WO2022035918A1 - Anti-sars-cov-2 antibodies - Google Patents

Anti-sars-cov-2 antibodies Download PDF

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Publication number
WO2022035918A1
WO2022035918A1 PCT/US2021/045466 US2021045466W WO2022035918A1 WO 2022035918 A1 WO2022035918 A1 WO 2022035918A1 US 2021045466 W US2021045466 W US 2021045466W WO 2022035918 A1 WO2022035918 A1 WO 2022035918A1
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WIPO (PCT)
Prior art keywords
seq
antibody
nucleic acid
cov
sars
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PCT/US2021/045466
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French (fr)
Inventor
Barrett Allan
Arezoo BIDSHAHRI
Thomas Peter CUJEC
Stephen John DEMAREST
Daniel Scott GIRARD
Donmienne Doen Mun Leung
Jingxing Li
Bo Ma
Aoise O'NEILL
Ying Tang
Ping XIANG
Stefanie ŽENTELIS
Valentine Julie Layla BERTRAND DE PUYRAIMOND
Original Assignee
Eli Lilly And Company
Abcellera Biologics Inc.
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Publication of WO2022035918A1 publication Critical patent/WO2022035918A1/en

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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
    • C07K16/08—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from viruses
    • C07K16/10—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from viruses from RNA viruses
    • C07K16/1002—Coronaviridae
    • C07K16/1003—Severe acute respiratory syndrome coronavirus 2 [SARS‐CoV‐2 or Covid-19]
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/12—Antivirals
    • A61P31/14—Antivirals for RNA viruses
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/20—Immunoglobulins specific features characterized by taxonomic origin
    • C07K2317/21—Immunoglobulins specific features characterized by taxonomic origin from primates, e.g. man
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/50—Immunoglobulins specific features characterized by immunoglobulin fragments
    • C07K2317/56—Immunoglobulins specific features characterized by immunoglobulin fragments variable (Fv) region, i.e. VH and/or VL
    • C07K2317/565—Complementarity determining region [CDR]
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/70—Immunoglobulins specific features characterized by effect upon binding to a cell or to an antigen
    • C07K2317/76—Antagonist effect on antigen, e.g. neutralization or inhibition of binding
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/90—Immunoglobulins specific features characterized by (pharmaco)kinetic aspects or by stability of the immunoglobulin
    • C07K2317/92—Affinity (KD), association rate (Ka), dissociation rate (Kd) or EC50 value

Definitions

  • the present disclosure relates to antibodies that bind SARS-CoV-2 spike protein (“anti- SARS-CoV-2 antibodies”), compositions comprising such anti-SARS-CoV-2 antibodies, and methods of using such anti-SARS-CoV-2 antibodies.
  • Coronaviruses are a large family of viruses that infect mammals and birds, with four endemic strains that circulate commonly in humans, including human coronavirus 229E, OC43, NL63 and HKU1. These endemic strains generally cause mild flu-like symptoms, but can also cause more serious pneumonia in vulnerable populations. However, when new coronaviruses jump from zoonotic hosts (e.g., bats, birds, camels) to humans, they can result in a much more severe respiratory disease that can spread quickly through the population.
  • zoonotic hosts e.g., bats, birds, camels
  • SARS severe Acute Respiratory Syndrome
  • SARS-CoV SARS-associated coronavirus
  • MERS Middle Eastern Respiratory Syndrome
  • SARS-CoV-2 The new coronavirus of 2019, named SARS-CoV-2, causing a severe respiratory disease referred to as COVID-19, has spread rapidly on a global scale, with over 20 million confirmed cases in more than 216 countries and regions to date and led to over 700,000 deaths. SARS-CoV-2 is the most serious coronavirus outbreak in history. There is an urgent need for antibody-based therapeutics, prophylactics, and diagnostics to treat, prevent, and detect SARS- CoV-2 infection and COVID- 19.
  • the anti-SARS-CoV-2 antibodies are engineered antibodies that have one or more of the following properties: 1) bind to SARS-CoV-2 spike protein with desirable binding affinities and/or association and dissociation rates, 2) neutralize SARS-CoV-2 virus with desired potency, and/or 3) have a long half-life and good stability, solubility, viscosity, and/or pharmacokinetic characteristics for development and use in the treatment of COVID-19.
  • the cryo-electron microscopy structure of the SARS-CoV-2 S protein has been described recently, and shows the SARS-CoV-2 S protein is a trimeric class I fusion protein that exists in a metastable pre-fusion conformation and undergoes a substantial structural rearrangement to fuse the viral membrane with the host cell membrane (Wrapp et al., Science 367,
  • SARS-CoV-2 spike (S) protein has been described before, for example, GenBank Accession No: YP 009724390.1 provides such an exemplary sequence, as shown below:
  • MFVFLVLLPLVS SQC VNLTTRTQLPPAYTNSFTRGVYYPDK VFRS S VLHSTQDLFLPFF S NVTWFHAIHVSGTNGTKRFDNPVLPFNDGVYFASTEKSNIIRGWIFGTTLDSKTQSLLIV NNATNVVIKVCEFQFCNDPFLGVYYHKNNKSWMESEFRVYSSANNCTFEYVSQPFLMD LEGKQGNFKNLREFVFKNIDGYFKIYSKHTPINLVRDLPQGFSALEPLVDLPIGINITRFQT LLALHRSYLTPGDSSSGWTAGAAAYYVGYLQPRTFLLKYNENGTITDAVDCALDPLSET KCTLKSFTVEKGIYQTSNFRVQPTESIVRFPNITNLCPFGEVFNATRFASVYAWNRKRISN CVADYSVLYNSASFSTFKCYGVSPTKLNDLCFTNVYADSFVIRGDEVRQIAPGQTGKIA DYNY
  • the receptor-binding domain comprises amino acids 319 to 592 of the SARS- CoV-2 S protein, e.g., amino acids 319 to 592 of SEQ ID NO: 130 or a variant comprising one or more mutations;
  • the N-terminal domain comprises amino acids 16-307 of the SARS-CoV- 2 S protein, e.g., amino acids 16-307 of SEQ ID NO: 130 or a variant comprising one or more mutations;
  • S2 domain comprises amino acids 686-1213 of the SARS-CoV-2 S protein, e.g., amino acids 686-1213 of SEQ ID NO: 130 or a variant comprising one or more mutations (see Wrapp et al., Science 367, 1260-1263, 2020).
  • anti -SARS-CoV-2 antibodies comprising a heavy chain variable region (VH) and a light chain variable region (VL), wherein the VH comprises heavy chain complementarity determining regions HCDR1, HCDR2, and HCDR3, and the VL comprises light chain complementarity determining regions LCDR1, LCDR2, and LCDR3, wherein the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 are selected from the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 in Table 1.
  • anti-SARS-CoV-2 antibodies that comprise a VH domain comprising the same three HCDRs as comprised in the VH domain of any anti-SARS-CoV-2 antibody in Table 1.
  • anti-SARS-CoV-2 antibodies that comprise a VL domain comprising the same three LCDRs as the VL domain of any anti-SARS-CoV-2 antibody in Table 1.
  • anti-SARS-CoV-2 antibodies that comprise a VH domain comprising the same three HCDRs as comprised in the VH domain of any anti-SARS- CoV-2 antibody in Table 1, and a VL domain comprising the same three LCDRs as the VL domain of any anti-SARS-CoV-2 antibody in Table 1.
  • anti- SARS-CoV-2 antibodies that comprise the same VH and/or the VL of any anti-SARS-CoV-2 antibody in Table 1.
  • anti-SARS-CoV-2 antibodies that comprise a VH having at least 95% sequence identity to any of the VHs in Table 1, and a VL having at least 95% sequence identity to any of the VLs in Table 1 .
  • the anti-SARS-CoV-2 antibodies described herein have IgGl isotype.
  • anti-SARS-CoV-2 antibodies comprising a heavy chain (HC) selected from any HC in Table 1, and/or a light chain (LC) selected from any LC in Table 1.
  • anti-SARS-CoV-2 antibodies that comprise a HC having at least 95% sequence identity to any of the HCs in Table 1, and a LC having at least 95% sequence identity to any of the LCs in Table 1.
  • an antibody that binds SARS-CoV-2 spike protein wherein the antibody comprises a VH and a VL, wherein the VH comprises HCDR1, HCDR2, and HCDR3, and the VL comprises LCDR1, LCDR2, and LCDR3, wherein the antibody comprises: HCDR1 comprising SEQ ID NO: 3, HCDR2 comprising SEQ ID NO: 4, HCDR3 comprising SEQ ID NO: 5, 18, or 26, LCDR1 comprising SEQ ID NO: 6, LCDR2 comprising SEQ ID NO: 7 or 13, and LCDR3 comprising SEQ ID NO: 8, 14, or 19.
  • an antibody that binds SARS-CoV-2 spike protein wherein the antibody comprises a VH and a VL, wherein the VH comprises HCDR1, HCDR2, and HCDR3, and the VL comprises LCDR1, LCDR2, and LCDR3, wherein the antibody comprises: HCDR1 comprising SEQ ID NO: 3, HCDR2 comprising SEQ ID NO: 4, HCDR3 comprising SEQ ID NO: 5, LCDR1 comprising SEQ ID NO: 6, LCDR2 comprising SEQ ID NO: 7, and LCDR3 comprising SEQ ID NO: 8.
  • an antibody that binds SARS-CoV-2 spike protein wherein the antibody comprises a VH and a VL, wherein the VH comprises HCDR1, HCDR2, and HCDR3, and the VL comprises LCDR1, LCDR2, and LCDR3, wherein the antibody comprises: HCDR1 comprising SEQ ID NO: 3, HCDR2 comprising SEQ ID NO: 4, HCDR3 comprising SEQ ID NO: 5, LCDR1 comprising SEQ ID NO: 6, LCDR2 comprising SEQ ID NO: 13, and LCDR3 comprising SEQ ID NO: 14.
  • an antibody that binds SARS-CoV-2 spike protein wherein the antibody comprises a VH and a VL, wherein the VH comprises HCDR1, HCDR2, and HCDR3, and the VL comprises LCDR1, LCDR2, and LCDR3, wherein the antibody comprises: HCDR1 comprising SEQ ID NO: 3, HCDR2 comprising SEQ ID NO: 4, HCDR3 comprising SEQ ID NO: 18, LCDR1 comprising SEQ ID NO: 6, LCDR2 comprising SEQ ID NO: 7, and LCDR3 comprising SEQ ID NO: 19.
  • an antibody that binds SARS-CoV-2 spike protein wherein the antibody comprises a VH and a VL, wherein the VH comprises HCDR1, HCDR2, and HCDR3, and the VL comprises LCDR1, LCDR2, and LCDR3, wherein the antibody comprises: HCDR1 comprising SEQ ID NO: 3, HCDR2 comprising SEQ ID NO: 4, HCDR3 comprising SEQ ID NO: 5, LCDR1 comprising SEQ ID NO: 6, LCDR2 comprising SEQ ID NO: 13, and LCDR3 comprising SEQ ID NO: 19.
  • an antibody that binds SARS-CoV-2 spike protein wherein the antibody comprises a VH and a VL, wherein the VH comprises HCDR1, HCDR2, and HCDR3, and the VL comprises LCDR1, LCDR2, and LCDR3, wherein the antibody comprises: HCDR1 comprising SEQ ID NO: 3, HCDR2 comprising SEQ ID NO: 4, HCDR3 comprising SEQ ID NO: 26, LCDR1 comprising SEQ ID NO: 6, LCDR2 comprising SEQ ID NO: 7, and LCDR3 comprising SEQ ID NO: 8.
  • an antibody that binds SARS-CoV-2 spike protein wherein the antibody comprises a VH and a VL, wherein the VH comprises HCDR1, HCDR2, and HCDR3, and the VL comprises LCDR1, LCDR2, and LCDR3, wherein the antibody comprises: HCDR1 comprising SEQ ID NO: 3, HCDR2 comprising SEQ ID NO: 4, HCDR3 comprising SEQ ID NO: 18, LCDR1 comprising SEQ ID NO: 6, LCDR2 comprising SEQ ID NO: 7, and LCDR3 comprising SEQ ID NO: 8.
  • an antibody that binds SARS-CoV-2 spike protein wherein the antibody comprises a VH and a VL, wherein the VH comprises HCDR1, HCDR2, and HCDR3, and the VL comprises LCDR1, LCDR2, and LCDR3, wherein the antibody comprises: HCDR1 comprising SEQ ID NO: 31, HCDR2 comprising SEQ ID NO: 32, HCDR3 comprising SEQ ID NO: 33, 44, or 50, LCDR1 comprising SEQ ID NO: 34, LCDR2 comprising SEQ ID NO: 35, and LCDR3 comprising SEQ ID NO: 36 or 45.
  • an antibody that binds SARS-CoV-2 spike protein wherein the antibody comprises a VH and a VL, wherein the VH comprises HCDR1, HCDR2, and HCDR3, and the VL comprises LCDR1, LCDR2, and LCDR3, wherein the antibody comprises: HCDR1 comprising SEQ ID NO: 31, HCDR2 comprising SEQ ID NO: 32, HCDR3 comprising SEQ ID NO: 33, LCDR1 comprising SEQ ID NO: 34, LCDR2 comprising SEQ ID NO: 35, and LCDR3 comprising SEQ ID NO: 36.
  • an antibody that binds SARS-CoV-2 spike protein wherein the antibody comprises a VH and a VL, wherein the VH comprises HCDR1, HCDR2, and HCDR3, and the VL comprises LCDR1, LCDR2, and LCDR3, wherein the antibody comprises: HCDR1 comprising SEQ ID NO: 31, HCDR2 comprising SEQ ID NO: 32, HCDR3 comprising SEQ ID NO: 44, LCDR1 comprising SEQ ID NO: 34, LCDR2 comprising SEQ ID NO: 35, and LCDR3 comprising SEQ ID NO: 45.
  • an antibody that binds SARS-CoV-2 spike protein wherein the antibody comprises a VH and a VL, wherein the VH comprises HCDR1, HCDR2, and HCDR3, and the VL comprises LCDR1, LCDR2, and LCDR3, wherein the antibody comprises: HCDR1 comprising SEQ ID NO: 31, HCDR2 comprising SEQ ID NO: 32, HCDR3 comprising SEQ ID NO: 50, LCDR1 comprising SEQ ID NO: 34, LCDR2 comprising SEQ ID NO: 35, and LCDR3 comprising SEQ ID NO: 36.
  • an antibody that binds SARS-CoV-2 spike protein wherein the antibody comprises a VH and a VL, wherein the VH comprises HCDR1, HCDR2, and HCDR3, and the VL comprises LCDR1, LCDR2, and LCDR3, wherein the antibody comprises: HCDR1 comprising SEQ ID NO: 55, HCDR2 comprising SEQ ID NO: 56 or 73, HCDR3 comprising SEQ ID NO: 57, LCDR1 comprising SEQ ID NO: 58, 65, 69, or 74, LCDR2 comprising SEQ ID NO: 59 or 66, and LCDR3 comprising SEQ ID NO: 60.
  • an antibody that binds SARS-CoV-2 spike protein wherein the antibody comprises a VH and a VL, wherein the VH comprises HCDR1, HCDR2, and HCDR3, and the VL comprises LCDR1, LCDR2, and LCDR3, wherein the antibody comprises: HCDR1 comprising SEQ ID NO: 55, HCDR2 comprising SEQ ID NO: 56, HCDR3 comprising SEQ ID NO: 57, LCDR1 comprising SEQ ID NO: 58, LCDR2 comprising SEQ ID NO: 59, and LCDR3 comprising SEQ ID NO: 60.
  • an antibody that binds SARS-CoV-2 spike protein wherein the antibody comprises a VH and a VL, wherein the VH comprises HCDR1, HCDR2, and HCDR3, and the VL comprises LCDR1, LCDR2, and LCDR3, wherein the antibody comprises: HCDR1 comprising SEQ ID NO: 55, HCDR2 comprising SEQ ID NO: 56, HCDR3 comprising SEQ ID NO: 57, LCDR1 comprising SEQ ID NO: 65, LCDR2 comprising SEQ ID NO: 66, and LCDR3 comprising SEQ ID NO: 60.
  • an antibody that binds SARS-CoV-2 spike protein wherein the antibody comprises a VH and a VL, wherein the VH comprises HCDR1, HCDR2, and HCDR3, and the VL comprises LCDR1, LCDR2, and LCDR3, wherein the antibody comprises: HCDR1 comprising SEQ ID NO: 55, HCDR2 comprising SEQ ID NO: 56, HCDR3 comprising SEQ ID NO: 57, LCDR1 comprising SEQ ID NO: 69, LCDR2 comprising SEQ ID NO: 59, and LCDR3 comprising SEQ ID NO: 60.
  • an antibody that binds SARS-CoV-2 spike protein wherein the antibody comprises a VH and a VL, wherein the VH comprises HCDR1, HCDR2, and HCDR3, and the VL comprises LCDR1, LCDR2, and LCDR3, wherein the antibody comprises: HCDR1 comprising SEQ ID NO: 55, HCDR2 comprising SEQ ID NO: 73, HCDR3 comprising SEQ ID NO: 57, LCDR1 comprising SEQ ID NO: 74, LCDR2 comprising SEQ ID NO: 59, and LCDR3 comprising SEQ ID NO: 60.
  • an antibody that binds SARS-CoV-2 spike protein wherein the antibody comprises a VH and a VL, wherein the VH comprises HCDR1, HCDR2, and HCDR3, and the VL comprises LCDR1, LCDR2, and LCDR3, wherein the antibody comprises: HCDR1 comprising SEQ ID NO: 80, 89, or 95, HCDR2 comprising SEQ ID NO: 81, 1 HCDR3 comprising SEQ ID NO: 82 or 90, LCDR1 comprising SEQ ID NO: 83, LCDR2 comprising SEQ ID NO: 13, and LCDR3 comprising SEQ ID NO: 84 or 96.
  • an antibody that binds SARS-CoV-2 spike protein wherein the antibody comprises a VH and a VL, wherein the VH comprises HCDR1, HCDR2, and HCDR3, and the VL comprises LCDR1, LCDR2, and LCDR3, wherein the antibody comprises: HCDR1 comprising SEQ ID NO: 80, HCDR2 comprising SEQ ID NO: 81, HCDR3 comprising SEQ ID NO: 82, LCDR1 comprising SEQ ID NO: 83, LCDR2 comprising SEQ ID NO: 13, and LCDR3 comprising SEQ ID NO: 84.
  • an antibody that binds SARS-CoV-2 spike protein wherein the antibody comprises a VH and a VL, wherein the VH comprises HCDR1, HCDR2, and HCDR3, and the VL comprises LCDR1, LCDR2, and LCDR3, wherein the antibody comprises: HCDR1 comprising SEQ ID NO: 89, HCDR2 comprising SEQ ID NO: 81, HCDR3 comprising SEQ ID NO: 90, LCDR1 comprising SEQ ID NO: 83, LCDR2 comprising SEQ ID NO: 13, and LCDR3 comprising SEQ ID NO: 84.
  • an antibody that binds SARS-CoV-2 spike protein wherein the antibody comprises a VH and a VL, wherein the VH comprises HCDR1, HCDR2, and HCDR3, and the VL comprises LCDR1, LCDR2, and LCDR3, wherein the antibody comprises: HCDR1 comprising SEQ ID NO: 95, HCDR2 comprising SEQ ID NO: 81, HCDR3 comprising SEQ ID NO: 82, LCDR1 comprising SEQ ID NO: 83, LCDR2 comprising SEQ ID NO: 13, and LCDR3 comprising SEQ ID NO: 96.
  • an antibody that binds SARS-CoV-2 spike protein wherein the antibody comprises a VH and a VL, wherein the VH comprises HCDR1, HCDR2, and HCDR3, and the VL comprises LCDR1, LCDR2, and LCDR3, wherein the antibody comprises: HCDR1 comprising SEQ ID NO: 80, HCDR2 comprising SEQ ID NO: 81, HCDR3 comprising SEQ ID NO: 82, LCDR1 comprising SEQ ID NO: 83, LCDR2 comprising SEQ ID NO: 13, and LCDR3 comprising SEQ ID NO: 96.
  • an antibody that binds SARS-CoV-2 spike protein wherein the antibody comprises a VH and a VL, wherein the VH comprises HCDR1, HCDR2, and HCDR3, and the VL comprises LCDR1, LCDR2, and LCDR3, wherein the antibody comprises: HCDR1 comprising SEQ ID NO: 102 or 112, HCDR2 comprising SEQ ID NO: 103 or 119, HCDR3 comprising SEQ ID NO: 104, 120, or 127, LCDR1 comprising SEQ ID NO: 105 or 116, LCDR2 comprising SEQ ID NO: 106, and LCDR3 comprising SEQ ID NO: 107.
  • an antibody that binds SARS-CoV-2 spike protein wherein the antibody comprises a VH and a VL, wherein the VH comprises HCDR1, HCDR2, and HCDR3, and the VL comprises LCDR1, LCDR2, and LCDR3, wherein the antibody comprises: HCDR1 comprising SEQ ID NO: 102, HCDR2 comprising SEQ ID NO: 103, HCDR3 comprising SEQ ID NO: 104, LCDR1 comprising SEQ ID NO: 105, LCDR2 comprising SEQ ID NO: 106, and LCDR3 comprising SEQ ID NO: 107.
  • an antibody that binds SARS-CoV-2 spike protein wherein the antibody comprises a VH and a VL, wherein the VH comprises HCDR1, HCDR2, and HCDR3, and the VL comprises LCDR1, LCDR2, and LCDR3, wherein the antibody comprises: HCDR1 comprising SEQ ID NO: 112, HCDR2 comprising SEQ ID NO: 103, HCDR3 comprising SEQ ID NO: 104, LCDR1 comprising SEQ ID NO: 105, LCDR2 comprising SEQ ID NO: 106, and LCDR3 comprising SEQ ID NO: 107.
  • an antibody that binds SARS-CoV-2 spike protein wherein the antibody comprises a VH and a VL, wherein the VH comprises HCDR1, HCDR2, and HCDR3, and the VL comprises LCDR1, LCDR2, and LCDR3, wherein the antibody comprises: HCDR1 comprising SEQ ID NO: 112, HCDR2 comprising SEQ ID NO: 103, HCDR3 comprising SEQ ID NO: 104, LCDR1 comprising SEQ ID NO: 116, LCDR2 comprising SEQ ID NO: 106, and LCDR3 comprising SEQ ID NO: 107.
  • an antibody that binds SARS-CoV-2 spike protein wherein the antibody comprises a VH and a VL, wherein the VH comprises HCDR1, HCDR2, and HCDR3, and the VL comprises LCDR1, LCDR2, and LCDR3, wherein the antibody comprises: HCDR1 comprising SEQ ID NO: 112, HCDR2 comprising SEQ ID NO: 119, HCDR3 comprising SEQ ID NO: 120, LCDR1 comprising SEQ ID NO: 116, LCDR2 comprising SEQ ID NO: 106, and LCDR3 comprising SEQ ID NO: 107.
  • an antibody that binds SARS-CoV-2 spike protein wherein the antibody comprises a VH and a VL, wherein the VH comprises HCDR1, HCDR2, and HCDR3, and the VL comprises LCDR1, LCDR2, and LCDR3, wherein the antibody comprises: HCDR1 comprising SEQ ID NO: 112, HCDR2 comprising SEQ ID NO: 119, HCDR3 comprising SEQ ID NO: 104, LCDR1 comprising SEQ ID NO: 116, LCDR2 comprising SEQ ID NO: 106, and LCDR3 comprising SEQ ID NO: 107.
  • an antibody that binds SARS-CoV-2 spike protein wherein the antibody comprises a VH and a VL, wherein the VH comprises HCDR1, HCDR2, and HCDR3, and the VL comprises LCDR1, LCDR2, and LCDR3, wherein the antibody comprises: HCDR1 comprising SEQ ID NO: 112, HCDR2 comprising SEQ ID NO: 119, HCDR3 comprising SEQ ID NO: 127, LCDR1 comprising SEQ ID NO: 116, LCDR2 comprising SEQ ID NO: 106, and LCDR3 comprising SEQ ID NO: 107.
  • an antibody that binds SARS-CoV-2 spike protein wherein the antibody comprises a VH and a VL, wherein the VH comprises heavy chain complementarity determining regions HCDR1, HCDR2, andHCDR3, and the VL comprises light chain complementarity determining regions LCDR1, LCDR2, and LCDR3, wherein the antibody comprises: HCDR1 comprising SEQ ID NO: 137, HCDR2 comprising SEQ ID NO: 138, HCDR3 comprising SEQ ID NO: 139, LCDR1 comprising SEQ ID NO: 140, LCDR2 comprising SEQ ID NO: 141, and LCDR3 comprising SEQ ID NO: 142.
  • an antibody that binds SARS-CoV-2 spike protein wherein the antibody comprises a VH comprising SEQ ID NO: 1, 16 or 25, and a VL comprising SEQ ID NO: 2, 12, 17, or 23.
  • an antibody that binds SARS-CoV-2 spike protein wherein the antibody comprises a VH comprising SEQ ID NO: 1, and a VL comprising SEQ ID NO: 2.
  • an antibody that binds SARS-CoV-2 spike protein wherein the antibody comprises a VH comprising SEQ ID NO: 1, and a VL comprising SEQ ID NO: 12.
  • an antibody that binds SARS-CoV-2 spike protein wherein the antibody comprises a VH comprising SEQ ID NO: 16, and a VL comprising SEQ ID NO: 17.
  • an antibody that binds SARS-CoV-2 spike protein wherein the antibody comprises a VH comprising SEQ ID NO: 1, and a VL comprising SEQ ID NO: 23.
  • an antibody that binds SARS-CoV-2 spike protein wherein the antibody comprises a VH comprising SEQ ID NO: 25, and a VL comprising SEQ ID NO: 2.
  • an antibody that binds SARS-CoV-2 spike protein wherein the antibody comprises a VH comprising SEQ ID NO: 16, and a VL comprising SEQ ID NO: 2.
  • an antibody that binds SARS-CoV-2 spike protein wherein the antibody comprises a VH comprising SEQ ID NO: 29, 42, or 49, and a VL comprising SEQ ID NO: 30, 40, 43.
  • an antibody that binds SARS-CoV-2 spike protein wherein the antibody comprises a VH comprising SEQ ID NO: 29, and a VL comprising SEQ ID NO: 30.
  • an antibody that binds SARS-CoV-2 spike protein wherein the antibody comprises a VH comprising SEQ ID NO: 29, and a VL comprising SEQ ID NO: 40.
  • an antibody that binds SARS-CoV-2 spike protein wherein the antibody comprises a VH comprising SEQ ID NO: 42, and a VL comprising SEQ ID NO: 43.
  • an antibody that binds SARS-CoV-2 spike protein wherein the antibody comprises a VH comprising SEQ ID NO: 49, and a VL comprising SEQ ID NO: 40.
  • an antibody that binds SARS-CoV-2 spike protein wherein the antibody comprises a VH comprising SEQ ID NO: 53 or 71, and a VL comprising SEQ ID NO: 54, 64, 68, or 72.
  • an antibody that binds SARS-CoV-2 spike protein wherein the antibody comprises a VH comprising SEQ ID NO: 53, and a VL comprising SEQ ID NO: 54.
  • an antibody that binds SARS-CoV-2 spike protein wherein the antibody comprises a VH comprising SEQ ID NO: 53, and a VL comprising SEQ ID NO: 64.
  • an antibody that binds SARS-CoV-2 spike protein wherein the antibody comprises a VH comprising SEQ ID NO: 53, and a VL comprising SEQ ID NO: 68.
  • an antibody that binds SARS-CoV-2 spike protein wherein the antibody comprises a VH comprising SEQ ID NO: 71, and a VL comprising SEQ ID NO: 72.
  • an antibody that binds SARS-CoV-2 spike protein wherein the antibody comprises a VH comprising SEQ ID NO: 78, 88 or 93, and a VL comprising SEQ ID NO: 79 or 94.
  • an antibody that binds SARS-CoV-2 spike protein wherein the antibody comprises a VH comprising SEQ ID NO: 78, and a VL comprising SEQ ID NO: 79.
  • an antibody that binds SARS-CoV-2 spike protein wherein the antibody comprises a VH comprising SEQ ID NO: 88, and a VL comprising SEQ ID NO: 79.
  • an antibody that binds SARS-CoV-2 spike protein wherein the antibody comprises a VH comprising SEQ ID NO: 93, and a VL comprising SEQ ID NO: 94.
  • an antibody that binds SARS-CoV-2 spike protein wherein the antibody comprises a VH comprising SEQ ID NO: 78, and a VL comprising SEQ ID NO: 94.
  • an antibody that binds SARS-CoV-2 spike protein wherein the antibody comprises a VH comprising SEQ ID NO: 100, 111, 118, 123, or 126, and a VL comprising SEQ ID NO: 101 or 115.
  • an antibody that binds SARS-CoV-2 spike protein wherein the antibody comprises a VH comprising SEQ ID NO: 100, and a VL comprising SEQ ID NO: 101.
  • an antibody that binds SARS-CoV-2 spike protein wherein the antibody comprises a VH comprising SEQ ID NO: 111, and a VL comprising SEQ ID NO: 101.
  • an antibody that binds SARS-CoV-2 spike protein wherein the antibody comprises a VH comprising SEQ ID NO: 111, and a VL comprising SEQ ID NO: 115.
  • an antibody that binds SARS-CoV-2 spike protein wherein the antibody comprises a VH comprising SEQ ID NO: 118, and a VL comprising SEQ ID NO: 115.
  • an antibody that binds SARS-CoV-2 spike protein wherein the antibody comprises a VH comprising SEQ ID NO: 123, and a VL comprising SEQ ID NO: 115.
  • an antibody that binds SARS-CoV-2 spike protein wherein the antibody comprises a VH comprising SEQ ID NO: 126, and a VL comprising SEQ ID NO: 115.
  • an antibody that binds SARS-CoV-2 spike protein wherein the antibody comprises a VH comprising SEQ ID NO: 135, and a VL comprising SEQ ID NO: 136.
  • an antibody that binds SARS-CoV-2 spike protein wherein the antibody comprises a HC comprising SEQ ID NO: 9, 10, 20, 21, 27, or 28, and a LC comprising SEQ ID NO: 11, 15, 22, or 24.
  • an antibody that binds SARS-CoV-2 spike protein wherein the antibody comprises a HC comprising SEQ ID NO: 9, 20, or 27, and a LC comprising SEQ ID NO: 11, 15, 22, or 24.
  • an antibody that binds SARS-CoV-2 spike protein wherein the antibody comprises a HC comprising SEQ ID NO: 10, 21, or 28, and a LC comprising SEQ ID NO: 11, 15, 22, or 24.
  • an antibody that binds SARS-CoV-2 spike protein wherein the antibody comprises a HC comprising SEQ ID NO: 9, and a LC comprising SEQ ID NO: 11.
  • an antibody that binds SARS-CoV-2 spike protein wherein the antibody comprises a HC comprising SEQ ID NO: 10, and a LC comprising SEQ ID NO: 11.
  • an antibody that binds SARS-CoV-2 spike protein wherein the antibody comprises a HC comprising SEQ ID NO: 9, and a LC comprising SEQ ID NO: 15.
  • an antibody that binds SARS-CoV-2 spike protein wherein the antibody comprises a HC comprising SEQ ID NO: 10, and a LC comprising SEQ ID NO: 15.
  • an antibody that binds SARS-CoV-2 spike protein wherein the antibody comprises a HC comprising SEQ ID NO: 9, and a LC comprising SEQ ID NO: 24.
  • an antibody that binds SARS-CoV-2 spike protein wherein the antibody comprises a HC comprising SEQ ID NO: 10, and a LC comprising SEQ ID NO: 24.
  • an antibody that binds SARS-CoV-2 spike protein wherein the antibody comprises a HC comprising SEQ ID NO: 20, and a LC comprising SEQ ID NO: 11.
  • an antibody that binds SARS-CoV-2 spike protein wherein the antibody comprises a HC comprising SEQ ID NO: 21, and a LC comprising SEQ ID NO: 11.
  • an antibody that binds SARS-CoV-2 spike protein wherein the antibody comprises a HC comprising SEQ ID NO: 20, and a LC comprising SEQ ID NO: 22.
  • an antibody that binds SARS-CoV-2 spike protein wherein the antibody comprises a HC comprising SEQ ID NO: 21, and a LC comprising SEQ ID NO: 22.
  • an antibody that binds SARS-CoV-2 spike protein wherein the antibody comprises a HC comprising SEQ ID NO: 27, and a LC comprising SEQ ID NO: 11.
  • an antibody that binds SARS-CoV-2 spike protein wherein the antibody comprises a HC comprising SEQ ID NO: 28, and a LC comprising SEQ ID NO: 11.
  • an antibody that binds SARS-CoV-2 spike protein wherein the antibody comprises a HC comprising SEQ ID NO: 37, 38, 46, 47, 51, or 52, and a LC comprising SEQ ID NO: 39, 41, or 48.
  • an antibody that binds SARS-CoV-2 spike protein wherein the antibody comprises a HC comprising SEQ ID NO: 37, 46, or 51, and a LC comprising SEQ ID NO: 39, 41, or 48.
  • an antibody that binds SARS-CoV-2 spike protein wherein the antibody comprises a HC comprising SEQ ID NO: 38, 47, or 52, and a LC comprising SEQ ID NO: 39, 41, or 48.
  • an antibody that binds SARS-CoV-2 spike protein wherein the antibody comprises a HC comprising SEQ ID NO: 37, and a LC comprising SEQ ID NO: 39.
  • an antibody that binds SARS-CoV-2 spike protein wherein the antibody comprises a HC comprising SEQ ID NO: 38, and a LC comprising SEQ ID NO: 39.
  • an antibody that binds SARS-CoV-2 spike protein wherein the antibody comprises a HC comprising SEQ ID NO: 37, and a LC comprising SEQ ID NO: 41.
  • an antibody that binds SARS-CoV-2 spike protein wherein the antibody comprises a HC comprising SEQ ID NO: 38, and a LC comprising SEQ ID NO: 41.
  • an antibody that binds SARS-CoV-2 spike protein wherein the antibody comprises a HC comprising SEQ ID NO: 51, and a LC comprising SEQ ID NO: 41.
  • an antibody that binds SARS-CoV-2 spike protein wherein the antibody comprises a HC comprising SEQ ID NO: 52, and a LC comprising SEQ ID NO: 41.
  • an antibody that binds SARS-CoV-2 spike protein wherein the antibody comprises a HC comprising SEQ ID NO: 46, and a LC comprising SEQ ID NO: 48.
  • an antibody that binds SARS-CoV-2 spike protein wherein the antibody comprises a HC comprising SEQ ID NO: 47, and a LC comprising SEQ ID NO: 48.
  • an antibody that binds SARS-CoV-2 spike protein wherein the antibody comprises a HC comprising SEQ ID NO: 61, 62, 75 or 76, and a LC comprising SEQ ID NO: 63, 67, 70, or 77.
  • an antibody that binds SARS-CoV-2 spike protein wherein the antibody comprises a HC comprising SEQ ID NO: 61, or 75, and a LC comprising SEQ ID NO: 63, 67, 70, or 77.
  • an antibody that binds SARS-CoV-2 spike protein wherein the antibody comprises aHC comprising SEQ ID NO: 62, or 76, and aLC comprising SEQ ID NO: 63, 67, 70, or 77.
  • an antibody that binds SARS-CoV-2 spike protein wherein the antibody comprises a HC comprising SEQ ID NO: 61, and a LC comprising SEQ ID NO: 63.
  • an antibody that binds SARS-CoV-2 spike protein wherein the antibody comprises a HC comprising SEQ ID NO: 62, and a LC comprising SEQ ID NO: 63.
  • an antibody that binds SARS-CoV-2 spike protein wherein the antibody comprises a HC comprising SEQ ID NO: 61, and a LC comprising SEQ ID NO: 67.
  • an antibody that binds SARS-CoV-2 spike protein wherein the antibody comprises a HC comprising SEQ ID NO: 62, and a LC comprising SEQ ID NO: 67.
  • an antibody that binds SARS-CoV-2 spike protein wherein the antibody comprises a HC comprising SEQ ID NO: 61, and a LC comprising SEQ ID NO: 70.
  • an antibody that binds SARS-CoV-2 spike protein wherein the antibody comprises aHC comprising SEQ ID NO: 62, and a LC comprising SEQ ID NO: 70.
  • an antibody that binds SARS-CoV-2 spike protein wherein the antibody comprises a HC comprising SEQ ID NO: 75, and a LC comprising SEQ ID NO: 77.
  • an antibody that binds SARS-CoV-2 spike protein wherein the antibody comprises a HC comprising SEQ ID NO: 76, and a LC comprising SEQ ID NO: 77.
  • an antibody that binds SARS-CoV-2 spike protein wherein the antibody comprises a HC comprising SEQ ID NO: 85, 86, 91, 92, 97, or 98, and a LC comprising SEQ ID NO: 87 or 99.
  • an antibody that binds SARS-CoV-2 spike protein wherein the antibody comprises a HC comprising SEQ ID NO: 85, 91, or 97, and a LC comprising SEQ ID NO: 87 or 99.
  • an antibody that binds SARS-CoV-2 spike protein wherein the antibody comprises a HC comprising SEQ ID NO: 86, 92, or 98, and a LC comprising SEQ ID NO: 87 or 99.
  • an antibody that binds SARS-CoV-2 spike protein wherein the antibody comprises a HC comprising SEQ ID NO: 85, and a LC comprising SEQ ID NO: 87.
  • an antibody that binds SARS-CoV-2 spike protein wherein the antibody comprises a HC comprising SEQ ID NO: 86, and a LC comprising SEQ ID NO: 87.
  • an antibody that binds SARS-CoV-2 spike protein wherein the antibody comprises a HC comprising SEQ ID NO: 85, and a LC comprising SEQ ID NO: 99.
  • an antibody that binds SARS-CoV-2 spike protein wherein the antibody comprises a HC comprising SEQ ID NO: 86, and a LC comprising SEQ ID NO: 99.
  • an antibody that binds SARS-CoV-2 spike protein wherein the antibody comprises a HC comprising SEQ ID NO: 91, and a LC comprising SEQ ID NO: 87.
  • an antibody that binds SARS-CoV-2 spike protein wherein the antibody comprises a HC comprising SEQ ID NO: 92, and a LC comprising SEQ ID NO: 87.
  • an antibody that binds SARS-CoV-2 spike protein wherein the antibody comprises a HC comprising SEQ ID NO: 97, and a LC comprising SEQ ID NO: 99.
  • an antibody that binds SARS-CoV-2 spike protein wherein the antibody comprises a HC comprising SEQ ID NO: 98, and a LC comprising SEQ ID NO: 99.
  • an antibody that binds SARS-CoV-2 spike protein wherein the antibody comprises a HC comprising SEQ ID NO: 108, 109, 113, 114, 121, 122, 124, 125, 128, or 129, and a LC comprising SEQ ID NO: 110 or 117.
  • the antibody comprises a HC comprising SEQ ID NO: 108, 113, 121, 124, or 128, and a LC comprising SEQ ID NO: 110 or 117.
  • an antibody that binds SARS-CoV- 2 spike protein wherein the antibody comprises a HC comprising SEQ ID NO: 109, 114, 122, 125, or 129, and a LC comprising SEQ ID NO: 110 or 117.
  • an antibody that binds SARS-CoV-2 spike protein wherein the antibody comprises a HC comprising SEQ ID NO: 108, and a LC comprising SEQ ID NO: 110.
  • an antibody that binds SARS-CoV-2 spike protein wherein the antibody comprises a HC comprising SEQ ID NO: 109, and a LC comprising SEQ ID NO: 110.
