primate (e.g., cows, pigs, horses, cats, dogs, rats, etc.) or a primate (e.g., monkey and human). In certain embodiments, the term “subject,” as used herein, refers to a vertebrate, such as a mammal. Mammals include, without limitation, humans, non-human primates, wild animals, feral animals, farm animals, sport animals, and pets. Any living organism in which an immune response can be elicited may be a subject or patient. In certain exemplary embodiments, a subject is a human.

As used herein to “treat” a disease as the term is used herein, means to reduce the frequency or severity of at least one sign or symptom of a disease or disorder experienced by a subject. In one aspect, the term “treat” excludes propylaxis.

As used herein, “treating” or “treatment” of a disease in a subject refers to (1) preventing the symptoms or disease from occurring in a subject that is predisposed or does not yet display symptoms of the disease; (2) inhibiting the disease or arresting its development; or (3) ameliorating or causing regression of the disease or the symptoms of the disease. As understood in the art, “treatment” is an approach for obtaining beneficial or desired results, including clinical results. For the purposes of the present technology, beneficial or desired results can include one or more, but are not limited to, alleviation or amelioration of one or more symptoms, diminishment of extent of a condition (including a disease), stabilized (i.e., not worsening) state of a condition (including disease), delay or slowing of condition (including disease), progression, amelioration or palliation of the condition (including disease), states and remission (whether partial or total), whether detectable or undetectable.

As used herein, the term “therapy” refers to any protocol, method and/or agent (e.g., prophylactic, therapeutic, adjuvant or vaccine) that can be used in the prevention, management, treatment and/or amelioration of a disease or a symptom related thereto.

As used herein, the term “vector” is a composition of matter that comprises an isolated nucleic acid and which can be used to deliver the isolated nucleic acid to the interior of a cell. Numerous vectors are known in the art including, but not limited to, linear polynucleotides, polynucleotides associated with ionic or amphiphilic compounds, plasmids, and viruses. Thus, the term “vector” includes an autonomously replicating plasmid or a virus. The term should also be construed to include non-plasmid and non-viral compounds which facilitate transfer of nucleic acid into cells, such as, for example, polylysine compounds, liposomes, and the like. Examples of viral vectors include, but are not limited to, Sendai viral vectors, adenoviral vectors, adeno-associated virus vectors, retroviral vectors, lentiviral vectors, and the like.

Ranges: throughout this disclosure, various aspects of the invention can be presented in a range format. It should be understood that the description in range format is merely for convenience and brevity and should not be construed as an inflexible limitation on the scope of the invention. Accordingly, the description of a range should be considered to have specifically disclosed all the possible subranges as well as individual numerical values within that range. For example, description of a range such as from 1 to 6 should be considered to have specifically disclosed subranges such as from 1 to 3, from 1 to 4, from 1 to 5, from 2 to 4, from 2 to 6, from 3 to 6 etc., as well as individual numbers within that range, for example, 1, 2, 2.7, 3, 4, 5, 5.3, and 6. This applies regardless of the breadth of the range.

Isolated Antibodies

One aspect of the present disclosure provides an isolated antibody or antibody fragment thereof that binds to a SARS-CoV-2 surface protein. In one aspect, the isolated antibody or fragment thereof has improved binding and/or neutralization characteristics against a broad range of SARS-CoV-2 strains when compared to a monoclonal antibody selected from sotrovimab (S309), COV2-2381, cilgavimab (COV2-2130 or AZD1061), and/or tixagevimab (COV2-2196 or AZD8895). In some embodiments, the SARS-CoV-2 surface protein is a spike (S) glycoprotein.

Coronaviruses

Coronaviruses constitute the subfamily Orthocoronavirinae, in the family Coronaviridae, order Nidovirales, and realm Riboviria. They are enveloped viruses with a positive-sense single-stranded RNA genome and a nucleocapsid of helical symmetry. The genome size of coronaviruses ranges from approximately 26 to 32 kilobases, one of the largest among RNA viruses. They have characteristic club-shaped spikes that project from their surface, which in electron micrographs create an image reminiscent of the solar corona, from which their name derives.

