Serological Assays Estimate Highly Variable SARS-CoV-2 Neutralizing Antibody Activity in Recovered COVID-19 Patients

  1. Larry L. Luchsinger
  2. Brett P. Ransegnola
  3. Daniel K. Jin
  4. Frauke Muecksch
  5. Yiska Weisblum
  6. Weili Bao
  7. Parakkal Jovvian George
  8. Marilis Rodriguez
  9. Nancy Tricoche
  10. Fabian Schmidt
  11. Chengjie Gao
  12. Shabnam Jawahar
  13. Mouli Pal
  14. Emily Schnall
  15. Huan Zhang
  16. Donna Strauss
  17. Karina Yazdanbakhsh
  18. Christopher D. Hillyer
  19. Paul D. Bieniasz
  20. Theodora Hatziioannou

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Abstract

The development of neutralizing antibodies (NAbs) against severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) following infection or vaccination is likely to be critical for the development of sufficient population immunity to drive cessation of the coronavirus disease of 2019 (COVID-19) pandemic. A large number of serologic tests, platforms, and methodologies are being employed to determine seroprevalence in populations to select convalescent plasma samples for therapeutic trials and to guide policies about reopening.

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  1. Version published to 10.1128/jcm.02005-20
  2. Version published to 10.1101/2020.06.08.20124792v2 on medRxiv
  3. ScreenIT

    SciScore for 10.1101/2020.06.08.20124792: (What is this?)

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

    Table 1: Rigor

    Institutional Review Board Statementnot detected.
    Randomizationnot detected.
    Blindingnot detected.
    Power Analysisnot detected.
    Sex as a biological variablenot detected.
    Cell Line Authenticationnot detected.

    Table 2: Resources

    Antibodies
    SentencesResources
    Neutralization assays: To measure neutralizing antibody activity in convalescent plasma, five-fold serial dilutions of plasma were incubated for 1 hour at 37°C in 96-well plates with an aliquot of HIV-1 or VSV-based SARS-CoV-2 pseudotyped virus containing approximately 1×103 infectious units.
    HIV-1
    suggested: (Advanced Biotechnologies Cat# 13-103-100, RRID:AB_1929216)
    Standard curves for both S1 and RBD assays were generated by using mouse anti-SARS-CoV spike protein monoclonal antibody (clone [3A2], ABIN2452119, Antibodies-Online) as the standard.
    anti-SARS-CoV spike protein
    suggested: None
    Plates were then washed three times with PBST and incubated for 1 hr with ELISA assay buffer containing Goat anti-Human IgA, IgG, IgM (Heavy & Light Chain) Antibody-HRP (Cat. No. ABIN100792, Antibodies-Online) and Goat anti-Mouse IgG2b (Heavy Chain) Antibody-HRP (Cat. No. ABIN376251, Antibodies-Online) at 1:30000 and 1:3000 dilutions, respectively.
    anti-Human IgA , IgG
    suggested: None
    Antibodies-Online
    suggested: (Antibodies-Online Cat# ABIN376251, RRID:AB_10763156)
    anti-Mouse IgG2b
    suggested: (Antibodies-Online Cat# ABIN376251, RRID:AB_10763156)
    Antibody-HRP
    suggested: (Bioworld Technology Cat# MB001H, RRID:AB_2857326)
    Experimental Models: Cell Lines
    SentencesResources
    Cell lines: Huh7.5 cells were a gift from Charles Rice(33).
    Huh7.5
    suggested: RRID:CVCL_7927)
    To generate (VSV/NG/NanoLuc)-SARS-CoV-2 pseudotype particles, 293T cells were infected with recombinant T7-expressing vaccinia virus (vTF7-3) and transfected with rVSVΔG/NG/NanoLuc, pBS-N, pBS-P, pBS-L, and pBS-G (PMID: 20709108).
    293T
    suggested: None
    Software and Algorithms
    SentencesResources
    LFA Densitometry Analysis: Relative quantification of anti-SARS-CoV-2 IgG and IgM in convalescent plasma samples was performed using built-in gel analysis macros in FIJI (https://fiji.sc/).
    FIJI
    suggested: (Fiji, RRID:SCR_002285)
    Standard curves were constructed in Prism 8.4 (Graphpad Software Inc.) using a Sigmoidal 4PL Non-Linear Regression (curve fit) model.
    Prism
    suggested: (PRISM, RRID:SCR_005375)
    Graphpad
    suggested: (GraphPad, RRID:SCR_000306)
    Samples were analyzed using the Abbott SARS-CoV-2 IgG chemiluminescent microparticle immunoassay with the Abbott Architect i2000SR (Abbott Core Laboratories), as well as the VITROS Immunodiagnostic Products Anti-SARS-CoV-2 Total Test and the Anti-SARS-CoV-2 IgG Test with the VITROS 5600 (Ortho Clinical Diagnostics).
    Abbott
    suggested: (Abbott, RRID:SCR_010477)
    Abbott Architect
    suggested: (Abbott ARCHITECT i1000sr System, RRID:SCR_019328)

    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: We detected the following sentences addressing limitations in the study:
    Demographic limitations of the CP donor population: Recent studies have noted a disproportion in COVID19 morbidity and mortality among minority communities.(16) In this study, of the 370 CP samples analyzed, only 204 donors (55%) elected to identify ethnicity, representing the least reported demographic category we collected. Nevertheless, we did not observe a significant difference in nAb or serology results as a function of any demographic metric, including ethnicity. Although we showed that the CP donor samples analyzed in this study comprised a relatively normal distribution of demographic indicators, based on the U.S. census data, we acknowledge that some factors, including ethnicity, are underrepresented in this cohort and limit the interpretation of the study beyond the population aggregate. The potential explanations of this phenomenon are complex and extend beyond the scope of this study.(17) The blood banking community is continuously working to recruit minority donors, who are consistently underrepresented amongst regular blood donors.(18) Efforts to increase public participation in local blood and CP donor programs would both improve blood product diversity of transfusion products and strengthen the rigor of epidemiological disease. Thus, studies designed to characterize serological responses to COVID19 specifically in minority groups are warranted and necessary to augment our current understanding of the pandemic. Seroconversion assays of the population: Quantifi...

    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.

  4. Version published to 10.1101/2020.06.08.20124792v1 on medRxiv