  • an antibody that binds SARS-CoV-2 spike protein wherein the antibody comprises a HC comprising SEQ ID NO: 113, and a LC comprising SEQ ID NO: 110.
  • an antibody that binds SARS-CoV-2 spike protein wherein the antibody comprises a HC comprising SEQ ID NO: 114, and a LC comprising SEQ ID NO: 110.
  • an antibody that binds SARS-CoV-2 spike protein wherein the antibody comprises a HC comprising SEQ ID NO: 113, and a LC comprising SEQ ID NO: 117.
  • an antibody that binds SARS-CoV-2 spike protein wherein the antibody comprises a HC comprising SEQ ID NO: 114, and a LC comprising SEQ ID NO: 117.
  • an antibody that binds SARS-CoV-2 spike protein wherein the antibody comprises a HC comprising SEQ ID NO: 121, and a LC comprising SEQ ID NO: 117.
  • an antibody that binds SARS-CoV-2 spike protein wherein the antibody comprises a HC comprising SEQ ID NO: 122, and a LC comprising SEQ ID NO: 117.
  • an antibody that binds SARS-CoV-2 spike protein wherein the antibody comprises a HC comprising SEQ ID NO: 124, and a LC comprising SEQ ID NO: 117.
  • an antibody that binds SARS-CoV-2 spike protein wherein the antibody comprises a HC comprising SEQ ID NO: 125, and a LC comprising SEQ ID NO: 117.
  • an antibody that binds SARS-CoV-2 spike protein wherein the antibody comprises a HC comprising SEQ ID NO: 128, and a LC comprising SEQ ID NO: 117.
  • an antibody that binds SARS-CoV-2 spike protein wherein the antibody comprises a HC comprising SEQ ID NO: 129, and a LC comprising SEQ ID NO: 117.
  • an antibody that binds SARS-CoV-2 spike protein wherein the antibody comprises a HC comprising SEQ ID NO: 143 or 144, and a LC comprising SEQ ID NO: 145.
  • an antibody that binds SARS-CoV-2 spike protein wherein the antibody comprises a HC comprising SEQ ID NO: 143, and a LC comprising SEQ ID NO: 145.
  • an antibody that binds SARS-CoV-2 spike protein wherein the antibody comprises a HC comprising SEQ ID NO: 144, and a LC comprising SEQ ID NO: 145.
  • the anti-SARS-CoV-2 antibodies or antigen binding fragments provided herein bind an epitope in the RBD domain of the SARS-CoV-2 S protein, e.g., 555-com- Al, 555-com-D2, 555-com-H3, 555-com-D7, 555-com-H7, 555-com-A8, 488-2A4, 488-2A6, 488-2C5, 488-2G1, 481-L1H1, 481-L1H4, 481-L5H2, 481-L5H1.
  • the anti-SARS-CoV-2 antibodies or antigen binding fragments provided herein bind an epitope in the NTD domain of the SARS-CoV-2 S protein, e.g., 419-WT- 3, 419-FrF-10, 419-WT-15, 419-FrF-15, 419-HC2.1/LC56, 373-L1H21, 373-L1H24, 373-L2H24, 373-L2H10, 373-L2H20, 373-L2H61.
  • the anti-SARS-CoV-2 antibodies or antigen binding fragments provided herein are antibodies that block SARS-CoV-2 binding to ACE2, i.e., ACE2 blockers, e.g., 555-com-Al, 555-com-D2, 555-com-H3, 555-com-D7, 555-com-H7, 555-com-A8, 488-2A4, 488-2A6, 488-2C5, 488-2G1, 481-L1H1, 481-L1H4, 481-L5H2, 481-L5H1, 419-WT-3, 419-FrF- 10, 419-WT-15, 419-FrF-15, 419-HC2.1/LC56, 373-L1H21, 373-L1H24, 373-L2H24, 373- L2H10, 373-L2H20, 373-L2H61.
  • ACE2 blockers e.g., 555-com-Al,
  • the anti-SARS-CoV-2 antibodies disclosed herein are neutralizing antibodies.
  • Neutralization can be tested by various neutralization assays, e.g., a conventional neutralization assay based on the inhibition of a virus cytopathic effect (CPE) on cells in culture.
  • CPE virus cytopathic effect
  • neutralization may be tested by reducing or blocking formation of CPE in cells infected with SARS-CoV-2.
  • Virus and antibody may be premixed before addition to cells, followed by measuring blocking of virus entry.
  • neutralization can be tested by a pseudoneutralization assay or a live virus neutralization assay as describe in the Examples.
  • the anti-SARS-CoV-2 antibodies disclosed herein may contain one or more amino acid residue substitutions, mutations and/or modifications, which result in an antibody with preferred characteristics including, but not limited to: altered pharmacokinetics, increased serum half-life, increase binding affinity, reduced immunogenicity, increased production, altered Fc ligand binding, enhanced or reduced ADCC or CDC activity, altered glycosylation and/or disulfide bonds and/or modified binding specificity.
  • the anti-SARS-CoV-2 antibodies disclosed herein can encompass an Fc region with modifications that improve their half-lives (e.g., serum half-lives) in a human.
  • Fc engineering can enhance antibody binding to neonatal Fc receptor (FcRn) and elongate antibody half-lives.
  • the increased half-lives of the antibodies can result in a higher serum titer of antibodies, and thus, reduces the frequency of the administration of antibodies and/or reduces the concentration of antibodies or antibody fragments to be administered.
  • N434A/Y436T/Q438R/S440E ACT1
  • N434A/Y436V/Q438R/S440E ACT2
  • M428L/N434A/Y436T/Q438R/S440E ACT3
  • M428L/N434A/Y436V/Q438R/S440E ACT4
  • M428L/N434A/Q438R/S440E e.g., N434A/Y436T/Q438R/S440E (ACT1)
  • N434A/Y436V/Q438R/S440E ACT2
  • M428L/N434A/Y436T/Q438R/S440E ACT3
  • M428L/N434A/Y436V/Q438R/S440E ACT4
  • M428L/N434A/Q438R/S440E ACT5
  • the anti-SARS-CoV-2 antibodies disclosed herein comprise an Fc region comprising any set of mutations selected from ACT1, ACT2, ACT3, ACT4, ACT5. In some embodiments, the anti-SARS-CoV-2 antibodies disclosed herein comprise an Fc region comprising the ACT5 mutations: M428L/N434A/Q438R/S440E (positions numbered according to EU Index numbering).
  • the anti-SARS-CoV-2 antibodies disclosed herein can comprise an Fc region with altered (e.g., enhanced or reduced) effector functions, including but not limited to, Clq binding; Fc receptor binding; complement dependent cytotoxicity (CDC); antibodydependent cell-mediated cytotoxicity (ADCC); phagocytosis; down regulation of cell surface receptors.
  • CDC refers to the lysing of a target cell in the presence of complement.
  • the complement activation pathway is initiated by the binding of the first component of the complement system (Clq) to a molecule, an antibody for example, complexed with a cognate antigen.
  • a CDC assay may be performed.
  • ADCC refers to a form of cytotoxicity in which secreted Ig bound onto Fc receptors (FcRs) present on certain cytotoxic cells (e.g., Natural Killer (NK) cells, neutrophils, and macrophages) enables these cytotoxic effector cells to bind specifically to an antigen-bearing target cell and subsequently kill the target cell with cytotoxins.
  • FcRs Fc receptors
  • cytotoxic cells e.g., Natural Killer (NK) cells, neutrophils, and macrophages
  • NK Natural Killer
  • the anti-SARS-CoV-2 antibodies disclosed herein have enhanced ADCC activities.
  • the anti-SARS-CoV-2 antibodies disclosed herein have reduced ADCC activities.
  • the anti-SARS-CoV-2 antibodies described herein comprise a modified IgGl Fc region having reduced Fc effector functions.
  • the anti-SARS-CoV- 2 antibodies can comprise an IgGl Fc region comprising amino acid substitutions L234A and L235A (all residues are numbered according to the EU Index numbering).
  • the anti-SARS-CoV-2 antibodies can comprise an IgGl Fc region comprising amino acid substitutions L234A, L235E, G237A, A330S and P331S (all residues are numbered according to the EU Index numbering).
  • glycosylation patterns of the anti-SARS-CoV-2 antibodies disclosed herein can be modified.
  • Engineered glycoforms may be useful for a variety of purposes, including but not limited to enhancing or reducing effector function, increasing the affinity of the antibody for a target antigen or facilitating production of the antibody.
  • Such carbohydrate modifications can be accomplished by, for example, altering one or more sites of glycosylation within the antibody sequence. That is, one or more amino acid substitutions can be made that result in elimination of one or more variable region framework glycosylation sites to thereby eliminate glycosylation at that site.
  • effector functions or improved binding can also be modified by engineering in one or more additional glycosylation sites.
  • the anti-SARS-CoV-2 antibodies can comprise an Fc region that has an altered glycosylation composition, such as a hypofucosylated antibody having reduced amounts of fucosyl residues or an antibody having increased bisecting GlcNAc structures. These and similar altered glycosylation patterns have been demonstrated to increase the ADCC ability of antibodies.
  • Engineered glycoforms may be generated by any method known to one skilled in the art, for example by using engineered or variant expression strains, by co-expression with one or more enzymes (for example N-acetylglucosaminyltransferase III (GnTIl 1)), by expressing a molecule comprising an Fc region in various organisms or cell lines from various organisms or by modifying carbohydrate(s) after the molecule comprising Fc region has been expressed.
  • the anti-SARS-CoV-2 antibodies described herein have reduced fucosylation.
  • the anti-SARS-CoV-2 antibodies described herein comprise an Fc region comprising N-glycoside-linked sugar chains bound to the Fc region, wherein the sugar chains do not contain fucose. In some embodiments, the anti-SARS-CoV-2 antibodies described herein have increased ADCC activities, compared to the same antibodies comprising N-glycoside-linked sugar chains that comprise fucose.
  • anti-SARS-CoV-2 antibodies may be used in conjugated or unconjugated form. That is, the anti-SARS-CoV-2 antibody may be associated with or conjugated to (e.g. covalently or non-covalently) pharmaceutically active compounds, biological response modifiers, diagnostic moieties, or biocompatible modifiers.
  • conjugates may comprise peptides, polypeptides, proteins, fusion proteins, nucleic acid molecules, small molecules, mimetic agents, synthetic drugs, inorganic molecules, organic molecules and radioisotopes.
  • a conjugate may be covalently or non- covalently linked to the anti-SARS-CoV-2 antibody in various molar ratios depending, at least in part, on the method used for the conjugation.
  • the anti-SARS-CoV-2 antibodies disclosed herein are conjugated to a diagnostic or detectable agent, marker or reporter, which may be a biological molecule (e.g., a peptide or nucleotide), a small molecule, fluorophore, or radioisotope.
  • a diagnostic or detectable agent e.g., a peptide or nucleotide
  • labeled modulators can be useful for monitoring the development or progression of SARS-CoV-2 infection or as part of a clinical testing procedure to determine the efficacy of a particular therapy including anti-SARS-CoV-2 antibodies disclosed herein (i.e., theragnostics), or to determine a future course of treatment.
  • markers or reporters may also be useful in purifying anti-SARS-CoV-2 antibodies disclosed herein.
  • Diagnosis and detection can be accomplished by coupling the modulator to detectable substances including, but not limited to, various enzymes comprising for example horseradish peroxidase, alkaline phosphatase, beta-galactosidase, or acetylcholinesterase; prosthetic groups, such as but not limited to streptavidinlbiotin and avidin/biotin; fluorescent materials, such as but not limited to, umbelliferone, fluorescein, fluorescein isothiocynate, rhodamine, dichlorotriazinylamine fluorescein, dansyl chloride or phycoerythrin; luminescent materials, such as but not limited to, luminol; bioluminescent materials, such as but not limited to, luciferase, luciferin, and aequorin; radioactive materials, such as but not limited to iodine ( 131 I, 125 1, 123 1, 121 I,), carbon ( 14
  • the anti-SARS-CoV-2 antibodies disclosed herein can be fused to marker sequences, such as a peptide or fluorophore to facilitate purification or diagnostic procedures such as immunohistochemistry or FACs.
  • the marker amino acid sequence is a hexa-histidine peptide, such as the tag provided in a pQE vector, among others, many of which are commercially available.
  • Other peptide tags useful for purification include, but are not limited to, the hemagglutinin “HA” tag, and the “FLAG” tag.
  • nucleic acids encoding a heavy chain or light chain, or a VH or VL, of the anti-SARS-CoV-2 antibodies disclosed herein, and vectors comprising one or more such nucleic acids.
  • vectors comprising a first nucleic acid sequence encoding an antibody heavy chain and a second nucleic acid sequence encoding an antibody light chain.
  • compositions comprising a first vector comprising a nucleic acid sequence encoding an antibody heavy chain, and a second vector comprising a nucleic acid sequence encoding an antibody light chain.
  • nucleic acid sequences encoding an antibody heavy chain or light chain described herein are provided in SEQ ID NOs: 146-160.
  • nucleic acids comprising a sequence encoding an antibody heavy chain e.g., a nucleic acid comprising a sequence encoding SEQ ID NO: 9, 10, 20, 21, 27, or 28.
  • nucleic acids comprising a sequence encoding an antibody light chain e.g., a nucleic acid comprising a sequence encoding SEQ ID NO: 11, 15, 22, or 24.
  • vectors comprising a nucleic acid comprising a sequence encoding SEQ ID NO: 9, 10, 20, 21, 27, or 28, or a nucleic acid comprising a sequence encoding SEQ ID NO: 11, 15, 22, or 24.
  • the nucleic acids or vectors can comprise SEQ ID NO: 149, 150, or 151.
  • vectors comprising a first nucleic acid comprising a sequence encoding SEQ ID NO: 9,
  • compositions comprising a first vector comprising a sequence encoding SEQ ID NO: 9, 10, 20, 21, 27, or 28, and a second vector comprising a sequence encoding SEQ ID NO:
  • compositions can comprise a first vector comprising SEQ ID NO: 149 or 150, and a second vector comprising SEQ ID NO: 151 .
  • nucleic acids comprising a sequence encoding an antibody heavy chain, e.g., a nucleic acid comprising a sequence encoding SEQ ID NO: 37, 38, 46, 47, 51, or 52.
  • nucleic acids comprising a sequence encoding an antibody light chain, e.g., a nucleic acid comprising a sequence encoding SEQ ID NO: 39, 41, or 48.
  • vectors comprising a nucleic acid comprising a sequence encoding SEQ ID NO: 37, 38, 46, 47, 51, or 52, or a nucleic acid comprising a sequence encoding SEQ ID NO: 39, 41, or 48.
  • vectors comprising a first nucleic acid comprising a sequence encoding SEQ ID NO: 37, 38, 46, 47, 51, or 52, and a second nucleic acid comprising a sequence encoding SEQ ID NO: 39, 41, or 48.
  • compositions comprising a first vector comprising a sequence encoding SEQ ID NO: 37, 38, 46, 47, 51, or 52, and a second vector comprising a sequence encoding SEQ ID NO: 39, 41, or 48.
  • nucleic acids comprising a sequence encoding an antibody heavy chain, e.g., a nucleic acid comprising a sequence encoding SEQ ID NO: 61, 62, 75, or 76.
  • nucleic acids comprising a sequence encoding an antibody light chain, e.g., a nucleic acid comprising a sequence encoding SEQ ID NO: 63, 67, 70, or 77.
  • vectors comprising a nucleic acid comprising a sequence encoding SEQ ID NO: 61, 62, 75, or 76, or a nucleic acid comprising a sequence encoding SEQ ID NO: 63, 67, 70, or 77.
  • the nucleic acids or vectors can comprise SEQ ID NO: 152, 153, or 154.
  • vectors comprising a first nucleic acid comprising a sequence encoding SEQ ID NO: 61, 62, 75, or 76, and a second nucleic acid comprising a sequence encoding SEQ ID NO: 63, 67, 70, or 77.
  • the vectors can comprise a first nucleic acid comprising SEQ ID NO: 152 or 153, and a second nucleic acid comprising SEQ ID NO: 154.
  • compositions comprising a first vector comprising a sequence encoding SEQ ID NO: 61, 62, 75, or 76, and a second vector comprising a sequence encoding SEQ ID NO: 63, 67, 70, or 77.
  • the compositions can comprise a first vector comprising SEQ ID NO: 152 or 153, and a second vector comprising SEQ ID NO: 154.
  • nucleic acids comprising a sequence encoding an antibody heavy chain, e.g., a nucleic acid comprising a sequence encoding SEQ ID NO: 85, 86, 91, 92, 97, or 98.
  • nucleic acids comprising a sequence encoding an antibody light chain, e.g., a nucleic acid comprising a sequence encoding SEQ ID NO: 87 or 99.
  • vectors comprising a nucleic acid comprising a sequence encoding SEQ ID NO: 85, 86, 91, 92, 97, or 98, or a nucleic acid comprising a sequence encoding SEQ ID NO: 87 or 99.
  • the nucleic acids or vectors can comprise SEQ ID NO: 155, 156, or 157.
  • vectors comprising a first nucleic acid comprising a sequence encoding SEQ ID NO: 85, 86, 91, 92, 97, or 98, and a second nucleic acid comprising a sequence encoding SEQ ID NO: 87 or 99.
  • the vectors can comprise a first nucleic acid comprising SEQ ID NO: 155 or 156, and a second nucleic acid comprising SEQ ID NO: 157.
  • compositions comprising a first vector comprising a sequence encoding SEQ ID NO: 85, 86, 91, 92, 97, or 98, and a second vector comprising a sequence encoding SEQ ID NO: 87 or 99.
  • the compositions can comprise a first vector comprising SEQ ID NO: 155 or 156, and a second vector comprising SEQ ID NO: 157.
  • nucleic acids comprising a sequence encoding an antibody heavy chain, e.g., a nucleic acid comprising a sequence encoding SEQ ID NO: 108, 109, 113, 114, 121, 122, 124, 125, 128, or 129.
  • nucleic acids comprising a sequence encoding an antibody light chain, e.g., a nucleic acid comprising a sequence encoding SEQ ID NO: 110 or 117.
  • vectors comprising a nucleic acid comprising a sequence encoding SEQ ID NO: 108, 109, 113, 114, 121, 122, 124, 125, 128, or 129, or a nucleic acid comprising a sequence encoding SEQ ID NO: 110 or 117.
  • the nucleic acids or vectors can comprise SEQ ID NO: 158, 159, or 160.
  • vectors comprising a first nucleic acid comprising a sequence encoding SEQ ID NO: 108, 109, 113, 114, 121, 122, 124, 125, 128, or 129, and a second nucleic acid comprising a sequence encoding SEQ ID NO: 110 or 117.
  • the vectors can comprise a first nucleic acid comprising SEQ ID NO: 158 or 159, and a second nucleic acid comprising SEQ ID NO: 160.
  • compositions comprising a first vector comprising a sequence encoding SEQ ID NO: 108, 109, 113, 114, 121, 122, 124, 125, 128, or 129, and a second vector comprising a sequence encoding SEQ ID NO: 110 or 117.
  • the compositions can comprise a first vector comprising SEQ ID NO: 158 or 159, and a second vector comprising SEQ ID NO: 160.
  • nucleic acids comprising a sequence encoding an antibody heavy chain, e.g., a nucleic acid comprising a sequence encoding SEQ ID NO: 143 or 144.
  • the nucleic acids can comprise SEQ ID NO: 146 or 147.
  • nucleic acids comprising a sequence encoding an antibody light chain e.g., a nucleic acid comprising a sequence encoding SEQ ID NO: 145.
  • the nucleic acids can comprise SEQ ID NO: 148.
  • vectors comprising a nucleic acid comprising a sequence encoding SEQ ID NO: 143 or 144, or a nucleic acid comprising a sequence encoding SEQ ID NO: 145.
  • the vectors can comprise a nucleic acid comprising SEQ ID NO: 146, 147, or 148.
  • vectors comprising a first nucleic acid comprising a sequence encoding SEQ ID NO: 143 or 144, and a second nucleic acid comprising a sequence encoding SEQ ID NO: 145.
  • the vectors can comprise a first nucleic acid comprising SEQ ID NO: 146 or 147, and a second nucleic acid comprising SEQ ID NO: 148.
  • compositions comprising a first vector comprising a sequence encoding SEQ ID NO: 143 or 144, and a second vector comprising a sequence encoding SEQ ID NO: 145.
  • the compositions can comprise a first vector comprising SEQ ID NO: 146 or 147, and a second vector comprising SEQ ID NO: 148.
  • Nucleic acids of the present disclosure may be expressed in a host cell, for example, after the nucleic acids have been operably linked to an expression control sequence.
  • Expression control sequences capable of expression of nucleic acids to which they are operably linked are well known in the art.
  • An expression vector may include a sequence that encodes one or more signal peptides that facilitate secretion of the polypeptide(s) from a host cell.
  • Expression vectors containing a nucleic acid of interest e.g., a nucleic acid encoding a heavy chain or light chain of an antibody
  • expression vectors may contain one or more selection markers, e.g., tetracycline, neomycin, and dihydrofolate reductase, to aide in detection of host cells transformed with the desired nucleic acid sequences.
  • cells e.g., host cells, comprising the nucleic acids, vectors, or nucleic acid compositions described herein.
  • a host cell may be a cell stably or transiently transfected, transformed, transduced or infected with one or more expression vectors expressing all or a portion of an antibody described herein.
  • a host cell may be stably or transiently transfected, transformed, transduced or infected with an expression vector expressing HC and LC polypeptides of an antibody of the present disclosure.
  • a host cell may be stably or transiently transfected, transformed, transduced or infected with a first vector expressing HC polypeptides and a second vector expressing LC polypeptides of an antibody described herein.
  • Such host cells e.g., mammalian host cells, can express the anti-SARS-CoV-2 antibodies described herein.
  • Mammalian host cells known to be capable of expressing antibodies include CHO cells, HEK293 cells, COS cells, and NS0 cells.
  • the present disclosure further provides a process for producing an anti-SARS-CoV-2 antibody described herein by culturing the host cell described above, e.g., a mammalian host cell, under conditions such that the antibody is expressed and recovering the expressed antibody from the culture medium.
  • the culture medium, into which an antibody has been secreted may be purified by conventional techniques. Various methods of protein purification may be employed, and such methods are known in the art and described, for example, in Deutscher, Methods in Enzymology 182: 83-89 (1990) and Scopes, Protein Purification: Principles and Practice, 3rd Edition, Springer, NY (1994).
  • compositions comprising an anti-SARS-CoV-2 antibody described herein.
  • the compositions further comprise at least one additional antibody that binds the SARS-CoV-2 spike protein.
  • compositions e.g., pharmaceutical compositions
  • compositions comprising two or three anti-SARS- CoV-2 antibodies, wherein at least one antibody is an antibody described herein.
  • compositions e.g., pharmaceutical compositions
  • Such pharmaceutical compositions can also comprise one or more pharmaceutically acceptable excipient, diluent or carrier.
  • Pharmaceutical compositions can be prepared by methods well known in the art (e.g., Remington: The Science and Practice of Pharmacy, 22nd ed. (2012), A. Loyd et al., Pharmaceutical Press).
  • compositions comprising two or three anti-SARS-CoV-2 antibodies that bind different epitopes of the SARS-CoV-2 S protein.
  • compositions comprising a first anti-SARS-CoV-2 antibody that binds a first epitope in the RBD domain of the SARS-CoV-2 S protein, and a second anti-SARS-CoV-2 antibody that binds a second epitope of the SARS-CoV-2 S protein, wherein the second epitope is different from the first epitope.
  • the second anti-SARS-CoV-2 antibody also binds an epitope in the RBD domain, but does not overlap with the epitope of the first anti- SARS-CoV-2 antibody. In some embodiments, the second anti-SARS-CoV-2 antibody binds an epitope in the NTD domain.
  • compositions comprising two or three anti-SARS-CoV-2 antibodies, wherein at least one antibody binds the RBD domain of the SARS-CoV-2 S protein.
  • compositions comprising two or three anti-SARS- CoV-2 antibodies, wherein at least one antibody binds the NTD domain of the SARS-CoV-2 S protein.
  • compositions comprising two or three anti -SARS-CoV-2 antibodies, wherein at least one antibody binds the RBD domain of the SARS-CoV-2 S protein, and at least one antibody binds the NTD domain of the SARS-CoV-2 S protein.
  • the pharmaceutical compositions described herein comprise one or more anti -SARS-CoV-2 antibodies and one or more of the following excipients: histidine, sodium chloride, sucrose, polysorbate 80.
  • the pharmaceutical compositions comprise at least one anti-SARS-CoV-2 antibody, histidine, sodium chloride, sucrose, polysorbate 80.
  • the pharmaceutical compositions have a pH of about 6.0.
  • the pharmaceutical composition comprises at least one anti-SARS-CoV-2 antibody, 5 mM histidine, 50 mM NaCl, 6% sucrose, and 0.05% polysorbate 80 and has a pH of about 6.0.
  • the present disclosure also encompass in vitro or in vivo methods for detecting, diagnosing or monitoring coronavirus infections and methods of screening cells from a patient to identify coronavirus infected cells, including cells from a patient who is currently infected with SARS-CoV-2, or cells from a patient who is recovered from a past SARS-CoV-2 infection.
  • Such methods include identifying an individual infected with coronavirus for treatment, monitoring progression of a coronavirus infection by contacting the patient or a sample obtained from a patient with one or more anti-SARS-CoV-2 antibodies disclosed herein, and detecting the presence or absence, or level of association of the antibody to a coronavirus antigen in the sample.
  • one or more anti-SARS-CoV-2 antibodies disclosed herein may be used to detect and quantify coronavirus levels in a patient sample (e.g., plasma or blood).
  • methods of identifying a SARS-CoV-2- infected cell comprise: contacting a cell with an anti-SARS-CoV-2 antibody described herein, conjugated to a detectable agent; and detecting specific binding of the antibody to the cell.
  • methods of diagnosing a SARS-CoV-2 infection in a patient comprise: contacting a sample obtained from a patient with an anti-SARS-CoV-2 antibody described herein, conjugated to a detectable agent; and detecting specific binding of the antibody to a SARS-CoV-2 antigen present in the sample.
  • Detection of a coronavirus antigen in the sample likely suggests that the individual may be effectively treated with one or more anti-SARS-CoV-2 antibodies disclosed herein.
  • the methods may further comprise a step of comparing the level of binding to a control.
  • Other diagnostic or theragnostic methods compatible with the teachings herein are well known in the art and can be practiced using commercial materials such as dedicated reporting systems.
  • Exemplary compatible assay methods include radioimmunoassays, enzyme immunoassays, competitive - binding assays, fluorescent immunoassay, immunoblot assays, Western Blot analysis, flow cytometry assays, and ELISA assays (Enzyme-Linked ImmunoSorbent Assay).
  • More generally detection of coronavirus in a biological sample may be accomplished using any art-known assay.
  • Compatible in vivo theragnostics or diagnostics may comprise art recognized imaging or monitoring techniques such as magnetic resonance imaging (MRI), computerized tomography (e.g. CAT scan), positron tomography (e.g., PET scan) radiography, ultrasound, etc.
  • MRI magnetic resonance imaging
  • CAT scan computerized tomography
  • positron tomography e.g., PET scan
  • ultrasound etc.
  • the disclosure provides methods of analyzing coronavirus infection progression and/or pathogenesis in vivo, using one or more anti-SARS-CoV-2 antibodies disclosed herein.
  • kits for detecting, monitoring, or diagnosing a coronavirus infection, identifying an individual having a coronavirus infection for possible treatment or monitoring progression (or regression) of the infection in a patient wherein the kit comprises an anti-SARS-CoV-2 antibody as described herein, and reagents for detecting the effect of the anti-SARS-CoV-2 antibody on a sample from the patient.
  • Anti-SARS-CoV-2 antibodies disclosed herein and cells, cultures, populations and compositions comprising the same, including progeny thereof, can also be used to screen for or identify compounds or agents (e.g., drugs) that affect a function or activity of coronavirus virions or coronavirus infected cells or progeny thereof by binding to an antigen present on the surface of the virion or infected cell.
  • compounds or agents e.g., drugs
  • the disclosure therefore encompasses systems and methods for evaluation or identification of a compound or agent that can affect a function or activity of the coronavirus virus.
  • Such compounds and agents can be drug candidates that are screened for the treatment of coronavirus infection, for example.
  • a system or method comprises coronavirus virions and/or coronavirus infected cells and a compound or agent (e.g., drug), wherein the virions/cells and compound or agent (e.g., drug) are in contact with each other.
  • exemplary activity or function that can be modulated include changes in cell morphology, expression of a marker, differentiation or de-differentiation, maturation, proliferation, viability, apoptosis or cell death neuronal progenitor cells or progeny thereof.
  • kits for treating or preventing COVID- 19 by administering to a patient a therapeutically effective amount of one or more anti-SARS-CoV-2 antibodies described herein or a pharmaceutical composition comprising one or more anti-SARS- CoV-2 antibody described herein.
  • kits for treating or preventing COVID- 19 comprising: contacting a sample obtained from a patient with an antibody described herein, conjugated to a detectable agent; detecting specific binding of the antibody to a SARS-CoV-2 antigen present in the sample; and administering to the patient a therapeutically effective amount of an anti-SARS-CoV-2 antibody described herein or a pharmaceutical composition comprising an anti-SARS-CoV-2 antibody described herein.
  • the patient has moderate to severe COVID-19.
  • the patient has mild to moderate COVID- 19.
  • mild COVID- 19 patients can include individuals who have any of various signs and symptoms, e.g., fever, cough, sore throat, malaise, headache, muscle pain, without shortness of breath, dyspnea, or abnormal imaging.
  • Moderate COVID-19 patients can include individuals who have evidence of lower respiratory disease by clinical assessment or imaging and a saturation of oxygen (SaO2) greater than (>)93 percent (%) on room air at sea level.
  • the patient has not developed any symptoms of COVID-19.
  • the patient is at risk of developing COVID-19.
  • the patient is at risk of infection by SARS-CoV-2.
  • anti-SARS-CoV-2 antibodies or pharmaceutical compositions comprising one or more (e.g., two or three) anti-SARS-CoV-2 antibodies for use in therapy.
  • anti-SARS-CoV-2 antibodies or pharmaceutical compositions comprise one or more (e.g., two or three) anti-SARS-CoV-2 antibodies, for use in the treatment or prevention of COVID-19.
  • the anti-SARS-CoV-2 antibody may be administered to a patient, by various routes, including, but not limited to, intravenous or subcutaneous.
  • antibody refers to an immunoglobulin molecule that binds an antigen.
  • Embodiments of an antibody include a monoclonal antibody, polyclonal antibody, human antibody, humanized antibody, chimeric antibody, or conjugated antibody.
  • the antibodies can be of any class (e.g., IgG, IgE, IgM, IgD, IgA) and any subclass (e.g., IgGl, IgG2, IgG3, IgG4).
  • An exemplary antibody is an immunoglobulin G (IgG) type antibody comprised of four polypeptide chains: two heavy chains (HC) and two light chains (LC) that are cross-linked via inter-chain disulfide bonds.
  • the amino-terminal portion of each of the four polypeptide chains includes a variable region of about 100-125 or more amino acids primarily responsible for antigen recognition.
  • the carboxyl -terminal portion of each of the four polypeptide chains contains a constant region primarily responsible for effector function.
  • Each heavy chain is comprised of a heavy chain variable region (VH) and a heavy chain constant region.
  • Each light chain is comprised of a light chain variable region (VL) and a light chain constant region.
  • the IgG isotype may be further divided into subclasses (e.g., IgGl, IgG2, IgG3, and IgG4).
  • VH and VL regions can be further subdivided into regions of hyper-variability, termed complementarity determining regions (CDRs), interspersed with regions that are more conserved, termed framework regions (FR).
  • CDRs complementarity determining regions
  • FR framework regions
  • the CDRs are exposed on the surface of the protein and are important regions of the antibody for antigen binding specificity.
  • Each VH and VL is composed of three CDRs and four FRs, arranged from amino-terminus to carboxyl -terminus in the following order: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4.
  • the three CDRs of the heavy chain are referred to as “HCDR1, HCDR2, and HCDR3” and the three CDRs of the light chain are referred to as “LCDR1, LCDR2 and LCDR3”.
  • the CDRs contain most of the residues that form specific interactions with the antigen. Assignment of amino acid residues to the CDRs may be done according to the well-known schemes, including those described in Kabat (Kabat et al., “Sequences of Proteins of Immunological Interest,” National Institutes of Health, Bethesda, Md.
  • the present disclosure also include antibody fragments or antigen-binding fragments, which , as used herein, comprise at least a portion of an antibody retaining the ability to specifically interact with an antigen, such as Fab, Fab’, F(ab’)2, Fv fragments, scFv, scFab, disulfide-linked Fvs (sdFv), a Fd fragment and linear antibodies.
  • an antigen such as Fab, Fab’, F(ab’)2, Fv fragments, scFv, scFab, disulfide-linked Fvs (sdFv), a Fd fragment and linear antibodies.
  • bind and “binds” as used herein are intended to mean, unless indicated otherwise, the ability of a protein or molecule to form a chemical bond or attractive interaction with another protein or molecule, which results in proximity of the two proteins or molecules as determined by common methods known in the art.
  • Fc region refers to a region of an antibody, which comprises the CH2 and CH3 domains of the antibody heavy chain.
  • the Fc region may include a portion of the hinge region or the entire hinge region of the antibody heavy chain.
  • neutralizing antibody refers to an antibody that binds to or interacts with a virion and prevents binding or association of the virion with a host cell and/or entry into a host cell or acts as an egress inhibitor insofar as the antibody may not appear to be a neutralizing antibody in a conventional in vitro neutralization assay, but the antibody still inhibits propagation of the viral infection.
  • nucleic acid or “polynucleotide”, as used interchangeably herein, refer to polymers of nucleotides, including single-stranded and / or double-stranded nucleotide-containing molecules, such as DNA, cDNA and RNA molecules, incorporating native, modified, and / or analogs of, nucleotides.
  • Polynucleotides of the present disclosure may also include substrates incorporated therein, for example, by DNA or RNA polymerase or a synthetic reaction.
  • prevention As used herein, “prevention”, “prevent”, and / or “preventing”, which are used interchangeably herein, are intended to refer to all processes wherein there may be a slowing, interrupting, arresting, controlling, stopping, alleviating symptoms or complications or reversing of the progression of a disease, for example, caused by an injury, insult such as SARS-CoV-2 viral infection or disease such as COVID-19 disease. As used herein, prevention is not intended to necessarily indicate a total elimination of all disorder symptoms.
  • subject refers to a mammal, including, but are not limited to, a human, chimpanzee, ape, monkey, cattle, horse, sheep, goat, swine, rabbit, dog, cat, rat, mouse, guinea pig, and the like.
  • the subject is a human.
  • a therapeutically effective amount refers to an amount of a protein or nucleic acid or vector or composition that will elicit the biological or medical response of a subject, for example, reduction or inhibition of an enzyme or a protein activity, or ameliorate symptoms, alleviate conditions, slow or delay disease progression, or prevent a disease, etc.
  • a therapeutically effective amount refers to the amount of a protein or nucleic acid or vector or composition that, when administered to a subject, is effective to at least partially alleviate, inhibit, prevent and/or ameliorate a condition, or a disorder or a disease.
  • treatment refers to all processes wherein there may be a slowing, controlling, delaying or stopping of the progression of the disorders or disease disclosed herein, or ameliorating disorder or disease symptoms, but does not necessarily indicate a total elimination of all disorder or disease symptoms.
  • Treatment includes administration of a protein or nucleic acid or vector or composition for treatment of a disease or condition in a patient, particularly in a human.
  • FIGs. 1A-1G are bar graphs showing the results of the bead-based binding assays.
  • FIGs. 1 A-1B show the binding of the tested anti-SARS-CoV-2 antibodies to the full-length SARS- CoV-2 spike protein (FIG. 1A) or the SI subunit of SARS-CoV-2 spike protein (FIG. IB).
  • FIGs. 1C-1F show the binding of the tested anti-SARS-CoV-2 antibodies to the spike protein of SARS- CoV-1 (FIG. 1C), MERS (FIG. ID), WIV1 (FIG. IE), HKU1 (FIG. IF).
  • FIG. 1G shows none of the tested anti-SARS-CoV-2 antibodies bind to the negative control beads.
  • FIGs. 2A-2E are bar graphs showing the results of the cell-based binding assays.
  • FIG. 2A shows the binding of the tested anti-SARS-CoV-2 antibodies to SARS-CoV-2 full-length wildtype spike protein displayed on cells.
  • FIGs. 2B-2D show the binding of the tested anti-SARS- CoV-2 antibodies to the reported SARS-CoV-2 spike protein mutant: S50L (FIG. 2B), G476S (FIG. 2C) or D614G (FIG. 2D), displayed on cells.
  • FIG. 2E shows none of the tested anti-SARS- CoV-2 antibodies bind to the untransfected control cells.
  • Example 1 Generation of the engineered anti-SARS-CoV-2 antibodies
  • Anti-SARS-CoV-2 antibodies are identified from blood samples obtained from convalescent human patients with confirmed SARS-CoV-2 infection. After determining the VH and VL sequences of the antibodies that recognize SARS-CoV-2, they are converted to full-length IgGl antibody sequences for further characterization and engineering.
  • Lead antibodies 555, 419, 488, 481, and 373 are thoroughly characterized, as described in a separate patent application. Based on the specific profile of each antibody, specific engineering strategies and processes are used.
  • Antibodies 555 and 488 are engineered using a phage system.
  • the VH and VL sequences of the antibodies are cloned into an internal phage system vector for expression of Fabs in E. coli, construction of CDR mutant libraries by Kunkel mutagenesis, and screen of affinity improved variants by filter lift assay. All CDR residues are scanned with random single mutation libraries by the filter lift assays specifically optimized for hits that improve k on (association rate constant) and/or k O ff (dissociate rate constant). All hits are picked and verified by DNA sequencing. Clones containing unique CDR mutations are expressed in E. coli. Variant Fab proteins are extracted from E. coli periplasm.
  • the crude extracts or Fab proteins purified by Nickel -NTA bead are analyzed for affinity improvements using Octet BLI and the standard ELISA Fab titration.
  • a selected set of beneficial k on and k O ff mutations are combined in a total combinatorial library and screened by the same assays described above under more stringent conditions.
  • the final combinatorial (combi.) variants are converted into IgGl for further characterization.
  • the CDR mutagenesis of 78 positions is used to enhance the affinity and potency of the parental 419 Fab.
  • Libraries are created using NNK random codon-based mutagenic oligonucleotides incorporated using Kunkel mutagenesis into an uracil-containing single-stranded DNA template encoding the original parental 419 Fab with the selected CDR sequences deleted.
  • Phage-expressed Fabs are screened by capture lift assay using in-house expressed biotinylated human SARS-CoV2 SI spike trimer antigen.
  • Affinity enhanced mutations identified in this manner are DNA sequenced and unique clones expressed as periplasmic Fab in E. coli and crude supernatants analyzed by titration ELISA.
  • This process identifies multiple CDR amino acid substitutions with increased affinity.
  • Beneficial CDR mutations are combined in a library allowing all the beneficial mutations to be randomly combined or remain wild type sequence.
  • Combinatorial libraries are screened under more stringent conditions including lower antigen concentration and increased washing at 37°C.
  • Hits are once again DNA sequenced and unique variants screened by ELISA.
  • the ELISA includes a one-hour binding step followed by a three-hour 37°C wash step to magnify differences in dissociation rates.
  • the final combi Fab variants are converted into IgGl for further characterization. Following expression and purification, several different ELISA formats are utilized to differentiate IgG based on relative affinities.