In some embodiments, the coronavirus as used herein refers to a severe acute respiratory syndrome (SARS) associated coronavirus (SARS-CoV). In some embodiments, the coronavirus is either or both of SARS-CoV-1 and SARS-CoV-2. In some embodiments, the coronavirus comprises a virus selected from the group consisting of an Alphacoronavirus; a Colacovirus such as Bat coronavirus CDPHE15; a Decacovirus such as Bat coronavirus HKU10 or Rhinolophus ferrumequinum alphacoronavirus HuB-2013; a Duvinacovirus such as Human coronavirus 229E; a Luchacovirus such as Lucheng Rn rat coronavirus; a Minacovirus such as a Ferret coronavirus or Mink coronavirus 1; a Minunacovirus such as Miniopterus bat coronavirus 1 or Miniopterus bat coronavirus HKU8; a Myotacovirus such as Myotis ricketti alphacoronavirus Sax-2011; a nyctacovirus such as Nyctalus velutinus alphacoronavirus SC-2013; a Pedacovirus such as Porcine epidemic diarrhea virus or Scotophilus bat coronavirus 512; a Rhinacovirus such as Rhinolophus bat coronavirus HKU2; a Setracovirus such as Human coronavirus NL63 or NL63-related bat coronavirus strain BtKYNL63-9b; a Tegacovirus such as Alphacoronavirus 1; a Betacoronavirus; a Embecovirus such as Betacoronavirus 1, Human coronavirus OC43, China Rattus coronavirus HKU24, Human coronavirus HKU1 or Murine coronavirus; a Hibecovirus such as Bat Hp-betacoronavirus Zhejiang2013; a Merbecovirus such as Hedgehog coronavirus 1, Middle East respiratory syndrome-related coronavirus (MERS-CoV), Pipistrellus bat coronavirus HKU5 or Tylonycteris bat coronavirus HKU4; a Nobecovirus such as Rousettus bat coronavirus GCCDC1 or Rousettus bat coronavirus HKU9, a Sarbecovirus such as a Severe acute respiratory syndrome-related coronavirus, Severe acute respiratory syndrome coronavirus (SARS-CoV) or Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2, COVID-19); a Deltacoronavirus; an Andecovirus such as Wigeon coronavirus HKU20; a Buldecovirus such as Bulbul coronavirus HKU11, Porcine coronavirus HKU15, Munia coronavirus HKU13 or White-eye coronavirus HKU16; a Herdecovirus such as Night Heron coronavirus HKU19; a Moordecovirus such as Common moorhen coronavirus HKU21; a Gammacoronavirus; a Cegacovirus such as Beluga Whale coronavirus SW1; and an Igacovirus such as Avian coronavirus.

Symptoms of a coronavirus infection include, but are not limited to, mild symptoms, such as fatigues, tingling, tingling or numbness in the hands and feet, dizziness, confusion, brain fog, body ache, chills, loss of appetite, nausea, vomiting, abdominal pain or discomfort, loss of smell, inability to taste, muscle weakness, photophobia, adenopathy, headaches, cough, dry cough, shortness of breath, sore throat, lower extremity weakness/numbness, diarrhea, low blood O2, sneezing, runny nose or post-nasal drip; severe symptoms, such as ventilatory use, high fever, severe cough, delirium, seizures, stroke, systematic inflammation, cytokine storm; and other symptoms, such as fever, swollen adenoids, pneumonia, bronchitis, and Dyspnea.

SARS-CoV-2

SARS-CoV-2 is a contagious virus that causes the acute respiratory disease designated coronavirus disease 2019 (COVID-19). All viruses, including SARS-CoV-2, change over time. Most changes have little to no impact on the virus' properties. However, some changes may affect the virus's properties, such as how easily it spreads, the associated disease severity, or the performance of vaccines, therapeutic medicines, diagnostic tools, or other public health and social measures. During the ongoing COVID-19 outbreak (December 2019 to present), SARS-CoV-2 has continuously evolved into more dangerous strains. During late 2020, the emergence of variants that posed an increased risk to global public health prompted the characterization of specific Variants of Interest (VOIs) and Variants of Concern (VOCs). See e.g., Center for Disease Control and Prevention, cdc.gov/coronavirus/2019-ncov/variants/variant-classifications.html#anchor_1632154493691 (Updated Apr. 26, 2022); World Health Organization, who.int/en/activities/tracking-SARS-CoV-2-variants (updated on 25 Apr. 2022).

