SARS-CoV-2 neutralization and serology testing of COVID-19 convalescent plasma from donors with non-severe disease

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Abstract

We determined the antigen binding activity of convalescent plasma units from 47 individuals with a history of non-severe COVID-19 using three clinical diagnostic serology assays (Beckman, DiaSorin, and Roche) with different SARS-CoV-2 targets. We compared these results with functional neutralization activity using a fluorescent reporter strain of SARS-CoV-2 in a microwell assay. This revealed positive correlations of varying strength (Spearman r = 0.37-0.52) between binding and neutralization. Donors age 48-75 had the highest neutralization activity. Units in the highest tertile of binding activity for each assay were enriched (75-82%) for those with the highest levels of neutralization.

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  1. SciScore for 10.1101/2020.08.07.242271: (What is this?)

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

    Table 1: Rigor

    Institutional Review Board StatementIRB: This program and associated human subject research, performed in in accordance with the ethical standards of the Helsinki Declaration, were approved by the NorthShore University HealthSystem Institutional Review Board.
    Consent: All potential donors provided written consent for the study and provided information about their COVID-19 disease history and demographics.
    RandomizationSamples were divided into tertiles based on the Roche assay results, then 47 were randomly selected to equally sample each tertile.
    Blindingnot detected.
    Power Analysisnot detected.
    Sex as a biological variablenot detected.
    Cell Line Authenticationnot detected.

    Table 2: Resources

    Antibodies
    SentencesResources
    SARS-CoV-2 S1 RBD Ig ELISA: The anti-SARS-CoV-2 S1 RBD total Ig assay employed a standard indirect enzyme immunoassay technique (ELISA), described in detail elsewhere, using a secondary antibody recognizing all human immunoglobulin isotypes (goat anti-human IgG H+L-HRP, Invitrogen/ThermoFisher) [12,13]
    anti-SARS-CoV-2
    suggested: None
    anti-human IgG H+L-HRP
    suggested: None
    Experimental Models: Cell Lines
    SentencesResources
    Live SARS-CoV-2 Virus Neutralization Assay: Vero E6 cells (2.5×104) were seeded in each well of a 96-well Black/Clear Flat Bottom TC-treated plate (Falcon) and incubated in DMEM overnight at 37°C with 5% CO2 prior to infection.
    Vero E6
    suggested: None
    Software and Algorithms
    SentencesResources
    A nonlinear regression method was used to determine the dilution that neutralized 50% of mNeonGreen fluorescence (NT50) by using Prism 8 (GraphPad).
    GraphPad
    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.
    • No funding statement was detected.
    • 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. SciScore for 10.1101/2020.08.07.242271: (What is this?)

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

    Table 1: Rigor

    Institutional Review Board StatementThis program and associated human subject research, performed in in accordance with the ethical standards of the Helsinki Declaration, were approved by the NorthShore University HealthSystem Institutional Review Board.RandomizationSamples were divided into tertiles based on the Roche assay results, then 47 were randomly selected to equally sample each tertile.Blindingnot detected.Power Analysisnot detected.Sex as a biological variablenot detected.Cell Line Authenticationnot detected.

    Table 2: Resources

    Antibodies
    SentencesResources
    SARS-CoV-2 S1 RBD Ig ELISA The anti-SARS-CoV-2 S1 RBD total Ig assay employed a standard indirect enzyme immunoassay technique (ELISA), described in detail elsewhere, using a secondary antibody recognizing all human immunoglobulin isotypes (goat anti-human IgG H+L-HRP, Invitrogen/ThermoFisher) [12,13]
    anti-SARS-CoV-2
    suggested: None
    anti-human IgG H+L-HRP
    suggested: None
    Experimental Models: Cell Lines
    SentencesResources
    Live SARS-CoV-2 Virus Neutralization Assay Vero E6 cells (2.5×104) were seeded in each well of a 96-well Black/Clear Flat Bottom TC-treated plate (Falcon) and incubated in DMEM overnight at 37°C with 5% CO2 prior to infection.
    Vero E6
    suggested: None
    Software and Algorithms
    SentencesResources
    A nonlinear regression method was used to determine the dilution that neutralized 50% of mNeonGreen fluorescence (NT50) by using Prism 8 (GraphPad).
    GraphPad
    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 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.


    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.