  • IgG is captured using an anti-Fc coated plate.
  • Monomeric NTD domain of SARS-CoV-2 spike protein with an Avi-tag fusion to enable in vivo biotinylation is subsequently titrated and binding detected using an alkaline phosphatase conjugated neutravidin reagent.
  • This process identifies several higher affinity IgG variants containing multiple CDR residue changes.
  • a SARS-CoV-2 spike trimeric antigen is used to coat an ELISA plate. IgG variants are titrated and detected with an anti -human Fc alkaline phosphatase conjugate.
  • binding is detected but the off-rate differences are decreased due to avidity.
  • This process leads to the identification of four higher affinity variants termed, 419-WT-3, 419-WT-15, 419- FrF-10 and 419-FrF-10.
  • Antibodies 481 and 373 are engineered as full-length antibodies in mammalian cell expression vectors using a high throughput site-specific mutagenesis protocol to find mutations that improve affinity and potentially mitigate developability issues (expression, chemical and biophysical stability, solubility) without impacting potency or leading to further developability issues based on amino acid property and percent exposure to solvent using molecular models. Briefly, every amino acid in the CDRs of both the VL and VH chains is mutated in individual mutagenesis reactions to the 18 variants (minus Cys), as well as back to the original residue (embedded wild-type, WT), using a series of forward and reverse oligo’ s arrayed in a 384 microtiter plate.
  • Site-directed mutagenesis reactions are carried out according to established procedures and digestion of the WT plasmid accomplished by incubation with the Dpnl restriction enzyme. Digestion products are transformed into E. coll and DNA isolated from the bulk transformants following incubation overnight at 37°C. DNA from each individual VL and VH mutagenesis reaction is mixed with the appropriate WT antibody variable region and expressed in CHO cells in a 96-deep well plate. Secreted antibodies are quantified and normalized to a consistent titer prior to screening in downstream assays.
  • the WT 481 mAb has a low affinity for RBD (for a monoclonal antibody) with a KD in the 10-7 M range.
  • the WT 481 antibody also contains an asparagine in LCDR3 (N92) that deamidates under stressful conditions.
  • the saturation mutagenesis and screening indicate only N92I mutation eliminates the deamidation without dramatically reducing the affinity of the mAb. Mutation at LCDR3 position 93 to residues known to inhibit deamidation also lead to loss of binding, thus, N92I is the best choice to eliminate any deamidation.
  • the parental 373 mAb has relatively low affinity (>10-6 KD).
  • the only developability issues within the 373 parental mAb are (i) high hydrophobicity as measured by interactions with hydrophobic chromatography resins due to a hydrophobic cluster in the HCDR3 of parental antibody 373 (...FWSGYF%), and (ii) potential for oxidation at HCRD3 MIOOi.
  • the saturation mutagenesis screening provides the team with information to reduce the hydrophobicity and eliminate the potential for methionine oxidation while significantly improving the affinity.
  • All the engineered 373 variants contain mutations that address both the developability and improve the potency.
  • the optimized 481 and 373 variants are also measured for non-specific binding and found they have very low nonspecific binding potential.
  • the optimized 555, 488, 419, 481, and 373 VH domains are all cloned onto a human IgGl backbone or a human IgGl-ACT5 backbone containing mutations that increase the binding affinity to the neonatal Fc receptor (FcRn), which have been shown to improve the in vivo half-life of human IgGl (Maeda A, et al., 2017. MAbs 9, 844-53).
  • FcRn neonatal Fc receptor
  • the anti-SARS-CoV-2 antibodies are produced by expression in Chinese hamster ovary (CHO) cells and purified using standard antibody purification techniques.
  • a ForteBio Octet® RED384 Bio-Layer Interferometry system (Octet RED384, ForteBio, CA) is used to screen antigen binding kinetics of antibody variants at 30 °C with a shake speed of 1000 rpm. Briefly, biotinylated SARS-CoV-2 spike SI protein or the receptor binding domain (RBD) (Sino Biologicals, Cat#40592-V08H) is diluted to 1 pg/mL in the assay buffer (PBS with 0.5% BSA and 0.05% Tween-20). Antibody proteins are diluted 1: 1 in the same assay buffer.
  • antibody affinity can be measured by capturing Fab protein on FAB2G tip (ForteBio, Catalog 18-5125) and dipping into antigen solution to obtain the binding kinetics.
  • MSD MA6000 384 well plate: MSD, Cat#L21XA-4) are prepared by adding 35 pL/well of PBS overnight at 4°C. Plates are washed 3 times with PBST (PBS + 0.1% Tween 20). Next, PBS is added to the plates for 0.5-1 hours at room temperature. The PBS solution is then removed and 20 mL of each test mAh (4 replicates per sample) is diluted to 5 or 10 mg/mL in PBS either directly from cell culture supernatants or from purified material and added to the MSD plate.
  • PBST PBS + 0.1% Tween 20
  • the affinity and binding kinetics of selected anti-SARS-CoV-2 antibodies to the receptor binding domain (RBD) or N-terminal binding domain (NTD) of the SARS-CoV-2 spike protein, or the full-length SARS-CoV-2 spike protein, is determined by SPR using Biacore 8K (GE Healthcare). The procedure generally follows the “Instrument Handbook”. The amino acid sequences of the RBD, NTD, or full-length SARS-CoV-2 spike protein, used for these experiments are provided in Table 2.
  • Antibody is first captured on Biacore Protein A chip followed by flowing a serial concentration of RBD or NTD from 100 nM down to 0.391 nM in 2-fold serial dilution in PBS- P20-BSA (0.005% surfactant P20, 0. Img/mL BSA).
  • polyclonal goat anti -human IgG-Fc Jackson Immunology cat#109005098
  • goat anti -kappa and goat anti-lambda are directly immobilized onto Series S CM5 sensor chips (GE Healthcare Cat.: #29104988) using standard amine coupling (NHS/EDC/ethanolamine) protocols provided by the manufacturer’s protocols and a 10 mM Acetate pH 4.5 binding buffer/HBS-EP running buffer. Immobilization levels are between 8-10K resonance units (RUs).
  • All kinetic binding experiments are performed at 37 °C.
  • Multi-cycle kinetics setting that runs each analyte concentration in a separate cycle regenerating the surface after each sample injection is used.
  • the test articles (optimized 555, 488, 481, 419, and 373 mAbs) are diluted to 10-100 pg/mL in PBS pH 7.4 and captured onto these chip surfaces to a level of 50-200 RU by injection at 10 pL/min using an HBS-EP running buffer.
  • recombinantly-derived SARS-Cov-2 receptor binding domain (RBD) or N-terminal binding domain (NTD) antigens are diluted into HBS-EP at 200, 100, 50, 25, 12.5, 6.25, 3.13 and 0 nM and injected (180-300 s) in succession over the sensor chip surfaces containing the captured mAb variants. Dissociation of the antigen is allowed to occur between 600 and 1800 s depending on the off rate (kd). Between each antigen injection, mAb and non-dissociated antigen are removed from the sensor chip surface with injection of 10 mM glycine, pH 1.5, buffer (regeneration). The regeneration is optimized to maintain consistent surface properties from cycle to cycle. The data is fit to a 1: 1 binding model to derive ka and kd, and to calculate KD (Table 3).
  • Each cycle starts with injection of antibody at 50 pl/min flow rate for 3 minutes and a dissociation in PBS-P20 for 15 minutes.
  • the chip surface is then regenerated with injection of 25 pl of 10 mM glycine buffer, pH 1.5, at 50 pl/min flow rate for three times.
  • the experiments are carried out at 37 °C.
  • the data is fit to a bivalent binding model to derive the following parameters (shown in Table 4).
  • Table 3 Binding Kinetics and Affinity of the engineered anti-SARS-CoV-2 Antibodies to RBD or NTD of anti-SARS-CoV-2 spike protein
  • Engineered mAbs are subjected to several biophysical characterizations including analytical size exclusion chromatography (SEC-HPLC), hydrophobic interaction chromatography - high performance liquid chromatography (HIC-HPLC), heparin chromatography (Heparin-HPLC), cross-interaction chromatography (CIC), and affinity-capture self-interaction nanoparticle spectroscopy (AC-SINS). See Table 5.
  • SEC-HPLC analytical size exclusion chromatography
  • HIC-HPLC hydrophobic interaction chromatography - high performance liquid chromatography
  • Heparin-HPLC heparin chromatography
  • CIC cross-interaction chromatography
  • AC-SINS affinity-capture self-interaction nanoparticle spectroscopy
  • CIC is performed as described previously (Jacobs, S.A., et al., Pharm. Res. 27, 65- 71, 2010).
  • the CIC column is prepared by coupling ⁇ 30 mg of human serum polyclonal antibodies (14506; Sigma) to a 1-mL HITRAP® column (17-0716-01; GE Healthcare), followed by quenching with ethanolamine.
  • the blank column is prepared in the same except without human serum IgGs.
  • Approximately 20 pg of each antibody is tested at a flow rate of 0.2 mL/min using lOmM sodium citrate, lOmM NaCl, pH 6.5 as a mobile phase on an Agilent 1260 series HPLC system. Retention times obtained by both IgG and blank columns are used to calculate k’ .
  • peak width at 50% height is also obtained to monitor non-specific interaction of test antibodies.
  • the AC-SINS assay is performed as described previously with modifications (Wu, J. et al., Protein Eng. Des. Sei. 28, 403-14, 2015).
  • gold nanoparticles (15705; Ted Pella Inc.) are coated with 100% capturing anti-human goat IgG Fc (109-005-008; Jackson ImmunoResearch).
  • the conjugation reaction is quenched with I g/ml polyethylene glycol (PEG) and eluted into 0.25 x PBS. PEG is added to the conjugated gold mixture at a concentration of 0.2 pg/ml prior to use.
  • the antibodies of interest are then incubated with the coated gold particles for 1 h at room temperature and the wavelength shift is measured using Tecan M1000 Pro Plate Reader within the range of 475-625 nm, in increments of 1 nm.
  • Test antibodies are diluted in either PBS or 10 mM histidine, pH 6.0 prior to incubation. Delta plasma wavelength shift in comparison with buffer control is reported.
  • the self-interacting antibodies show a higher wavelength shift away from the buffer controls.
  • Binding of the anti-SARS-CoV-2 antibodies to full-length wildtype SARS-CoV-2 spike protein or the reported viral mutants (with S50L, G476S or D614G mutation) displayed on cells is tested using a high throughput flow cytometry.
  • CHO cells are transiently transfected to express the full length spike protein of either wild type SARS-CoV-2 or mutant SARS-CoV-2 (with S50L, G476S, or D614G mutation) on the cell surface.
  • Purified antibodies are incubated with the readout cells, and an untransfected control CHO line at 50 nM antibody concentration for 30 minutes at 4°C.
  • CHO cells are washed; and the binding is detected by using a fluorescently labeled anti-human secondary antibody. Fluorescence is measured using high throughput platebased flow cytometry. Benchmark antibodies identified for SARS-CoV-1 are used as positive controls due to similarity in spike sequences between SARS-CoV-1 and SARS-CoV-2; human IgG isotype and an irrelevant antibody are used as negative controls.
  • Binding of the anti-SARS-CoV-2 antibodies to SARS-CoV-2 spike protein, the SI subunit of SARS-CoV-2 spike protein, or the spike protein of other coronaviruses SARS-CoV-1, MERS, HKU1, WIV1, is tested using a multiplexed bead assay on high throughput flow cytometry.
  • Different optically encoded bead types are conjugated to one of the following unique antigens: full length spike protein of SARS-CoV-2, MERS, SARS-CoV-1, HKU1, WIV1 or the SI subunit of SARS-CoV-2 spike protein.
  • Purified antibodies are incubated with the multiplexed beads, and negative control beads conjugated to BSA-His or FoldOn-His at 50 nM antibody concentration for 30 minutes at room temperature. Beads are washed and the binding is detected by using a fluorescently labeled anti-human secondary antibody. Fluorescence is measured using high throughput plate-based flow cytometry. Benchmark antibodies identified for SARS-CoV-1 are used as positive controls due to similarity in spike sequences between SARS-CoV-1 and SARS-CoV-2; human IgG isotype and an irrelevant antibody are used as negative controls.
  • FIGs. 1A-1G show the results of the bead-based binding assays.
  • FIGs. 2A-2E show the results of the cell-based binding assays. All tested antibodies show binding to the wildtype SARS-CoV-2 spike protein and the selected SARS-CoV-2 spike protein mutants (S50L, G476S, D614G) displayed on cells (FIGs. 2A-2D). All tested antibodies also show binding to the SARS- CoV-2 spike protein and the SI subunit of SARS-CoV-2 spike protein on beads (FIGs. 1A and IB). None of the tested antibodies show binding to the untransfected control cells (FIG. 2E) or the negative control beads (FIG. 1G).
  • the anti-SARS-CoV-2 antibodies 555-com-A8, 555-com- D2, 555-com-H3 and 555-com-H7 show weak cross-reactivity to WIV1 spike proteins on beads (FIG. IE).
  • the anti-SARS-CoV-2 antibodies 419-FrF-15, 419-WT-3 and 419-WT-15 show over 100-fold binding to HKU1 spike protein on beads, indicating cross reactivity to HKU1 (FIG. IF).
  • Vero E6 cells African green monkey kidney; ATCC Cat. #CRL-1586. These cells are known to be highly susceptible to infection by SARS-CoV-2. Cells are maintained according to standard ATCC protocols. Briefly, Vero E6 cells are grown in minimal essential medium (MEM) supplemented with 10% heat-inactivated fetal bovine serum (FBS), 2 mM L-glutamine, and 1% MEM nonessential amino acid (NEAA) solution (Fisher Cat. #MT25025CI). Cell cultures are grown in 75 or 150 cm 2 flasks at 37 °C in 5% CO2 and passaged 2 to 3 times per week using trypsin-EDTA. Cell cultures used for virus testing are prepared as subconfluent monolayers.
  • MEM minimal essential medium
  • FBS heat-inactivated fetal bovine serum
  • NEAA nonessential amino acid
  • Assays are conducted using virus produced by infecting cultured Vero E6 cells with the SARS-CoV-2 clinical isolate USA/WA/1/2020 (BEI resources number NR52281) or the Italy- FNMI1 isolate of SARS-CoV-2 (European Virus Archive - Global, ref #008V-03893), and incubating at 37 °C until cytopathology is evident (typically 48 to 72 hours). Expansion is limited to 1 to 2 passages in cell culture to retain integrity of the original viral sequence. The virus stock is quantified by standard plaque assay, and aliquots are stored at -80°C. A freshly-thawed aliquot is used for each neutralization experiment.
  • the anti -SARS-CoV-2 antibodies and an IgGl isotype control are prepared in phosphate-buffered saline (PBS) at 1 mg/mL. Plaque reduction assays are performed in 6-well plates. Vero E6 cells are seeded at a concentration of approximately 10 6 cells/well and grown overnight at 37°C in 5% CO2 to reach 95% confluency. The next day, serial three-fold dilutions of anti-SARS-CoV-2 antibodies or an IgGl isotype control are prepared in Eagle’s minimal essential medium, mixed with approximately 100 pfu of SARS-CoV-2, and incubated for 1 to 2 hours on ice or at 37°C.
  • PBS phosphate-buffered saline
  • the mAb/virus mixture is inoculated directly onto the cells and allowed to adsorb for 1 hour at 37 °C with 5% CO2, with rocking at 15 minute intervals.
  • An overlay media composed of 1.25% Avicel RC-581 (FMC Biopolymer) in Eagles minimum essentials medium (MEM) with 5% FBS is added, and plates are incubated for 48 hours at 37°C with 5% CO2 for virus plaques to develop. After incubation, overlays are removed by aspiration and the cells are fixed with 10% buffered formalin containing crystal violet stain for 1 hour. Plaques are counted manually, and plaque forming units are determined by averaging technical replicates per sample.
  • Percent neutralization is calculated relative to the signals produced by an IgGl isotype control or a no-virus control, and the data are plotted using nonlinear regression with a four- parameter fit analysis (GraphPad Prism v8.0.0).
  • IC50 is estimated using a meta-analysis using a random effects model (Berkey, et al., Stat Med. 1995; 14(4):395-411) and the R package meta for (Viechtbauer, J. Stat. Software 2010;36(3): l-48).
  • SEQ ID NO: 1 Heavy chain variable region of Antibody 555-com-Al
  • SEQ ID NO: 2 Light chain variable region of Antibody 555-com-Al
  • SEQ ID NO: 3 Heavy chain complementarity determining region 1 of Antibody 555-com-Al KASGGTFDNYAIS
  • SEQ ID NO: 4 Heavy chain complementarity determining region 2 of Antibody 555-com-Al RIIPILGIAN
  • SEQ ID NO: 5 Heavy chain complementarity determining region 3 of Antibody 555-com-Al ARTYYEARHYYYYYAMDV
  • SEQ ID NO: 6 Light chain complementarity determining region 1 of Antibody 555-com-Al RASQSISSYLS
  • SEQ ID NO: 7 Light chain complementarity determining region 2 of Antibody 555-com-Al YAASDLQS
  • SEQ ID NO: 8 Light chain complementarity determining region 3 of Antibody 555-com-Al QQSYSSPRT
  • SEQ ID NO: 9 Heavy chain 1 - hlgGl of Antibody 555-com-Al
  • SEQ ID NO: 10 Heavy chain 2 - h!gGl-ACT5 of Antibody 555-com-Al
  • SEQ ID NO: 11 Light chain of Antibody 555-com-Al
  • SEQ ID NO: 12 Light chain variable region of Antibody 555-com-D2
  • SEQ ID NO: 13 Light chain complementarity determining region 2 of Antibody 555-com-D2 YAASSLQS
  • SEQID NO: 14 Light chain complementarity determining region 3 of Antibody 555-com-D2 QQSHSPPRT
  • SEQ ID NO: 15 Light chain of Antibody 555-com-D2
  • SEQ ID NO: 16 Heavy chain variable region of Antibody 555-com-H3
  • SEQ ID NO: 17 Light chain variable region of Antibody 555-com-H3
  • SEQ ID NO: 18 Heavy chain complementarity determining region 3 of Antibody 555-com-H3
  • SEQ ID NO: 19 Light chain complementarity determining region 3 of Antibody 555-com-H3 QQSHSSPRT
  • SEQ ID NO: 20 Heavy chain 1 - hlgGl of Antibody 555-com-H3
  • SEQ ID NO: 21 Heavy chain 2 - h!gGl-ACT5 of Antibody 555-com-H3
  • SEQ ID NO: 22 Light chain of Antibody 555-com-H3
  • SEQ ID NO: 23 Light chain variable region of Antibody 555-com-D7
  • SEQ ID NO: 24 Light chain of Antibody 555-com-D7
  • SEQ ID NO: 25 Heavy chain variable region of Antibody 555-com-H7
  • SEQ ID NO: 26 Heavy chain complementarity determining region 3 of Antibody 555-com-H7 ARTYYEARHYYYVYAMDV
  • SEQ ID NO: 27 Heavy chain 1 - hlgGl of Antibody 555-com-H7
  • SEQ ID NO: 28 Heavy chain 2 - hIgGl-ACT5 of Antibody 555-com-H7
  • SEQ ID NO: 29 Heavy chain variable region of Antibody 419-Wild Type-3
  • SEQ ID NO: 30 Light chain variable region of Antibody 419-Wild Type-3
  • SEQ ID NO: 31 Heavy chain complementarity determining region 1 of Antibody 419-Wild Type-3 KVSGYTFTSRYMH
  • SEQ ID NO: 32 Heavy chain complementarity determining region 2 of Antibody 419-Wild Type-3 IINPSSGSTSYAQKFQG
  • SEQ ID NO: 33 Heavy chain complementarity determining region 3 of Antibody 419-Wild Type-3 ARDWTQSSGYDYYYGLDV
  • SEQ ID NO: 34 Light chain complementarity determining region 1 of Antibody 419-Wild Type-3 SGDALPKQYAY
  • SEQ ID NO: 35 Light chain complementarity determining region 2 of Antibody 419-Wild Type-3 KDSERYS
  • SEQ ID NO: 36 Light chain complementarity determining region 3 of Antibody 419-Wild Type-3 QSADSSGTYLV
  • SEQ ID NO: 37 Heavy chain 1 - hlgGl of Antibody 419-Wild Type-3
  • SEQ ID NO: 38 Heavy chain 2 - hlgGl -ACT5 of Antibody 419-Wild Type -3
  • SEQ ID NO: 39 Light chain of Antibody 419-Wild Type -3
  • SEQ ID NO: 40 Light chain variable region of Antibody 419-FrF-10
  • SEQ ID NO: 42 Heavy chain variable region of Antibody 419-Wild Type-15
  • SEQ ID NO: 43 Light chain variable region of Antibody 419-Wild Type-15
  • SEQ ID NO: 44 Heavy chain complementarity determining region 3 of Antibody 419-Wild Type-15 ARDWTQTSGYDYYYGLDV
  • SEQ ID NO: 45 Light chain complementarity determining region 3 of Antibody 419-Wild Type-15 QSADSSGTYVV
  • SEQ ID NO: 46 Heavy chain 1 - hlgGl of Antibody 419-Wild Type-15
  • SEQ ID NO: 47 Heavy chain 2 - h!gGl-ACT5 of Antibody 419-Wild Type-15
  • SEQ ID NO: 48 Light chain of Antibody 419-Wild Type-15
  • SEQ ID NO: 49 Heavy chain variable region of Antibody 419-FrF-15
  • SEQ ID NO: 50 Light chain complementarity determining region 3 of Antibody 419-FrF-15 ARDLTRSSGYDYYYGLDV
  • SEQ ID NO: 51 Heavy chain 1 - hlgGl of Antibody 419-FrF- 15
  • SEQ ID NO: 52 Heavy chain 2 - h!gGl-ACT5 of Antibody 419-FrF-15
  • SEQ ID NO: 53 Heavy chain variable region of Antibody 488-2A4
  • SEQ ID NO: 54 Light chain variable region of Antibody 488-2A4
  • SEQ ID NO: 55 Heavy chain complementarity determining region 1 of Antibody 488-2A4
  • SEQ ID NO: 56 Heavy chain complementarity determining region 2 of Antibody 488-2A4
  • SEQ ID NO: 57 Heavy chain complementarity determining region 3 of Antibody 488-2A4
  • SEQ ID NO: 58 Light chain complementarity determining region 1 of Antibody 488-2A4
  • SEQ ID NO: 59 Light chain complementarity determining region 2 of Antibody 488-2A4
  • SEQ ID NO: 60 Light chain complementarity determining region 3 of Antibody 488-2A4
  • SEQ ID NO: 61 Heavy chain 1 - hlgGl of Antibody 488-2A4 EVQLVESGGGLIQPGGSLRLSCAASGLTVSSNYMSWVRQAPGKGLEWVSVIYPGGSTYYADSVK
  • SEQ ID NO: 62 Heavy chain 2 - hIgGl-ACT5 of Antibody 488-2A4
  • SEQ ID NO: 63 Light chain of Antibody 488-2A4
  • SEQ ID NO: 64 Light chain variable region of Antibody 488-2A6
  • SEQ ID NO: 65 Light chain complementarity determining region 1 of Antibody 488-2A6 QASQDTMNYLN
  • SEQ ID NO: 66 Light chain complementarity determining region 2 of Antibody 488-2A6
  • SEQ ID NO: 67 Light chain of Antibody 488-2A6 DIQMTQSPSSLSASVGDRVTITCQASQDTMNYLNWYQQKPGKAPKLLIYRASNLETGVPSRFSGS GSGTDFTFTISSLQPEDIATYYCQQYDNLPITFGQGTRLEIKRTVAAPSVFIFPPSDEQLKSGTASVV CLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEV THQGLSSPVTKSFNRGEC
  • SEQ ID NO: 68 Light chain variable region of Antibody 488-2C5
  • SEQ ID NO: 69 Light chain complementarity determining region 1 of Antibody 488-2C5 QASQDIMNYLN
  • SEQ ID NO: 70 Light chain of Antibody 488-2C5
  • SEQ ID NO: 71 Heavy chain variable region of Antibody 488-2G1
  • SEQ ID NO: 72 Light chain variable region of Antibody 488-2G1
  • SEQ ID NO: 73 Heavy chain complementarity determining region 2 of Antibody 488-2G1 TIYSGGSTY
  • SEQ ID NO: 74 Light chain complementarity determining region 1 of Antibody 488-2G1 QASQDTQNYLN
  • SEQ ID NO: 75 Heavy chain 1 - hlgGl of Antibody 488-2G1 EVQLVESGGGLIQPGGSLRLSCAASGLTVSSNYMSWVRQAPGKGLEWVSTIYSGGSTYYADSVK GRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARSPYGGNVWGQGTLVTVSSASTKGPSVFPLAP SSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGT QTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTC VVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCK
  • SEQ ID NO: 76 Heavy chain 2 - hIgGl-ACT5 of Antibody 488-2G1
  • SEQ ID NO: 77 Light chain of Antibody 488-2G1
  • SEQ ID NO: 78 Heavy chain variable region of Antibody 481-L1H1
  • SEQ ID NO: 79 Light chain variable region of Antibody 481-L1H1
  • SEQ ID NO: 80 Heavy chain complementarity determining region 1 of Antibody 481-L1H1
  • AASVFTVSYNYMS SEQ ID NO: 81: Heavy chain complementarity determining region 2 of Antibody 481-L1H1
  • SEQ ID NO: 82 Heavy chain complementarity determining region 3 of Antibody 481-L1H1 AREVAGTYDY
  • SEQ ID NO: 83 Light chain complementarity determining region 1 of Antibody 481-L1H1 RASQGISSWLA
  • SEQ ID NO: 84 Light chain complementarity determining region 3 of Antibody 481-L1H1 QDAISFPGGT
  • SEQ ID NO: 85 Heavy chain 1 - hlgGl of Antibody 481-L1H1
  • SEQ ID NO: 86 Heavy chain 2 - h!gGl-ACT5 of Antibody 481-L1H1
  • SEQ ID NO: 87 Light chain of Antibody 481-L1H1
  • SEQ ID NO: 88 Heavy chain variable region of Antibody 481-L1H4
  • SEQ ID NO: 89 Heavy chain complementarity determining region 1 of Antibody 481-L1H4 AASGITVSSNYMS
  • SEQ ID NO: 90 Heavy chain complementarity determining region 3 of Antibody 481-L1H4 AREVYGTYDY
  • SEQ ID NO: 91 Heavy chain 1 - hlgGl of Antibody 481-L1H4
  • SEQ ID NO: 92 Heavy chain 2 - hIgGl-ACT5 of Antibody 481-L1H4
  • SEQ ID NO: 93 Heavy chain variable region of Antibody 481-L5H2
  • GRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAREVAGTYDYWGQGTLVTVSS SEQ ID NO: 94: Light chain variable region of Antibody 481-L5H2
  • SEQ ID NO: 95 Heavy chain complementarity determining region 1 of Antibody 481-L2H5 AASVFTVSSNYMS
  • SEQ ID NO: 96 Light chain complementarity determining region 3 of Antibody 481-L2H5 QQTISFPGGT
  • SEQ ID NO: 97 Heavy chain 1 - hlgGl of Antibody 481-L2H5
  • SEQ ID NO: 98 Heavy chain 2 - hIgGl-ACT5 of Antibody 481-L2H5
  • SEQ ID NO: 99 Light chain of Antibody 481-L2H5
  • SEQ ID NO: 101 Light chain variable region of Antibody 373-L1H21
  • SEQ ID NO: 102 Heavy chain complementarity determining region 1 of Antibody 373-L1H21 SGGSISVYHWE
  • SEQ ID NO: 103 Heavy chain complementarity determining region 2 of Antibody 373-L1H21 YIKYSGSTNYNP
  • SEQ ID NO: 104 Heavy chain complementarity determining region 3 of Antibody 373-L1H21
  • SEQ ID NO: 105 Light chain complementarity determining region 1 of Antibody 373-L1H21 RASQSVESSYLA
  • SEQ ID NO: 106 Light chain complementarity determining region 2 of Antibody 373-L1H21 GASSRAT
  • SEQ ID NO: 107 Light chain complementarity determining region 3 of Antibody 373-L1H21 QQYGSSTT
  • SEQ ID NO: 108 Heavy chain 1 - hlgGl of Antibody 373-L1H21
  • SEQ ID NO: 109 Heavy chain 2 - h!gGl-ACT5 of Antibody 373-L1H21
  • SEQ ID NO: 110 Light chain of Antibody 373-L1H21
  • SEQ ID NO: 111 Heavy chain variable region of Antibody 373-L1H24
  • SEQ ID NO: 112 Heavy chain complementarity determining region 1 of Antibody 373-L1H24 SGHSISSYHWE
  • SEQ ID NO: 113 Heavy chain 1 - hlgGl of Antibody 373-L1H24
  • SEQ ID NO: 114 Heavy chain 2 - h!gGl-ACT5 of Antibody 373-L1H24
  • SEQ ID NO: 115 Light chain variable region of Antibody 373-L2H24
  • SEQ ID NO: 116 Light chain complementarity determining region 1 of Antibody 373-L2H24 RASQSVDTSYLA
  • SEQ ID NO: 117 Light chain of Antibody 373-L2H24
  • SEQ ID NO: 118 Heavy chain variable region of Antibody 373-L2H10
  • SEQ ID NO: 119 Heavy chain complementarity determining region 2 of Antibody 373-L2H10
  • YIYYSGSTNYNP SEQ ID NO: 120 Heavy chain complementarity determining region 3 of Antibody 373-L2H10
  • SEQ ID NO: 121 Heavy chain 1 - hlgGl of Antibody 373-L2H10
  • SEQ ID NO: 122 Heavy chain 2 - hlgGl -ACT5 of Antibody 373-L2H10
  • SEQ ID NO: 123 Heavy chain variable region of Antibody 373-L2H20
  • SEQ ID NO: 124 Heavy chain 1 - hlgGl of Antibody 373-L2H20
  • SEQ ID NO: 125 Heavy chain 2 - hIgGl-ACT5 of Antibody 373-L2H20
  • SEQ ID NO: 126 Heavy chain variable region of Antibody 373-L2H61
  • SEQ ID NO: 127 Heavy chain complementarity determining region 3 of Antibody 373-L2H61 ARGPDWYDFWAGYYYGTDV
  • SEQ ID NO: 128 Heavy chain 1 - hlgGl of Antibody 373-L2H61
  • SEQ ID NO: 129 Heavy chain 2 - hlgGl -ACT5 of Antibody 373-L2H61
  • SEQ ID NO: 130 SARS-CoV-2 spike (S) protein (GenBank Accession No: YP_009724390.1) MFVFLVLLPLVSSQCVNLTTRTQLPPAYTNSFTRGVYYPDKVFRSSVLHSTQDLFLPFFSNVTWF HAIHVSGTNGTKRFDNPVLPFNDGVYFASTEKSNIIRGWIFGTTLDSKTQSLLIVNNATNVVIKVC EFQFCNDPFLGVYYHKNNKSWMESEFRVYSSANNCTFEYVSQPFLMDLEGKQGNFKNLREFVF KNIDGYFKIYSKHTPINLVRDLPQGFSALEPLVDLPIGINITRFQTLLALHRSYLTPGDSSSGWTAG AAAYYVGYLQPRTFLLKYNENGTITDAVDCALDPLSETKCTLKSFTVEKGIYQTSNFRVQPTESI VRFPNITNLCPFGEVFNATRFASVYAWNRKRISNCVADYSVLYNSASFSTFK
  • SEQ ID NO: 131 Receptor binding domain- AviHis tag antigen
  • SEQ ID NO: 133 N-terminal binding domain -A viHis tag antigen
  • SEQ ID NO: 134 Full-length SARS-CoV-2 spike protein-A viHis tag

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Abstract

The present disclosure relates to antibodies that bind SARS-CoV-2 spike protein (anti-SARS-CoV-2 antibodies), compositions comprising such anti-SARS-CoV-2 antibodies, and methods of using such anti-SARS-CoV-2 antibodies.

Description

ANTI-SARS-COV-2 ANTIBODIES
FIELD OF THE INVENTION
[0001] The present disclosure relates to antibodies that bind SARS-CoV-2 spike protein (“anti- SARS-CoV-2 antibodies”), compositions comprising such anti-SARS-CoV-2 antibodies, and methods of using such anti-SARS-CoV-2 antibodies.
BACKGROUND
[0002] Coronaviruses are a large family of viruses that infect mammals and birds, with four endemic strains that circulate commonly in humans, including human coronavirus 229E, OC43, NL63 and HKU1. These endemic strains generally cause mild flu-like symptoms, but can also cause more serious pneumonia in vulnerable populations. However, when new coronaviruses jump from zoonotic hosts (e.g., bats, birds, camels) to humans, they can result in a much more severe respiratory disease that can spread quickly through the population. Examples of such outbreaks include SARS (Severe Acute Respiratory Syndrome) in 2003, caused by SARS- associated coronavirus (SARS-CoV), which resulted in an outbreak that infected over 8,000 people worldwide and was -10% fatal, and MERS (Middle Eastern Respiratory Syndrome) in 2012, caused by MERS-CoV, which to date has infected over 2,500 people with approximately 35% fatalities.
[0003] The new coronavirus of 2019, named SARS-CoV-2, causing a severe respiratory disease referred to as COVID-19, has spread rapidly on a global scale, with over 20 million confirmed cases in more than 216 countries and regions to date and led to over 700,000 deaths. SARS-CoV-2 is the most serious coronavirus outbreak in history. There is an urgent need for antibody-based therapeutics, prophylactics, and diagnostics to treat, prevent, and detect SARS- CoV-2 infection and COVID- 19.
DETAILED DESCRIPTION
[0004] In one aspect, provided herein are antibodies that bind the spike (S) protein of SARS- CoV-2. In some embodiments, the anti-SARS-CoV-2 antibodies are engineered antibodies that have one or more of the following properties: 1) bind to SARS-CoV-2 spike protein with desirable binding affinities and/or association and dissociation rates, 2) neutralize SARS-CoV-2 virus with desired potency, and/or 3) have a long half-life and good stability, solubility, viscosity, and/or pharmacokinetic characteristics for development and use in the treatment of COVID-19.
[0005] The cryo-electron microscopy structure of the SARS-CoV-2 S protein has been described recently, and shows the SARS-CoV-2 S protein is a trimeric class I fusion protein that exists in a metastable pre-fusion conformation and undergoes a substantial structural rearrangement to fuse the viral membrane with the host cell membrane (Wrapp et al., Science 367,
1260-1263, 2020). This process is triggered when the receptor-binding domain (RBD) of the SI subunit of the SARS-CoV-2 S protein binds to the host cell receptor angiotensin-converting enzyme 2 (ACE2); and receptor binding destabilizes the pre-fusion trimer, resulting in shedding of the SI subunit and transition of the S2 subunit to a stable post-fusion conformation (Wrapp et al., Science 367, 1260-1263, 2020).
[0006] The amino acid sequence of SARS-CoV-2 spike (S) protein has been described before, for example, GenBank Accession No: YP 009724390.1 provides such an exemplary sequence, as shown below:
MFVFLVLLPLVS SQC VNLTTRTQLPPAYTNSFTRGVYYPDK VFRS S VLHSTQDLFLPFF S NVTWFHAIHVSGTNGTKRFDNPVLPFNDGVYFASTEKSNIIRGWIFGTTLDSKTQSLLIV NNATNVVIKVCEFQFCNDPFLGVYYHKNNKSWMESEFRVYSSANNCTFEYVSQPFLMD LEGKQGNFKNLREFVFKNIDGYFKIYSKHTPINLVRDLPQGFSALEPLVDLPIGINITRFQT LLALHRSYLTPGDSSSGWTAGAAAYYVGYLQPRTFLLKYNENGTITDAVDCALDPLSET KCTLKSFTVEKGIYQTSNFRVQPTESIVRFPNITNLCPFGEVFNATRFASVYAWNRKRISN CVADYSVLYNSASFSTFKCYGVSPTKLNDLCFTNVYADSFVIRGDEVRQIAPGQTGKIA DYNYKLPDDFTGCVIAWNSNNLDSKVGGNYNYLYRLFRKSNLKPFERDISTEIYQAGST PCNGVEGFNCYFPLQSYGFQPTNGVGYQPYRVWLSFELLHAPATVCGPKKSTNLVKN KCVNFNFNGLTGTGVLTESNKKFLPFQQFGRDIADTTDAVRDPQTLEILDITPCSFGGVS VITPGTNTSNQVAVLYQDVNCTEVPVAIHADQLTPTWRVYSTGSNVFQTRAGCLIGAEH VNNSYECDIPIGAGICASYQTQTNSPRRARSVASQSIIAYTMSLGAENSVAYSNNSIAIPT NFTISVTTEILPVSMTKTSVDCTMYICGDSTECSNLLLQYGSFCTQLNRALTGIAVEQDK NTQEVFAQVKQIYKTPPIKDFGGFNFSQILPDPSKPSKRSFIEDLLFNKVTLADAGFIKQY GDCLGDIAARDLICAQKFNGLTVLPPLLTDEMIAQYTSALLAGTITSGWTFGAGAALQIP FAMQMAYRFNGIGVTQNVLYENQKLIANQFNSAIGKIQDSLSSTASALGKLQDVVNQN AQALNTLVKQLS SNFGAIS SVLNDILSRLDKVEAEVQIDRLITGRLQ SLQTYVTQQLIRAA EIRASANLAATKMSECVLGQSKRVDFCGKGYHLMSFPQSAPHGVVFLHVTYVPAQEKN FTTAPAICHDGKAHFPREGVFVSNGTHWFVTQRNFYEPQIITTDNTFVSGNCDWIGIVN NT VYDPLQPELD SFKEELDK YFKNHT SPD VDLGDI SGI N AS V VNIQ KE IDRLNE VAKNLN ESLID LQELGKYEQYIKWPWYIWLGFIAGLIAIVMVTIMLCCMTSCCSCLKGCCSCGSCC KFDEDDSEPVLKGVKLHYT (SEQ ID NO: 130).
[0007] The receptor-binding domain (RBD) comprises amino acids 319 to 592 of the SARS- CoV-2 S protein, e.g., amino acids 319 to 592 of SEQ ID NO: 130 or a variant comprising one or more mutations; the N-terminal domain (NTD) comprises amino acids 16-307 of the SARS-CoV- 2 S protein, e.g., amino acids 16-307 of SEQ ID NO: 130 or a variant comprising one or more mutations; S2 domain comprises amino acids 686-1213 of the SARS-CoV-2 S protein, e.g., amino acids 686-1213 of SEQ ID NO: 130 or a variant comprising one or more mutations (see Wrapp et al., Science 367, 1260-1263, 2020).
[0008] In some embodiments, provided herein are anti -SARS-CoV-2 antibodies comprising a heavy chain variable region (VH) and a light chain variable region (VL), wherein the VH comprises heavy chain complementarity determining regions HCDR1, HCDR2, and HCDR3, and the VL comprises light chain complementarity determining regions LCDR1, LCDR2, and LCDR3, wherein the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 are selected from the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 in Table 1. In some embodiments, provided herein are anti-SARS-CoV-2 antibodies that comprise a VH domain comprising the same three HCDRs as comprised in the VH domain of any anti-SARS-CoV-2 antibody in Table 1. In some embodiments, provided herein are anti-SARS-CoV-2 antibodies that comprise a VL domain comprising the same three LCDRs as the VL domain of any anti-SARS-CoV-2 antibody in Table 1. In some embodiments, provided herein are anti-SARS-CoV-2 antibodies that comprise a VH domain comprising the same three HCDRs as comprised in the VH domain of any anti-SARS- CoV-2 antibody in Table 1, and a VL domain comprising the same three LCDRs as the VL domain of any anti-SARS-CoV-2 antibody in Table 1. In some embodiments, provided herein are anti- SARS-CoV-2 antibodies that comprise the same VH and/or the VL of any anti-SARS-CoV-2 antibody in Table 1. In some embodiments, provided herein are anti-SARS-CoV-2 antibodies that comprise a VH having at least 95% sequence identity to any of the VHs in Table 1, and a VL having at least 95% sequence identity to any of the VLs in Table 1 .