A SARS-CoV-2 variant that meets the definition of a VOI (see below) and, through a comparative assessment, has been demonstrated to be associated with one or more of the following changes at a degree of global public health significance: (1) increase in transmissibility or detrimental change in COVID-19 epidemiology; (2) increase in virulence or change in clinical disease presentation; and/or decrease in effectiveness of public health and social measures or available diagnostics, vaccines, therapeutics. Variants that do not otherwise meet all these criteria may be designated as VOCs/VOIs/VUMs, and those posing a diminishing risk relative to other circulating variants may be reclassified.

As of On Apr. 14, 2022, the U.S. government SARS-CoV-2 Interagency Group (SIG) recognizes the following defined four classes of SARS-CoV-2 variants: (1) Variant Being Monitored (VBM); Variant of Interest (VOI); Variant of Concern (VOC); and Variant of High Consequence (VOHC). As of that date, no SARS-CoV-2 variants are designated as VOI or VOHC. See e.g., Center for Disease Control and Prevention, cdc.gov/coronavirus/2019-ncov/variants/variant-classifications.html#anchor_1632154493691 (Updated Apr. 26, 2022).

The Variant Being Monitored (VBM) are selected from Alpha (B.1.1.7 and Q lineages), Beta (B.1.351 and descendent lineages), Gamma (P.1 and descendent lineages), Delta (B.1.617.2 and AY lineages), Epsilon (B.1.427 and B.1.429), Eta (B.1.525), Iota (B.1.526), Kappa (B.1.617.1), 1.617.3, Mu (B.1.621, B.1.621.1), Zeta (P.2). See e.g., Center for Disease Control and Prevention, cdc.gov/coronavirus/2019-ncov/variants/variant-classifications.html#anchor_1632154493691 (Updated Apr. 26, 2022).

The Variant of Concern (VOC) is Omicron (B.1.1.529, BA.1, BA.1.1, BA.2, BA.3, BA.4 and BA.5 lineages). The Omicron variant, which was first identified in South Africa shows evidence of an increase in transmissibility, more severe disease (for example, increased hospitalizations or deaths), significant reduction in neutralization by antibodies generated during previous infection or vaccination, reduced effectiveness of treatments or vaccines, or diagnostic detection failures.

The Omicron variant and its lineages comprise mutations selected from deletions, insertions and substitutions in the SARS-CoV-2's Spike Protein. The substitutions are selected from A67V, T95I, Y145D, L212I, G339D, S371L, S373P, S375F, K417N, N440K, G4465, S477N, T478K, E484A, Q493R, G496S, Q498R, N501Y, Y505H, T547K, D614G, H655Y, N679K, P681H, N764K, D796Y, N856K, Q954H, N969K, and/or L981F. The deletions are selected from de169-70, de1142-144, and/or de1211; and the insertion is ins214EPE. See e.g., Center for Disease Control and Prevention, cdc.gov/coronavirus/2019-ncov/variants/variant-classifications.html#anchor_1632154493691 (Updated Apr. 26, 2022)

Given the continuous evolution of the virus that leads to SARS-CoV-2 and the constant developments in our understanding of the impacts of variants, there is a need for novel prophylactic monoclonal antibodies that can bind to and neutralize old and emerging strains of SARS-CoV-2, such as the Omicron variants.

In some embodiments, the antibody or antibody fragment disclosed herein binds a SARS-CoV-2 variant selected from Alpha (B.1.1.7 and Q lineages); Beta (B.1.351 and descendent lineages); Gamma (P.1 and descendent lineages); Delta (B.1.617.2 and AY lineages); Epsilon (B.1.427 and B.1.429); Eta (B.1.525); Iota (B.1.526); Kappa (B.1.617.1); 1.617.3; Mu (B.1.621, B.1.621.1); Zeta (P.2); or Omicron (B.1.1.529, BA.1, BA.1.1, BA.2, BA.3, BA.4 and BA.5 lineages). In some embodiments, the monoclonal antibody or antibody fragment binds to and neutralizes SARS-CoV-2 Omicron (B.1.1.529, BA.1, BA.1.1, BA.2, BA.3, BA.4 and BA.5 lineages).

