The receptor binding domain of SARS-CoV-2 spike is the key target of neutralizing antibody in human polyclonal sera

  1. Tara L. Steffen
  2. E. Taylor Stone
  3. Mariah Hassert
  4. Elizabeth Geerling
  5. Brian T. Grimberg
  6. Ana M. Espino
  7. Petraleigh Pantoja
  8. Consuelo Climent
  9. Daniel F. Hoft
  10. Sarah L. George
  11. Carlos A. Sariol
  12. Amelia K. Pinto
  13. James D. Brien

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Abstract

Natural infection of SARS-CoV-2 in humans leads to the development of a strong neutralizing antibody response, however the immunodominant targets of the polyclonal neutralizing antibody response are still unknown. Here, we functionally define the role SARS-CoV-2 spike plays as a target of the human neutralizing antibody response. In this study, we identify the spike protein subunits that contain antigenic determinants and examine the neutralization capacity of polyclonal sera from a cohort of patients that tested qRT-PCR-positive for SARS-CoV-2. Using an ELISA format, we assessed binding of human sera to spike subunit 1 (S1), spike subunit 2 (S2) and the receptor binding domain (RBD) of spike. To functionally identify the key target of neutralizing antibody, we depleted sera of subunit-specific antibodies to determine the contribution of these individual subunits to the antigen-specific neutralizing antibody response. We show that epitopes within RBD are the target of a majority of the neutralizing antibodies in the human polyclonal antibody response . These data provide critical information for vaccine development and development of sensitive and specific serological testing.

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  1. ScreenIT

    SciScore for 10.1101/2020.08.21.261727: (What is this?)

    Please note, not all rigor criteria are appropriate for all manuscripts.

    Table 1: Rigor

    Institutional Review Board StatementIRB: All collection, processing and archiving of human specimens was performed under approval from the University Institutional Review
    Randomizationnot detected.
    Blindingnot detected.
    Power Analysisnot detected.
    Sex as a biological variablenot detected.
    Cell Line Authenticationnot detected.

    Table 2: Resources

    Experimental Models: Cell Lines
    SentencesResources
    Viruses and cells: SARS-CoV-2 was obtained from Biodefense and Emerging Infections (BEI) Research Resources Repository and passaged once in Vero E6 cells purchased from American Type Culture Collection (ATCC CCL-81)
    Vero E6
    suggested: None
    (FRNT50 post control depletion))*100. Focus Reduction Neutralization Test: Four-fold serial dilutions of human serum were mixed with ~100 focus-forming units (FFU) of virus, incubated at 37°C for 1 h, and added to Vero WHO monolayers in 96-well plates for 1 h at 37°C to allow virus adsorption.
    Vero
    suggested: None
    Software and Algorithms
    SentencesResources
    Sequences were trimmed and aligned with Geneious 7 and single amino acid variants were identified and quantified using the analyze sequence variation tool of the NIAID Virus Pathogen Database and Analysis Resource (VIPR) and shannon entropy calculations.
    Geneious
    suggested: (Geneious, RRID:SCR_010519)
    Display of the aa variation on the cryo-em structure was completed using pymol v2.40
    pymol
    suggested: (PyMOL, RRID:SCR_000305)
    FRNT curves were generated by log-transformation of the x axis followed by non-linear curve fit regression analysis using Graphpad Prism 8.
    Graphpad Prism
    suggested: (GraphPad Prism, RRID:SCR_002798)

    Results from OddPub: We did not detect open data. We also did not detect open code. Researchers are encouraged to share open data when possible (see Nature blog).

    Results from LimitationRecognizer: An explicit section about the limitations of the techniques employed in this study was not found. We encourage authors to address study limitations.

    Results from TrialIdentifier: No clinical trial numbers were referenced.

    Results from Barzooka: We did not find any issues relating to the usage of bar graphs.

    Results from JetFighter: We did not find any issues relating to colormaps.

    Results from rtransparent:
    • Thank you for including a conflict of interest statement. Authors are encouraged to include this statement when submitting to a journal.
    • Thank you for including a funding statement. Authors are encouraged to include this statement when submitting to a journal.
    • No protocol registration statement was detected.

    About SciScore

    SciScore is an automated tool that is designed to assist expert reviewers by finding and presenting formulaic information scattered throughout a paper in a standard, easy to digest format. SciScore checks for the presence and correctness of RRIDs (research resource identifiers), and for rigor criteria such as sex and investigator blinding. For details on the theoretical underpinning of rigor criteria and the tools shown here, including references cited, please follow this link.

  2. Version published to 10.1101/2020.08.21.261727 on bioRxiv