[0009] In some embodiments, the anti-SARS-CoV-2 antibodies described herein have IgGl isotype. In some embodiments, provided herein are anti-SARS-CoV-2 antibodies comprising a heavy chain (HC) selected from any HC in Table 1, and/or a light chain (LC) selected from any LC in Table 1. In some embodiments, provided herein are anti-SARS-CoV-2 antibodies that comprise a HC having at least 95% sequence identity to any of the HCs in Table 1, and a LC having at least 95% sequence identity to any of the LCs in Table 1.
Table 1. Sequences of Exemplary Anti-SARS-CoV-2 Antibodies
Figure imgf000006_0001
Figure imgf000007_0001
Figure imgf000008_0001
Figure imgf000009_0001
Figure imgf000010_0001
Figure imgf000011_0001
Figure imgf000012_0001
Figure imgf000013_0001
Figure imgf000014_0001
Figure imgf000015_0001
Figure imgf000016_0001
Figure imgf000017_0001
Figure imgf000018_0001
Figure imgf000019_0001
Figure imgf000020_0001
Figure imgf000021_0001
Figure imgf000022_0001
Figure imgf000023_0001
Figure imgf000024_0001
Figure imgf000025_0001
Figure imgf000026_0001
[0010] In some embodiments, provided herein is an antibody that binds SARS-CoV-2 spike protein, wherein the antibody comprises a VH and a VL, wherein the VH comprises HCDR1, HCDR2, and HCDR3, and the VL comprises LCDR1, LCDR2, and LCDR3, wherein the antibody comprises: HCDR1 comprising SEQ ID NO: 3, HCDR2 comprising SEQ ID NO: 4, HCDR3 comprising SEQ ID NO: 5, 18, or 26, LCDR1 comprising SEQ ID NO: 6, LCDR2 comprising SEQ ID NO: 7 or 13, and LCDR3 comprising SEQ ID NO: 8, 14, or 19.
[0011] In some embodiments, provided herein is an antibody that binds SARS-CoV-2 spike protein, wherein the antibody comprises a VH and a VL, wherein the VH comprises HCDR1, HCDR2, and HCDR3, and the VL comprises LCDR1, LCDR2, and LCDR3, wherein the antibody comprises: HCDR1 comprising SEQ ID NO: 3, HCDR2 comprising SEQ ID NO: 4, HCDR3 comprising SEQ ID NO: 5, LCDR1 comprising SEQ ID NO: 6, LCDR2 comprising SEQ ID NO: 7, and LCDR3 comprising SEQ ID NO: 8.
[0012] In some embodiments, provided herein is an antibody that binds SARS-CoV-2 spike protein, wherein the antibody comprises a VH and a VL, wherein the VH comprises HCDR1, HCDR2, and HCDR3, and the VL comprises LCDR1, LCDR2, and LCDR3, wherein the antibody comprises: HCDR1 comprising SEQ ID NO: 3, HCDR2 comprising SEQ ID NO: 4, HCDR3 comprising SEQ ID NO: 5, LCDR1 comprising SEQ ID NO: 6, LCDR2 comprising SEQ ID NO: 13, and LCDR3 comprising SEQ ID NO: 14.
[0013] In some embodiments, provided herein is an antibody that binds SARS-CoV-2 spike protein, wherein the antibody comprises a VH and a VL, wherein the VH comprises HCDR1, HCDR2, and HCDR3, and the VL comprises LCDR1, LCDR2, and LCDR3, wherein the antibody comprises: HCDR1 comprising SEQ ID NO: 3, HCDR2 comprising SEQ ID NO: 4, HCDR3 comprising SEQ ID NO: 18, LCDR1 comprising SEQ ID NO: 6, LCDR2 comprising SEQ ID NO: 7, and LCDR3 comprising SEQ ID NO: 19.
[0014] In some embodiments, provided herein is an antibody that binds SARS-CoV-2 spike protein, wherein the antibody comprises a VH and a VL, wherein the VH comprises HCDR1, HCDR2, and HCDR3, and the VL comprises LCDR1, LCDR2, and LCDR3, wherein the antibody comprises: HCDR1 comprising SEQ ID NO: 3, HCDR2 comprising SEQ ID NO: 4, HCDR3 comprising SEQ ID NO: 5, LCDR1 comprising SEQ ID NO: 6, LCDR2 comprising SEQ ID NO: 13, and LCDR3 comprising SEQ ID NO: 19.
[0015] In some embodiments, provided herein is an antibody that binds SARS-CoV-2 spike protein, wherein the antibody comprises a VH and a VL, wherein the VH comprises HCDR1, HCDR2, and HCDR3, and the VL comprises LCDR1, LCDR2, and LCDR3, wherein the antibody comprises: HCDR1 comprising SEQ ID NO: 3, HCDR2 comprising SEQ ID NO: 4, HCDR3 comprising SEQ ID NO: 26, LCDR1 comprising SEQ ID NO: 6, LCDR2 comprising SEQ ID NO: 7, and LCDR3 comprising SEQ ID NO: 8.
[0016] In some embodiments, provided herein is an antibody that binds SARS-CoV-2 spike protein, wherein the antibody comprises a VH and a VL, wherein the VH comprises HCDR1, HCDR2, and HCDR3, and the VL comprises LCDR1, LCDR2, and LCDR3, wherein the antibody comprises: HCDR1 comprising SEQ ID NO: 3, HCDR2 comprising SEQ ID NO: 4, HCDR3 comprising SEQ ID NO: 18, LCDR1 comprising SEQ ID NO: 6, LCDR2 comprising SEQ ID NO: 7, and LCDR3 comprising SEQ ID NO: 8.
[0017] In some embodiments, provided herein is an antibody that binds SARS-CoV-2 spike protein, wherein the antibody comprises a VH and a VL, wherein the VH comprises HCDR1, HCDR2, and HCDR3, and the VL comprises LCDR1, LCDR2, and LCDR3, wherein the antibody comprises: HCDR1 comprising SEQ ID NO: 31, HCDR2 comprising SEQ ID NO: 32, HCDR3 comprising SEQ ID NO: 33, 44, or 50, LCDR1 comprising SEQ ID NO: 34, LCDR2 comprising SEQ ID NO: 35, and LCDR3 comprising SEQ ID NO: 36 or 45.
[0018] In some embodiments, provided herein is an antibody that binds SARS-CoV-2 spike protein, wherein the antibody comprises a VH and a VL, wherein the VH comprises HCDR1, HCDR2, and HCDR3, and the VL comprises LCDR1, LCDR2, and LCDR3, wherein the antibody comprises: HCDR1 comprising SEQ ID NO: 31, HCDR2 comprising SEQ ID NO: 32, HCDR3 comprising SEQ ID NO: 33, LCDR1 comprising SEQ ID NO: 34, LCDR2 comprising SEQ ID NO: 35, and LCDR3 comprising SEQ ID NO: 36.
[0019] In some embodiments, provided herein is an antibody that binds SARS-CoV-2 spike protein, wherein the antibody comprises a VH and a VL, wherein the VH comprises HCDR1, HCDR2, and HCDR3, and the VL comprises LCDR1, LCDR2, and LCDR3, wherein the antibody comprises: HCDR1 comprising SEQ ID NO: 31, HCDR2 comprising SEQ ID NO: 32, HCDR3 comprising SEQ ID NO: 44, LCDR1 comprising SEQ ID NO: 34, LCDR2 comprising SEQ ID NO: 35, and LCDR3 comprising SEQ ID NO: 45.
[0020] In some embodiments, provided herein is an antibody that binds SARS-CoV-2 spike protein, wherein the antibody comprises a VH and a VL, wherein the VH comprises HCDR1, HCDR2, and HCDR3, and the VL comprises LCDR1, LCDR2, and LCDR3, wherein the antibody comprises: HCDR1 comprising SEQ ID NO: 31, HCDR2 comprising SEQ ID NO: 32, HCDR3 comprising SEQ ID NO: 50, LCDR1 comprising SEQ ID NO: 34, LCDR2 comprising SEQ ID NO: 35, and LCDR3 comprising SEQ ID NO: 36.
[0021] In some embodiments, provided herein is an antibody that binds SARS-CoV-2 spike protein, wherein the antibody comprises a VH and a VL, wherein the VH comprises HCDR1, HCDR2, and HCDR3, and the VL comprises LCDR1, LCDR2, and LCDR3, wherein the antibody comprises: HCDR1 comprising SEQ ID NO: 55, HCDR2 comprising SEQ ID NO: 56 or 73, HCDR3 comprising SEQ ID NO: 57, LCDR1 comprising SEQ ID NO: 58, 65, 69, or 74, LCDR2 comprising SEQ ID NO: 59 or 66, and LCDR3 comprising SEQ ID NO: 60. [0022] In some embodiments, provided herein is an antibody that binds SARS-CoV-2 spike protein, wherein the antibody comprises a VH and a VL, wherein the VH comprises HCDR1, HCDR2, and HCDR3, and the VL comprises LCDR1, LCDR2, and LCDR3, wherein the antibody comprises: HCDR1 comprising SEQ ID NO: 55, HCDR2 comprising SEQ ID NO: 56, HCDR3 comprising SEQ ID NO: 57, LCDR1 comprising SEQ ID NO: 58, LCDR2 comprising SEQ ID NO: 59, and LCDR3 comprising SEQ ID NO: 60.
[0023] In some embodiments, provided herein is an antibody that binds SARS-CoV-2 spike protein, wherein the antibody comprises a VH and a VL, wherein the VH comprises HCDR1, HCDR2, and HCDR3, and the VL comprises LCDR1, LCDR2, and LCDR3, wherein the antibody comprises: HCDR1 comprising SEQ ID NO: 55, HCDR2 comprising SEQ ID NO: 56, HCDR3 comprising SEQ ID NO: 57, LCDR1 comprising SEQ ID NO: 65, LCDR2 comprising SEQ ID NO: 66, and LCDR3 comprising SEQ ID NO: 60.
[0024] In some embodiments, provided herein is an antibody that binds SARS-CoV-2 spike protein, wherein the antibody comprises a VH and a VL, wherein the VH comprises HCDR1, HCDR2, and HCDR3, and the VL comprises LCDR1, LCDR2, and LCDR3, wherein the antibody comprises: HCDR1 comprising SEQ ID NO: 55, HCDR2 comprising SEQ ID NO: 56, HCDR3 comprising SEQ ID NO: 57, LCDR1 comprising SEQ ID NO: 69, LCDR2 comprising SEQ ID NO: 59, and LCDR3 comprising SEQ ID NO: 60.
[0025] In some embodiments, provided herein is an antibody that binds SARS-CoV-2 spike protein, wherein the antibody comprises a VH and a VL, wherein the VH comprises HCDR1, HCDR2, and HCDR3, and the VL comprises LCDR1, LCDR2, and LCDR3, wherein the antibody comprises: HCDR1 comprising SEQ ID NO: 55, HCDR2 comprising SEQ ID NO: 73, HCDR3 comprising SEQ ID NO: 57, LCDR1 comprising SEQ ID NO: 74, LCDR2 comprising SEQ ID NO: 59, and LCDR3 comprising SEQ ID NO: 60.
[0026] In some embodiments, provided herein is an antibody that binds SARS-CoV-2 spike protein, wherein the antibody comprises a VH and a VL, wherein the VH comprises HCDR1, HCDR2, and HCDR3, and the VL comprises LCDR1, LCDR2, and LCDR3, wherein the antibody comprises: HCDR1 comprising SEQ ID NO: 80, 89, or 95, HCDR2 comprising SEQ ID NO: 81, 1 HCDR3 comprising SEQ ID NO: 82 or 90, LCDR1 comprising SEQ ID NO: 83, LCDR2 comprising SEQ ID NO: 13, and LCDR3 comprising SEQ ID NO: 84 or 96.
[0027] In some embodiments, provided herein is an antibody that binds SARS-CoV-2 spike protein, wherein the antibody comprises a VH and a VL, wherein the VH comprises HCDR1, HCDR2, and HCDR3, and the VL comprises LCDR1, LCDR2, and LCDR3, wherein the antibody comprises: HCDR1 comprising SEQ ID NO: 80, HCDR2 comprising SEQ ID NO: 81, HCDR3 comprising SEQ ID NO: 82, LCDR1 comprising SEQ ID NO: 83, LCDR2 comprising SEQ ID NO: 13, and LCDR3 comprising SEQ ID NO: 84.
[0028] In some embodiments, provided herein is an antibody that binds SARS-CoV-2 spike protein, wherein the antibody comprises a VH and a VL, wherein the VH comprises HCDR1, HCDR2, and HCDR3, and the VL comprises LCDR1, LCDR2, and LCDR3, wherein the antibody comprises: HCDR1 comprising SEQ ID NO: 89, HCDR2 comprising SEQ ID NO: 81, HCDR3 comprising SEQ ID NO: 90, LCDR1 comprising SEQ ID NO: 83, LCDR2 comprising SEQ ID NO: 13, and LCDR3 comprising SEQ ID NO: 84.
[0029] In some embodiments, provided herein is an antibody that binds SARS-CoV-2 spike protein, wherein the antibody comprises a VH and a VL, wherein the VH comprises HCDR1, HCDR2, and HCDR3, and the VL comprises LCDR1, LCDR2, and LCDR3, wherein the antibody comprises: HCDR1 comprising SEQ ID NO: 95, HCDR2 comprising SEQ ID NO: 81, HCDR3 comprising SEQ ID NO: 82, LCDR1 comprising SEQ ID NO: 83, LCDR2 comprising SEQ ID NO: 13, and LCDR3 comprising SEQ ID NO: 96.
[0030] In some embodiments, provided herein is an antibody that binds SARS-CoV-2 spike protein, wherein the antibody comprises a VH and a VL, wherein the VH comprises HCDR1, HCDR2, and HCDR3, and the VL comprises LCDR1, LCDR2, and LCDR3, wherein the antibody comprises: HCDR1 comprising SEQ ID NO: 80, HCDR2 comprising SEQ ID NO: 81, HCDR3 comprising SEQ ID NO: 82, LCDR1 comprising SEQ ID NO: 83, LCDR2 comprising SEQ ID NO: 13, and LCDR3 comprising SEQ ID NO: 96.
[0031] In some embodiments, provided herein is an antibody that binds SARS-CoV-2 spike protein, wherein the antibody comprises a VH and a VL, wherein the VH comprises HCDR1, HCDR2, and HCDR3, and the VL comprises LCDR1, LCDR2, and LCDR3, wherein the antibody comprises: HCDR1 comprising SEQ ID NO: 102 or 112, HCDR2 comprising SEQ ID NO: 103 or 119, HCDR3 comprising SEQ ID NO: 104, 120, or 127, LCDR1 comprising SEQ ID NO: 105 or 116, LCDR2 comprising SEQ ID NO: 106, and LCDR3 comprising SEQ ID NO: 107.
[0032] In some embodiments, provided herein is an antibody that binds SARS-CoV-2 spike protein, wherein the antibody comprises a VH and a VL, wherein the VH comprises HCDR1, HCDR2, and HCDR3, and the VL comprises LCDR1, LCDR2, and LCDR3, wherein the antibody comprises: HCDR1 comprising SEQ ID NO: 102, HCDR2 comprising SEQ ID NO: 103, HCDR3 comprising SEQ ID NO: 104, LCDR1 comprising SEQ ID NO: 105, LCDR2 comprising SEQ ID NO: 106, and LCDR3 comprising SEQ ID NO: 107.
[0033] In some embodiments, provided herein is an antibody that binds SARS-CoV-2 spike protein, wherein the antibody comprises a VH and a VL, wherein the VH comprises HCDR1, HCDR2, and HCDR3, and the VL comprises LCDR1, LCDR2, and LCDR3, wherein the antibody comprises: HCDR1 comprising SEQ ID NO: 112, HCDR2 comprising SEQ ID NO: 103, HCDR3 comprising SEQ ID NO: 104, LCDR1 comprising SEQ ID NO: 105, LCDR2 comprising SEQ ID NO: 106, and LCDR3 comprising SEQ ID NO: 107.
[0034] In some embodiments, provided herein is an antibody that binds SARS-CoV-2 spike protein, wherein the antibody comprises a VH and a VL, wherein the VH comprises HCDR1, HCDR2, and HCDR3, and the VL comprises LCDR1, LCDR2, and LCDR3, wherein the antibody comprises: HCDR1 comprising SEQ ID NO: 112, HCDR2 comprising SEQ ID NO: 103, HCDR3 comprising SEQ ID NO: 104, LCDR1 comprising SEQ ID NO: 116, LCDR2 comprising SEQ ID NO: 106, and LCDR3 comprising SEQ ID NO: 107.
[0035] In some embodiments, provided herein is an antibody that binds SARS-CoV-2 spike protein, wherein the antibody comprises a VH and a VL, wherein the VH comprises HCDR1, HCDR2, and HCDR3, and the VL comprises LCDR1, LCDR2, and LCDR3, wherein the antibody comprises: HCDR1 comprising SEQ ID NO: 112, HCDR2 comprising SEQ ID NO: 119, HCDR3 comprising SEQ ID NO: 120, LCDR1 comprising SEQ ID NO: 116, LCDR2 comprising SEQ ID NO: 106, and LCDR3 comprising SEQ ID NO: 107. [0036] In some embodiments, provided herein is an antibody that binds SARS-CoV-2 spike protein, wherein the antibody comprises a VH and a VL, wherein the VH comprises HCDR1, HCDR2, and HCDR3, and the VL comprises LCDR1, LCDR2, and LCDR3, wherein the antibody comprises: HCDR1 comprising SEQ ID NO: 112, HCDR2 comprising SEQ ID NO: 119, HCDR3 comprising SEQ ID NO: 104, LCDR1 comprising SEQ ID NO: 116, LCDR2 comprising SEQ ID NO: 106, and LCDR3 comprising SEQ ID NO: 107.
[0037] In some embodiments, provided herein is an antibody that binds SARS-CoV-2 spike protein, wherein the antibody comprises a VH and a VL, wherein the VH comprises HCDR1, HCDR2, and HCDR3, and the VL comprises LCDR1, LCDR2, and LCDR3, wherein the antibody comprises: HCDR1 comprising SEQ ID NO: 112, HCDR2 comprising SEQ ID NO: 119, HCDR3 comprising SEQ ID NO: 127, LCDR1 comprising SEQ ID NO: 116, LCDR2 comprising SEQ ID NO: 106, and LCDR3 comprising SEQ ID NO: 107.
[0038] In some embodiments, provided herein is an antibody that binds SARS-CoV-2 spike protein, wherein the antibody comprises a VH and a VL, wherein the VH comprises heavy chain complementarity determining regions HCDR1, HCDR2, andHCDR3, and the VL comprises light chain complementarity determining regions LCDR1, LCDR2, and LCDR3, wherein the antibody comprises: HCDR1 comprising SEQ ID NO: 137, HCDR2 comprising SEQ ID NO: 138, HCDR3 comprising SEQ ID NO: 139, LCDR1 comprising SEQ ID NO: 140, LCDR2 comprising SEQ ID NO: 141, and LCDR3 comprising SEQ ID NO: 142.
[0039] In some embodiments, provided herein is an antibody that binds SARS-CoV-2 spike protein, wherein the antibody comprises a VH comprising SEQ ID NO: 1, 16 or 25, and a VL comprising SEQ ID NO: 2, 12, 17, or 23.
[0040] In some embodiments, provided herein is an antibody that binds SARS-CoV-2 spike protein, wherein the antibody comprises a VH comprising SEQ ID NO: 1, and a VL comprising SEQ ID NO: 2. In some embodiments, provided herein is an antibody that binds SARS-CoV-2 spike protein, wherein the antibody comprises a VH comprising SEQ ID NO: 1, and a VL comprising SEQ ID NO: 12. In some embodiments, provided herein is an antibody that binds SARS-CoV-2 spike protein, wherein the antibody comprises a VH comprising SEQ ID NO: 16, and a VL comprising SEQ ID NO: 17. In some embodiments, provided herein is an antibody that binds SARS-CoV-2 spike protein, wherein the antibody comprises a VH comprising SEQ ID NO: 1, and a VL comprising SEQ ID NO: 23. In some embodiments, provided herein is an antibody that binds SARS-CoV-2 spike protein, wherein the antibody comprises a VH comprising SEQ ID NO: 25, and a VL comprising SEQ ID NO: 2. In some embodiments, provided herein is an antibody that binds SARS-CoV-2 spike protein, wherein the antibody comprises a VH comprising SEQ ID NO: 16, and a VL comprising SEQ ID NO: 2.
[0041] In some embodiments, provided herein is an antibody that binds SARS-CoV-2 spike protein, wherein the antibody comprises a VH comprising SEQ ID NO: 29, 42, or 49, and a VL comprising SEQ ID NO: 30, 40, 43.
[0042] In some embodiments, provided herein is an antibody that binds SARS-CoV-2 spike protein, wherein the antibody comprises a VH comprising SEQ ID NO: 29, and a VL comprising SEQ ID NO: 30. In some embodiments, provided herein is an antibody that binds SARS-CoV-2 spike protein, wherein the antibody comprises a VH comprising SEQ ID NO: 29, and a VL comprising SEQ ID NO: 40. In some embodiments, provided herein is an antibody that binds SARS-CoV-2 spike protein, wherein the antibody comprises a VH comprising SEQ ID NO: 42, and a VL comprising SEQ ID NO: 43. In some embodiments, provided herein is an antibody that binds SARS-CoV-2 spike protein, wherein the antibody comprises a VH comprising SEQ ID NO: 49, and a VL comprising SEQ ID NO: 40.
[0043] In some embodiments, provided herein is an antibody that binds SARS-CoV-2 spike protein, wherein the antibody comprises a VH comprising SEQ ID NO: 53 or 71, and a VL comprising SEQ ID NO: 54, 64, 68, or 72.
[0044] In some embodiments, provided herein is an antibody that binds SARS-CoV-2 spike protein, wherein the antibody comprises a VH comprising SEQ ID NO: 53, and a VL comprising SEQ ID NO: 54. In some embodiments, provided herein is an antibody that binds SARS-CoV-2 spike protein, wherein the antibody comprises a VH comprising SEQ ID NO: 53, and a VL comprising SEQ ID NO: 64. In some embodiments, provided herein is an antibody that binds SARS-CoV-2 spike protein, wherein the antibody comprises a VH comprising SEQ ID NO: 53, and a VL comprising SEQ ID NO: 68. In some embodiments, provided herein is an antibody that binds SARS-CoV-2 spike protein, wherein the antibody comprises a VH comprising SEQ ID NO: 71, and a VL comprising SEQ ID NO: 72.
[0045] In some embodiments, provided herein is an antibody that binds SARS-CoV-2 spike protein, wherein the antibody comprises a VH comprising SEQ ID NO: 78, 88 or 93, and a VL comprising SEQ ID NO: 79 or 94.
[0046] In some embodiments, provided herein is an antibody that binds SARS-CoV-2 spike protein, wherein the antibody comprises a VH comprising SEQ ID NO: 78, and a VL comprising SEQ ID NO: 79. In some embodiments, provided herein is an antibody that binds SARS-CoV-2 spike protein, wherein the antibody comprises a VH comprising SEQ ID NO: 88, and a VL comprising SEQ ID NO: 79. In some embodiments, provided herein is an antibody that binds SARS-CoV-2 spike protein, wherein the antibody comprises a VH comprising SEQ ID NO: 93, and a VL comprising SEQ ID NO: 94. In some embodiments, provided herein is an antibody that binds SARS-CoV-2 spike protein, wherein the antibody comprises a VH comprising SEQ ID NO: 78, and a VL comprising SEQ ID NO: 94.
[0047] In some embodiments, provided herein is an antibody that binds SARS-CoV-2 spike protein, wherein the antibody comprises a VH comprising SEQ ID NO: 100, 111, 118, 123, or 126, and a VL comprising SEQ ID NO: 101 or 115.
[0048] In some embodiments, provided herein is an antibody that binds SARS-CoV-2 spike protein, wherein the antibody comprises a VH comprising SEQ ID NO: 100, and a VL comprising SEQ ID NO: 101. In some embodiments, provided herein is an antibody that binds SARS-CoV-2 spike protein, wherein the antibody comprises a VH comprising SEQ ID NO: 111, and a VL comprising SEQ ID NO: 101. In some embodiments, provided herein is an antibody that binds SARS-CoV-2 spike protein, wherein the antibody comprises a VH comprising SEQ ID NO: 111, and a VL comprising SEQ ID NO: 115. In some embodiments, provided herein is an antibody that binds SARS-CoV-2 spike protein, wherein the antibody comprises a VH comprising SEQ ID NO: 118, and a VL comprising SEQ ID NO: 115. In some embodiments, provided herein is an antibody that binds SARS-CoV-2 spike protein, wherein the antibody comprises a VH comprising SEQ ID NO: 123, and a VL comprising SEQ ID NO: 115. In some embodiments, provided herein is an antibody that binds SARS-CoV-2 spike protein, wherein the antibody comprises a VH comprising SEQ ID NO: 126, and a VL comprising SEQ ID NO: 115.
[0049] In some embodiments, provided herein is an antibody that binds SARS-CoV-2 spike protein, wherein the antibody comprises a VH comprising SEQ ID NO: 135, and a VL comprising SEQ ID NO: 136.
[0050] In some embodiments, provided herein is an antibody that binds SARS-CoV-2 spike protein, wherein the antibody comprises a HC comprising SEQ ID NO: 9, 10, 20, 21, 27, or 28, and a LC comprising SEQ ID NO: 11, 15, 22, or 24. In some embodiments, provided herein is an antibody that binds SARS-CoV-2 spike protein, wherein the antibody comprises a HC comprising SEQ ID NO: 9, 20, or 27, and a LC comprising SEQ ID NO: 11, 15, 22, or 24. In some embodiments, provided herein is an antibody that binds SARS-CoV-2 spike protein, wherein the antibody comprises a HC comprising SEQ ID NO: 10, 21, or 28, and a LC comprising SEQ ID NO: 11, 15, 22, or 24.
[0051] In some embodiments, provided herein is an antibody that binds SARS-CoV-2 spike protein, wherein the antibody comprises a HC comprising SEQ ID NO: 9, and a LC comprising SEQ ID NO: 11. In some embodiments, provided herein is an antibody that binds SARS-CoV-2 spike protein, wherein the antibody comprises a HC comprising SEQ ID NO: 10, and a LC comprising SEQ ID NO: 11. In some embodiments, provided herein is an antibody that binds SARS-CoV-2 spike protein, wherein the antibody comprises a HC comprising SEQ ID NO: 9, and a LC comprising SEQ ID NO: 15. In some embodiments, provided herein is an antibody that binds SARS-CoV-2 spike protein, wherein the antibody comprises a HC comprising SEQ ID NO: 10, and a LC comprising SEQ ID NO: 15. In some embodiments, provided herein is an antibody that binds SARS-CoV-2 spike protein, wherein the antibody comprises a HC comprising SEQ ID NO: 9, and a LC comprising SEQ ID NO: 24. In some embodiments, provided herein is an antibody that binds SARS-CoV-2 spike protein, wherein the antibody comprises a HC comprising SEQ ID NO: 10, and a LC comprising SEQ ID NO: 24. In some embodiments, provided herein is an antibody that binds SARS-CoV-2 spike protein, wherein the antibody comprises a HC comprising SEQ ID NO: 20, and a LC comprising SEQ ID NO: 11. In some embodiments, provided herein is an antibody that binds SARS-CoV-2 spike protein, wherein the antibody comprises a HC comprising SEQ ID NO: 21, and a LC comprising SEQ ID NO: 11. In some embodiments, provided herein is an antibody that binds SARS-CoV-2 spike protein, wherein the antibody comprises a HC comprising SEQ ID NO: 20, and a LC comprising SEQ ID NO: 22. In some embodiments, provided herein is an antibody that binds SARS-CoV-2 spike protein, wherein the antibody comprises a HC comprising SEQ ID NO: 21, and a LC comprising SEQ ID NO: 22. In some embodiments, provided herein is an antibody that binds SARS-CoV-2 spike protein, wherein the antibody comprises a HC comprising SEQ ID NO: 27, and a LC comprising SEQ ID NO: 11. In some embodiments, provided herein is an antibody that binds SARS-CoV-2 spike protein, wherein the antibody comprises a HC comprising SEQ ID NO: 28, and a LC comprising SEQ ID NO: 11.
[0052] In some embodiments, provided herein is an antibody that binds SARS-CoV-2 spike protein, wherein the antibody comprises a HC comprising SEQ ID NO: 37, 38, 46, 47, 51, or 52, and a LC comprising SEQ ID NO: 39, 41, or 48. In some embodiments, provided herein is an antibody that binds SARS-CoV-2 spike protein, wherein the antibody comprises a HC comprising SEQ ID NO: 37, 46, or 51, and a LC comprising SEQ ID NO: 39, 41, or 48. In some embodiments, provided herein is an antibody that binds SARS-CoV-2 spike protein, wherein the antibody comprises a HC comprising SEQ ID NO: 38, 47, or 52, and a LC comprising SEQ ID NO: 39, 41, or 48.
[0053] In some embodiments, provided herein is an antibody that binds SARS-CoV-2 spike protein, wherein the antibody comprises a HC comprising SEQ ID NO: 37, and a LC comprising SEQ ID NO: 39. In some embodiments, provided herein is an antibody that binds SARS-CoV-2 spike protein, wherein the antibody comprises a HC comprising SEQ ID NO: 38, and a LC comprising SEQ ID NO: 39. In some embodiments, provided herein is an antibody that binds SARS-CoV-2 spike protein, wherein the antibody comprises a HC comprising SEQ ID NO: 37, and a LC comprising SEQ ID NO: 41. In some embodiments, provided herein is an antibody that binds SARS-CoV-2 spike protein, wherein the antibody comprises a HC comprising SEQ ID NO: 38, and a LC comprising SEQ ID NO: 41. In some embodiments, provided herein is an antibody that binds SARS-CoV-2 spike protein, wherein the antibody comprises a HC comprising SEQ ID NO: 51, and a LC comprising SEQ ID NO: 41. In some embodiments, provided herein is an antibody that binds SARS-CoV-2 spike protein, wherein the antibody comprises a HC comprising SEQ ID NO: 52, and a LC comprising SEQ ID NO: 41. In some embodiments, provided herein is an antibody that binds SARS-CoV-2 spike protein, wherein the antibody comprises a HC comprising SEQ ID NO: 46, and a LC comprising SEQ ID NO: 48. In some embodiments, provided herein is an antibody that binds SARS-CoV-2 spike protein, wherein the antibody comprises a HC comprising SEQ ID NO: 47, and a LC comprising SEQ ID NO: 48.
[0054] In some embodiments, provided herein is an antibody that binds SARS-CoV-2 spike protein, wherein the antibody comprises a HC comprising SEQ ID NO: 61, 62, 75 or 76, and a LC comprising SEQ ID NO: 63, 67, 70, or 77. In some embodiments, provided herein is an antibody that binds SARS-CoV-2 spike protein, wherein the antibody comprises a HC comprising SEQ ID NO: 61, or 75, and a LC comprising SEQ ID NO: 63, 67, 70, or 77. In some embodiments, provided herein is an antibody that binds SARS-CoV-2 spike protein, wherein the antibody comprises aHC comprising SEQ ID NO: 62, or 76, and aLC comprising SEQ ID NO: 63, 67, 70, or 77.
[0055] In some embodiments, provided herein is an antibody that binds SARS-CoV-2 spike protein, wherein the antibody comprises a HC comprising SEQ ID NO: 61, and a LC comprising SEQ ID NO: 63. In some embodiments, provided herein is an antibody that binds SARS-CoV-2 spike protein, wherein the antibody comprises a HC comprising SEQ ID NO: 62, and a LC comprising SEQ ID NO: 63. In some embodiments, provided herein is an antibody that binds SARS-CoV-2 spike protein, wherein the antibody comprises a HC comprising SEQ ID NO: 61, and a LC comprising SEQ ID NO: 67. In some embodiments, provided herein is an antibody that binds SARS-CoV-2 spike protein, wherein the antibody comprises a HC comprising SEQ ID NO: 62, and a LC comprising SEQ ID NO: 67. In some embodiments, provided herein is an antibody that binds SARS-CoV-2 spike protein, wherein the antibody comprises a HC comprising SEQ ID NO: 61, and a LC comprising SEQ ID NO: 70. In some embodiments, provided herein is an antibody that binds SARS-CoV-2 spike protein, wherein the antibody comprises aHC comprising SEQ ID NO: 62, and a LC comprising SEQ ID NO: 70. In some embodiments, provided herein is an antibody that binds SARS-CoV-2 spike protein, wherein the antibody comprises a HC comprising SEQ ID NO: 75, and a LC comprising SEQ ID NO: 77. In some embodiments, provided herein is an antibody that binds SARS-CoV-2 spike protein, wherein the antibody comprises a HC comprising SEQ ID NO: 76, and a LC comprising SEQ ID NO: 77. [0056] In some embodiments, provided herein is an antibody that binds SARS-CoV-2 spike protein, wherein the antibody comprises a HC comprising SEQ ID NO: 85, 86, 91, 92, 97, or 98, and a LC comprising SEQ ID NO: 87 or 99. In some embodiments, provided herein is an antibody that binds SARS-CoV-2 spike protein, wherein the antibody comprises a HC comprising SEQ ID NO: 85, 91, or 97, and a LC comprising SEQ ID NO: 87 or 99. In some embodiments, provided herein is an antibody that binds SARS-CoV-2 spike protein, wherein the antibody comprises a HC comprising SEQ ID NO: 86, 92, or 98, and a LC comprising SEQ ID NO: 87 or 99.
[0057] In some embodiments, provided herein is an antibody that binds SARS-CoV-2 spike protein, wherein the antibody comprises a HC comprising SEQ ID NO: 85, and a LC comprising SEQ ID NO: 87. In some embodiments, provided herein is an antibody that binds SARS-CoV-2 spike protein, wherein the antibody comprises a HC comprising SEQ ID NO: 86, and a LC comprising SEQ ID NO: 87. In some embodiments, provided herein is an antibody that binds SARS-CoV-2 spike protein, wherein the antibody comprises a HC comprising SEQ ID NO: 85, and a LC comprising SEQ ID NO: 99. In some embodiments, provided herein is an antibody that binds SARS-CoV-2 spike protein, wherein the antibody comprises a HC comprising SEQ ID NO: 86, and a LC comprising SEQ ID NO: 99. In some embodiments, provided herein is an antibody that binds SARS-CoV-2 spike protein, wherein the antibody comprises a HC comprising SEQ ID NO: 91, and a LC comprising SEQ ID NO: 87. In some embodiments, provided herein is an antibody that binds SARS-CoV-2 spike protein, wherein the antibody comprises a HC comprising SEQ ID NO: 92, and a LC comprising SEQ ID NO: 87. In some embodiments, provided herein is an antibody that binds SARS-CoV-2 spike protein, wherein the antibody comprises a HC comprising SEQ ID NO: 97, and a LC comprising SEQ ID NO: 99. In some embodiments, provided herein is an antibody that binds SARS-CoV-2 spike protein, wherein the antibody comprises a HC comprising SEQ ID NO: 98, and a LC comprising SEQ ID NO: 99.
[0058] In some embodiments, provided herein is an antibody that binds SARS-CoV-2 spike protein, wherein the antibody comprises a HC comprising SEQ ID NO: 108, 109, 113, 114, 121, 122, 124, 125, 128, or 129, and a LC comprising SEQ ID NO: 110 or 117. In some embodiments, provided herein is an antibody that binds SARS-CoV-2 spike protein, wherein the antibody comprises a HC comprising SEQ ID NO: 108, 113, 121, 124, or 128, and a LC comprising SEQ ID NO: 110 or 117. In some embodiments, provided herein is an antibody that binds SARS-CoV- 2 spike protein, whereinthe antibody comprises a HC comprising SEQ ID NO: 109, 114, 122, 125, or 129, and a LC comprising SEQ ID NO: 110 or 117.
[0059] In some embodiments, provided herein is an antibody that binds SARS-CoV-2 spike protein, wherein the antibody comprises a HC comprising SEQ ID NO: 108, and a LC comprising SEQ ID NO: 110. In some embodiments, provided herein is an antibody that binds SARS-CoV-2 spike protein, wherein the antibody comprises a HC comprising SEQ ID NO: 109, and a LC comprising SEQ ID NO: 110. In some embodiments, provided herein is an antibody that binds SARS-CoV-2 spike protein, wherein the antibody comprises a HC comprising SEQ ID NO: 113, and a LC comprising SEQ ID NO: 110. In some embodiments, provided herein is an antibody that binds SARS-CoV-2 spike protein, wherein the antibody comprises a HC comprising SEQ ID NO: 114, and a LC comprising SEQ ID NO: 110. In some embodiments, provided herein is an antibody that binds SARS-CoV-2 spike protein, wherein the antibody comprises a HC comprising SEQ ID NO: 113, and a LC comprising SEQ ID NO: 117. In some embodiments, provided herein is an antibody that binds SARS-CoV-2 spike protein, wherein the antibody comprises a HC comprising SEQ ID NO: 114, and a LC comprising SEQ ID NO: 117. In some embodiments, provided herein is an antibody that binds SARS-CoV-2 spike protein, wherein the antibody comprises a HC comprising SEQ ID NO: 121, and a LC comprising SEQ ID NO: 117. In some embodiments, provided herein is an antibody that binds SARS-CoV-2 spike protein, wherein the antibody comprises a HC comprising SEQ ID NO: 122, and a LC comprising SEQ ID NO: 117. In some embodiments, provided herein is an antibody that binds SARS-CoV-2 spike protein, wherein the antibody comprises a HC comprising SEQ ID NO: 124, and a LC comprising SEQ ID NO: 117. In some embodiments, provided herein is an antibody that binds SARS-CoV-2 spike protein, wherein the antibody comprises a HC comprising SEQ ID NO: 125, and a LC comprising SEQ ID NO: 117. In some embodiments, provided herein is an antibody that binds SARS-CoV-2 spike protein, wherein the antibody comprises a HC comprising SEQ ID NO: 128, and a LC comprising SEQ ID NO: 117. In some embodiments, provided herein is an antibody that binds SARS-CoV-2 spike protein, wherein the antibody comprises a HC comprising SEQ ID NO: 129, and a LC comprising SEQ ID NO: 117.
[0060] In some embodiments, provided herein is an antibody that binds SARS-CoV-2 spike protein, wherein the antibody comprises a HC comprising SEQ ID NO: 143 or 144, and a LC comprising SEQ ID NO: 145. In some embodiments, provided herein is an antibody that binds SARS-CoV-2 spike protein, wherein the antibody comprises a HC comprising SEQ ID NO: 143, and a LC comprising SEQ ID NO: 145. In some embodiments, provided herein is an antibody that binds SARS-CoV-2 spike protein, wherein the antibody comprises a HC comprising SEQ ID NO: 144, and a LC comprising SEQ ID NO: 145.
[0061] In some embodiments, the anti-SARS-CoV-2 antibodies or antigen binding fragments provided herein bind an epitope in the RBD domain of the SARS-CoV-2 S protein, e.g., 555-com- Al, 555-com-D2, 555-com-H3, 555-com-D7, 555-com-H7, 555-com-A8, 488-2A4, 488-2A6, 488-2C5, 488-2G1, 481-L1H1, 481-L1H4, 481-L5H2, 481-L5H1.