In some embodiments, the antibody or antibody fragment disclosed herein binds a SARS-CoV-2 variant comprising a mutation in the spike (S) glycoprotein. In one embodiment, the mutation is a substitution selected from D614G, G339D, S371L, S373P, S375F, K417N, N440K, G4465, S477N, T478K, E484A, Q493R, G496S, Q498R, N501Y, Y505H, T547K, H655Y, N764K, D796Y, N856K, Q954H, N969K, L981F, or a combination thereof. In one embodiment, the substitution is selected from A67V, T95I, Y145D, L212I, G339D, S371L, S373P, S375F, K417N, N440K, G4465, S477N, T478K, E484A, Q493R, G496S, Q498R, N501Y, Y505H, T547K, D614G, H655Y, N679K, P681H, N764K, D796Y, N856K, Q954H, N969K, and/or L981F. In some embodiments, the SARS-CoV-2 variant comprises a D614G substitution. In some embodiments, the mutation comprises deletion of amino acids 143-145 (143-145del). In some embodiments, the mutation is in the receptor-binding domain (RBD) or the N-terminal domain of the spike glycoprotein.

In some embodiments, the antibody or antibody fragment disclosed herein is capable of neutralizing from about 80 to about 100% of the SARS-CoV-2 virus with a concentration of about 0.5 ng/ml to about 0.01 ng/ml, 0.75 ng/ml to about 0.01 ng/ml, 1 ng/ml to about 0.01 ng/ml, 1.6 ng/ml to about 0.01 ng/ml, 2 ng/ml to about 0.01 ng/ml, 5 ng/ml to about 0.01 ng/ml, 8 ng/ml to about 0.01 ng/ml, 10 ng/ml to about 0.01 ng/ml, 12 ng/ml to about 0.01 ng/ml, 15 ng/ml to about 0.01 ng/ml, 16 ng/ml to about 0.01 ng/ml, about 20 ng/ml to about 0.01 ng/ml, about 30 ng/ml to about 0.01 ng/ml, about 40 ng/ml to about 0.01 ng/ml, about 50 ng/ml to about 0.01 ng/ml, about 75 ng/ml to about 0.01 ng/ml, about 100 ng/ml to about 0.01 ng/ml, about 125 ng/ml to about 0.01 ng/ml, 150 ng/ml to about 0.01 ng/ml, about 175 ng/ml to about 0.01 ng/ml, about 190 ng/ml to about 0.01 ng/ml, about 200 ng/ml to about 0.01 ng/ml, about 220 ng/ml to about 0.01 ng/ml, about 235 ng/ml to about 0.01 ng/ml, about 240 ng/ml to about 0.01 ng/ml, or about 250 ng/ml to about 0.01 ng/ml.

In some embodiments, the antibody or antibody fragment disclosed herein is capable of neutralizing from about 80 to about 100% of the SARS-CoV-2 virus with a concentration from about 0.01 ng/ml to about 1.0 ng/ml; from about 0.5 ng/ml to about 1.5 ng/ml; from about 1 ng/ml to about 2.0 ng/ml; from about 0.01 ng/ml to about 1.9 ng/ml; from about 1 ng/ml to about 5.0 ng/ml; or from about 5.0 ng/ml to about 10.0 ng/ml.

In some embodiments, the antibody or antibody fragment inhibits or blocks the binding of the SARS-CoV-2 virus to a human receptor angiotensin converting enzyme 2 (hACE2). In some embodiments, the antibody or antibody fragment inhibits or blocks the binding to hACE2 with an IC₅₀of about 0.1 ng/ml, 0.5 ng/ml to about 0.1 ng/ml, 0.8 ng/ml to about 0.1 ng/ml, 1 ng/ml to about 0.1 ng/ml, 2 ng/ml to about 0.1 ng/ml, 5 ng/ml to about 0.1 ng/ml, 10 ng/ml to about 0.1 ng/ml, 15 ng/ml to about 0.1 ng/ml, about 20 ng/ml to about 0.1 ng/ml, about 30 ng/ml to about 0.1 ng/ml, about 40 ng/ml to about 0.1 ng/ml, about 50 ng/ml to about 0.1 ng/ml, about 75 ng/ml to about 0.1 ng/ml, about 100 ng/ml to about 0.1 ng/ml, about 110 ng/ml to about 0.1 ng/ml, about 120 ng/ml to about 0.1 ng/ml, about 130 ng/ml to about 0.1 ng/ml, about 140 ng/ml to about 0.1 ng/ml, or about 150 ng/ml to about 0.1 ng/ml, as measured by Biacore™ assay systems, or any methods known to those of skill in the art. See e.g., Casper et al., Anal Biochem, 325(1):126-36 (2004); Jason-Moller et al., Curr Protoc Protein Sci, Chapter 19: Unit 19.13 (2006); giffordbioscience.com/spr-biacore-services/; Zost et al., Nature 584: 443-449 (2020). In some embodiments, the antibody or antibody fragment reduces (e.g., blocks) S protein binding to a cell that expresses hACE2 with an IC₅₀of less than about 1.5 nM, or about 1 nM, or about 0.8 nM, or about 0.6 nM, or about 0.4 nM, or about 0.2 nM, or about 0.1 nM, e.g., between about 0.2 nM and about 0.1 nM, e.g., about 0.15 nM to about 0.1 nM, e.g., about 0.145 nM. In some embodiments, the antibody or antibody fragment reduces (e.g., blocks) Spike protein binding to a cell that expresses hACE2 with an IC₅₀of less than about 2 nM, or less than about 1.5 nM, or less than about 1 nM, or less than about 0.5 nM, or less than about 0.2 nM, e.g., between about 0.5 nM and about 0.01 nM, or about 0.2 nM to 0.05 nM, e.g., about 0.1 nM.

