Exploring beyond clinical routine SARS-CoV-2 serology using MultiCoV-Ab to evaluate endemic coronavirus cross-reactivity

  1. Matthias Becker
  2. Monika Strengert
  3. Daniel Junker
  4. Philipp D. Kaiser
  5. Tobias Kerrinnes
  6. Bjoern Traenkle
  7. Heiko Dinter
  8. Julia Häring
  9. Stéphane Ghozzi
  10. Anne Zeck
  11. Frank Weise
  12. Andreas Peter
  13. Sebastian Hörber
  14. Simon Fink
  15. Felix Ruoff
  16. Alex Dulovic
  17. Tamam Bakchoul
  18. Armin Baillot
  19. Stefan Lohse
  20. Markus Cornberg
  21. Thomas Illig
  22. Jens Gottlieb
  23. Sigrun Smola
  24. André Karch
  25. Klaus Berger
  26. Hans-Georg Rammensee
  27. Katja Schenke-Layland
  28. Annika Nelde
  29. Melanie Märklin
  30. Jonas S. Heitmann
  31. Juliane S. Walz
  32. Markus Templin
  33. Thomas O. Joos
  34. Ulrich Rothbauer
  35. Gérard Krause
  36. Nicole Schneiderhan-Marra

Reviewed by

Read the full article See related articles

Listed in

Log in to save this article

Abstract

The humoral immune response to SARS-CoV-2 is a benchmark for immunity and detailed analysis is required to understand the manifestation and progression of COVID-19, monitor seroconversion within the general population, and support vaccine development. The majority of currently available commercial serological assays only quantify the SARS-CoV-2 antibody response against individual antigens, limiting our understanding of the immune response. To overcome this, we have developed a multiplex immunoassay (MultiCoV-Ab) including spike and nucleocapsid proteins of SARS-CoV-2 and the endemic human coronaviruses. Compared to three broadly used commercial in vitro diagnostic tests, our MultiCoV-Ab achieves a higher sensitivity and specificity when analyzing a well-characterized sample set of SARS-CoV-2 infected and uninfected individuals. We find a high response against endemic coronaviruses in our sample set, but no consistent cross-reactive IgG response patterns against SARS-CoV-2. Here we show a robust, high-content-enabled, antigen-saving multiplex assay suited to both monitoring vaccination studies and facilitating epidemiologic screenings for humoral immunity towards pandemic and endemic coronaviruses.

Article activity feed

  1. ScreenIT

    SciScore for 10.1101/2020.07.17.20156000: (What is this?)

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

    Table 1: Rigor

    Institutional Review Board StatementIRB: Ethical approval was granted from the Ethics Committee of Hannover Medical School (#9122_BO_K2020).
    RandomizationThe majority of non-SARS-COV-2 infected samples was randomly selected and consistent of prepandemic blood donors, commercially available (Central BioHub GmbH, Berlin, Germany and BBI Solutions, Crumlin, UK) or bio-banked specimens.
    Blindingnot detected.
    Power Analysisnot detected.
    Sex as a biological variablenot detected.
    Cell Line Authenticationnot detected.

    Table 2: Resources

    Antibodies
    SentencesResources
    All purified proteins were analyzed via standard SDS-PAGE followed by staining with InstantBlue Coomassie stain (Expedeon) and immunoblotting using an anti-His antibody (Penta-His Antibody, #34660, Qiagen) in combination with a donkey-anti-mouse antibody labeled with AlexaFluor647 (Invitrogen) on a Typhoon Trio (GE-Healthcare, Freiburg, Germany; excitation 633 nm, emission filter settings 670 nm BP 30) to confirm protein integrity.
    anti-His
    suggested: None
    donkey-anti-mouse
    suggested: (Jackson ImmunoResearch Labs Cat# 715-065-151, RRID:AB_2340785)
    Bound antibodies were detected with R-phycoerythrin labeled goat-anti-human IgG or IgA antibodies (incubation for 45 min at 21°C).
    R-phycoerythrin labeled goat-anti-human IgG
    suggested: None
    IgA
    suggested: None
    Experimental Models: Cell Lines
    SentencesResources
    The viral S1-domains, SARS-CoV-2 RBD, and the stabilized trimeric SARS-CoV-2 Spike protein were expressed in Expi293 cells following the protocol as described in Stadlbauer et al.8.
    Expi293
    suggested: RRID:CVCL_D615)

    Results from OddPub: Thank you for sharing your data.

    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 10.1038/s41467-021-20973-3
  3. Version published to 10.1101/2020.07.17.20156000v2 on medRxiv
  4. Version published to 10.1101/2020.07.17.20156000v1 on medRxiv