[0062] In some embodiments, the anti-SARS-CoV-2 antibodies or antigen binding fragments provided herein bind an epitope in the NTD domain of the SARS-CoV-2 S protein, e.g., 419-WT- 3, 419-FrF-10, 419-WT-15, 419-FrF-15, 419-HC2.1/LC56, 373-L1H21, 373-L1H24, 373-L2H24, 373-L2H10, 373-L2H20, 373-L2H61.
[0063] In some embodiments, the anti-SARS-CoV-2 antibodies or antigen binding fragments provided herein are antibodies that block SARS-CoV-2 binding to ACE2, i.e., ACE2 blockers, e.g., 555-com-Al, 555-com-D2, 555-com-H3, 555-com-D7, 555-com-H7, 555-com-A8, 488-2A4, 488-2A6, 488-2C5, 488-2G1, 481-L1H1, 481-L1H4, 481-L5H2, 481-L5H1, 419-WT-3, 419-FrF- 10, 419-WT-15, 419-FrF-15, 419-HC2.1/LC56, 373-L1H21, 373-L1H24, 373-L2H24, 373- L2H10, 373-L2H20, 373-L2H61.
[0064] In some embodiments, the anti-SARS-CoV-2 antibodies disclosed herein are neutralizing antibodies. Neutralization can be tested by various neutralization assays, e.g., a conventional neutralization assay based on the inhibition of a virus cytopathic effect (CPE) on cells in culture. For example, neutralization may be tested by reducing or blocking formation of CPE in cells infected with SARS-CoV-2. Virus and antibody may be premixed before addition to cells, followed by measuring blocking of virus entry. In some embodiments, neutralization can be tested by a pseudoneutralization assay or a live virus neutralization assay as describe in the Examples. [0065] The anti-SARS-CoV-2 antibodies disclosed herein may contain one or more amino acid residue substitutions, mutations and/or modifications, which result in an antibody with preferred characteristics including, but not limited to: altered pharmacokinetics, increased serum half-life, increase binding affinity, reduced immunogenicity, increased production, altered Fc ligand binding, enhanced or reduced ADCC or CDC activity, altered glycosylation and/or disulfide bonds and/or modified binding specificity.
[0066] In some embodiments, the anti-SARS-CoV-2 antibodies disclosed herein can encompass an Fc region with modifications that improve their half-lives (e.g., serum half-lives) in a human. Fc engineering can enhance antibody binding to neonatal Fc receptor (FcRn) and elongate antibody half-lives. The increased half-lives of the antibodies can result in a higher serum titer of antibodies, and thus, reduces the frequency of the administration of antibodies and/or reduces the concentration of antibodies or antibody fragments to be administered. Studies have shown that some Fc mutation that enhances FcRn binding results in increased binding to rheumatoid factor (RF), whereas some Fc mutation combinations enhance FcRn binding and prolong antibody half-life without increased binding to RF, e.g., N434A/Y436T/Q438R/S440E (ACT1), N434A/Y436V/Q438R/S440E (ACT2), M428L/N434A/Y436T/Q438R/S440E (ACT3), M428L/N434A/Y436V/Q438R/S440E (ACT4), M428L/N434A/Q438R/S440E (ACT5) (Maeda, et al., MABS 2017, 9(5): 844-853, positions numbered according to EU Index numbering). In some embodiments, the anti-SARS-CoV-2 antibodies disclosed herein comprise an Fc region comprising any set of mutations selected from ACT1, ACT2, ACT3, ACT4, ACT5. In some embodiments, the anti-SARS-CoV-2 antibodies disclosed herein comprise an Fc region comprising the ACT5 mutations: M428L/N434A/Q438R/S440E (positions numbered according to EU Index numbering).
[0067] In some embodiments, the anti-SARS-CoV-2 antibodies disclosed herein can comprise an Fc region with altered (e.g., enhanced or reduced) effector functions, including but not limited to, Clq binding; Fc receptor binding; complement dependent cytotoxicity (CDC); antibodydependent cell-mediated cytotoxicity (ADCC); phagocytosis; down regulation of cell surface receptors. CDC refers to the lysing of a target cell in the presence of complement. The complement activation pathway is initiated by the binding of the first component of the complement system (Clq) to a molecule, an antibody for example, complexed with a cognate antigen. To assess complement activation, a CDC assay may be performed. ADCC refers to a form of cytotoxicity in which secreted Ig bound onto Fc receptors (FcRs) present on certain cytotoxic cells (e.g., Natural Killer (NK) cells, neutrophils, and macrophages) enables these cytotoxic effector cells to bind specifically to an antigen-bearing target cell and subsequently kill the target cell with cytotoxins. In some embodiments, the anti-SARS-CoV-2 antibodies disclosed herein have enhanced ADCC activities. In some embodiments, the anti-SARS-CoV-2 antibodies disclosed herein have reduced ADCC activities.
[0068] In some embodiments, the anti-SARS-CoV-2 antibodies described herein comprise a modified IgGl Fc region having reduced Fc effector functions. For example, the anti-SARS-CoV- 2 antibodies can comprise an IgGl Fc region comprising amino acid substitutions L234A and L235A (all residues are numbered according to the EU Index numbering). In some embodiments, the anti-SARS-CoV-2 antibodies can comprise an IgGl Fc region comprising amino acid substitutions L234A, L235E, G237A, A330S and P331S (all residues are numbered according to the EU Index numbering).
[0069] In some embodiments, glycosylation patterns of the anti-SARS-CoV-2 antibodies disclosed herein can be modified. Engineered glycoforms may be useful for a variety of purposes, including but not limited to enhancing or reducing effector function, increasing the affinity of the antibody for a target antigen or facilitating production of the antibody. Such carbohydrate modifications can be accomplished by, for example, altering one or more sites of glycosylation within the antibody sequence. That is, one or more amino acid substitutions can be made that result in elimination of one or more variable region framework glycosylation sites to thereby eliminate glycosylation at that site. Conversely, effector functions or improved binding can also be modified by engineering in one or more additional glycosylation sites.
[0070] Additionally, or alternatively, the anti-SARS-CoV-2 antibodies can comprise an Fc region that has an altered glycosylation composition, such as a hypofucosylated antibody having reduced amounts of fucosyl residues or an antibody having increased bisecting GlcNAc structures. These and similar altered glycosylation patterns have been demonstrated to increase the ADCC ability of antibodies. Engineered glycoforms may be generated by any method known to one skilled in the art, for example by using engineered or variant expression strains, by co-expression with one or more enzymes (for example N-acetylglucosaminyltransferase III (GnTIl 1)), by expressing a molecule comprising an Fc region in various organisms or cell lines from various organisms or by modifying carbohydrate(s) after the molecule comprising Fc region has been expressed. In some embodiments, the anti-SARS-CoV-2 antibodies described herein have reduced fucosylation. In some embodiments, the anti-SARS-CoV-2 antibodies described herein comprise an Fc region comprising N-glycoside-linked sugar chains bound to the Fc region, wherein the sugar chains do not contain fucose. In some embodiments, the anti-SARS-CoV-2 antibodies described herein have increased ADCC activities, compared to the same antibodies comprising N-glycoside-linked sugar chains that comprise fucose.
[0071] Those skilled in the art will appreciate that anti-SARS-CoV-2 antibodies may be used in conjugated or unconjugated form. That is, the anti-SARS-CoV-2 antibody may be associated with or conjugated to (e.g. covalently or non-covalently) pharmaceutically active compounds, biological response modifiers, diagnostic moieties, or biocompatible modifiers. In this respect it will be understood that such conjugates may comprise peptides, polypeptides, proteins, fusion proteins, nucleic acid molecules, small molecules, mimetic agents, synthetic drugs, inorganic molecules, organic molecules and radioisotopes. Moreover, a conjugate may be covalently or non- covalently linked to the anti-SARS-CoV-2 antibody in various molar ratios depending, at least in part, on the method used for the conjugation.
[0072] In some embodiments, the anti-SARS-CoV-2 antibodies disclosed herein are conjugated to a diagnostic or detectable agent, marker or reporter, which may be a biological molecule (e.g., a peptide or nucleotide), a small molecule, fluorophore, or radioisotope. Labeled modulators can be useful for monitoring the development or progression of SARS-CoV-2 infection or as part of a clinical testing procedure to determine the efficacy of a particular therapy including anti-SARS-CoV-2 antibodies disclosed herein (i.e., theragnostics), or to determine a future course of treatment. Such markers or reporters may also be useful in purifying anti-SARS-CoV-2 antibodies disclosed herein.
[0073] Diagnosis and detection can be accomplished by coupling the modulator to detectable substances including, but not limited to, various enzymes comprising for example horseradish peroxidase, alkaline phosphatase, beta-galactosidase, or acetylcholinesterase; prosthetic groups, such as but not limited to streptavidinlbiotin and avidin/biotin; fluorescent materials, such as but not limited to, umbelliferone, fluorescein, fluorescein isothiocynate, rhodamine, dichlorotriazinylamine fluorescein, dansyl chloride or phycoerythrin; luminescent materials, such as but not limited to, luminol; bioluminescent materials, such as but not limited to, luciferase, luciferin, and aequorin; radioactive materials, such as but not limited to iodine (131I, 1251, 1231, 121I,), carbon (14C), sulfur (35S), tritium (3H), indium (115In, 113In, 112In, inIn,), and technetium ("Tc), thallium (201Ti), gallium (68Ga, 67Ga), palladium (103Pd), molybdenum (99Mo), xenon (133Xe), fluorine (18F), 153Sm, 177Lu, > 59Gd, 49Pm, 140La, 175Yb, 166Ho, 90Y, 47Sc, 186Re, 188Re, 142Pr, 105Rh, 97Ru, 68Ge, 57Co, 65Zn, 85 Sr, 32P, 153Gd, 169Yb, 51Cr, 54Mn, 75 Se, 1 13 Sn, and 1 17Tin; positron emitting metals using various positron emission tomographies, noradioactive paramagnetic metal ions, and molecules that are radiolabeled or conjugated to specific radioisotopes. In such embodiments appropriate detection methodology is well known in the art and readily available from numerous commercial sources.
[0074] In other embodiments, the anti-SARS-CoV-2 antibodies disclosed herein can be fused to marker sequences, such as a peptide or fluorophore to facilitate purification or diagnostic procedures such as immunohistochemistry or FACs. In some embodiments, the marker amino acid sequence is a hexa-histidine peptide, such as the tag provided in a pQE vector, among others, many of which are commercially available. Other peptide tags useful for purification include, but are not limited to, the hemagglutinin “HA” tag, and the “FLAG” tag.
[0075] Provided herein are also nucleic acids encoding a heavy chain or light chain, or a VH or VL, of the anti-SARS-CoV-2 antibodies disclosed herein, and vectors comprising one or more such nucleic acids. Provided herein are also vectors comprising a first nucleic acid sequence encoding an antibody heavy chain and a second nucleic acid sequence encoding an antibody light chain. Also provided are compositions comprising a first vector comprising a nucleic acid sequence encoding an antibody heavy chain, and a second vector comprising a nucleic acid sequence encoding an antibody light chain.
[0076] Exemplary nucleic acid sequences encoding an antibody heavy chain or light chain described herein are provided in SEQ ID NOs: 146-160. [0077] In some embodiments, provided herein are nucleic acids comprising a sequence encoding an antibody heavy chain, e.g., a nucleic acid comprising a sequence encoding SEQ ID NO: 9, 10, 20, 21, 27, or 28. In some embodiments, provided herein are nucleic acids comprising a sequence encoding an antibody light chain, e.g., a nucleic acid comprising a sequence encoding SEQ ID NO: 11, 15, 22, or 24. In some embodiments, provided herein are vectors comprising a nucleic acid comprising a sequence encoding SEQ ID NO: 9, 10, 20, 21, 27, or 28, or a nucleic acid comprising a sequence encoding SEQ ID NO: 11, 15, 22, or 24. For example, the nucleic acids or vectors can comprise SEQ ID NO: 149, 150, or 151. In some embodiments, provided herein are vectors comprising a first nucleic acid comprising a sequence encoding SEQ ID NO: 9,
10, 20, 21, 27, or 28, and a second nucleic acid comprising a sequence encoding SEQ ID NO: 11, 15, 22, or 24. For example, the vectors can comprise a first nucleic acid comprising SEQ ID NO: 149 or 150, and a second nucleic acid comprising SEQ ID NO: 151. In some embodiments, provided herein are compositions comprising a first vector comprising a sequence encoding SEQ ID NO: 9, 10, 20, 21, 27, or 28, and a second vector comprising a sequence encoding SEQ ID NO:
11, 15, 22, or 24. For example, the compositions can comprise a first vector comprising SEQ ID NO: 149 or 150, and a second vector comprising SEQ ID NO: 151 .
[0078] In some embodiments, provided herein are nucleic acids comprising a sequence encoding an antibody heavy chain, e.g., a nucleic acid comprising a sequence encoding SEQ ID NO: 37, 38, 46, 47, 51, or 52. In some embodiments, provided herein are nucleic acids comprising a sequence encoding an antibody light chain, e.g., a nucleic acid comprising a sequence encoding SEQ ID NO: 39, 41, or 48. In some embodiments, provided herein are vectors comprising a nucleic acid comprising a sequence encoding SEQ ID NO: 37, 38, 46, 47, 51, or 52, or a nucleic acid comprising a sequence encoding SEQ ID NO: 39, 41, or 48. In some embodiments, provided herein are vectors comprising a first nucleic acid comprising a sequence encoding SEQ ID NO: 37, 38, 46, 47, 51, or 52, and a second nucleic acid comprising a sequence encoding SEQ ID NO: 39, 41, or 48. In some embodiments, provided herein are compositions comprising a first vector comprising a sequence encoding SEQ ID NO: 37, 38, 46, 47, 51, or 52, and a second vector comprising a sequence encoding SEQ ID NO: 39, 41, or 48.
[0079] In some embodiments, provided herein are nucleic acids comprising a sequence encoding an antibody heavy chain, e.g., a nucleic acid comprising a sequence encoding SEQ ID NO: 61, 62, 75, or 76. In some embodiments, provided herein are nucleic acids comprising a sequence encoding an antibody light chain, e.g., a nucleic acid comprising a sequence encoding SEQ ID NO: 63, 67, 70, or 77. In some embodiments, provided herein are vectors comprising a nucleic acid comprising a sequence encoding SEQ ID NO: 61, 62, 75, or 76, or a nucleic acid comprising a sequence encoding SEQ ID NO: 63, 67, 70, or 77. For example, the nucleic acids or vectors can comprise SEQ ID NO: 152, 153, or 154. In some embodiments, provided herein are vectors comprising a first nucleic acid comprising a sequence encoding SEQ ID NO: 61, 62, 75, or 76, and a second nucleic acid comprising a sequence encoding SEQ ID NO: 63, 67, 70, or 77. For example, the vectors can comprise a first nucleic acid comprising SEQ ID NO: 152 or 153, and a second nucleic acid comprising SEQ ID NO: 154. In some embodiments, provided herein are compositions comprising a first vector comprising a sequence encoding SEQ ID NO: 61, 62, 75, or 76, and a second vector comprising a sequence encoding SEQ ID NO: 63, 67, 70, or 77. For example, the compositions can comprise a first vector comprising SEQ ID NO: 152 or 153, and a second vector comprising SEQ ID NO: 154.
[0080] In some embodiments, provided herein are nucleic acids comprising a sequence encoding an antibody heavy chain, e.g., a nucleic acid comprising a sequence encoding SEQ ID NO: 85, 86, 91, 92, 97, or 98. In some embodiments, provided herein are nucleic acids comprising a sequence encoding an antibody light chain, e.g., a nucleic acid comprising a sequence encoding SEQ ID NO: 87 or 99. In some embodiments, provided herein are vectors comprising a nucleic acid comprising a sequence encoding SEQ ID NO: 85, 86, 91, 92, 97, or 98, or a nucleic acid comprising a sequence encoding SEQ ID NO: 87 or 99. For example, the nucleic acids or vectors can comprise SEQ ID NO: 155, 156, or 157. In some embodiments, provided herein are vectors comprising a first nucleic acid comprising a sequence encoding SEQ ID NO: 85, 86, 91, 92, 97, or 98, and a second nucleic acid comprising a sequence encoding SEQ ID NO: 87 or 99. For example, the vectors can comprise a first nucleic acid comprising SEQ ID NO: 155 or 156, and a second nucleic acid comprising SEQ ID NO: 157. In some embodiments, provided herein are compositions comprising a first vector comprising a sequence encoding SEQ ID NO: 85, 86, 91, 92, 97, or 98, and a second vector comprising a sequence encoding SEQ ID NO: 87 or 99. For example, the compositions can comprise a first vector comprising SEQ ID NO: 155 or 156, and a second vector comprising SEQ ID NO: 157. [0081] In some embodiments, provided herein are nucleic acids comprising a sequence encoding an antibody heavy chain, e.g., a nucleic acid comprising a sequence encoding SEQ ID NO: 108, 109, 113, 114, 121, 122, 124, 125, 128, or 129. In some embodiments, provided herein are nucleic acids comprising a sequence encoding an antibody light chain, e.g., a nucleic acid comprising a sequence encoding SEQ ID NO: 110 or 117. In some embodiments, provided herein are vectors comprising a nucleic acid comprising a sequence encoding SEQ ID NO: 108, 109, 113, 114, 121, 122, 124, 125, 128, or 129, or a nucleic acid comprising a sequence encoding SEQ ID NO: 110 or 117. For example, the nucleic acids or vectors can comprise SEQ ID NO: 158, 159, or 160. In some embodiments, provided herein are vectors comprising a first nucleic acid comprising a sequence encoding SEQ ID NO: 108, 109, 113, 114, 121, 122, 124, 125, 128, or 129, and a second nucleic acid comprising a sequence encoding SEQ ID NO: 110 or 117. For example, the vectors can comprise a first nucleic acid comprising SEQ ID NO: 158 or 159, and a second nucleic acid comprising SEQ ID NO: 160. In some embodiments, provided herein are compositions comprising a first vector comprising a sequence encoding SEQ ID NO: 108, 109, 113, 114, 121, 122, 124, 125, 128, or 129, and a second vector comprising a sequence encoding SEQ ID NO: 110 or 117. For example, the compositions can comprise a first vector comprising SEQ ID NO: 158 or 159, and a second vector comprising SEQ ID NO: 160.
[0082] In some embodiments, provided herein are nucleic acids comprising a sequence encoding an antibody heavy chain, e.g., a nucleic acid comprising a sequence encoding SEQ ID NO: 143 or 144. For example, the nucleic acids can comprise SEQ ID NO: 146 or 147. In some embodiments, provided herein are nucleic acids comprising a sequence encoding an antibody light chain, e.g., a nucleic acid comprising a sequence encoding SEQ ID NO: 145. For example, the nucleic acids can comprise SEQ ID NO: 148. In some embodiments, provided herein are vectors comprising a nucleic acid comprising a sequence encoding SEQ ID NO: 143 or 144, or a nucleic acid comprising a sequence encoding SEQ ID NO: 145. For example, the vectors can comprise a nucleic acid comprising SEQ ID NO: 146, 147, or 148. In some embodiments, provided herein are vectors comprising a first nucleic acid comprising a sequence encoding SEQ ID NO: 143 or 144, and a second nucleic acid comprising a sequence encoding SEQ ID NO: 145. For example, the vectors can comprise a first nucleic acid comprising SEQ ID NO: 146 or 147, and a second nucleic acid comprising SEQ ID NO: 148. In some embodiments, provided herein are compositions comprising a first vector comprising a sequence encoding SEQ ID NO: 143 or 144, and a second vector comprising a sequence encoding SEQ ID NO: 145. For example, the compositions can comprise a first vector comprising SEQ ID NO: 146 or 147, and a second vector comprising SEQ ID NO: 148.
[0083] Nucleic acids of the present disclosure may be expressed in a host cell, for example, after the nucleic acids have been operably linked to an expression control sequence. Expression control sequences capable of expression of nucleic acids to which they are operably linked are well known in the art. An expression vector may include a sequence that encodes one or more signal peptides that facilitate secretion of the polypeptide(s) from a host cell. Expression vectors containing a nucleic acid of interest (e.g., a nucleic acid encoding a heavy chain or light chain of an antibody) may be transferred into a host cell by well-known methods, e.g., stable or transient transfection, transformation, transduction or infection. Additionally, expression vectors may contain one or more selection markers, e.g., tetracycline, neomycin, and dihydrofolate reductase, to aide in detection of host cells transformed with the desired nucleic acid sequences.
[0084] In another aspect, provided herein are cells, e.g., host cells, comprising the nucleic acids, vectors, or nucleic acid compositions described herein. A host cell may be a cell stably or transiently transfected, transformed, transduced or infected with one or more expression vectors expressing all or a portion of an antibody described herein. In some embodiments, a host cell may be stably or transiently transfected, transformed, transduced or infected with an expression vector expressing HC and LC polypeptides of an antibody of the present disclosure. In some embodiments, a host cell may be stably or transiently transfected, transformed, transduced or infected with a first vector expressing HC polypeptides and a second vector expressing LC polypeptides of an antibody described herein. Such host cells, e.g., mammalian host cells, can express the anti-SARS-CoV-2 antibodies described herein. Mammalian host cells known to be capable of expressing antibodies include CHO cells, HEK293 cells, COS cells, and NS0 cells.
[0085] The present disclosure further provides a process for producing an anti-SARS-CoV-2 antibody described herein by culturing the host cell described above, e.g., a mammalian host cell, under conditions such that the antibody is expressed and recovering the expressed antibody from the culture medium. The culture medium, into which an antibody has been secreted, may be purified by conventional techniques. Various methods of protein purification may be employed, and such methods are known in the art and described, for example, in Deutscher, Methods in Enzymology 182: 83-89 (1990) and Scopes, Protein Purification: Principles and Practice, 3rd Edition, Springer, NY (1994).
[0086] Also provided are antibodies produced by any of the processes described herein.
[0087] In another aspect, provided herein are compositions (e.g., pharmaceutical compositions) comprising an anti-SARS-CoV-2 antibody described herein. In some embodiments, the compositions (e.g., pharmaceutical compositions) further comprise at least one additional antibody that binds the SARS-CoV-2 spike protein. In some embodiments, provided herein are compositions (e.g., pharmaceutical compositions) comprising two or three anti-SARS- CoV-2 antibodies, wherein at least one antibody is an antibody described herein. In some embodiments, provided herein are compositions (e.g., pharmaceutical compositions) comprising two or three anti-SARS-CoV-2 antibodies described herein. Such pharmaceutical compositions can also comprise one or more pharmaceutically acceptable excipient, diluent or carrier. Pharmaceutical compositions can be prepared by methods well known in the art (e.g., Remington: The Science and Practice of Pharmacy, 22nd ed. (2012), A. Loyd et al., Pharmaceutical Press).
[0088] In some embodiments, provided herein are compositions (e.g., pharmaceutical compositions) comprising two or three anti-SARS-CoV-2 antibodies that bind different epitopes of the SARS-CoV-2 S protein. In some embodiments, provided herein are compositions (e.g., pharmaceutical compositions) comprising a first anti-SARS-CoV-2 antibody that binds a first epitope in the RBD domain of the SARS-CoV-2 S protein, and a second anti-SARS-CoV-2 antibody that binds a second epitope of the SARS-CoV-2 S protein, wherein the second epitope is different from the first epitope. In some embodiments, the second anti-SARS-CoV-2 antibody also binds an epitope in the RBD domain, but does not overlap with the epitope of the first anti- SARS-CoV-2 antibody. In some embodiments, the second anti-SARS-CoV-2 antibody binds an epitope in the NTD domain.
[0089] In some embodiments, provided herein are compositions (e.g., pharmaceutical compositions) comprising two or three anti-SARS-CoV-2 antibodies, wherein at least one antibody binds the RBD domain of the SARS-CoV-2 S protein. In some embodiments, provided herein are compositions (e.g., pharmaceutical compositions) comprising two or three anti-SARS- CoV-2 antibodies, wherein at least one antibody binds the NTD domain of the SARS-CoV-2 S protein. In some embodiments, provided herein are compositions (e.g., pharmaceutical compositions) comprising two or three anti -SARS-CoV-2 antibodies, wherein at least one antibody binds the RBD domain of the SARS-CoV-2 S protein, and at least one antibody binds the NTD domain of the SARS-CoV-2 S protein.
[0090] In some embodiments, the pharmaceutical compositions described herein comprise one or more anti -SARS-CoV-2 antibodies and one or more of the following excipients: histidine, sodium chloride, sucrose, polysorbate 80. In some embodiments, the pharmaceutical compositions comprise at least one anti-SARS-CoV-2 antibody, histidine, sodium chloride, sucrose, polysorbate 80. In some embodiments, the pharmaceutical compositions have a pH of about 6.0. In some embodiments, the pharmaceutical composition comprises at least one anti-SARS-CoV-2 antibody, 5 mM histidine, 50 mM NaCl, 6% sucrose, and 0.05% polysorbate 80 and has a pH of about 6.0.
[0091] The present disclosure also encompass in vitro or in vivo methods for detecting, diagnosing or monitoring coronavirus infections and methods of screening cells from a patient to identify coronavirus infected cells, including cells from a patient who is currently infected with SARS-CoV-2, or cells from a patient who is recovered from a past SARS-CoV-2 infection. Such methods include identifying an individual infected with coronavirus for treatment, monitoring progression of a coronavirus infection by contacting the patient or a sample obtained from a patient with one or more anti-SARS-CoV-2 antibodies disclosed herein, and detecting the presence or absence, or level of association of the antibody to a coronavirus antigen in the sample. In some embodiments, one or more anti-SARS-CoV-2 antibodies disclosed herein may be used to detect and quantify coronavirus levels in a patient sample (e.g., plasma or blood).
[0092] In some embodiments, provided herein are methods of identifying a SARS-CoV-2- infected cell; such methods comprise: contacting a cell with an anti-SARS-CoV-2 antibody described herein, conjugated to a detectable agent; and detecting specific binding of the antibody to the cell. In some embodiments, provided herein are methods of diagnosing a SARS-CoV-2 infection in a patient; such methods comprise: contacting a sample obtained from a patient with an anti-SARS-CoV-2 antibody described herein, conjugated to a detectable agent; and detecting specific binding of the antibody to a SARS-CoV-2 antigen present in the sample. [0093] Detection of a coronavirus antigen in the sample likely suggests that the individual may be effectively treated with one or more anti-SARS-CoV-2 antibodies disclosed herein. The methods may further comprise a step of comparing the level of binding to a control. Other diagnostic or theragnostic methods compatible with the teachings herein are well known in the art and can be practiced using commercial materials such as dedicated reporting systems. Exemplary compatible assay methods include radioimmunoassays, enzyme immunoassays, competitive - binding assays, fluorescent immunoassay, immunoblot assays, Western Blot analysis, flow cytometry assays, and ELISA assays (Enzyme-Linked ImmunoSorbent Assay). More generally detection of coronavirus in a biological sample may be accomplished using any art-known assay. Compatible in vivo theragnostics or diagnostics may comprise art recognized imaging or monitoring techniques such as magnetic resonance imaging (MRI), computerized tomography (e.g. CAT scan), positron tomography (e.g., PET scan) radiography, ultrasound, etc. Those skilled in the art will readily be able to recognize and implement appropriate detection, monitoring or imaging techniques (often comprising commercially available sources) based on the etiology, pathological manifestation, or clinical progression of the disorder.
[0094] In another embodiment, the disclosure provides methods of analyzing coronavirus infection progression and/or pathogenesis in vivo, using one or more anti-SARS-CoV-2 antibodies disclosed herein.
[0095] In another aspect, and as discussed in more detail below, the inventions disclosed herein also encompass kits for detecting, monitoring, or diagnosing a coronavirus infection, identifying an individual having a coronavirus infection for possible treatment or monitoring progression (or regression) of the infection in a patient, wherein the kit comprises an anti-SARS-CoV-2 antibody as described herein, and reagents for detecting the effect of the anti-SARS-CoV-2 antibody on a sample from the patient.
[0096] Anti-SARS-CoV-2 antibodies disclosed herein and cells, cultures, populations and compositions comprising the same, including progeny thereof, can also be used to screen for or identify compounds or agents (e.g., drugs) that affect a function or activity of coronavirus virions or coronavirus infected cells or progeny thereof by binding to an antigen present on the surface of the virion or infected cell. The disclosure therefore encompasses systems and methods for evaluation or identification of a compound or agent that can affect a function or activity of the coronavirus virus. Such compounds and agents can be drug candidates that are screened for the treatment of coronavirus infection, for example. In one embodiment, a system or method comprises coronavirus virions and/or coronavirus infected cells and a compound or agent (e.g., drug), wherein the virions/cells and compound or agent (e.g., drug) are in contact with each other. Exemplary activity or function that can be modulated include changes in cell morphology, expression of a marker, differentiation or de-differentiation, maturation, proliferation, viability, apoptosis or cell death neuronal progenitor cells or progeny thereof.
[0097] In another aspect, provided herein are methods of treating or preventing COVID- 19 by administering to a patient a therapeutically effective amount of one or more anti-SARS-CoV-2 antibodies described herein or a pharmaceutical composition comprising one or more anti-SARS- CoV-2 antibody described herein.
[0098] In some embodiments, provided herein are methods of treating or preventing COVID- 19 comprising: contacting a sample obtained from a patient with an antibody described herein, conjugated to a detectable agent; detecting specific binding of the antibody to a SARS-CoV-2 antigen present in the sample; and administering to the patient a therapeutically effective amount of an anti-SARS-CoV-2 antibody described herein or a pharmaceutical composition comprising an anti-SARS-CoV-2 antibody described herein.
[0099] In some embodiments, the patient has moderate to severe COVID-19. In some embodiments, the patient has mild to moderate COVID- 19. For example, mild COVID- 19 patients can include individuals who have any of various signs and symptoms, e.g., fever, cough, sore throat, malaise, headache, muscle pain, without shortness of breath, dyspnea, or abnormal imaging. Moderate COVID-19 patients can include individuals who have evidence of lower respiratory disease by clinical assessment or imaging and a saturation of oxygen (SaO2) greater than (>)93 percent (%) on room air at sea level. In some embodiments, the patient has not developed any symptoms of COVID-19. In some embodiments, the patient is at risk of developing COVID-19. In some embodiments, the patient is at risk of infection by SARS-CoV-2.
[00100] Also provided are anti-SARS-CoV-2 antibodies or pharmaceutical compositions comprising one or more (e.g., two or three) anti-SARS-CoV-2 antibodies for use in therapy. In some embodiments anti-SARS-CoV-2 antibodies or pharmaceutical compositions comprise one or more (e.g., two or three) anti-SARS-CoV-2 antibodies, for use in the treatment or prevention of COVID-19. Further provided herein are uses of anti-SARS-CoV-2 antibodies described herein in the manufacture of a medicament for the treatment or prevention of COVID- 19.
[00101] The anti-SARS-CoV-2 antibody may be administered to a patient, by various routes, including, but not limited to, intravenous or subcutaneous.
[00102] As used herein, the term “a,” “an,” “the” and similar terms used in the context of the present disclosure (especially in the context of the claims) are to be construed to cover both the singular and plural unless otherwise indicated herein or clearly contradicted by the context.
[00103] The term “antibody,” as used herein, refers to an immunoglobulin molecule that binds an antigen. Embodiments of an antibody include a monoclonal antibody, polyclonal antibody, human antibody, humanized antibody, chimeric antibody, or conjugated antibody. The antibodies can be of any class (e.g., IgG, IgE, IgM, IgD, IgA) and any subclass (e.g., IgGl, IgG2, IgG3, IgG4).
[00104] An exemplary antibody is an immunoglobulin G (IgG) type antibody comprised of four polypeptide chains: two heavy chains (HC) and two light chains (LC) that are cross-linked via inter-chain disulfide bonds. The amino-terminal portion of each of the four polypeptide chains includes a variable region of about 100-125 or more amino acids primarily responsible for antigen recognition. The carboxyl -terminal portion of each of the four polypeptide chains contains a constant region primarily responsible for effector function. Each heavy chain is comprised of a heavy chain variable region (VH) and a heavy chain constant region. Each light chain is comprised of a light chain variable region (VL) and a light chain constant region. The IgG isotype may be further divided into subclasses (e.g., IgGl, IgG2, IgG3, and IgG4).
[00105] The VH and VL regions can be further subdivided into regions of hyper-variability, termed complementarity determining regions (CDRs), interspersed with regions that are more conserved, termed framework regions (FR). The CDRs are exposed on the surface of the protein and are important regions of the antibody for antigen binding specificity. Each VH and VL is composed of three CDRs and four FRs, arranged from amino-terminus to carboxyl -terminus in the following order: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4. Herein, the three CDRs of the heavy chain are referred to as “HCDR1, HCDR2, and HCDR3” and the three CDRs of the light chain are referred to as “LCDR1, LCDR2 and LCDR3”. The CDRs contain most of the residues that form specific interactions with the antigen. Assignment of amino acid residues to the CDRs may be done according to the well-known schemes, including those described in Kabat (Kabat et al., “Sequences of Proteins of Immunological Interest,” National Institutes of Health, Bethesda, Md. (1991)), Chothia (Chothia et al., “Canonical structures for the hypervariable regions of immunoglobulins”, Journal of Molecular Biology, 196, 901-917 (1987); Al-Lazikani et al., “Standard conformations for the canonical structures of immunoglobulins”, Journal of Molecular Biology, 273, 927-948 (1997)), North (North et al., “A New Clustering of Antibody CDR Loop Conformations”, Journal of Molecular Biology, 406, 228-256 (2011)), or IMGT (the international ImMunoGeneTics database available on at www.imgt.org; see Lefranc et al., Nucleic Acids Res. 1999; 27:209-212). The North CDR definitions are used for the anti-SARS-CoV-2 antibodies described herein.
[00106] The present disclosure also include antibody fragments or antigen-binding fragments, which , as used herein, comprise at least a portion of an antibody retaining the ability to specifically interact with an antigen, such as Fab, Fab’, F(ab’)2, Fv fragments, scFv, scFab, disulfide-linked Fvs (sdFv), a Fd fragment and linear antibodies.
[00107] The terms “bind” and “binds” as used herein are intended to mean, unless indicated otherwise, the ability of a protein or molecule to form a chemical bond or attractive interaction with another protein or molecule, which results in proximity of the two proteins or molecules as determined by common methods known in the art.
[00108] The term “Fc region” as used herein refers to a region of an antibody, which comprises the CH2 and CH3 domains of the antibody heavy chain. Optionally, the Fc region may include a portion of the hinge region or the entire hinge region of the antibody heavy chain.
[00109] The term “neutralizing antibody” as used herein refers to an antibody that binds to or interacts with a virion and prevents binding or association of the virion with a host cell and/or entry into a host cell or acts as an egress inhibitor insofar as the antibody may not appear to be a neutralizing antibody in a conventional in vitro neutralization assay, but the antibody still inhibits propagation of the viral infection.
[00110] The terms “nucleic acid” or “polynucleotide”, as used interchangeably herein, refer to polymers of nucleotides, including single-stranded and / or double-stranded nucleotide-containing molecules, such as DNA, cDNA and RNA molecules, incorporating native, modified, and / or analogs of, nucleotides. Polynucleotides of the present disclosure may also include substrates incorporated therein, for example, by DNA or RNA polymerase or a synthetic reaction.
[00111] As used herein, “prevention”, “prevent”, and / or “preventing”, which are used interchangeably herein, are intended to refer to all processes wherein there may be a slowing, interrupting, arresting, controlling, stopping, alleviating symptoms or complications or reversing of the progression of a disease, for example, caused by an injury, insult such as SARS-CoV-2 viral infection or disease such as COVID-19 disease. As used herein, prevention is not intended to necessarily indicate a total elimination of all disorder symptoms.
[00112] The term “subject”, as used herein, refers to a mammal, including, but are not limited to, a human, chimpanzee, ape, monkey, cattle, horse, sheep, goat, swine, rabbit, dog, cat, rat, mouse, guinea pig, and the like. Preferably the subject is a human.
[00113] The term “therapeutically effective amount,” as used herein, refers to an amount of a protein or nucleic acid or vector or composition that will elicit the biological or medical response of a subject, for example, reduction or inhibition of an enzyme or a protein activity, or ameliorate symptoms, alleviate conditions, slow or delay disease progression, or prevent a disease, etc. In a non-limiting embodiment, the term “a therapeutically effective amount” refers to the amount of a protein or nucleic acid or vector or composition that, when administered to a subject, is effective to at least partially alleviate, inhibit, prevent and/or ameliorate a condition, or a disorder or a disease.
[00114] As used herein, “treatment” or “treating” refers to all processes wherein there may be a slowing, controlling, delaying or stopping of the progression of the disorders or disease disclosed herein, or ameliorating disorder or disease symptoms, but does not necessarily indicate a total elimination of all disorder or disease symptoms. Treatment includes administration of a protein or nucleic acid or vector or composition for treatment of a disease or condition in a patient, particularly in a human.
BRIEF DESCRIPTION OF THE DRAWINGS
[00115] FIGs. 1A-1G are bar graphs showing the results of the bead-based binding assays. FIGs. 1 A-1B show the binding of the tested anti-SARS-CoV-2 antibodies to the full-length SARS- CoV-2 spike protein (FIG. 1A) or the SI subunit of SARS-CoV-2 spike protein (FIG. IB). FIGs. 1C-1F show the binding of the tested anti-SARS-CoV-2 antibodies to the spike protein of SARS- CoV-1 (FIG. 1C), MERS (FIG. ID), WIV1 (FIG. IE), HKU1 (FIG. IF). FIG. 1G shows none of the tested anti-SARS-CoV-2 antibodies bind to the negative control beads.
[00116] FIGs. 2A-2E are bar graphs showing the results of the cell-based binding assays. FIG. 2A shows the binding of the tested anti-SARS-CoV-2 antibodies to SARS-CoV-2 full-length wildtype spike protein displayed on cells. FIGs. 2B-2D show the binding of the tested anti-SARS- CoV-2 antibodies to the reported SARS-CoV-2 spike protein mutant: S50L (FIG. 2B), G476S (FIG. 2C) or D614G (FIG. 2D), displayed on cells. FIG. 2E shows none of the tested anti-SARS- CoV-2 antibodies bind to the untransfected control cells.
EXAMPLES
[00117] The following examples are offered to illustrate, but not to limit, the claimed invention.
[00118] Example 1. Generation of the engineered anti-SARS-CoV-2 antibodies
[00119] Anti-SARS-CoV-2 antibodies are identified from blood samples obtained from convalescent human patients with confirmed SARS-CoV-2 infection. After determining the VH and VL sequences of the antibodies that recognize SARS-CoV-2, they are converted to full-length IgGl antibody sequences for further characterization and engineering.
[00120] Lead antibodies 555, 419, 488, 481, and 373 are thoroughly characterized, as described in a separate patent application. Based on the specific profile of each antibody, specific engineering strategies and processes are used.
[00121] Antibodies 555 and 488 are engineered using a phage system. The VH and VL sequences of the antibodies are cloned into an internal phage system vector for expression of Fabs in E. coli, construction of CDR mutant libraries by Kunkel mutagenesis, and screen of affinity improved variants by filter lift assay. All CDR residues are scanned with random single mutation libraries by the filter lift assays specifically optimized for hits that improve kon (association rate constant) and/or kOff (dissociate rate constant). All hits are picked and verified by DNA sequencing. Clones containing unique CDR mutations are expressed in E. coli. Variant Fab proteins are extracted from E. coli periplasm. The crude extracts or Fab proteins purified by Nickel -NTA bead are analyzed for affinity improvements using Octet BLI and the standard ELISA Fab titration. A selected set of beneficial kon and kOff mutations are combined in a total combinatorial library and screened by the same assays described above under more stringent conditions. The final combinatorial (combi.) variants are converted into IgGl for further characterization.