In some embodiments, the antibody or antibody fragment binds to a target antigen (e.g., SARS-CoV-2 spike protein receptor binding domain) with a binding affinity (K_D) of about 0.01 nM to about 0.1 nM, about 0.03 nM to about 0.1 nM, about 0.05 nM to about 0.1 nM, about 0.07 nM to about 0.1 nM, about 0.1 nM to about 0.5 nM, about 0.25 nM to about 0.5 nM, about 0.5 nM to about 0.5 nM, about 0.75 nM to about 0.5 nM, about 1.0 nM to about 0.5 nM, about 1.25 nM to about 0.5 nM, about 1.5 nM to about 0.5 nM, about 1.75 nM to about 0.5 nM, about 2.0 nM to about 0.5 nM, about 2.5 nM to about 0.5 nM, from about 10 nM to about 120 nM, from 30 nM to about 120 nM, from 50 nM to about 120 nM, from about 60 nM to about 120 nM, from about 80 nM to about 105 nM, from about 63 nM to about 85 nM, or from about 50 nM to about 85 nM, as measured by Biacore™ assay systems or any methods known to those of skill in the art. See e.g., Casper et al., Anal Biochem, 325(1):126-36 (2004); Jason-Moller et al., Curr Protoc Protein Sci, Chapter 19: Unit 19.13 (2006); giffordbioscience.com/spr-biacore-services/; Zost et al., Nature 584: 443-449 (2020).

In some embodiments, the antibody or antibody fragment binds to a target antigen (e.g., SARS-CoV-2 spike protein receptor binding domain) with a K_Dslower than 5×10⁻⁴, 1×10⁻⁴, 5×10⁻⁵, 1×10⁻⁵, 5×10⁻⁶or 1×10⁻⁶s⁻¹, e.g., about 6.33×10⁻⁵s⁻¹, as measured by Biacore™ assay systems. In some embodiments, the antibody or antibody fragment binds to a target antigen (e.g., hACE2) with a KA faster than 1×10⁴, 5×10⁴, 1×10⁵, 5×10⁵or 1×10⁶M⁻¹s⁻¹, e.g., about 3.07×10⁴M⁻¹s⁻¹, as measured by Biacore™ assay systems, or any methods known to those of skill in the art. See e.g., Casper et al., Anal Biochem, 325(1):126-36 (2004); Jason-Moller et al., Curr Protoc Protein Sci, Chapter 19: Unit 19.13 (2006); giffordbioscience.com/spr-biacore-services/; Zost et al., Nature 584: 443-449 (2020).

