Multiplexed, microscale, microarray-based serological assay for antibodies against all human-relevant coronaviruses

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Abstract

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  1. SciScore for 10.1101/2020.09.03.20179598: (What is this?)

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

    Table 1: Rigor

    Institutional Review Board StatementConsent: Deidentified specimens from Mount Sinai Icanh School of Medicine (New York, NY) were obtained under a license agreement which indicates appropriate IRB approval and informed consent for the specimens provided.
    Randomizationnot detected.
    BlindingTesting personnel were blinded to these orthogonal results prior to completing VaxArray CoV SeroAssay analysis.
    Power Analysisnot detected.
    Sex as a biological variablenot detected.

    Table 2: Resources

    Antibodies
    SentencesResources
    2.2 Linear Dynamic Range and Lower Limit of Quantification Analysis: A study to determine the lower limit of quantification and linear dynamic range of the different capture antigens represented was executed using monoclonal antibodies that target the spike proteins of SARS-CoV-1 (MRO-1214LC, CR3022
    MRO-1214LC
    suggested: None
    The CR3022 antibody targeting SARS-CoV-1 is known to bind the the nCoV(ii) RBD antigen, the SARS antigen (and the nCoV(i) full-length spike antigen to a much weaker extent), and the SARS-CoV-2 Genetex antibody is known to bind the nCoV(i) full-length spike antigen (and the nCoV(iii) S2 antigen to a much weaker extent).
    CR3022
    suggested: None
    S2
    suggested: None
    The four antibodies were mixed, and a 13-point serial dilution in Protein Blocking Buffer and three blank wells containing Protein Blocking Buffer without antibody were prepared, with each sample subsequently analyzed on the VaxArray CoV SeroAssay according to the operation manual with one exception: because the anti-SARS-CoV1 antibodies are human antibodies and the other three antibodies are mouse antibodies, antibodies were detected with a mixture of anti-mouse and anti-human IgG secondary antibody labels (VXCV-7620 and VXCV-7326, InDevR, Inc., respectively).
    anti-SARS-CoV1
    suggested: None
    anti-mouse
    suggested: None
    anti-human IgG
    suggested: None
    Because monoclonal antibodies binding to the OC43, NL63, or 229E capture antigens were not available at the time of testing, linearity and limit of detection for these 3 capture antigens was explored using a limiting endpoint dilution series of a pooled human serum sample known to be positive for antibodies to all four human CoVs: OC43, NL63, HKU1, and 229E.
    HKU1
    suggested: None
    No monoclonal antibodies were available at the time of testing that target OC43, NL63, or 229E, and so specificity for these capture antigens was not assessed.
    NL63
    suggested: None
    A total of 132 serum samples known to be negative for the presence of antibodies to SARS-CoV-2 based on date of collection prior to December 2019, including 33 specimens from pediatric donors age 2-16, were analyzed via the standard VaxArray CoV SeroAssay procedure at a 1:100 dilution in PBB. 2.4 Reproducibility and Accuracy: To assess reproducibility and accuracy, a pooled human serum sample known to be positive for antibodies to SARS-CoV-2 (and known to bind all 3 SARS-CoV-2 antigens on the microarray) and all 4 of the endemic coronaviruses (HKU1, OC43, NL63, and 229E) was prepared in adequate volume to run a large number of replicates.
    SARS-CoV-2
    suggested: None

    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.