[00122] The CDR mutagenesis of 78 positions is used to enhance the affinity and potency of the parental 419 Fab. Libraries are created using NNK random codon-based mutagenic oligonucleotides incorporated using Kunkel mutagenesis into an uracil-containing single-stranded DNA template encoding the original parental 419 Fab with the selected CDR sequences deleted. Phage-expressed Fabs are screened by capture lift assay using in-house expressed biotinylated human SARS-CoV2 SI spike trimer antigen. Affinity enhanced mutations identified in this manner are DNA sequenced and unique clones expressed as periplasmic Fab in E. coli and crude supernatants analyzed by titration ELISA. This process identifies multiple CDR amino acid substitutions with increased affinity. Beneficial CDR mutations are combined in a library allowing all the beneficial mutations to be randomly combined or remain wild type sequence. Combinatorial libraries are screened under more stringent conditions including lower antigen concentration and increased washing at 37°C. Hits are once again DNA sequenced and unique variants screened by ELISA. To enable differentiation based on relative affinity, the ELISA includes a one-hour binding step followed by a three-hour 37°C wash step to magnify differences in dissociation rates. The final combi Fab variants are converted into IgGl for further characterization. Following expression and purification, several different ELISA formats are utilized to differentiate IgG based on relative affinities. In one example, IgG is captured using an anti-Fc coated plate. Monomeric NTD domain of SARS-CoV-2 spike protein with an Avi-tag fusion to enable in vivo biotinylation is subsequently titrated and binding detected using an alkaline phosphatase conjugated neutravidin reagent. This process identifies several higher affinity IgG variants containing multiple CDR residue changes. To confirm that the IgG variants bind SARS- CoV-2 spike protein, a SARS-CoV-2 spike trimeric antigen is used to coat an ELISA plate. IgG variants are titrated and detected with an anti -human Fc alkaline phosphatase conjugate. In this case, binding is detected but the off-rate differences are decreased due to avidity. This process leads to the identification of four higher affinity variants termed, 419-WT-3, 419-WT-15, 419- FrF-10 and 419-FrF-10.
[00123] Antibodies 481 and 373 are engineered as full-length antibodies in mammalian cell expression vectors using a high throughput site-specific mutagenesis protocol to find mutations that improve affinity and potentially mitigate developability issues (expression, chemical and biophysical stability, solubility) without impacting potency or leading to further developability issues based on amino acid property and percent exposure to solvent using molecular models. Briefly, every amino acid in the CDRs of both the VL and VH chains is mutated in individual mutagenesis reactions to the 18 variants (minus Cys), as well as back to the original residue (embedded wild-type, WT), using a series of forward and reverse oligo’ s arrayed in a 384 microtiter plate. Site-directed mutagenesis reactions are carried out according to established procedures and digestion of the WT plasmid accomplished by incubation with the Dpnl restriction enzyme. Digestion products are transformed into E. coll and DNA isolated from the bulk transformants following incubation overnight at 37°C. DNA from each individual VL and VH mutagenesis reaction is mixed with the appropriate WT antibody variable region and expressed in CHO cells in a 96-deep well plate. Secreted antibodies are quantified and normalized to a consistent titer prior to screening in downstream assays.
[00124] Hits are confirmed by ELISA titrations, Octet, or Biacore8K. The WT 481 mAb has a low affinity for RBD (for a monoclonal antibody) with a KD in the 10-7 M range. The WT 481 antibody also contains an asparagine in LCDR3 (N92) that deamidates under stressful conditions. The saturation mutagenesis and screening indicate only N92I mutation eliminates the deamidation without dramatically reducing the affinity of the mAb. Mutation at LCDR3 position 93 to residues known to inhibit deamidation also lead to loss of binding, thus, N92I is the best choice to eliminate any deamidation. All other 481 mutations in the 481 variants increase the affmity/potency. The parental 373 mAb has relatively low affinity (>10-6 KD). The only developability issues within the 373 parental mAb are (i) high hydrophobicity as measured by interactions with hydrophobic chromatography resins due to a hydrophobic cluster in the HCDR3 of parental antibody 373 (...FWSGYF...), and (ii) potential for oxidation at HCRD3 MIOOi. The saturation mutagenesis screening provides the team with information to reduce the hydrophobicity and eliminate the potential for methionine oxidation while significantly improving the affinity. All the engineered 373 variants contain mutations that address both the developability and improve the potency. The optimized 481 and 373 variants are also measured for non-specific binding and found they have very low nonspecific binding potential.
[00125] For the experimental work, the optimized 555, 488, 419, 481, and 373 VH domains are all cloned onto a human IgGl backbone or a human IgGl-ACT5 backbone containing mutations that increase the binding affinity to the neonatal Fc receptor (FcRn), which have been shown to improve the in vivo half-life of human IgGl (Maeda A, et al., 2017. MAbs 9, 844-53).
[00126] The anti-SARS-CoV-2 antibodies are produced by expression in Chinese hamster ovary (CHO) cells and purified using standard antibody purification techniques.
[00127] Measurement of Antigen Binding Affinity and Kinetics using Bio-Layer Interferometry (BLI)
[00128] A ForteBio Octet® RED384 Bio-Layer Interferometry system (Octet RED384, ForteBio, CA) is used to screen antigen binding kinetics of antibody variants at 30 °C with a shake speed of 1000 rpm. Briefly, biotinylated SARS-CoV-2 spike SI protein or the receptor binding domain (RBD) (Sino Biologicals, Cat#40592-V08H) is diluted to 1 pg/mL in the assay buffer (PBS with 0.5% BSA and 0.05% Tween-20). Antibody proteins are diluted 1: 1 in the same assay buffer. In the loading step, dip streptavidin (SA) tip (ForteBio, Catalog 18-5019) in 80 pL of antigen solution for 30 min to obtain 3 nm signal shift. Then dip the antigen tip into antibody sample for 120 seconds to obtain its association curve, followed by dipping into assay buffer to obtain its dissociation curve. Double referencing is performed in all experiments and data analysis, where signal from bare biosensor and buffer references are subtracted from sample data. Sample curves are calculated using ForteBio data analysis software and 1: 1 fitting to generate associate rate constant (ka) and dissociate rate constant (kd). Antigen binding affinity equals kd/ka (M).
[00129] Alternatively, antibody affinity can be measured by capturing Fab protein on FAB2G tip (ForteBio, Catalog 18-5125) and dipping into antigen solution to obtain the binding kinetics.
[00130] Non-Specific Binding Assay
[00131] Mesoscale Discovery (MSD) plates (MSD MA6000 384 well plate: MSD, Cat#L21XA-4) are prepared by adding 35 pL/well of PBS overnight at 4°C. Plates are washed 3 times with PBST (PBS + 0.1% Tween 20). Next, PBS is added to the plates for 0.5-1 hours at room temperature. The PBS solution is then removed and 20 mL of each test mAh (4 replicates per sample) is diluted to 5 or 10 mg/mL in PBS either directly from cell culture supernatants or from purified material and added to the MSD plate. In this 20 mL solution, 2 mg/mL of the secondary, labeled detection mAh (MSD anti-Human Antibody (Goat) Sulfo-TAG Labeled, 50pg R32AJ-5) is also simultaneously added. The test mAb and secondary mAb should be pre-complexed in solution for 30 minutes at room temperature prior to adding to the MSD plate. Incubation with of the test mAb/ secondary mAb on the MSD plate is allowed to proceed for 1 hr. The MSD plates are then washed 3 times with PBST. Read buffer (MSD Read Buffer T with surfactant (4x), Cat#R92TC-3) is diluted 4-fold and 35 mL are added to each well. Read the plate using the MSD reader. The electrochemiluminescence (ECL) signal is converted to log scale. Samples with signals over 3.4 or 3.5 are considered to have an increased risk of non-specific binding leading to faster non-specific clearance in vivo.
[00132] Example 2. Characterization of the engineered anti-SARS-CoV-2 antibodies
[00133] Surface Plasmon Resonance (SPR) —based Binding Experiments
[00134] The affinity and binding kinetics of selected anti-SARS-CoV-2 antibodies to the receptor binding domain (RBD) or N-terminal binding domain (NTD) of the SARS-CoV-2 spike protein, or the full-length SARS-CoV-2 spike protein, is determined by SPR using Biacore 8K (GE Healthcare). The procedure generally follows the “Instrument Handbook”. The amino acid sequences of the RBD, NTD, or full-length SARS-CoV-2 spike protein, used for these experiments are provided in Table 2.
[00135] Antibody is first captured on Biacore Protein A chip followed by flowing a serial concentration of RBD or NTD from 100 nM down to 0.391 nM in 2-fold serial dilution in PBS- P20-BSA (0.005% surfactant P20, 0. Img/mL BSA).
[00136] In some experiments, polyclonal goat anti -human IgG-Fc (Jackson Immunology cat#109005098), goat anti -kappa and goat anti-lambda (SouthernBiotech, Cat. #2060-01 and Cat. #2070-01, respectively) are directly immobilized onto Series S CM5 sensor chips (GE Healthcare Cat.: #29104988) using standard amine coupling (NHS/EDC/ethanolamine) protocols provided by the manufacturer’s protocols and a 10 mM Acetate pH 4.5 binding buffer/HBS-EP running buffer. Immobilization levels are between 8-10K resonance units (RUs). [00137] All kinetic binding experiments are performed at 37 °C. Multi-cycle kinetics setting that runs each analyte concentration in a separate cycle regenerating the surface after each sample injection is used. To measure binding kinetics, the test articles (optimized 555, 488, 481, 419, and 373 mAbs) are diluted to 10-100 pg/mL in PBS pH 7.4 and captured onto these chip surfaces to a level of 50-200 RU by injection at 10 pL/min using an HBS-EP running buffer. Next, recombinantly-derived SARS-Cov-2 receptor binding domain (RBD) or N-terminal binding domain (NTD) antigens are diluted into HBS-EP at 200, 100, 50, 25, 12.5, 6.25, 3.13 and 0 nM and injected (180-300 s) in succession over the sensor chip surfaces containing the captured mAb variants. Dissociation of the antigen is allowed to occur between 600 and 1800 s depending on the off rate (kd). Between each antigen injection, mAb and non-dissociated antigen are removed from the sensor chip surface with injection of 10 mM glycine, pH 1.5, buffer (regeneration). The regeneration is optimized to maintain consistent surface properties from cycle to cycle. The data is fit to a 1: 1 binding model to derive ka and kd, and to calculate KD (Table 3).
[00138] The affinity and binding kinetics of selected anti-SARS-CoV-2 antibodies to the full length SARS-CoV-2 spike protein fused to an AviTag (MW 420 kDa) are measured by Biacore 8K in a reversed setting. Briefly, neutravidin (NA) (Thermo Scientific # 31000) is first immortalized on Biacore® CM5 sensor chip via amine coupling and followed by injecting 2 nM antigen protein at 10 pl/min flow rate for 3 minutes. Antibody binding kinetics is measured by flowing a serial concentration of antibody from 300 nM in 2-fold serial dilution down to 2.34 nM. Each cycle starts with injection of antibody at 50 pl/min flow rate for 3 minutes and a dissociation in PBS-P20 for 15 minutes. The chip surface is then regenerated with injection of 25 pl of 10 mM glycine buffer, pH 1.5, at 50 pl/min flow rate for three times. The experiments are carried out at 37 °C. The data is fit to a bivalent binding model to derive the following parameters (shown in Table 4).
Table 2. Antigens used for anti-SARS-CoV-2 antibody binding kinetic and affinity studies
Figure imgf000061_0001
Figure imgf000062_0001
Figure imgf000063_0001
Table 3: Binding Kinetics and Affinity of the engineered anti-SARS-CoV-2 Antibodies to RBD or NTD of anti-SARS-CoV-2 spike protein
Figure imgf000063_0002
Figure imgf000064_0001
Table 4. Binding kinetics and affinity of selected anti-SARS-CoV-2 Antibodies to the full- length anti-SARS-CoV-2 spike protein based on a bivalent binding model*
Figure imgf000064_0002
Figure imgf000065_0001
[00139] Biophysical Characterization of the anti-SARS-CoV-2 antibodies
[00140] Engineered mAbs are subjected to several biophysical characterizations including analytical size exclusion chromatography (SEC-HPLC), hydrophobic interaction chromatography - high performance liquid chromatography (HIC-HPLC), heparin chromatography (Heparin-HPLC), cross-interaction chromatography (CIC), and affinity-capture self-interaction nanoparticle spectroscopy (AC-SINS). See Table 5.
SEC-HPLC
[00141] 4 pg of samples are injected onto a Waters BEH200 SEC, 4.6 x 150mm, 1.7pm column (Waters Cat#l 86005225). A flow rate of 0.3 mL/min with the running buffer containing 50mM sodium phosphate, pH6.8, 0.3M NaCl, 0.005% sodium azide is used. UV absorbance is monitored at 280nm using an Agilent 1260 HPLC. Retention time (RT) of main peak and percentage of monomer are reported.
HIC-HPLC
[00142] 20 pg IgG samples (1 mg/mL) are diluted 1 : 1 with 2x Buffer A concentrate (2 M ammonium sulfate, 0.1 M sodium phosphate at pH 6.8) to achieve a final ammonium sulfate concentration of 1 M before analysis. A TSKGEL® butyl-NPR (4.6mm ID x 10cm, 2.5um, Tosoh # 42168) column is used with a linear gradient (0-100% buffer B) of mobile phase A (IM ammonium sulfate, 50mM sodium phosphate, pH 6.8) and mobile phase B solution (50 mM sodium phosphate, pH 6.8) over 23 min at a flow rate of 1 mL/min with UV absorbance monitoring at 280 nm. Retention time of main peak is reported.
Heparin-HPLC [00143] 20 pg of IgG samples (Img/ml) in PBS is injected onto a POROS™ Heparin 50 um
(4.6 x 50mm, 0.8 ml, Thermo Scientific #4333412 column). Flow is kept at 1.5ml/ml with an initial linear gradient from 0% buffer B to 40% buffer B in 6 minutes then up to 60% B in 2 minutes, followed by a 1 minute gradient increase to 100% B. Gradient is kept at 100% for an additional minute to remove any remaining protein. Retention time of main peak is reported.
CIC
[00144] CIC is performed as described previously (Jacobs, S.A., et al., Pharm. Res. 27, 65- 71, 2010). In brief, the CIC column is prepared by coupling ~30 mg of human serum polyclonal antibodies (14506; Sigma) to a 1-mL HITRAP® column (17-0716-01; GE Healthcare), followed by quenching with ethanolamine. The blank column is prepared in the same except without human serum IgGs. Approximately 20 pg of each antibody is tested at a flow rate of 0.2 mL/min using lOmM sodium citrate, lOmM NaCl, pH 6.5 as a mobile phase on an Agilent 1260 series HPLC system. Retention times obtained by both IgG and blank columns are used to calculate k’ . In addition, due to peak tailing in some samples, peak width at 50% height is also obtained to monitor non-specific interaction of test antibodies.
AC-SINS
[00145] The AC-SINS assay is performed as described previously with modifications (Wu, J. et al., Protein Eng. Des. Sei. 28, 403-14, 2015). In short, gold nanoparticles (15705; Ted Pella Inc.) are coated with 100% capturing anti-human goat IgG Fc (109-005-008; Jackson ImmunoResearch). The conjugation reaction is quenched with I g/ml polyethylene glycol (PEG) and eluted into 0.25 x PBS. PEG is added to the conjugated gold mixture at a concentration of 0.2 pg/ml prior to use. The antibodies of interest are then incubated with the coated gold particles for 1 h at room temperature and the wavelength shift is measured using Tecan M1000 Pro Plate Reader within the range of 475-625 nm, in increments of 1 nm. Test antibodies are diluted in either PBS or 10 mM histidine, pH 6.0 prior to incubation. Delta plasma wavelength shift in comparison with buffer control is reported. The self-interacting antibodies show a higher wavelength shift away from the buffer controls.
Table 5. Biophysical Properties of the anti-SARS-CoV-2 Antibodies
Figure imgf000067_0001
[00146] Example 3. Binding of the anti-SARS-COV-2 antibodies to the wildtype or variant SARS-COV-2 spike protein and cross-reactivity to other coronaviruses SARS-CoV-1, MERS, HKU1, WIV1
Cell-based Binding Assays
[00147] Full-length spike protein sequences of either wildtype SARS-CoV-2 or the reported mutants of SARS-CoV-2 (with S50L, G476S or D614G mutation) and GFP inserts are cloned into the vector backbone pCDNA3.1. Suspension CHO cells are transiently transfected with the plasmid using electroporation. Full length native conformation spike protein expression is confirmed by testing with benchmark antibodies discovered against SARS-CoV-1 (aka SARS- CoV) that target different stalk and head domains using flow cytometry.
[00148] Binding of the anti-SARS-CoV-2 antibodies to full-length wildtype SARS-CoV-2 spike protein or the reported viral mutants (with S50L, G476S or D614G mutation) displayed on cells is tested using a high throughput flow cytometry. CHO cells are transiently transfected to express the full length spike protein of either wild type SARS-CoV-2 or mutant SARS-CoV-2 (with S50L, G476S, or D614G mutation) on the cell surface. Purified antibodies are incubated with the readout cells, and an untransfected control CHO line at 50 nM antibody concentration for 30 minutes at 4°C. CHO cells are washed; and the binding is detected by using a fluorescently labeled anti-human secondary antibody. Fluorescence is measured using high throughput platebased flow cytometry. Benchmark antibodies identified for SARS-CoV-1 are used as positive controls due to similarity in spike sequences between SARS-CoV-1 and SARS-CoV-2; human IgG isotype and an irrelevant antibody are used as negative controls.
Bead-based Binding Assays
[00149] Binding of the anti-SARS-CoV-2 antibodies to SARS-CoV-2 spike protein, the SI subunit of SARS-CoV-2 spike protein, or the spike protein of other coronaviruses SARS-CoV-1, MERS, HKU1, WIV1, is tested using a multiplexed bead assay on high throughput flow cytometry. Different optically encoded bead types are conjugated to one of the following unique antigens: full length spike protein of SARS-CoV-2, MERS, SARS-CoV-1, HKU1, WIV1 or the SI subunit of SARS-CoV-2 spike protein. Purified antibodies are incubated with the multiplexed beads, and negative control beads conjugated to BSA-His or FoldOn-His at 50 nM antibody concentration for 30 minutes at room temperature. Beads are washed and the binding is detected by using a fluorescently labeled anti-human secondary antibody. Fluorescence is measured using high throughput plate-based flow cytometry. Benchmark antibodies identified for SARS-CoV-1 are used as positive controls due to similarity in spike sequences between SARS-CoV-1 and SARS-CoV-2; human IgG isotype and an irrelevant antibody are used as negative controls.
Data Analysis
[00150] Median fluorescence intensity of each antibody is normalized over the median fluorescence intensity of the human isotype control for respective antigens. The mean fold over isotype values from different validation experiments are plotted. Mean and standard deviation is calculated where applicable and the values plotted as a column bar graph with median fold over isotype on Y-axis and the different antibodies represented along the X-axis. Antibodies with binding values greater than 5-fold over isotype to a given antigen with less than 2.5 -fold over isotype binding to negative control cells or beads are considered as binders. The cut-off value is determined based on the binding to the negative controls.
[00151] FIGs. 1A-1G show the results of the bead-based binding assays. FIGs. 2A-2E show the results of the cell-based binding assays. All tested antibodies show binding to the wildtype SARS-CoV-2 spike protein and the selected SARS-CoV-2 spike protein mutants (S50L, G476S, D614G) displayed on cells (FIGs. 2A-2D). All tested antibodies also show binding to the SARS- CoV-2 spike protein and the SI subunit of SARS-CoV-2 spike protein on beads (FIGs. 1A and IB). None of the tested antibodies show binding to the untransfected control cells (FIG. 2E) or the negative control beads (FIG. 1G). The anti-SARS-CoV-2 antibodies 555-com-A8, 555-com- D2, 555-com-H3 and 555-com-H7 show weak cross-reactivity to WIV1 spike proteins on beads (FIG. IE). The anti-SARS-CoV-2 antibodies 419-FrF-15, 419-WT-3 and 419-WT-15 show over 100-fold binding to HKU1 spike protein on beads, indicating cross reactivity to HKU1 (FIG. IF).
[00152] Example 4. Live virus neutralization assay of the anti-SARS-COV-2 antibodies
[00153] The efficacy of some anti-SARS-CoV-2 antibodies is measured by detecting the neutralization of infectious virus in a dose-response mode using cultured Vero E6 cells (African green monkey kidney; ATCC Cat. #CRL-1586). These cells are known to be highly susceptible to infection by SARS-CoV-2. Cells are maintained according to standard ATCC protocols. Briefly, Vero E6 cells are grown in minimal essential medium (MEM) supplemented with 10% heat-inactivated fetal bovine serum (FBS), 2 mM L-glutamine, and 1% MEM nonessential amino acid (NEAA) solution (Fisher Cat. #MT25025CI). Cell cultures are grown in 75 or 150 cm2 flasks at 37 °C in 5% CO2 and passaged 2 to 3 times per week using trypsin-EDTA. Cell cultures used for virus testing are prepared as subconfluent monolayers.
[00154] Assays are conducted using virus produced by infecting cultured Vero E6 cells with the SARS-CoV-2 clinical isolate USA/WA/1/2020 (BEI resources number NR52281) or the Italy- FNMI1 isolate of SARS-CoV-2 (European Virus Archive - Global, ref #008V-03893), and incubating at 37 °C until cytopathology is evident (typically 48 to 72 hours). Expansion is limited to 1 to 2 passages in cell culture to retain integrity of the original viral sequence. The virus stock is quantified by standard plaque assay, and aliquots are stored at -80°C. A freshly-thawed aliquot is used for each neutralization experiment.
[00155] The anti -SARS-CoV-2 antibodies and an IgGl isotype control are prepared in phosphate-buffered saline (PBS) at 1 mg/mL. Plaque reduction assays are performed in 6-well plates. Vero E6 cells are seeded at a concentration of approximately 106 cells/well and grown overnight at 37°C in 5% CO2 to reach 95% confluency. The next day, serial three-fold dilutions of anti-SARS-CoV-2 antibodies or an IgGl isotype control are prepared in Eagle’s minimal essential medium, mixed with approximately 100 pfu of SARS-CoV-2, and incubated for 1 to 2 hours on ice or at 37°C. The mAb/virus mixture is inoculated directly onto the cells and allowed to adsorb for 1 hour at 37 °C with 5% CO2, with rocking at 15 minute intervals. An overlay media composed of 1.25% Avicel RC-581 (FMC Biopolymer) in Eagles minimum essentials medium (MEM) with 5% FBS is added, and plates are incubated for 48 hours at 37°C with 5% CO2 for virus plaques to develop. After incubation, overlays are removed by aspiration and the cells are fixed with 10% buffered formalin containing crystal violet stain for 1 hour. Plaques are counted manually, and plaque forming units are determined by averaging technical replicates per sample.
[00156] Percent neutralization is calculated relative to the signals produced by an IgGl isotype control or a no-virus control, and the data are plotted using nonlinear regression with a four- parameter fit analysis (GraphPad Prism v8.0.0). IC50 is estimated using a meta-analysis using a random effects model (Berkey, et al., Stat Med. 1995; 14(4):395-411) and the R package meta for (Viechtbauer, J. Stat. Software 2010;36(3): l-48).
[00157] The results of the live virus neutralization assays are presented in Table 6.
Table 6. Results of Live Virus Neutralization Assays
Figure imgf000071_0001
SEQUENCE LISTING
SEQ ID NO: 1: Heavy chain variable region of Antibody 555-com-Al
QVQLVQSGAEVKKPGSSVKVSCKASGGTFDNYAISWVRQAPGQGLEWMGRIIPILGIANYAQKF QGRVHTADKSTSTAYMELSSLRSEDTAVYYCARTYYEARHYYYYYAMDVWGQGTAVTVSS
SEQ ID NO: 2: Light chain variable region of Antibody 555-com-Al
DIQMTQSPSSLSASVGDRVTITCRASQSISSYLSWYQQKPGKAPKLLIYAASDLQSGVPSRFSGSG SGTDFTLTITSLQPEDFATYYCQQSYSSPRTFGQGTKVEIK
SEQ ID NO: 3: Heavy chain complementarity determining region 1 of Antibody 555-com-Al KASGGTFDNYAIS
SEQ ID NO: 4: Heavy chain complementarity determining region 2 of Antibody 555-com-Al RIIPILGIAN
SEQ ID NO: 5: Heavy chain complementarity determining region 3 of Antibody 555-com-Al ARTYYEARHYYYYYAMDV
SEQ ID NO: 6: Light chain complementarity determining region 1 of Antibody 555-com-Al RASQSISSYLS
SEQ ID NO: 7: Light chain complementarity determining region 2 of Antibody 555-com-Al YAASDLQS
SEQ ID NO: 8: Light chain complementarity determining region 3 of Antibody 555-com-Al QQSYSSPRT
SEQ ID NO: 9: Heavy chain 1 - hlgGl of Antibody 555-com-Al
QVQLVQSGAEVKKPGSSVKVSCKASGGTFDNYAISWVRQAPGQGLEWMGRIIPILGIANYAQKF QGRVTITADKSTSTAYMELSSLRSEDTAVYYCARTYYEARHYYYYYAMDVWGQGTAVTVSSA STKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSS VVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDT LMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQD WLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDI
AVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKS
LSLSPGK
SEQ ID NO: 10: Heavy chain 2 - h!gGl-ACT5 of Antibody 555-com-Al
QVQLVQSGAEVKKPGSSVKVSCKASGGTFDNYAISWVRQAPGQGLEWMGRIIPILGIANYAQKF QGRVT1TADKSTSTAYMELSSLRSEDTAVYYCARTYYEARHYYYYYAMDVWGQGTAVTVSSA STKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSS VVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDT LMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQD
WLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDI AVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVLHEALHAHYTRKE LSLSPGK
SEQ ID NO: 11: Light chain of Antibody 555-com-Al
DIQMTQ SPSSL SASVGDRVTTTCRASQSIS SYLSWYQQKPGKAPKLLIYAASDLQ SGVPSRFSG SG SGTDFTLTITSLQPEDFATYYCQQSYSSPRTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVV CLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEV THQGLSSPVTKSFNRGEC
SEQ ID NO: 12: Light chain variable region of Antibody 555-com-D2
DIQMTQSPSSLSASVGDRVTITCRASQSISSYLSWYQQKPGKAPKLLIYAASSLQSGVPSRFSGSGS GTDFTLTITSLQPEDFATYYCQQSHSPPRTFGQGTKVEIK
SEQ ID NO: 13: Light chain complementarity determining region 2 of Antibody 555-com-D2 YAASSLQS
SEQID NO: 14: Light chain complementarity determining region 3 of Antibody 555-com-D2 QQSHSPPRT
SEQ ID NO: 15: Light chain of Antibody 555-com-D2
DIQMTQSPSSLSASVGDRVTITCRASQSISSYLSWYQQKPGKAPKLLIYAASSLQSGVPSRFSGSGS GTDFTLTITSLQPEDFATYYCQQSHSPPRTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVC LLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVT
HQGLSSPVTKSFNRGEC
SEQ ID NO: 16: Heavy chain variable region of Antibody 555-com-H3
QVQLVQSGAEVKKPGSSVKVSCKASGGTFDNYAISWVRQAPGQGLEWMGRIIPILGIANYAQKF
QGRVTITADKSTSTAYMELSSLRSEDTAVYYCARTPYEARHYYYYYAMDVWGQGTAVTVSS
SEQ ID NO: 17: Light chain variable region of Antibody 555-com-H3
DIQMTQSPSSLSASVGDRVTITCRASQSISSYLSWYQQKPGKAPKLLIYAASDLQSGVPSRFSGSG
SGTDFTLTITSLQPEDFATYYCQQSHSSPRTFGQGTKVEIK
SEQ ID NO: 18: Heavy chain complementarity determining region 3 of Antibody 555-com-H3
ARTPYEARHYYYYYAMDV
SEQ ID NO: 19: Light chain complementarity determining region 3 of Antibody 555-com-H3 QQSHSSPRT
SEQ ID NO: 20: Heavy chain 1 - hlgGl of Antibody 555-com-H3
QVQLVQSGAEVKKPGSSVKVSCKASGGTFDNYAISWVRQAPGQGLEWMGRIIPILGIANYAQKF
QGRVTITADKSTSTAYMELSSLRSEDTAVYYCARTPYEARHYYYYYAMDVWGQGTAVTVSSAS
TKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSS
VVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDT
LMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQD
WLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDI
AVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKS LSLSPGK
SEQ ID NO: 21: Heavy chain 2 - h!gGl-ACT5 of Antibody 555-com-H3
QVQLVQSGAEVKKPGSSVKVSCKASGGTFDNYAISWVRQAPGQGLEWMGRIIPILGIANYAQKF
QGRVTITADKSTSTAYMELSSLRSEDTAVYYCARTPYEARHYYYYYAMDVWGQGTAVTVSSAS
TKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSS
VVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDT
LMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQD
WLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDI AVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVLHEALHAHYTRKE
LSLSPGK
SEQ ID NO: 22: Light chain of Antibody 555-com-H3
DIQMTQSPSSLSASVGDRVT1TCRASQSISSYLSWYQQKPGKAPKLLIYAASDLQSGVPSRFSGSG SGTDFTLTITSLQPEDFATYYCQQSHSSPRTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVV CLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEV THQGLSSPVTKSFNRGEC
SEQ ID NO: 23: Light chain variable region of Antibody 555-com-D7
DIQMTQSPSSLSASVGDRVTITCRASQSISSYLSWYQQKPGKAPKLLIYAASSLQSGVPSRFSGSGS
GTDFTLTITSLQPEDFATYYCQQSHSSPRTFGQGTKVEIK
SEQ ID NO: 24: Light chain of Antibody 555-com-D7
DIQMTQSPSSLSASVGDRVTITCRASQSISSYLSWYQQKPGKAPKLLIYAASSLQSGVPSRFSGSGS GTDFTLTITSLQPEDFATYYCQQSHSSPRTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVC LLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVT HQGLSSPVTKSFNRGEC
SEQ ID NO: 25: Heavy chain variable region of Antibody 555-com-H7
QVQLVQSGAEVKKPGSSVKVSCKASGGTFDNYAISWVRQAPGQGLEWMGRIIPILGIANYAQKF QGRVTITADKSTSTAYMELSSLRSEDTAVYYCARTYYEARHYYYVYAMDVWGQGTAVTVSS
SEQ ID NO: 26: Heavy chain complementarity determining region 3 of Antibody 555-com-H7 ARTYYEARHYYYVYAMDV
SEQ ID NO: 27: Heavy chain 1 - hlgGl of Antibody 555-com-H7
QVQLVQSGAEVKKPGSSVKVSCKASGGTFDNYAISWVRQAPGQGLEWMGRIIPILGIANYAQKF
QGRVTITADKSTSTAYMELSSLRSEDTAVYYCARTYYEARHYYYVYAMDVWGQGTAVTVSSA STKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSS VVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDT LMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQD
WLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDI AVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKS LSLSPGK
SEQ ID NO: 28: Heavy chain 2 - hIgGl-ACT5 of Antibody 555-com-H7
QVQLVQSGAEVKKPGSSVKVSCKASGGTFDNYAISWVRQAPGQGLEWMGRIIPILGIANYAQKF
QGRVTITADKSTSTAYMELSSLRSEDTAVYYCARTYYEARHYYYVYAMDVWGQGTAVTVSSA
STKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSS
VVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDT
LMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQD
WLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDI
AVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVLHEALHAHYTRKE LSLSPGK
SEQ ID NO: 29: Heavy chain variable region of Antibody 419-Wild Type-3
QVQLVQSGAEVKKPGASVKVSCKVSGYTFTSRYMHWVRQAPGQGLEWMGIINPSSGSTSYAQK
FQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARDWTQSSGYDYYYGLDVWGQGTLVTVSS
SEQ ID NO: 30: Light chain variable region of Antibody 419-Wild Type-3
SYELTQPPSVSVSPGQTARITCSGDALPKQYAYWYQQKPGQAPVVVIYKDSERYSGIPERFSGSSS
GTTVTLTISGVQAEDEADYYCQSADSSGTYLVFGGGTKLTVL
SEQ ID NO: 31: Heavy chain complementarity determining region 1 of Antibody 419-Wild Type-3 KVSGYTFTSRYMH
SEQ ID NO: 32: Heavy chain complementarity determining region 2 of Antibody 419-Wild Type-3 IINPSSGSTSYAQKFQG
SEQ ID NO: 33: Heavy chain complementarity determining region 3 of Antibody 419-Wild Type-3 ARDWTQSSGYDYYYGLDV
SEQ ID NO: 34: Light chain complementarity determining region 1 of Antibody 419-Wild Type-3 SGDALPKQYAY
SEQ ID NO: 35: Light chain complementarity determining region 2 of Antibody 419-Wild Type-3 KDSERYS
SEQ ID NO: 36: Light chain complementarity determining region 3 of Antibody 419-Wild Type-3 QSADSSGTYLV
SEQ ID NO: 37: Heavy chain 1 - hlgGl of Antibody 419-Wild Type-3
QVQLVQSGAEVKKPGASVKVSCKVSGYTFTSRYMHWVRQAPGQGLEWMGIINPSSGSTSYAQK
FQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARDWTQSSGYDYYYGLDVWGQGTLVTVSS ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSL SSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPK
DTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQ DWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPS DIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQ KSLSLSPGK
SEQ ID NO: 38: Heavy chain 2 - hlgGl -ACT5 of Antibody 419-Wild Type -3
QVQLVQSGAEVKKPGASVKVSCKVSGYTFTSRYMHWVRQAPGQGLEWMGIINPSSGSTSYAQK
FQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARDWTQSSGYDYYYGLDVWGQGTLVTVSS
ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSL SSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPK DTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQ DWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPS
DIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVLHEALHAHYTRK ELSLSPGK
SEQ ID NO: 39: Light chain of Antibody 419-Wild Type -3
SYELTQPPSVSVSPGQTARITCSGDALPKQYAYWYQQKPGQAPVVVIYKDSERYSGIPERFSGSSS
GTTVTLTISGVQAEDEADYYCQSADSSGTYLVFGGGTKLTVLGQPKAAPSVTLFPPSSEELQANK ATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSC QVTHEGSTVEKTVAPTECS
SEQ ID NO: 40: Light chain variable region of Antibody 419-FrF-10
SYELTQPPSVSVSPGQTARITCSGDALPKQYAYWYQQKPGQAPVLVIYKDSERYSGIPERFSGSSS GTTVTLTISGVQAEDEADYYCQSADSSGTYLVFGGGTKLTVL SEQ ID NO: 41: Light chain of Antibody 419-FrF-10
SYELTQPPSVSVSPGQTARITCSGDALPKQYAYWYQQKPGQAPVLVIYKDSERYSGIPERFSGSSS GTTVTLTISGVQAEDEADYYCQSADSSGTYLVFGGGTKLTVLGQPKAAPSVTLFPPSSEELQANK ATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSC QVTHEGSTVEKTVAPTECS
SEQ ID NO: 42: Heavy chain variable region of Antibody 419-Wild Type-15
QVQLVQSGAEVKKPGASVKVSCKVSGYTFTSRYMHWVRQAPGQGLEWMGIINPSSGSTSYAQK FQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARDWTQTSGYDYYYGLDVWGQGTLVTVSS