In some embodiments, the antibody or antibody fragment (e.g., an isolated or recombinant antibody molecule) binds to a trimeric spike protein ectodomain and/or spike protein receptor binding domain (RBD) with high affinity. For example, the antibody or antibody fragment (e.g., an isolated or recombinant antibody molecule) binds to a trimeric spike protein ectodomain and/or spike protein receptor binding domain (RBD) with an affinity constant of at least about 10⁷M⁻¹, typically about 10⁸M⁻¹, and more typically, about 10⁹M⁻¹to 10¹⁰M⁻¹or stronger. In some embodiments, the antibody or antibody fragment binds specifically to an epitope on hACE2. In some embodiments, the antibody or antibody fragment binds specifically to the same or similar epitope as the epitope recognized by the monoclonal antibody COV2-2130 or a chimeric antibody thereof. In some embodiments, the antibody or antibody fragment binds specifically to the same or similar epitope as the epitope recognized by the monoclonal antibody COV2-2381 or a chimeric antibody thereof. In some embodiments, the antibody or antibody fragment binds specifically to the same or similar epitope as the epitope recognized by the monoclonal antibody COV2-2196 or a chimeric antibody thereof. In some embodiments, the antibody or antibody fragment binds to non-overlapping epitopes on the receptor-binding domain of SARS-CoV-2 spike protein when compared to the epitope bound by the monoclonal antibody or antibody fragment selected from sotrovimab (S309), COV2-2381, cilgavimab (COV2-2130 or AZD1061), or tixagevimab (COV2-2196 or AZD8895).

In some embodiments, the antibody or antibody fragment binds: a trimeric spike protein ectodomain, and/or a monomeric spike protein binding domain (RBD). In some embodiments, the binding affinity of the antibody or antibody fragment to the trimeric spike protein ectodomain and/or the monomeric spike protein RBD has an EC₅₀of about 1.0 pg/ml to 0.1 ng/ml, 1.0 pg/ml to 0.25 ng/ml, 1.0 pg/ml to 0.75 ng/ml, 1.0 pg/ml to 1.0 ng/ml, 1.0 pg/ml to 1.25 ng/ml, 1.0 pg/ml to 1.50 ng/ml, 1.0 pg/ml to 1.75 ng/ml, 1.0 pg/ml to about 2 ng/mL, 1.0 pg/ml to about 2.5 ng/mL, 1.0 pg/ml to about 3 ng/mL, 1.0 pg/ml to about 3.5 ng/mL, 1.0 pg/ml to about 4.0 ng/mL, 1.0 pg/ml to about 4.5 ng/mL, 1.0 pg/ml to about 5 ng/mL, or 1.0 pg/ml to about 10 ng/mL.

In some embodiments, the antibody or antibody fragment comprises a heavy chain complementary determining region 1 (CDRH1) comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 196, a CDRH2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 197, and a CDRH3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 202, and a light chain complementary determining region 1 (CDRL1) comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 226, a CDRL2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 242 or 243, and a CDRL3 comprising the amino acid sequence of SEQ ID NO: 251.

In some embodiments, the antibody or antibody fragment comprises a CDRH1 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 196, a CDRH2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 198, and a CDRH3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 203, and a CDRL1 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 227, a CDRL2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 242 or 243, and a CDRL3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 251.

In some embodiments, the antibody or antibody fragment comprises, or consisting essentially of, or consisting of a CDRH1 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 196, a CDRH2 comprising or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 199, and a CDRH3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 204, and a CDRL1 comprising or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 227, a CDRL2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 242 or 243, and a CDRL3 comprising or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 251.

In some embodiments, the antibody or antibody fragment comprises a CDRH1 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 196, a CDRH2 comprising or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 198, and a CDRH3 comprising or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 204, and a CDRL1 comprising or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 228, a CDRL2 comprising or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 242 or 243, and a CDRL3 comprising or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 251.

In some embodiments, the antibody or antibody fragment comprises a CDRH1 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 196, a CDRH2 comprising or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 198, and a CDRH3 comprising or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 204, and a CDRL1 comprising or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 229, a CDRL2 comprising or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 242 or 243, and a CDRL3 comprising or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 251.

In some embodiments, the antibody or antibody fragment comprises a CDRH1 comprising or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 196, a CDRH2 comprising or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 198, and a CDRH3 comprising or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 204, and a CDRL1 comprising or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 230, a CDRL2 comprising or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 242 or 244, and a CDRL3 comprising or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 251.

In some embodiments, the antibody or antibody fragment comprises a CDRH1 comprising or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 196, a CDRH2 comprising or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 198, and a CDRH3 comprising or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 204, and a CDRL1 comprising or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 231, a CDRL2 comprising or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 242 or 245, and a CDRL3 comprising or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 251.

In some embodiments, the antibody or antibody fragment comprises or consists essentially of, or consists of a CDRH1 comprising or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 196, a CDRH2 comprising or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 200, and a CDRH3 comprising or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 204, and a CDRL1 comprising or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO:232, a CDRL2 comprising or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 242 or 245, and a CDRL3 comprising or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 251.