SEQ ID NO: 43: Light chain variable region of Antibody 419-Wild Type-15
SYELTQPPSVSVSPGQTARITCSGDALPKQYAYWYQQKPGQAPVVVIYKDSERYSGIPERFSGSSS GTTVTLTISGVQAEDEADYYCQSADSSGTYVVFGGGTKLTVL
SEQ ID NO: 44: Heavy chain complementarity determining region 3 of Antibody 419-Wild Type-15 ARDWTQTSGYDYYYGLDV
SEQ ID NO: 45: Light chain complementarity determining region 3 of Antibody 419-Wild Type-15 QSADSSGTYVV
SEQ ID NO: 46: Heavy chain 1 - hlgGl of Antibody 419-Wild Type-15
QVQLVQSGAEVKKPGASVKVSCKVSGYTFTSRYMHWVRQAPGQGLEWMGIINPSGGSTSYAQ
KFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARDWTQTSGYDYYYGLDVWGQGTLVTVS SASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYS LSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKP
KDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLH QDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYP SDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYT QKSLSLSPGK
SEQ ID NO: 47: Heavy chain 2 - h!gGl-ACT5 of Antibody 419-Wild Type-15
QVQLVQSGAEVKKPGASVKVSCKVSGYTFTSRYMHWVRQAPGQGLEWMGIINPSSGSTSYAQK
FQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARDWTQTSGYDYYYGLDVWGQGTLVTVSS ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSL SSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPK DTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQ DWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPS DIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVLHEALHAHYTRK ELSLSPGK
SEQ ID NO: 48: Light chain of Antibody 419-Wild Type-15
SYELTQPPSVSVSPGQTARITCSGDALPKQYAYWYQQKPGQAPVVVIYKDSERYSGIPERFSGSSS GTTVTLTISGVQAEDEADYYCQSADSSGTYVVFGGGTKLTVLGQPKAAPSVTLFPPSSEELQANK ATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSC QVTHEGSTVEKTVAPTECS
SEQ ID NO: 49: Heavy chain variable region of Antibody 419-FrF-15
QVQLVQSGAEVKKPGASVKVSCKVSGYTFTSRYMHWVRQAPGQGLEWMGIINPSSGSTSYAQK FQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARDLTRSSGYDYYYGLDVWGQGTLVTVSS
SEQ ID NO: 50: Light chain complementarity determining region 3 of Antibody 419-FrF-15 ARDLTRSSGYDYYYGLDV
SEQ ID NO: 51: Heavy chain 1 - hlgGl of Antibody 419-FrF- 15
QVQLVQSGAEVKKPGASVKVSCKVSGYTFTSRYMHWVRQAPGQGLEWMGIINPSSGSTSYAQK FQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARDLTRSSGYDYYYGLDVWGQGTLVTVSSA STKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSS VVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDT LMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQD WLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDI AVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKS
LSLSPGK
SEQ ID NO: 52: Heavy chain 2 - h!gGl-ACT5 of Antibody 419-FrF-15
QVQLVQSGAEVKKPGASVKVSCKVSGYTFTSRYMHWVRQAPGQGLEWMGIINPSSGSTSYAQK FQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARDLTRSSGYDYYYGLDVWGQGTLVTVSSA STKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSS VVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDT
LMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQD
WLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDI
AVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVLHEALHAHYTRKE
LSLSPGK
SEQ ID NO: 53: Heavy chain variable region of Antibody 488-2A4
EVQLVESGGGLIQPGGSLRLSCAASGLTVSSNYMSWVRQAPGKGLEWVSVIYPGGSTYYADSVK
GRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARSPYGGNVWGQGTLVTVSS
SEQ ID NO: 54: Light chain variable region of Antibody 488-2A4
DIQMTQSPSSLSASVGDRVT1TCQASQDIQHYLNWYQQKPGKAPKLLIYDASNLETGVPSRFSGS
GSGTDFTFTISSLQPEDIATYYCQQYDNLPITFGQGTRLEIK
SEQ ID NO: 55: Heavy chain complementarity determining region 1 of Antibody 488-2A4
AASGLTVSSNYMS
SEQ ID NO: 56: Heavy chain complementarity determining region 2 of Antibody 488-2A4
VIYPGGSTY
SEQ ID NO: 57: Heavy chain complementarity determining region 3 of Antibody 488-2A4
ARSPYGGNV
SEQ ID NO: 58: Light chain complementarity determining region 1 of Antibody 488-2A4
QASQDIQHYLN
SEQ ID NO: 59: Light chain complementarity determining region 2 of Antibody 488-2A4
YDASNLET
SEQ ID NO: 60: Light chain complementarity determining region 3 of Antibody 488-2A4
QQYDNLPIT
SEQ ID NO: 61: Heavy chain 1 - hlgGl of Antibody 488-2A4 EVQLVESGGGLIQPGGSLRLSCAASGLTVSSNYMSWVRQAPGKGLEWVSVIYPGGSTYYADSVK
GRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARSPYGGNVWGQGTLVTVSSASTKGPSVFPLAP
SSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGT
QTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTC
VVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCK
VSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQP
ENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK
SEQ ID NO: 62: Heavy chain 2 - hIgGl-ACT5 of Antibody 488-2A4
EVQLVESGGGLIQPGGSLRLSCAASGLTVSSNYMSWVRQAPGKGLEWVSVIYPGGSTYYADSVK
GRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARSPYGGNVWGQGTLVTVSSASTKGPSVFPLAP
SSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGT
QTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTC
VVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCK
VSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQP
ENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVLHEALHAHYTRKELSLSPGK
SEQ ID NO: 63: Light chain of Antibody 488-2A4
DIQMTQSPSSLSASVGDRVT1TCQASQDIQHYLNWYQQKPGKAPKLLIYDASNLETGVPSRFSGS
GSGTDFTFTISSLQPEDIATYYCQQYDNLPITFGQGTRLEIKRTVAAPSVFIFPPSDEQLKSGTASVV
CLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEV THQGLSSPVTKSFNRGEC
SEQ ID NO: 64: Light chain variable region of Antibody 488-2A6
DIQMTQSPSSLSASVGDRVTITCQASQDTMNYLNWYQQKPGKAPKLLIYRASNLETGVPSRFSGS
GSGTDFTFTISSLQPEDIATYYCQQYDNLPITFGQGTRLEIK
SEQ ID NO: 65: Light chain complementarity determining region 1 of Antibody 488-2A6 QASQDTMNYLN
SEQ ID NO: 66: Light chain complementarity determining region 2 of Antibody 488-2A6
YRASNLET
SEQ ID NO: 67: Light chain of Antibody 488-2A6 DIQMTQSPSSLSASVGDRVTITCQASQDTMNYLNWYQQKPGKAPKLLIYRASNLETGVPSRFSGS GSGTDFTFTISSLQPEDIATYYCQQYDNLPITFGQGTRLEIKRTVAAPSVFIFPPSDEQLKSGTASVV CLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEV THQGLSSPVTKSFNRGEC
SEQ ID NO: 68: Light chain variable region of Antibody 488-2C5
DIQMTQSPSSLSASVGDRVTITCQASQDIMNYLNWYQQKPGKAPKLLIYDASNLETGVPSRFSGS
GSGTDFTFTISSLQPEDIATYYCQQYDNLPITFGQGTRLEIK
SEQ ID NO: 69: Light chain complementarity determining region 1 of Antibody 488-2C5 QASQDIMNYLN
SEQ ID NO: 70: Light chain of Antibody 488-2C5
DIQMTQSPSSLSASVGDRVT1TCQASQDIMNYLNWYQQKPGKAPKLLIYDASNLETGVPSRFSGS GSGTDFTFTISSLQPEDIATYYCQQYDNLPITFGQGTRLEIKRTVAAPSVFIFPPSDEQLKSGTASVV CLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEV THQGLSSPVTKSFNRGEC
SEQ ID NO: 71: Heavy chain variable region of Antibody 488-2G1
EVQLVESGGGLIQPGGSLRLSCAASGLTVSSNYMSWVRQAPGKGLEWVSTIYSGGSTYYADSVK GRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARSPYGGNVWGQGTLVTVSS
SEQ ID NO: 72: Light chain variable region of Antibody 488-2G1
DIQMTQSPSSLSASVGDRVTITCQASQDTQNYLNWYQQKPGKAPKLLIYDASNLETGVPSRFSGS GSGTDFTFTISSLQPEDIATYYCQQYDNLPITFGQGTRLEIK
SEQ ID NO: 73: Heavy chain complementarity determining region 2 of Antibody 488-2G1 TIYSGGSTY
SEQ ID NO: 74: Light chain complementarity determining region 1 of Antibody 488-2G1 QASQDTQNYLN
SEQ ID NO: 75: Heavy chain 1 - hlgGl of Antibody 488-2G1 EVQLVESGGGLIQPGGSLRLSCAASGLTVSSNYMSWVRQAPGKGLEWVSTIYSGGSTYYADSVK GRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARSPYGGNVWGQGTLVTVSSASTKGPSVFPLAP SSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGT QTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTC VVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCK
VSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQP ENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK
SEQ ID NO: 76: Heavy chain 2 - hIgGl-ACT5 of Antibody 488-2G1
EVQLVESGGGLIQPGGSLRLSCAASGLTVSSNYMSWVRQAPGKGLEWVSTIYSGGSTYYADSVK
GRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARSPYGGNVWGQGTLVTVSSASTKGPSVFPLAP SSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGT QTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTC VVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCK
VSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQP ENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVLHEALHAHYTRKELSLSPGK
SEQ ID NO: 77: Light chain of Antibody 488-2G1
DIQMTQSPSSLSASVGDRVT1TCQASQDTQNYLNWYQQKPGKAPKLLIYDASNLETGVPSRFSGS GSGTDFTFTISSLQPEDIATYYCQQYDNLPITFGQGTRLEIKRTVAAPSVFIFPPSDEQLKSGTASVV CLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEV THQGLSSPVTKSFNRGEC
SEQ ID NO: 78: Heavy chain variable region of Antibody 481-L1H1
EVQLVESGGGLIQPGGSLRLSCAASVFTVSYNYMSWVRQAPGKGLEWVSVIYPGGSTFYADSVK GRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAREVAGTYDYWGQGTLVTVSS
SEQ ID NO: 79: Light chain variable region of Antibody 481-L1H1
DIQMTQSPSSVSASVGDRVT1TCRASQGISSWLAWYQQKPGKAPKLLIYAASSLQSGVPSRFSGS GSGTDFTLTISSLQPEDFATYYCQDAISFPGGTFGPGTKVDIK
SEQ ID NO: 80: Heavy chain complementarity determining region 1 of Antibody 481-L1H1
AASVFTVSYNYMS SEQ ID NO: 81: Heavy chain complementarity determining region 2 of Antibody 481-L1H1
VIYPGGSTF
SEQ ID NO: 82: Heavy chain complementarity determining region 3 of Antibody 481-L1H1 AREVAGTYDY
SEQ ID NO: 83: Light chain complementarity determining region 1 of Antibody 481-L1H1 RASQGISSWLA
SEQ ID NO: 84: Light chain complementarity determining region 3 of Antibody 481-L1H1 QDAISFPGGT
SEQ ID NO: 85: Heavy chain 1 - hlgGl of Antibody 481-L1H1
EVQLVESGGGLIQPGGSLRLSCAASVFTVSYNYMSWVRQAPGKGLEWVSVIYPGGSTFYADSVK
GRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAREVAGTYDYWGQGTLVTVSSASTKGPSVFPL
APSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSL
GTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEV
TCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYK
CKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNG
QPENNYKITPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK
SEQ ID NO: 86: Heavy chain 2 - h!gGl-ACT5 of Antibody 481-L1H1
EVQLVESGGGLIQPGGSLRLSCAASVFTVSYNYMSWVRQAPGKGLEWVSVIYPGGSTFYADSVK
GRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAREVAGTYDYWGQGTLVTVSSASTKGPSVFPL
APSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSL
GTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEV
TCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYK
CKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNG
QPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVLHEALHAHYTRKELSLSPGK
SEQ ID NO: 87: Light chain of Antibody 481-L1H1
DIQMTQSPSSVSASVGDRVTITCRASQGISSWLAWYQQKPGKAPKLLIYAASSLQSGVPSRFSGS
GSGTDFTLTISSLQPEDFATYYCQDAISFPGGTFGPGTKVDIKRTVAAPSVFIFPPSDEQLKSGTAS VVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYAC
EVTHQGLSSPVTKSFNRGEC
SEQ ID NO: 88: Heavy chain variable region of Antibody 481-L1H4
EVQLVESGGGLIQPGGSLRLSCAASGITVSSNYMSWVRQAPGKGLEWVSVIYPGGSTFYADSVK
GRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAREVYGTYDYWGQGTLVTVSS
SEQ ID NO: 89: Heavy chain complementarity determining region 1 of Antibody 481-L1H4 AASGITVSSNYMS
SEQ ID NO: 90: Heavy chain complementarity determining region 3 of Antibody 481-L1H4 AREVYGTYDY
SEQ ID NO: 91: Heavy chain 1 - hlgGl of Antibody 481-L1H4
EVQLVESGGGLIQPGGSLRLSCAASGITVSSNYMSWVRQAPGKGLEWVSVIYPGGSTFYADSVK
GRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAREVYGTYDYWGQGTLVTVSSASTKGPSVFPL
APSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSL
GTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEV
TCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYK
CKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNG
QPENNYKITPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK
SEQ ID NO: 92: Heavy chain 2 - hIgGl-ACT5 of Antibody 481-L1H4
EVQLVESGGGLIQPGGSLRLSCAASGITVSSNYMSWVRQAPGKGLEWVSVIYPGGSTFYADSVK
GRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAREVYGTYDYWGQGTLVTVSSASTKGPSVFPL
APSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSL
GTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEV
TCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYK
CKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNG QPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVLHEALHAHYTRKELSLSPGK
SEQ ID NO: 93: Heavy chain variable region of Antibody 481-L5H2
EVQLVESGGGLIQPGGSLRLSCAASVFTVSSNYMSWVRQAPGKGLEWVSVIYPGGSTFYADSVK
GRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAREVAGTYDYWGQGTLVTVSS SEQ ID NO: 94: Light chain variable region of Antibody 481-L5H2
DIQMTQSPSSVSASVGDRVTITCRASQGISSWLAWYQQKPGKAPKLLIYAASSLQSGVPSRFSGS
GSGTDFTLTISSLQPEDFATYYCQQTISFPGGTFGPGTKVDIK
SEQ ID NO: 95: Heavy chain complementarity determining region 1 of Antibody 481-L2H5 AASVFTVSSNYMS
SEQ ID NO: 96: Light chain complementarity determining region 3 of Antibody 481-L2H5 QQTISFPGGT
SEQ ID NO: 97: Heavy chain 1 - hlgGl of Antibody 481-L2H5
EVQLVESGGGLIQPGGSLRLSCAASVFTVSSNYMSWVRQAPGKGLEWVSVIYPGGSTFYADSVK
GRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAREVAGTYDYWGQGTLVTVSSASTKGPSVFPL
APSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSL GTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEV TCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYK CKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNG QPENNYKITPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK
SEQ ID NO: 98: Heavy chain 2 - hIgGl-ACT5 of Antibody 481-L2H5
EVQLVESGGGLIQPGGSLRLSCAASVFTVSSNYMSWVRQAPGKGLEWVSVIYPGGSTFYADSVK
GRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAREVAGTYDYWGQGTLVTVSSASTKGPSVFPL
APSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSL GTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEV TCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYK CKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNG QPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVLHEALHAHYTRKELSLSPGK
SEQ ID NO: 99: Light chain of Antibody 481-L2H5
DIQMTQSPSSVSASVGDRVT1TCRASQGISSWLAWYQQKPGKAPKLLIYAASSLQSGVPSRFSGS
GSGTDFTLTISSLQPEDFATYYCQQTISFPGGTFGPGTKVDIKRTVAAPSVFIFPPSDEQLKSGTAS VVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYAC EVTHQGLSSPVTKSFNRGEC SEQ ID NO: 100: Heavy chain variable region of Antibody 373-L1H21
QVQLQESGPGLVKPSETLSLTCTVSGGSISVYHWEWIRQPPGKGLEWIGYIKYSGSTNYNPSLKS
RVTISVDTSKNQFSLKLSSVTAADTAVYYCARGPDYYDFWAGYYYGVDVWGQGTTVTVSS
SEQ ID NO: 101: Light chain variable region of Antibody 373-L1H21
EIVLTQSPGTLSLSPGERATLSCRASQSVESSYLAWYQQKPGQAPRLLIYGASSRATGIPDRFSGS
GSGTDFTLTISRLEPEDFAVYYCQQYGSSTTFGGGTKVEIK
SEQ ID NO: 102: Heavy chain complementarity determining region 1 of Antibody 373-L1H21 SGGSISVYHWE
SEQ ID NO: 103: Heavy chain complementarity determining region 2 of Antibody 373-L1H21 YIKYSGSTNYNP
SEQ ID NO: 104: Heavy chain complementarity determining region 3 of Antibody 373-L1H21
ARGPDYYDFWAGYYYGVDV
SEQ ID NO: 105: Light chain complementarity determining region 1 of Antibody 373-L1H21 RASQSVESSYLA
SEQ ID NO: 106: Light chain complementarity determining region 2 of Antibody 373-L1H21 GASSRAT
SEQ ID NO: 107: Light chain complementarity determining region 3 of Antibody 373-L1H21 QQYGSSTT
SEQ ID NO: 108: Heavy chain 1 - hlgGl of Antibody 373-L1H21
QVQLQESGPGLVKPSETLSLTCTVSGGSISVYHWEWIRQPPGKGLEWIGYIKYSGSTNYNPSLKS
RVTISVDTSKNQFSLKLSSVTAADTAVYYCARGPDYYDFWAGYYYGVDVWGQGTTVTVSSAST
KGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSV
VTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTL
MISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDW
LNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIA VEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSL
SLSPGK
SEQ ID NO: 109: Heavy chain 2 - h!gGl-ACT5 of Antibody 373-L1H21
QVQLQESGPGLVKPSETLSLTCTVSGGSISVYHWEWIRQPPGKGLEWIGYIKYSGSTNYNPSLKS
RVTISVDTSKNQFSLKLSSVTAADTAVYYCARGPDYYDFWAGYYYGVDVWGQGTTVTVSSAST
KGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSV
VTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTL
MISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDW
LNGKEYKCKVSNKALPAPIEKT1SKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIA
VEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVLHEALHAHYTRKELS LSPGK
SEQ ID NO: 110: Light chain of Antibody 373-L1H21
EIVLTQSPGTLSLSPGERATLSCRASQSVESSYLAWYQQKPGQAPRLLIYGASSRATGIPDRFSGS
GSGTDFTLTISRLEPEDFAVYYCQQYGSSTTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASV
VCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACE VTHQGLSSPVTKSFNRGEC
SEQ ID NO: 111: Heavy chain variable region of Antibody 373-L1H24
EIVLTQSPGTLSLSPGERATLSCRASQSVESSYLAWYQQKPGQAPRLLIYGASSRATGIPDRFSGS
GSGTDFTLTISRLEPEDFAVYYCQQYGSSTTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASV
VCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACE VTHQGLSSPVTKSFNRGEC
SEQ ID NO: 112: Heavy chain complementarity determining region 1 of Antibody 373-L1H24 SGHSISSYHWE
SEQ ID NO: 113: Heavy chain 1 - hlgGl of Antibody 373-L1H24
QVQLQESGPGLVKPSETLSLTCTVSGHSISSYHWEWIRQPPGKGLEWIGYIKYSGSTNYNPSLKSR
VTISVDTSKNQFSLKLSSVTAADTAVYYCARGPDYYDFWAGYYYGVDVWGQGTTVTVSSASTK
GPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVV
TVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLM
ISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLN GKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVE
WESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSL SPGK
SEQ ID NO: 114: Heavy chain 2 - h!gGl-ACT5 of Antibody 373-L1H24
QVQLQESGPGLVKPSETLSLTCTVSGHSISSYHWEWIRQPPGKGLEWIGYIKYSGSTNYNPSLKSR
VTISVDTSKNQFSLKLSSVTAADTAVYYCARGPDYYDFWAGYYYGVDVWGQGTTVTVSSASTK
GPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVV
TVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLM
ISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLN
GKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVE
WESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVLHEALHAHYTRKELSLS PGK
SEQ ID NO: 115: Light chain variable region of Antibody 373-L2H24
EIVLTQSPGTLSLSPGERATLSCRASQSVDTSYLAWYQQKPGQAPRLLIYGASSRATGIPDRFSGS
GSGTDFTLTISRLEPEDFAVYYCQQYGSSTTFGGGTKVEIK
SEQ ID NO: 116: Light chain complementarity determining region 1 of Antibody 373-L2H24 RASQSVDTSYLA
SEQ ID NO: 117: Light chain of Antibody 373-L2H24
EIVLTQSPGTLSLSPGERATLSCRASQSVDTSYLAWYQQKPGQAPRLLIYGASSRATGIPDRFSGS
GSGTDFTLTISRLEPEDFAVYYCQQYGSSTTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASV
VCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACE VTHQGLSSPVTKSFNRGEC
SEQ ID NO: 118: Heavy chain variable region of Antibody 373-L2H10
QVOLOESGPGLVKPSETLSLTCTVSGHSISSYHWEWIROPPGKGLEWIGYIYYSGSTNYNPSLKSR
VTISVDTSKNQFSLKLSSVTAADTAVYYCARGPDYYDYWAGYYYGVDVWGQGTTVTVSS
SEQ ID NO: 119: Heavy chain complementarity determining region 2 of Antibody 373-L2H10
YIYYSGSTNYNP SEQ ID NO: 120: Heavy chain complementarity determining region 3 of Antibody 373-L2H10
ARGPDYYDYWAGYYYGVDV
SEQ ID NO: 121: Heavy chain 1 - hlgGl of Antibody 373-L2H10
QVQLQESGPGLVKPSETLSLTCTVSGHSISSYHWEWIRQPPGKGLEWIGYIYYSGSTNYNPSLKSR
VTISVDTSKNQFSLKLSSVTAADTAVYYCARGPDYYDYWAGYYYGVDVWGQGTTVTVSSAST
KGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSV
VTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTL
MISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDW
LNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIA
VEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSL
SLSPGK
SEQ ID NO: 122: Heavy chain 2 - hlgGl -ACT5 of Antibody 373-L2H10
QVQLQESGPGLVKPSETLSLTCTVSGHSISSYHWEWIRQPPGKGLEWIGYIYYSGSTNYNPSLKSR
VTISVDTSKNQFSLKLSSVTAADTAVYYCARGPDYYDYWAGYYYGVDVWGQGTTVTVSSAST
KGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSV
VTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTL
MISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDW
LNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIA
VEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVLHEALHAHYTRKELS
LSPGK
SEQ ID NO: 123: Heavy chain variable region of Antibody 373-L2H20
QVQLQESGPGLVKPSETLSLTCTVSGHSISSYHWEWIRQPPGKGLEWIGYIYYSGSTNYNPSLKSR
VTISVDTSKNQFSLKLSSVTAADTAVYYCARGPDYYDFWAGYYYGVDVWGQGTTVTVSS
SEQ ID NO: 124: Heavy chain 1 - hlgGl of Antibody 373-L2H20
QVQLQESGPGLVKPSETLSLTCTVSGHSISSYHWEWIRQPPGKGLEWIGYIYYSGSTNYNPSLKSR
VTISVDTSKNQFSLKLSSVTAADTAVYYCARGPDYYDFWAGYYYGVDVWGQGTTVTVSSASTK
GPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVV
TVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLM
ISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLN
GKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVE WESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSL
SPGK
SEQ ID NO: 125: Heavy chain 2 - hIgGl-ACT5 of Antibody 373-L2H20
QVQLQESGPGLVKPSETLSLTCTVSGHSISSYHWEWIRQPPGKGLEWIGYIYYSGSTNYNPSLKSR
VTISVDTSKNQFSLKLSSVTAADTAVYYCARGPDYYDFWAGYYYGVDVWGQGTTVTVSSASTK
GPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVV
TVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLM
ISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLN
GKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVE
WESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVLHEALHAHYTRKELSLS PGK
SEQ ID NO: 126: Heavy chain variable region of Antibody 373-L2H61
QVQLQESGPGLVKPSETLSLTCTVSGHSISSYHWEWIRQPPGKGLEWIGYIYYSGSTNYNPSLKSR
VTISVDTSKNQFSLKLSSVTAADTAVYYCARGPDWYDFWAGYYYGTDVWGQGTTVTVSS
SEQ ID NO: 127: Heavy chain complementarity determining region 3 of Antibody 373-L2H61 ARGPDWYDFWAGYYYGTDV
SEQ ID NO: 128: Heavy chain 1 - hlgGl of Antibody 373-L2H61
QVQLQESGPGLVKPSETLSLTCTVSGHSISSYHWEWIRQPPGKGLEWIGYIYYSGSTNYNPSLKSR
VTISVDTSKNQFSLKLSSVTAADTAVYYCARGPDWYDFWAGYYYGTDVWGQGTTVTVSSAST
KGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSV
VTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTL
MISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDW
LNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIA
VEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSL SLSPGK
SEQ ID NO: 129: Heavy chain 2 - hlgGl -ACT5 of Antibody 373-L2H61
QVQLQESGPGLVKPSETLSLTCTVSGHSISSYHWEWIRQPPGKGLEWIGYIYYSGSTNYNPSLKSR
VTISVDTSKNQFSLKLSSVTAADTAVYYCARGPDWYDFWAGYYYGTDVWGQGTTVTVSSAST
KGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSV VTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTL MISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDW LNGKEYKCKVSNKALPAPIEKT1SKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIA VEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVLHEALHAHYTRKELS LSPGK
SEQ ID NO: 130: SARS-CoV-2 spike (S) protein (GenBank Accession No: YP_009724390.1) MFVFLVLLPLVSSQCVNLTTRTQLPPAYTNSFTRGVYYPDKVFRSSVLHSTQDLFLPFFSNVTWF HAIHVSGTNGTKRFDNPVLPFNDGVYFASTEKSNIIRGWIFGTTLDSKTQSLLIVNNATNVVIKVC EFQFCNDPFLGVYYHKNNKSWMESEFRVYSSANNCTFEYVSQPFLMDLEGKQGNFKNLREFVF KNIDGYFKIYSKHTPINLVRDLPQGFSALEPLVDLPIGINITRFQTLLALHRSYLTPGDSSSGWTAG AAAYYVGYLQPRTFLLKYNENGTITDAVDCALDPLSETKCTLKSFTVEKGIYQTSNFRVQPTESI VRFPNITNLCPFGEVFNATRFASVYAWNRKRISNCVADYSVLYNSASFSTFKCYGVSPTKLNDLC FTNVYADSFVIRGDEVRQIAPGQTGKIADYNYKLPDDFTGCVIAWNSNNLDSKVGGNYNYLYRL FRKSNLKPFERDISTEIYQAGSTPCNGVEGFNCYFPLQSYGFQPTNGVGYQPYRVVVLSFELLHAP ATVCGPKKSTNLVKNKCVNFNFNGLTGTGVLTESNKKFLPFQQFGRDIADTTDAVRDPQTLEILD ITPCSFGGVSVITPGTNTSNQVAVLYQDVNCTEVPVAIHADQLTPTWRVYSTGSNVFQTRAGCLI GAEHVNNSYECDIPIGAGICASYQTQTNSPRRARSVASQSIIAYTMSLGAENSVAYSNNSIAIPTNF TISVTTEILPVSMTKTSVDCTMYICGDSTECSNLLLQYGSFCTQLNRALTGIAVEQDKNTQEVFAQ VKQIYKTPPIKDFGGFNFSQILPDPSKPSKRSFIEDLLFNKVTLADAGFIKQYGDCLGDIAARDLIC
AQKFNGLTVLPPLLTDEMIAQYTSALLAGTITSGWTFGAGAALQIPFAMQMAYRFNGIGVTQNV LYENQKLIANQFNSAIGKIQDSLSSTASALGKLQDVVNQNAQALNTLVKQLSSNFGAISSVLNDIL SRLDKVEAEVQIDRLITGRLQSLQTYVTQQLIRAAEIRASANLAATKMSECVLGQSKRVDFCGKG YHLMSFPQSAPHGVVFLHVTYVPAQEKNFTTAPAICHDGKAHFPREGVFVSNGTHWFVTQRNF YEPQIITTDNTFVSGNCDVVIGIVNNTVYDPLQPELDSFKEELDKYFKNHTSPDVDLGDISGINAS VVNIQKEIDRLNEVAKNLNESLIDLQELGKYEQYIKWPWYIWLGFIAGLIAIVMVTIMLCCMTSC CSCLKGCCSCGSCCKFDEDDSEPVLKGVKLHYT
SEQ ID NO: 131: Receptor binding domain- AviHis tag antigen
RVQPTESIVRFPNITNLCPFGEVFNATRFASVYAWNRKRISNCVADYSVLYNSASFSTFKCYGVSP TKLNDLCFTNVYADSFVIRGDEVRQIAPGQTGKIADYNYKLPDDFTGCVIAWNSNNLDSKVGGN YNYLYRLFRKSNLKPFERDISTEIYQAGSTPCNGVEGFNCYFPLQSYGFQPTNGVGYQPYRVVVL SFELLHAPATVCGPKKSTNLVKNKCVNFNFNGLTGTGVLTESNKKFLPFQQFGRDIADTTDAVR DPQTLEILDITPCSFGGGGLNDIFEAQKIEWHEARAHHHHHH SEQ ID NO: 132: Receptor binding domain-His tag antigen
RVQPTESIVRFPNITNLCPFGEVFNATRFASVYAWNRKRISNCVADYSVLYNSASFSTFKCYGVSP TKLNDLCFTNVYADSFVIRGDEVRQIAPGQTGKIADYNYKLPDDFTGCVIAWNSNNLDSKVGGN YNYLYRLFRKSNLKPFERDISTEIYQAGSTPCNGVEGFNCYFPLQSYGFQPTNGVGYQPYRVVVL SFELLHAPATVCGPKKSTNLVKNKCVNFHHHHHH
SEQ ID NO: 133: N-terminal binding domain -A viHis tag antigen
VNLTTRTQLPPAYTNSFTRGVYYPDKVFRSSVLHSTQDLFLPFFSNVTWFHAIHVSGTNGTKRFD NPVLPFNDGVYFASTEKSNIIRGWIFGTTLDSKTQSLLIVNNATNVVIKVCEFQFCNDPFLGVYYH KNNKSWMESEFRVYSSANNCTFEYVSQPFLMDLEGKQGNFKNLREFVFKNIDGYFKIYSKHTPI NLVRDLPQGFSALEPLVDLPIGINITRFQTLLALHRSYLTPGDSSSGWTAGAAAYYVGYLQPRTFL LKYNENGTITDAVDCALDPLSETKCTLKSFTGGGGLNDIFEAQKIEWHEARAHHHHHH
SEQ ID NO: 134: Full-length SARS-CoV-2 spike protein-A viHis tag
VNLTTRTQLPPAYTNSFTRGVYYPDKVFRSSVLHSTQDLFLPFFSNVTWFHAIHVSGTNGTKRFD NPVLPFNDGVYFASTEKSNIIRGWIFGTTLDSKTQSLLIVNNATNVVIKVCEFQFCNDPFLGVYYH KNNKSWMESEFRVYSSANNCTFEYVSQPFLMDLEGKQGNFKNLREFVFKNIDGYFKIYSKHTPI
NLVRDLPQGFSALEPLVDLPIGINITRFQTLLALHRSYLTPGDSSSGWTAGAAAYYVGYLQPRTFL LKYNENGTITDAVDCALDPLSETKCTLKSFTVEKGIYQTSNFRVQPTESIVRFPNITNLCPFGEVFN ATRFASVYAWNRKRISNCVADYSVLYNSASFSTFKCYGVSPTKLNDLCFTNVYADSFVIRGDEV RQIAPGQTGKIADYNYKLPDDFTGCVIAWNSNNLDSKVGGNYNYLYRLFRKSNLKPFERDISTEI YQAGSTPCNGVEGFNCYFPLQSYGFQPTNGVGYQPYRVVVLSFELLHAPATVCGPKKSTNLVKN KCVNFNFNGLTGTGVLTESNKKFLPFQQFGRDIADTTDAVRDPQTLEILDITPCSFGGVSVITPGT NTSNQVAVLYQDVNCTEVPVAIHADQLTPTWRVYSTGSNVFQTRAGCLIGAEHVNNSYECDIPI GAGICASYQTQTNSPGSASSVASQSIIAYTMSLGAENSVAYSNNSIAIPTNFTISVTTEILPVSMTKT SVDCTMYICGDSTECSNLLLQYGSFCTQLNRALTGIAVEQDKNTQEVFAQVKQIYKTPPIKDFGG
FNFSQILPDPSKPSKRSFIEDLLFNKVTLADAGFIKQYGDCLGDIAARDLICAQKFNGLTVLPPLLT DEMIAQYTSALLAGT1TSGWTFGAGAALQIPFAMQMAYRFNGIGVTQNVLYENQKLIANQFNSA IGKIQDSLSSTASALGKLQDVVNQNAQALNTLVKQLSSNFGAISSVLNDILSRLDPPEAEVQIDRLI TGRLQSLQTYVTQQLIRAAEIRASANLAATKMSECVLGQSKRVDFCGKGYHLMSFPQSAPHGVV FLHVTYVPAQEKNFTTAPAICHDGKAHFPREGVFVSNGTHWFVTQRNFYEPQIITTDNTFVSGNC DVVIGIVNNTVYDPLQPELDSFKEELDKYFKNHTSPDVDLGDISGINASVVNIQKEIDRLNEVAKN LNESLIDLQELGKYEQGSGYIPEAPRDGQAYVRKDGEWVLLSTFLGGQGGQLNDIFEAQKIEWH
EARAGPAGHHHHHH
SEQ ID NO: 135 Heavy chain variable region of Antibody 419-HC2.1/LC56
QVQLVQSGAEVKKPGASVKVSCKASGYTFTSQYMHWVRQAPGQGLEWMGTINPSTGSTSYAQ
KYQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARDYTQTSGYDYYYGFDVWGQGTLVTVS S
SEQ ID NO: 136 Light chain variable region of Antibody 419-HC2.1/LC56
YELTQPPSVSVSPGQTARITCSGDALPKYYAWWYQQKPGQAPVVVIQKDSERPSGIPERFSGSSS
GTTVTLTISGVQAEDEADYYCQSADSSGSYVVFGGGTKLTVL
SEQ ID NO: 137 Heavy chain complementarity determining region 1 of Antibody 419-HC2.1/LC56
KASGYTFTSQYMH
SEQ ID NO: 138 Heavy chain complementarity determining region 2 of Antibody 419-HC2.1/LC56
TINPSTGSTSYAQKYQG
SEQ ID NO: 139 Heavy chain complementarity determining region 3 of Antibody 419-HC2.1/LC56
ARDYTQTSGYDYYYGFDV
SEQ ID NO: 140 Light chain complementarity determining region 1 of Antibody 419-HC2.1/LC56
SGDALPKYYAW
SEQ ID NO: 141 Light chain complementarity determining region 2 of Antibody 419-HC2.1/LC56
QKDSERPS
SEQ ID NO: 142 Light chain complementarity determining region 3 of Antibody 419-HC2.1/LC56
QSADSSGSYVV
SEQ ID NO: 143 Heavy chain 1- hlgGl of Antibody 419-HC2.1/LC56 QVQLVQSGAEVKKPGASVKVSCKASGYTFTSQYMHWVRQAPGQGLEWMGTINPSTGSTSYAQ
KYQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARDYTQTSGYDYYYGFDVWGQGTLVTVS
SASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYS
LSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKP
KDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLH
QDWLNGKEYKCKVSNKALPAPIEKT1SKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYP
SDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVLHEALHAHYTR
KELSLSPGK
SEQ ID NO: 144 Heavy chain 2 - IgGl-ACT-5 of Antibody 419-HC2.1/LC56
QVQLVQSGAEVKKPGASVKVSCKASGYTFTSQYMHWVRQAPGQGLEWMGTINPSTGSTSYAQ
KYQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARDYTQTSGYDYYYGFDVWGQGTLVTVS
SASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYS
LSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKP
KDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLH
QDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYP
SDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVLHEALHAHYTR
KELSLSPGK
SEQ ID NO: 145 Light chain of Antibody 419-HC2.1/LC56
YELTQPPSVSVSPGQTARITCSGDALPKYYAWWYQQKPGQAPVVVIQKDSERPSGIPERFSGSSS
GTTVTLTISGVQAEDEADYYCQSADSSGSYVVFGGGTKLTVLGQPKAAPSVTLFPPSSEELQANK
ATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSC
QVTHEGSTVEKTVAPTEC
SEQ ID NO: 146 Heavy Chain DNA Sequence of419-HC2.1/LC56- hlgGl
CAGGTGCAGCTGGTGCAGTCTGGGGCTGAGGTGAAGAAGCCTGGGGCCTCAGTGAAGGTTT
CCTGCAAGGCATCTGGATACACCTTCACCTCACAGTATATGCACTGGGTGCGACAGGCCCCT
GGACAAGGGCTTGAGTGGATGGGAACAATCAACCCTAGTACCGGTAGCACAAGCTACGCAC
AGAAGTACCAGGGCAGAGTCACCATGACCAGGGACACGTCCACGAGCACAGTCTACATGGA
GCTGAGCAGCCTGAGATCTGAGGACACGGCCGTGTATTACTGTGCGAGAGATTACACCCAA
ACCAGTGGCTACGATTACTACTACGGTTTCGACGTCTGGGGCCAAGGGACCCTGGTCACCGT
CTCCTCAGCTAGCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCT
CTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCAGTGACGGT
GTCGTGGAACTCAGGCGCACTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCT
CAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACC
TACATCTGCAACGTGAATCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTTGAGCCCA
AATCTTGTGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAACTCCTGGGGGGACCG
TCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTC
ACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTATGTGG
ACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGT
ACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAAGACTGGCTGAATGGCAAGGAGTACAA
GTGCAAGGTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAA GGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGAGGAGATGACCAAGA
ACCAAGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGG
GAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACG
GCTCCTTCTTCCTCTATTCCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTC
TTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCT
GTCTCCGGGCAAA
SEQ ID NO: 147 Heavy Chain DNA Sequence of419-HC2.1/LC56- h!gGl-ACT5
CAGGTGCAGCTGGTGCAGTCTGGGGCTGAGGTGAAGAAGCCTGGGGCCTCAGTGAAGGTTT
CCTGCAAGGCATCTGGATACACCTTCACCTCACAGTATATGCACTGGGTGCGACAGGCCCCT
GGACAAGGGCTTGAGTGGATGGGAACAATCAACCCTAGTACCGGTAGCACAAGCTACGCAC
AGAAGTACCAGGGCAGAGTCACCATGACCAGGGACACGTCCACGAGCACAGTCTACATGGA
GCTGAGCAGCCTGAGATCTGAGGACACGGCCGTGTATTACTGTGCGAGAGATTACACCCAA
ACCAGTGGCTACGATTACTACTACGGTTTCGACGTCTGGGGCCAAGGGACCCTGGTCACCGT
CTCCTCAGCCTCCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTC
TGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGT
CGTGGAACTCAGGCGCACTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCA
GGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTA
CATCTGCAACGTGAATCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTTGAGCCCAAA
TCTTGTGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAACTCCTGGGGGGACCGTC
AGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCA
CATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTATGTGGA
CGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTA
CCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAAGACTGGCTGAATGGCAAGGAGTACAAGT
GCAAGGTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGG
GCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGAGGAGATGACCAAGAAC
CAAGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGA
GAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGC
TCCTTCTTCCTCTATTCCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTT
CTCATGCTCCGTGCTGCATGAGGCTCTGCACGCTCACTACACGAGGAAGGAGCTCTCCCTGT
CTCCGGGCAAA
SEQ ID NO: 148 Light Chain DNA Sequence of 419-HC2.1/LC56
TATGAGCTGACACAGCCACCCTCGGTGTCAGTGTCCCCAGGACAGACGGCCAGGATCACCTG
CTCTGGAGATGCATTGCCAAAGTACTATGCTTGGTGGTACCAGCAGAAGCCAGGCCAGGCCC
CTGTGGTGGTGATACAGAAAGACAGTGAGAGGCCCTCAGGGATCCCTGAGCGATTCTCTGG
CTCCAGCTCAGGGACAACAGTCACGTTGACCATCAGTGGAGTCCAGGCAGAAGACGAGGCT
GACTATTACTGTCAATCAGCAGACAGCAGTGGTAGTTATGTCGTATTCGGCGGAGGGACCAA
GCTGACCGTCCTAGGCCAGCCCAAGGCTGCCCCCTCGGTCACTCTGTTCCCGCCCTCCTCTGA
GGAGCTTCAAGCCAACAAGGCCACACTGGTGTGTCTCATAAGTGACTTCTACCCGGGAGCCG
TGACAGTGGCCTGGAAGGCAGATAGCAGCCCCGTCAAGGCGGGAGTGGAGACAACCACACC
CTCCAAACAAAGCAACAACAAGTACGCGGCCAGCAGCTATCTGAGCCTGACGCCTGAGCAG
TGGAAGTCCCACAGAAGCTACAGCTGCCAGGTCACGCATGAAGGGAGCACCGTGGAGAAGA
CAGTGGCCCCTACAGAATGT SEQ ID NO: 149 Heavy Chain DNA Sequence of 555-com-H3 - hlgGl
CAGGTGCAGCTGGTGCAGTCTGGGGCTGAGGTGAAGAAGCCTGGGTCCTCGGTGAAGGTCT
CCTGCAAGGCTTCTGGAGGCACCTTCGACAACTATGCTATCAGCTGGGTGCGACAGGCCCCT
GGACAAGGGCTTGAGTGGATGGGAAGGATCATCCCTATCCTTGGTATAGCAAACTACGCAC
AGAAGTTCCAGGGCAGAGTCACGATTACCGCGGACAAATCCACGAGCACAGCCTACATGGA
GCTGAGCAGCCTGAGATCTGAGGACACGGCCGTGTATTACTGTGCGAGAACTCCTTACGAAG
CGAGGCATTACTACTACTACTACGCTATGGACGTCTGGGGCCAAGGGACCGCGGTCACCGTC
TCCTCAGCTAGCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTC
TGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCAGTGACGGTGT
CGTGGAACTCAGGCGCACTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCA
GGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTA
CATCTGCAACGTGAATCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTTGAGCCCAAA
TCTTGTGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAACTCCTGGGGGGACCGTC
AGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCA
CATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTATGTGGA
CGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTA
CCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAAGACTGGCTGAATGGCAAGGAGTACAAGT
GCAAGGTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGG
GCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGAGGAGATGACCAAGAAC
CAAGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGA
GAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGC
TCCTTCTTCCTCTATTCCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTT
CTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGT
CTCCGGGCAAA
SEQ ID NO: 150 Heavy Chain DNA Sequence of 555-com-H3 - hlgGl -ACT5
CAGGTGCAGCTGGTGCAGTCTGGGGCTGAGGTGAAGAAGCCTGGGTCCTCGGTGAAGGTCT CCTGCAAGGCTTCTGGAGGCACCTTCGACAACTATGCTATCAGCTGGGTGCGACAGGCCCCT GGACAAGGGCTTGAGTGGATGGGAAGGATCATCCCTATCCTTGGTATAGCAAACTACGCAC AGAAGTTCCAGGGCAGAGTCACGATTACCGCGGACAAATCCACGAGCACAGCCTACATGGA GCTGAGCAGCCTGAGATCTGAGGACACGGCCGTGTATTACTGTGCGAGAACTCCTTACGAAG CGAGGCATTACTACTACTACTACGCTATGGACGTCTGGGGCCAAGGGACCGCGGTCACCGTC TCCTCAGCCTCCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTCT GGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGT CGTGGAACTCAGGCGCACTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCA GGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTA CATCTGCAACGTGAATCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTTGAGCCCAAA TCTTGTGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAACTCCTGGGGGGACCGTC AGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCA CATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTATGTGGA
CGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTA CCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAAGACTGGCTGAATGGCAAGGAGTACAAGT GCAAGGTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGG GCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGAGGAGATGACCAAGAAC CAAGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGA GAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGC
TCCTTCTTCCTCTATTCCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTT
CTCATGCTCCGTGCTGCATGAGGCTCTGCACGCTCACTACACGAGGAAGGAGCTCTCCCTGT CTCCGGGCAAA
SEQ ID NO: 151 Light Chain DNA Sequence of 555-com-H3
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCAT
CACTTGCCGGGCAAGTCAGAGCATTAGCAGCTATTTAAGTTGGTATCAGCAGAAACCAGGG
AAAGCCCCTAAGCTCCTGATCTATGCTGCATCCGACTTGCAAAGTGGGGTCCCATCAAGGTT
CAGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCACCAGTCTGCAACCTGAAGATT
TTGCAACTTACTACTGTCAACAGAGTCACAGTTCCCCTCGCACGTTCGGCCAAGGGACCAAG
GTGGAAATCAAACGGACCGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCA
GTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCA
AAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGA
GCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGAC
TACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCA
CAAAGAGCTTCAACAGGGGAGAGTGC
SEQ ID NO: 152 Heavy Chain DNA Sequence of 488-2A6 - hlgGl
GAGGTGCAGCTGGTGGAGTCTGGAGGAGGCTTGATCCAGCCTGGGGGGTCCCTGAGACTCT
CCTGTGCAGCCTCTGGGTTAACCGTCAGTAGCAACTACATGAGCTGGGTCCGCCAGGCTCCA
GGGAAGGGGCTGGAGTGGGTCTCAGTTATTTATCCGGGTGGTAGCACATACTACGCAGACTC
CGTGAAGGGCCGATTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTATCTTCAAATGA
ACAGCCTGAGAGCCGAGGACACGGCCGTGTATTACTGTGCGAGATCCCCCTACGGTGGTAA
CGTCTGGGGCCAGGGAACCCTGGTCACCGTCTCCTCAGCTAGCACCAAGGGCCCATCGGTCT
TCCCCCTGGCACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTC
AAGGACTACTTCCCCGAACCAGTGACGGTGTCGTGGAACTCAGGCGCACTGACCAGCGGCG
TGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACC
GTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAA
CACCAAGGTGGACAAGAGAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCG
TGCCCAGCACCTGAACTCCTGGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGA
CACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAG
ACCCTGAGGTCAAGTTCAACTGGTATGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAA
GCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCAC
CAAGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCCCAGCCC
CCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCT
GCCCCCATCCCGGGAGGAGATGACCAAGAACCAAGTCAGCCTGACCTGCCTGGTCAAAGGC
TTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACA
AGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTATTCCAAGCTCACCGTGG
ACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCA
CAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGCAAA SEQ ID NO: 153 Heavy Chain DNA Sequence of 488-2A6 - h!gGl-ACT5
GAGGTGCAGCTGGTGGAGTCTGGAGGAGGCTTGATCCAGCCTGGGGGGTCCCTGAGACTCT
CCTGTGCAGCCTCTGGGTTAACCGTCAGTAGCAACTACATGAGCTGGGTCCGCCAGGCTCCA
GGGAAGGGGCTGGAGTGGGTCTCAGTTATTTATCCGGGTGGTAGCACATACTACGCAGACTC
CGTGAAGGGCCGATTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTATCTTCAAATGA
ACAGCCTGAGAGCCGAGGACACGGCCGTGTATTACTGTGCGAGATCCCCCTACGGTGGTAA
CGTCTGGGGCCAGGGAACCCTGGTCACCGTCTCCTCAGCCTCCACCAAGGGCCCATCGGTCT
TCCCCCTGGCACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTC
AAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCACTGACCAGCGGCG
TGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACC
GTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAA
CACCAAGGTGGACAAGAGAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCG
TGCCCAGCACCTGAACTCCTGGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGA
CACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAG
ACCCTGAGGTCAAGTTCAACTGGTATGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAA
GCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCAC
CAAGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCCCAGCCC
CCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCT
GCCCCCATCCCGGGAGGAGATGACCAAGAACCAAGTCAGCCTGACCTGCCTGGTCAAAGGC
TTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACA
AGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTATTCCAAGCTCACCGTGG
ACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGCTGCATGAGGCTCTGCAC
GCTCACTACACGAGGAAGGAGCTCTCCCTGTCTCCGGGCAAA
SEQ ID NO: 154 Light Chain DNA Sequence of 488-2A6
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCAT
CACTTGCCAGGCGAGTCAGGACACTATGAACTATTTAAATTGGTATCAGCAGAAACCAGGG
AAAGCCCCTAAGCTCCTGATCTACCGGGCATCCAATTTGGAAACAGGGGTCCCATCAAGGTT
CAGTGGAAGTGGATCTGGGACAGATTTTACTTTCACCATCAGCAGCCTGCAGCCTGAAGATA
TTGCAACATATTACTGTCAACAGTATGATAATCTCCCGATCACCTTCGGCCAAGGGACACGA
CTGGAGATTAAACGGACCGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCA
GTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCA
AAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGA
GCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGAC
TACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCA
CAAAGAGCTTCAACAGGGGAGAGTGC
SEQ ID NO: 155 Heavy Chain DNA Sequence of 481-L5H2- hlgGl
GAGGTGCAGCTGGTGGAGTCTGGAGGAGGCTTGATCCAGCCTGGGGGGTCCCTGAGACTCT
CCTGTGCAGCCTCTGTGTTCACCGTCAGTAGCAACTACATGAGCTGGGTCCGCCAGGCTCCA
GGGAAGGGGCTGGAGTGGGTCTCAGTTATTTATCCCGGTGGTAGCACATTCTACGCAGACTC
CGTGAAGGGCCGATTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTATCTTCAAATGA
ACAGCCTGAGAGCCGAGGACACGGCCGTGTATTACTGTGCGAGAGAGGTGGCTGGTACCTA TGACTACTGGGGCCAGGGAACCCTGGTCACCGTCTCCTCAGCCTCCACCAAGGGCCCATCGG
TCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTG
GTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCACTGACCAGCG
GCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTG
ACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAG
CAACACCAAGGTGGACAAGAGAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCA
CCGTGCCCAGCACCTGAACTCCTGGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAA
GGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACG
AAGACCCTGAGGTCAAGTTCAACTGGTATGTGGACGGCGTGGAGGTGCATAATGCCAAGAC
AAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTG
CACCAAGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCCCAG
CCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACAC
CCTGCCCCCATCCCGGGAGGAGATGACCAAGAACCAAGTCAGCCTGACCTGCCTGGTCAAA
GGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACT
ACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTATTCCAAGCTCACCG
TGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCT
GCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGCAAA
SEQ ID NO: 156 Heavy Chain DNA Sequence of481-L5H2- h!gGl-ACT5
GAGGTGCAGCTGGTGGAGTCTGGAGGAGGCTTGATCCAGCCTGGGGGGTCCCTGAGACTCT
CCTGTGCAGCCTCTGTGTTCACCGTCAGTAGCAACTACATGAGCTGGGTCCGCCAGGCTCCA
GGGAAGGGGCTGGAGTGGGTCTCAGTTATTTATCCCGGTGGTAGCACATTCTACGCAGACTC
CGTGAAGGGCCGATTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTATCTTCAAATGA
ACAGCCTGAGAGCCGAGGACACGGCCGTGTATTACTGTGCGAGAGAGGTGGCTGGTACCTA
TGACTACTGGGGCCAGGGAACCCTGGTCACCGTCTCCTCAGCCTCCACCAAGGGCCCATCGG
TCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTG
GTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCACTGACCAGCG
GCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTG
ACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAG
CAACACCAAGGTGGACAAGAGAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCA
CCGTGCCCAGCACCTGAACTCCTGGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAA
GGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACG
AAGACCCTGAGGTCAAGTTCAACTGGTATGTGGACGGCGTGGAGGTGCATAATGCCAAGAC
AAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTG
CACCAAGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCCCAG
CCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACAC
CCTGCCCCCATCCCGGGAGGAGATGACCAAGAACCAAGTCAGCCTGACCTGCCTGGTCAAA
GGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACT
ACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTATTCCAAGCTCACCG
TGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGCTGCATGAGGCTCTG
CACGCTCACTACACGAGGAAGGAGCTCTCCCTGTCTCCGGGCAAA
SEQ ID NO: 157 Light Chain DNA Sequence of 481-L5H2 GACATCCAGATGACCCAGTCTCCATCTTCTGTGTCTGCATCTGTAGGAGACAGAGTCACCAT
CACTTGTCGGGCGAGTCAGGGTATTAGCAGCTGGTTAGCCTGGTATCAGCAGAAACCAGGG
AAAGCCCCTAAACTCCTGATCTATGCTGCATCCAGTTTGCAAAGTGGGGTCCCATCAAGGTT
CAGCGGCAGTGGATCTGGGACAGATTTCACTCTCACTATCAGCAGCCTGCAGCCTGAAGATT
TTGCAACTTACTATTGTCAACAGACCATCAGTTTCCCTGGAGGCACTTTCGGCCCTGGGACC
AAAGTGGATATCAAAAGAACTGTGGCGGCGCCATCTGTCTTCATCTTCCCGCCATCTGATGA
GCAGTTGAAATCCGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGG
CCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCAC
AGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCA
GACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCG
TCACAAAGAGCTTCAACAGGGGAGAGTGC
SEQ ID NO: 158 Heavy Chain DNA Sequence of 373-L2H24 - hlgGl
CAGGTGCAGCTGCAGGAGTCGGGCCCAGGACTGGTGAAGCCTTCGGAGACCCTGTCCCTCA
CCTGCACTGTCTCTGGTCACTCCATCAGTAGTTACCACTGGGAGTGGATCCGGCAGCCCCCA
GGGAAGGGACTGGAGTGGATTGGGTATATCAAGTACAGTGGGAGCACCAACTACAACCCCT
CCCTCAAGAGTCGAGTCACCATATCAGTAGACACGTCCAAGAACCAGTTCTCCCTGAAGCTG
AGCTCTGTGACCGCTGCGGACACGGCCGTGTATTACTGTGCGAGAGGGCCCGACTATTACGA
TTTCTGGGCCGGTTATTACTACGGTGTGGACGTCTGGGGCCAAGGGACCACGGTCACCGTCT
CCTCAGCCTCCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTCT
GGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGT
CGTGGAACTCAGGCGCACTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCA
GGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTA
CATCTGCAACGTGAATCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTTGAGCCCAAA
TCTTGTGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAACTCCTGGGGGGACCGTC
AGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCA
CATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTATGTGGA
CGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTA
CCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAAGACTGGCTGAATGGCAAGGAGTACAAGT
GCAAGGTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGG
GCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGAGGAGATGACCAAGAAC
CAAGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGA
GAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGC
TCCTTCTTCCTCTATTCCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTT
CTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGT
CTCCGGGCAAA
SEQ ID NO: 159 Heavy Chain DNA Sequence of 373-L2H24 - hlgGl -ACT5
CAGGTGCAGCTGCAGGAGTCGGGCCCAGGACTGGTGAAGCCTTCGGAGACCCTGTCCCTCA
CCTGCACTGTCTCTGGTCACTCCATCAGTAGTTACCACTGGGAGTGGATCCGGCAGCCCCCA
GGGAAGGGACTGGAGTGGATTGGGTATATCAAGTACAGTGGGAGCACCAACTACAACCCCT
CCCTCAAGAGTCGAGTCACCATATCAGTAGACACGTCCAAGAACCAGTTCTCCCTGAAGCTG
AGCTCTGTGACCGCTGCGGACACGGCCGTGTATTACTGTGCGAGAGGGCCCGACTATTACGA
TTTCTGGGCCGGTTATTACTACGGTGTGGACGTCTGGGGCCAAGGGACCACGGTCACCGTCT CCTCAGCCTCCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTCT
GGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGT
CGTGGAACTCAGGCGCACTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCA
GGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTA
CATCTGCAACGTGAATCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTTGAGCCCAAA
TCTTGTGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAACTCCTGGGGGGACCGTC
AGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCA
CATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTATGTGGA
CGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTA
CCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAAGACTGGCTGAATGGCAAGGAGTACAAGT
GCAAGGTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGG
GCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGAGGAGATGACCAAGAAC
CAAGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGA
GAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGC
TCCTTCTTCCTCTATTCCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTT
CTCATGCTCCGTGCTGCATGAGGCTCTGCACGCTCACTACACGAGGAAGGAGCTCTCCCTGT
CTCCGGGCAAA
SEQ ID NO: 160 Light Chain DNA Sequence of 373-L2H24
GAAATTGTGTTGACGCAGTCTCCAGGCACCCTGTCTTTGTCTCCAGGGGAAAGAGCCACCCT
CTCCTGCAGGGCCAGTCAGAGTGTTGACACCAGCTACTTAGCCTGGTACCAGCAGAAACCTG
GCCAGGCTCCCAGGCTCCTCATCTATGGTGCATCCAGCAGGGCCACTGGCATCCCAGACAGG
TTCAGTGGCAGTGGGTCTGGGACAGACTTCACTCTCACCATCAGCAGACTGGAGCCTGAAGA
TTTTGCAGTGTATTACTGTCAGCAGTATGGTAGCTCAACCACTTTCGGCGGAGGGACCAAGG
TGGAGATCAAAAGAACTGTGGCGGCGCCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAG
TTGAAATCCGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAA
AGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAG
CAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACT
ACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCAC
AAAGAGCTTCAACAGGGGAGAGTGC

Claims

1. An antibody that binds SARS-CoV-2 spike protein, wherein the antibody comprises a heavy chain variable region (VH) and a light chain variable region (VL), wherein the VH comprises heavy chain complementarity determining regions HCDR1, HCDR2, and HCDR3, and the VL comprises light chain complementarity determining regions LCDR1, LCDR2, and LCDR3, wherein the antibody comprises:
HCDR1 comprising SEQ ID NO: 3,
HCDR2 comprising SEQ ID NO: 4,
HCDR3 comprising SEQ ID NO: 5, 18, or 26,
LCDR1 comprising SEQ ID NO: 6,
LCDR2 comprising SEQ ID NO: 7 or 13, and LCDR3 comprising SEQ ID NO: 8, 14, or 19.
2. The antibody of claim 1, wherein the antibody comprises:
(a) HCDR1 comprising SEQ ID NO: 3, HCDR2 comprising SEQ ID NO: 4, HCDR3 comprising SEQ ID NO: 5, LCDR1 comprising SEQ ID NO: 6, LCDR2 comprising SEQ ID NO: 7, and LCDR3 comprising SEQ ID NO: 8;
(b) HCDR1 comprising SEQ ID NO: 3, HCDR2 comprising SEQ ID NO: 4, HCDR3 comprising SEQ ID NO: 5, LCDR1 comprising SEQ ID NO: 6, LCDR2 comprising SEQ ID NO: 13, and LCDR3 comprising SEQ ID NO: 14;
(c) HCDR1 comprising SEQ ID NO: 3, HCDR2 comprising SEQ ID NO: 4, HCDR3 comprising SEQ ID NO: 18, LCDR1 comprising SEQ ID NO: 6, LCDR2 comprising SEQ ID NO: 7, and LCDR3 comprising SEQ ID NO: 19;
(d) HCDR1 comprising SEQ ID NO: 3, HCDR2 comprising SEQ ID NO: 4, HCDR3 comprising SEQ ID NO: 5, LCDR1 comprising SEQ ID NO: 6, LCDR2 comprising SEQ ID NO: 13, and LCDR3 comprising SEQ ID NO: 19;
(e) HCDR1 comprising SEQ ID NO: 3, HCDR2 comprising SEQ ID NO: 4, HCDR3 comprising SEQ ID NO: 26, LCDR1 comprising SEQ ID NO: 6, LCDR2 comprising SEQ ID NO: 7, and LCDR3 comprising SEQ ID NO: 8; or
(f) HCDR1 comprising SEQ ID NO: 3, HCDR2 comprising SEQ ID NO: 4, HCDR3 comprising SEQ ID NO: 18, LCDR1 comprising SEQ ID NO: 6, LCDR2 comprising SEQ ID NO: 7, and LCDR3 comprising SEQ ID NO: 8.
3. The antibody of claim 1, wherein the antibody comprises a VH comprising SEQ ID NO: 1, 16 or 25, and a VL comprising SEQ ID NO: 2, 12, 17, or 23.
4. The antibody of any one of claims 1-3, wherein the antibody comprises:
(a) a VH comprising SEQ ID NO: 1, and a VL comprising SEQ ID NO: 2;
(b) a VH comprising SEQ ID NO: 1, and a VL comprising SEQ ID NO: 12;
(c) a VH comprising SEQ ID NO: 16, and a VL comprising SEQ ID NO: 17;
(d) a VH comprising SEQ ID NO: 1, and a VL comprising SEQ ID NO: 23;
(e) a VH comprising SEQ ID NO: 25, and a VL comprising SEQ ID NO: 2; or
(f) a VH comprising SEQ ID NO: 16, and a VL comprising SEQ ID NO: 2.
5. The antibody of any one of claims 1-4, wherein the antibody comprises:
(a) a heavy chain (HC) comprising SEQ ID NO: 9 or 10, and a light chain (LC) comprising SEQ ID NO: 11;
(b) a HC comprising SEQ ID NO: 9 or 10, and a LC comprising SEQ ID NO: 15;
(c) a HC comprising SEQ ID NO: 20 or 21, and a LC comprising SEQ ID NO: 22;
(d) a HC comprising SEQ ID NO: 9 or 10, and a LC comprising SEQ ID NO: 24;
(e) a HC comprising SEQ ID NO: 27 or 28, and a LC comprising SEQ ID NO: 11; or
(f) a HC comprising SEQ ID NO: 20 or 21, and a LC comprising SEQ ID NO: 11.
6. A nucleic acid comprising a sequence encoding SEQ ID NO: 9, 10, 20, 21, 27, or 28.
7. A nucleic acid comprising a sequence encoding SEQ ID NO: 11, 15, 22, or 24.
8. A vector comprising the nucleic acid of claim 6, or the nucleic acid of claim 7, or both the nucleic acid of claim 6 and the nucleic acid of claim 7.
9. A cell comprising
(a) a vector comprising both the nucleic acid of claim 6 and the nucleic acid of claim 7; or
(b) a first vector comprising the nucleic acid of claim 6 and a second vector comprising the nucleic acid of claim 7.
10. A process of producing an antibody comprising culturing the cell of claim 9 under conditions such that the antibody is expressed and recovering the expressed antibody from the culture medium.
11. An antibody that binds SARS-CoV-2 spike protein, wherein the antibody comprises a VH and a VL, wherein the VH comprises heavy chain complementarity determining regions HCDR1, HCDR2, and HCDR3, and the VL comprises light chain complementarity determining regions LCDR1, LCDR2, and LCDR3, wherein the antibody comprises: HCDR1 comprising SEQ ID NO: 31,
HCDR2 comprising SEQ ID NO: 32,
HCDR3 comprising SEQ ID NO: 33, 44, or 50,
LCDR1 comprising SEQ ID NO: 34,
LCDR2 comprising SEQ ID NO: 35, and
LCDR3 comprising SEQ ID NO: 36 or 45. . The antibody of claim 11, wherein the antibody comprises:
(a) HCDR1 comprising SEQ ID NO: 31,
HCDR2 comprising SEQ ID NO: 32,
HCDR3 comprising SEQ ID NO: 33,
LCDR1 comprising SEQ ID NO: 34,
LCDR2 comprising SEQ ID NO: 35, and
LCDR3 comprising SEQ ID NO: 36;
(b) HCDR1 comprising SEQ ID NO: 31,
HCDR2 comprising SEQ ID NO: 32,
HCDR3 comprising SEQ ID NO: 44,
LCDR1 comprising SEQ ID NO: 34,
LCDR2 comprising SEQ ID NO: 35, and
LCDR3 comprising SEQ ID NO: 45; or
(c) HCDR1 comprising SEQ ID NO: 31,
HCDR2 comprising SEQ ID NO: 32,
HCDR3 comprising SEQ ID NO: 50,
LCDR1 comprising SEQ ID NO: 34,
LCDR2 comprising SEQ ID NO: 35, and
LCDR3 comprising SEQ ID NO: 36. . The antibody of claim 11, wherein the antibody comprises a VH comprising SEQ ID NO:, 42, or 49, and a VL comprising SEQ ID NO: 30, 40, 43. . The antibody of any one of claims 11-13, wherein the antibody comprises: (a) a VH comprising SEQ ID NO: 29, and a VL comprising SEQ ID NO: 30;
(b) a VH comprising SEQ ID NO: 29, and a VL comprising SEQ ID NO: 40;
(c) a VH comprising SEQ ID NO: 42, and a VL comprising SEQ ID NO: 43; or
(d) a VH comprising SEQ ID NO: 49, and a VL comprising SEQ ID NO: 40.
15. The antibody of any one of claims 11-14, wherein the antibody comprises:
(a) a HC comprising SEQ ID NO: 37 or 38, and a LC comprising SEQ ID NO: 39;
(b) a HC comprising SEQ ID NO: 37 or 38, and a LC comprising SEQ ID NO: 41;
(c) a HC comprising SEQ ID NO: 46 or 47, and a LC comprising SEQ ID NO: 48; or
(d) a HC comprising SEQ ID NO: 51 or 52, and a LC comprising SEQ ID NO: 41.
16. A nucleic acid comprising a sequence encoding SEQ ID NO: 37, 38, 46, 47, 51, or 52.
17. A nucleic acid comprising a sequence encoding SEQ ID NO: 39, 41, or 48.
18. A vector comprising the nucleic acid of claim 16, or the nucleic acid of claim 17, or both the nucleic acid of claim 16 and the nucleic acid of claim 17.
19. A cell comprising
(a) a vector comprising both the nucleic acid of claim 16 and the nucleic acid of claim 17; or
(b) a first vector comprising the nucleic acid of claim 16 and a second vector comprising the nucleic acid of claim 17.
20. A process of producing an antibody comprising culturing the cell of claim 19 under conditions such that the antibody is expressed and recovering the expressed antibody from the culture medium.
21. An antibody that binds SARS-CoV-2 spike protein, wherein the antibody comprises a VH and a VL, wherein the VH comprises heavy chain complementarity determining regions HCDR1, HCDR2, and HCDR3, and the VL comprises light chain complementarity determining regions LCDR1, LCDR2, and LCDR3, wherein the antibody comprises:
HCDR1 comprising SEQ ID NO: 55,
HCDR2 comprising SEQ ID NO: 56 or 73,
HCDR3 comprising SEQ ID NO: 57,
LCDR1 comprising SEQ ID NO: 58, 65, 69, or 74,
LCDR2 comprising SEQ ID NO: 59 or 66, and
LCDR3 comprising SEQ ID NO: 60.
22. The antibody of claim 21, wherein the antibody comprises:
(a) HCDR1 comprising SEQ ID NO: 55,
HCDR2 comprising SEQ ID NO: 56,
HCDR3 comprising SEQ ID NO: 57,
LCDR1 comprising SEQ ID NO: 58,
LCDR2 comprising SEQ ID NO: 59, and
LCDR3 comprising SEQ ID NO: 60;
(b) HCDR1 comprising SEQ ID NO: 55,
HCDR2 comprising SEQ ID NO: 56,
HCDR3 comprising SEQ ID NO: 57,
LCDR1 comprising SEQ ID NO: 65,
LCDR2 comprising SEQ ID NO: 66, and
LCDR3 comprising SEQ ID NO: 60;
(c) HCDR1 comprising SEQ ID NO: 55,
HCDR2 comprising SEQ ID NO: 56,
HCDR3 comprising SEQ ID NO: 57,
LCDR1 comprising SEQ ID NO: 69,
LCDR2 comprising SEQ ID NO: 59, and
LCDR3 comprising SEQ ID NO: 60; or
(d) HCDR1 comprising SEQ ID NO: 55,
HCDR2 comprising SEQ ID NO: 73,
HCDR3 comprising SEQ ID NO: 57, LCDR1 comprising SEQ ID NO: 74, LCDR2 comprising SEQ ID NO: 59, and LCDR3 comprising SEQ ID NO: 60.
23. The antibody of claim 21, wherein the antibody comprises a VH comprising SEQ ID NO: 53 or 71, and a VL comprising SEQ ID NO: 54, 64, 68, or 72.
24. The antibody of any one of claims 21-23, wherein the antibody comprises:
(a) a VH comprising SEQ ID NO: 53, and a VL comprising SEQ ID NO: 54;
(b) a VH comprising SEQ ID NO: 53, and a VL comprising SEQ ID NO: 64;
(c) a VH comprising SEQ ID NO: 53, and a VL comprising SEQ ID NO: 68; or
(d) a VH comprising SEQ ID NO: 71, and a VL comprising SEQ ID NO: 72.
25. The antibody of any one of claims 21-24, wherein the antibody comprises:
(a) a HC comprising SEQ ID NO: 61 or 62, and a LC comprising SEQ ID NO: 63;
(b) a HC comprising SEQ ID NO: 61 or 62, and a LC comprising SEQ ID NO: 67;
(c) a HC comprising SEQ ID NO: 61 or 62, and a LC comprising SEQ ID NO: 70; or
(d) a HC comprising SEQ ID NO: 75 or 76, and a LC comprising SEQ ID NO: 77.
26. A nucleic acid comprising a sequence encoding SEQ ID NO: 61, 62, 75, or 76.
27. A nucleic acid comprising a sequence encoding SEQ ID NO: 63, 67, 70, or 77.
28. A vector comprising the nucleic acid of claim 26, or the nucleic acid of claim 27, or both the nucleic acid of claim 26 and the nucleic acid of claim 27.
29. A cell comprising
(a) a vector comprising both the nucleic acid of claim 26 and the nucleic acid of claim 27; or
(b) a first vector comprising the nucleic acid of claim 26 and a second vector comprising the nucleic acid of claim 27.
30. A process of producing an antibody comprising culturing the cell of claim 29 under conditions such that the antibody is expressed and recovering the expressed antibody from the culture medium.
31. An antibody that binds SARS-CoV-2 spike protein, wherein the antibody comprises a VH and a VL, wherein the VH comprises heavy chain complementarity determining regions HCDR1, HCDR2, and HCDR3, and the VL comprises light chain complementarity determining regions LCDR1, LCDR2, and LCDR3, wherein the antibody comprises:
HCDR1 comprising SEQ ID NO: 80, 89, or 95,
HCDR2 comprising SEQ ID NO: 81,
HCDR3 comprising SEQ ID NO: 82 or 90,
LCDR1 comprising SEQ ID NO: 83,
LCDR2 comprising SEQ ID NO: 13, and LCDR3 comprising SEQ ID NO: 84 or 96.
32. The antibody of claim 31, wherein the antibody comprises:
(a) HCDR1 comprising SEQ ID NO: 80, HCDR2 comprising SEQ ID NO: 81, HCDR3 comprising SEQ ID NO: 82, LCDR1 comprising SEQ ID NO: 83, LCDR2 comprising SEQ ID NO: 13, and LCDR3 comprising SEQ ID NO: 84;
(b) HCDR1 comprising SEQ ID NO: 89,
HCDR2 comprising SEQ ID NO: 81,
HCDR3 comprising SEQ ID NO: 90,
LCDR1 comprising SEQ ID NO: 83,
LCDR2 comprising SEQ ID NO: 13, and LCDR3 comprising SEQ ID NO: 84;
(c) HCDR1 comprising SEQ ID NO: 95, HCDR2 comprising SEQ ID NO: 81, HCDR3 comprising SEQ ID NO: 82,
LCDR1 comprising SEQ ID NO: 83,
LCDR2 comprising SEQ ID NO: 13, and
LCDR3 comprising SEQ ID NO: 96; or
(d) HCDR1 comprising SEQ ID NO: 80,
HCDR2 comprising SEQ ID NO: 81,
HCDR3 comprising SEQ ID NO: 82,
LCDR1 comprising SEQ ID NO: 83,
LCDR2 comprising SEQ ID NO: 13, and
LCDR3 comprising SEQ ID NO: 96. . The antibody of claim 31, wherein the antibody comprises a VH comprising SEQ ID NO:, 88 or 93, and a VL comprising SEQ ID NO: 79 or 94. . The antibody of any one of claims 31-33, wherein the antibody comprises:
(a) a VH comprising SEQ ID NO: 78, and a VL comprising SEQ ID NO: 79;
(b) a VH comprising SEQ ID NO: 88, and a VL comprising SEQ ID NO: 79;
(c) a VH comprising SEQ ID NO: 93, and a VL comprising SEQ ID NO: 94; or
(d) a VH comprising SEQ ID NO: 78, and a VL comprising SEQ ID NO: 94. . The antibody of any one of claims 31-34, wherein the antibody comprises:
(a) a HC comprising SEQ ID NO: 85 or 86, and a LC comprising SEQ ID NO: 87;
(b) a HC comprising SEQ ID NO: 91 or 92, and a LC comprising SEQ ID NO: 87;
(c) a HC comprising SEQ ID NO: 97 or 98, and a LC comprising SEQ ID NO: 99; or
(d) a HC comprising SEQ ID NO: 85 or 86, and a LC comprising SEQ ID NO: 99. . A nucleic acid comprising a sequence encoding SEQ ID NO: 85, 86, 91, 92, 97, or 98. . A nucleic acid comprising a sequence encoding SEQ ID NO: 87 or 99.
38. A vector comprising the nucleic acid of claim 36, or the nucleic acid of claim 37, or both the nucleic acid of claim 36 and the nucleic acid of claim 37.
39. A cell comprising
(a) a vector comprising both the nucleic acid of claim 36 and the nucleic acid of claim 37; or
(b) a first vector comprising the nucleic acid of claim 36 and a second vector comprising the nucleic acid of claim 37.
40. A process of producing an antibody comprising culturing the cell of claim 39 under conditions such that the antibody is expressed and recovering the expressed antibody from the culture medium.
41. An antibody that binds SARS-CoV-2 spike protein, wherein the antibody comprises a VH and a VL, wherein the VH comprises heavy chain complementarity determining regions HCDR1, HCDR2, and HCDR3, and the VL comprises light chain complementarity determining regions LCDR1, LCDR2, and LCDR3, wherein the antibody comprises:
HCDR1 comprising SEQ ID NO: 102 or 112,
HCDR2 comprising SEQ ID NO: 103 or 119,
HCDR3 comprising SEQ ID NO: 104, 120, or 127,
LCDR1 comprising SEQ ID NO: 105 or 116,
LCDR2 comprising SEQ ID NO: 106, and LCDR3 comprising SEQ ID NO: 107.
42. The antibody of claim 41, wherein the antibody comprises:
(a) HCDR1 comprising SEQ ID NO: 102, HCDR2 comprising SEQ ID NO: 103, HCDR3 comprising SEQ ID NO: 104, LCDR1 comprising SEQ ID NO: 105, LCDR2 comprising SEQ ID NO: 106, and LCDR3 comprising SEQ ID NO: 107; (b) HCDR1 comprising SEQ ID NO: 112, HCDR2 comprising SEQ ID NO: 103, HCDR3 comprising SEQ ID NO: 104, LCDR1 comprising SEQ ID NO: 105, LCDR2 comprising SEQ ID NO: 106, and LCDR3 comprising SEQ ID NO: 107;
(c) HCDR1 comprising SEQ ID NO: 112, HCDR2 comprising SEQ ID NO: 103, HCDR3 comprising SEQ ID NO: 104, LCDR1 comprising SEQ ID NO: 116, LCDR2 comprising SEQ ID NO: 106, and LCDR3 comprising SEQ ID NO: 107;
(d) HCDR1 comprising SEQ ID NO: 112, HCDR2 comprising SEQ ID NO: 119, HCDR3 comprising SEQ ID NO: 120, LCDR1 comprising SEQ ID NO: 116, LCDR2 comprising SEQ ID NO: 106, and LCDR3 comprising SEQ ID NO: 107;
(e) HCDR1 comprising SEQ ID NO: 112, HCDR2 comprising SEQ ID NO: 119, HCDR3 comprising SEQ ID NO: 104, LCDR1 comprising SEQ ID NO: 116, LCDR2 comprising SEQ ID NO: 106, and LCDR3 comprising SEQ ID NO: 107; or
(f) HCDR1 comprising SEQ ID NO: 112, HCDR2 comprising SEQ ID NO: 119, HCDR3 comprising SEQ ID NO: 127, LCDR1 comprising SEQ ID NO: 116, LCDR2 comprising SEQ ID NO: 106, and LCDR3 comprising SEQ ID NO: 107.
43. The antibody of claim 41, wherein the antibody comprises a VH comprising SEQ ID NO: 100, 111, 118, 123, or 126, and a VL comprising SEQ ID NO: 101 or 115.
44. The antibody of any one of claims 41-43, wherein the antibody comprises:
(a) a VH comprising SEQ ID NO: 100, and a VL comprising SEQ ID NO: 101;
(b) a VH comprising SEQ ID NO: 111, and a VL comprising SEQ ID NO: 101;
(c) a VH comprising SEQ ID NO: 111, and a VL comprising SEQ ID NO: 115;
(d) a VH comprising SEQ ID NO: 118, and a VL comprising SEQ ID NO: 115;
(e) a VH comprising SEQ ID NO: 123, and a VL comprising SEQ ID NO: 115; or
(f) a VH comprising SEQ ID NO: 126, and a VL comprising SEQ ID NO: 115.
45. The antibody of any one of claims 41-44, wherein the antibody comprises:
(a) a HC comprising SEQ ID NO: 108 or 109, and a LC comprising SEQ ID NO: 110;
(b) a HC comprising SEQ ID NO: 113 or 114, and a LC comprising SEQ ID NO: 110;
(c) a HC comprising SEQ ID NO: 113 or 114, and a LC comprising SEQ ID NO: 117;
(d) a HC comprising SEQ ID NO: 121 or 122, and a LC comprising SEQ ID NO: 117;
(e) a HC comprising SEQ ID NO: 124 or 125, and a LC comprising SEQ ID NO: 117; or
(f) a HC comprising SEQ ID NO: 128 or 129, and a LC comprising SEQ ID NO: 117.
46. A nucleic acid comprising a sequence encoding SEQ ID NO: 108, 109, 113, 114, 121, 122, 124, 125, 128, or 129.
47. A nucleic acid comprising a sequence encoding SEQ ID NO: 110 or 117.
48. A vector comprising the nucleic acid of claim 46, or the nucleic acid of claim 47, or both the nucleic acid of claim 46 and the nucleic acid of claim 47.
49. A cell comprising
(a) a vector comprising both the nucleic acid of claim 46 and the nucleic acid of claim 47; or (b) a first vector comprising the nucleic acid of claim 46 and a second vector comprising the nucleic acid of claim 47.
50. A process of producing an antibody comprising culturing the cell of claim 49 under conditions such that the antibody is expressed and recovering the expressed antibody from the culture medium.
51. An antibody that binds SARS-CoV-2 spike protein, wherein the antibody comprises a VH and a VL, wherein the VH comprises heavy chain complementarity determining regions HCDR1, HCDR2, and HCDR3, and the VL comprises light chain complementarity determining regions LCDR1, LCDR2, and LCDR3, wherein the antibody comprises:
HCDR1 comprising SEQ ID NO: 137,
HCDR2 comprising SEQ ID NO: 138,
HCDR3 comprising SEQ ID NO: 139,
LCDR1 comprising SEQ ID NO: 140,
LCDR2 comprising SEQ ID NO: 141, and
LCDR3 comprising SEQ ID NO: 142.
52. The antibody of claim 51, wherein the antibody comprises a VH comprising SEQ ID NO: 135, and a VL comprising SEQ ID NO: 136.
53. The antibody of claim 51 or 52, wherein the antibody comprises a HC comprising SEQ ID NO: 143 or 144, and a LC comprising SEQ ID NO: 145.
54. A nucleic acid comprising a sequence encoding SEQ ID NO: 143 or 144.
55. A nucleic acid comprising a sequence encoding SEQ ID NO: 145.
56. A vector comprising the nucleic acid of claim 54, or the nucleic acid of claim 55, or both the nucleic acid of claim 54 and the nucleic acid of claim 55.
57. A cell comprising
(a) a vector comprising both the nucleic acid of claim 54 and the nucleic acid of claim 55; or
(b) a first vector comprising the nucleic acid of claim 54 and a second vector comprising the nucleic acid of claim 55.
58. A process of producing an antibody comprising culturing the cell of claim 57 under conditions such that the antibody is expressed and recovering the expressed antibody from the culture medium.
59. An antibody produced by the process of claim 10, 20, 30, 40, 50 or 58.
60. A pharmaceutical composition comprising the antibody of any one of claims 1-5, 11-15, 21- 25, 31-35, 41-45, 51-53, and 59, and one or more pharmaceutically acceptable carrier, diluent, or excipient.
61. The pharmaceutical composition of claim 60, wherein the pharmaceutical composition comprises at least one additional antibody that binds the SARS-CoV-2 spike protein.
62. A method of treating or preventing COVID-19 comprising administering to a patient a therapeutically effective amount of the antibody of any one of claims 1-5, 11-15, 21-25, 31-35, 41-45, 51-53, and 59, or the pharmaceutical composition of claim 60 or 61.
63. A method of treating or preventing COVID- 19 comprising: contacting a sample obtained from a patient with the antibody or antigen-binding fragment thereof of any one of claims 1-5, 11-15, 21-25, 31-35, 41-45, 51-53, and 59, conjugated to a detectable agent; detecting specific binding of the antibody or antigen-binding fragment thereof to a SARS- CoV-2 antigen present in the sample; and administering to the patient a therapeutically effective amount of the antibody of any one of claims 1-5, 11-15, 21-25, 31-35, 41-45, 51-53, and 59, or the pharmaceutical composition of claim 60 or 61.
64. A method of identifying a SARS-CoV-2-infected cell comprising: contacting a cell with the antibody of any one of claims 1-5, 11-15, 21-25, 31-35, 41-45, 51-53, and 59, conjugated to a detectable agent; and detecting specific binding of the antibody to the cell.
65. A method of diagnosing a SARS-CoV-2 infection in a patient comprising: contacting a sample obtained from a patient with the antibody of any one of claims 1-5, 11-15, 21-25, 31-35, 41-45, 51-53, and 59, conjugated to a detectable agent; and detecting specific binding of the antibody to a SARS-CoV-2 antigen present in the sample.
66. The antibody of any one of claims 1-5, 11-15, 21-25, 31-35, 41-45, 51-53, and 59, or the pharmaceutical composition of claim 60 or 61, for use in therapy.
67. The antibody of any one of claims 1-5, 11-15, 21-25, 31-35, 41-45, 51-53, and 59, or the pharmaceutical composition of claim 60 or 61, for use in the treatment or prevention of COVID- 19.
68. Use of the antibody of any one of claims 1-5, 11-15, 21-25, 31-35, 41-45, 51-53, and 59, in the manufacture of a medicament for the treatment or prevention of COVID- 19.
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