In some embodiments, the antibody or antibody fragment comprises a CDRH1 comprising or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 196, a CDRH2 comprising or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 198, and a CDRH3 comprising or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 205, and a CDRL1 comprising or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 233, a CDRL2 comprising or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 242 or 243, and a CDRL3 comprising or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 251.

In some embodiments, the antibody or antibody fragment comprises a CDRH1 comprising or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 196, a CDRH2 comprising or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 198, and a CDRH3 comprising or consisting essentially of, or consisting of the amino acid sequence of S SEQ ID NO: 205, and a CDRL1 comprising or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 234, a CDRL2 comprising or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 242 or 245, and a CDRL3 comprising or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 251.

In some embodiments, the antibody or antibody fragment comprises a CDRH1 comprising or consisting essentially of, or consisting of g the amino acid sequence of SEQ ID NO: 196, a CDRH2 comprising or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 198, and a CDRH3 comprising or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 206, and a CDRL1 comprising or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 235, a CDRL2 comprising or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 242 or 246, and a CDRL3 comprising or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 251.

In some embodiments, the antibody or antibody fragment comprises a CDRH1 comprising or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 196, a CDRH2 comprising or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 197, and a CDRH3 comprising or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 207, and a CDRL1 comprising or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 236, a CDRL2 comprising or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 242 or 243, and a CDRL3 comprising or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 251.

In some embodiments, the antibody or antibody fragment comprises a CDRH1 comprising or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 196, a CDRH2 comprising or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 201, and a CDRH3 comprising or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 208, and a CDRL1 comprising or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 228, a CDRL2 comprising or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 242 or 247, and a CDRL3 comprising or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 251.

In some embodiments, the antibody or antibody fragment comprises a CDRH1 comprising or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 196, a CDRH2 comprising or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 198, and a CDRH3 comprising or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 209, and a CDRL1 comprising or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 234, a CDRL2 comprising or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 242 or 245, and a CDRL3 comprising or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 251.

In some embodiments, the CDRs of the heavy chain variable domain or the light chain variable domain of the antibody or antibody fragment disclosed herein comprises the amino acid sequence disclosed in Tables 1, 2, or 3.

In some embodiments, the antibody or antibody fragment disclosed herein comprises: a heavy chain variable sequence having at least 70%, 80%, 90%, 95%, 96%, 97%, 98%, or 99% identity to the amino acid sequence of SEQ ID NO: 100, 101, 102, 103, 104, 105, 106, 107, 108, or 109; and/or a light chain variable sequences having at least 70%, 80%, 90%, 95%, 96%, 97%, 98%, or 99% identity to the amino acid sequence of SEQ ID NO: 155, 156, 157, 158, 159, 160, 161, 162, 163, 164, 165, or 166. In one embodiment, the CDRs of the heavy chain variable sequence or the light chain variable sequences are 100% identical to a CDR disclosed in Tables 1-4.

In some embodiments, the antibody or antibody fragment disclosed herein comprises: a heavy chain variable sequence having at least 70%, 80%, 90%, 95%, 96%, 97%, 98%, or 99% identity to the amino acid sequence disclosed in Table 4; and/or a light chain variable sequences having at least 70%, 80%, 90%, 95%, 96%, 97%, 98%, or 99% identity to the amino acid sequence disclosed in Table 5. In some embodiments, the CDRs of the heavy chain variable sequence or the light chain variable sequences are 100% identical to a CDR disclosed in Tables 1, 2, or 3.

In some embodiments, the antibody or antibody fragment comprises a CDRH1 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 196, a CDRH2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 198, and a CDRH3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 204, and a CDRL1 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 227, a CDRL2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 242 or 245, and a CDRL3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 251.

In some embodiments, the antibody or antibody fragment comprises a CDRH1 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 196, a CDRH2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 198, and a CDRH3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 204, and a CDRL1 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 227, a CDRL2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 242 or 244, and a CDRL3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 251.

In some embodiments, the antibody or antibody fragment comprises a CDRH1 comprising or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 196, a CDRH2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 197, and a CDRH3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 204, and a CDRL1 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 227, a CDRL2 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 242 or 243, and a CDRL3 comprising, or consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 251.