Peptide microarray‐based analysis of antibody responses to SARS‐CoV‐2 identifies unique epitopes with potential for diagnostic test development

  1. Pavlo Holenya
  2. Paul Joris Lange
  3. Ulf Reimer
  4. Wolfram Woltersdorf
  5. Thomas Panterodt
  6. Michael Glas
  7. Mark Wasner
  8. Maren Eckey
  9. Michael Drosch
  10. Jörg‐Michael Hollidt
  11. Michael Naumann
  12. Florian Kern
  13. Holger Wenschuh
  14. Robert Lange
  15. Karsten Schnatbaum
  16. Frank F. Bier

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Abstract

Humoral immunity to the Severe Adult Respiratory Syndrome (SARS) Coronavirus (CoV)‐2 is not fully understood yet but is a crucial factor of immune protection. The possibility of antibody cross‐reactivity between SARS‐CoV‐2 and other human coronaviruses (HCoVs) would have important implications for immune protection but also for the development of specific diagnostic ELISA tests. Using peptide microarrays, n = 24 patient samples and n = 12 control samples were screened for antibodies against the entire SARS‐CoV‐2 proteome as well as the Spike (S), Nucleocapsid (N), VME1 (V), R1ab, and Protein 3a (AP3A) of the HCoV strains SARS, MERS, OC43, and 229E. While widespread cross‐reactivity was revealed across several immunodominant regions of S and N, IgG binding to several SARS‐CoV‐2‐derived peptides provided statistically significant discrimination between COVID‐19 patients and controls. Selected target peptides may serve as capture antigens for future, highly COVID‐19‐specific diagnostic antibody tests.

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  1. Version published to 10.1002/eji.202049101
  2. ScreenIT

    SciScore for 10.1101/2020.11.24.20216663: (What is this?)

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

    Table 1: Rigor

    Institutional Review Board StatementConsent: Blood sampling: Following written informed consent peripheral blood samples were obtained by vene puncture / from periperal venous catheters.
    Randomizationnot detected.
    Blindingnot detected.
    Power Analysisnot detected.
    Sex as a biological variableSARS-CoV-2 infected group: COVID-19 patients (n=13 female, age range 23 to 70 years; n=11 male, age range 35 to 70 years) were recruited across the spectrum of clinical severity, ranging from asymptomatic/mild to severe/SARS.
    Cell Line Authenticationnot detected.

    Table 2: Resources

    Antibodies
    SentencesResources
    Microarray assay and data analysis: The peptide microarrays were incubated with sera (applied dilution 1:200) in an HS 4800 microarray processing station (Tecan) for two hours at 30°C, followed by incubation with 0.1 μg/mL fluorescently labelled anti human IgG deg/mL fluorescently labelled anti human IgG detection antibody (Jackson Immunoresearch, 109-605-098)
    anti human IgG
    suggested: None
    ELISA test: To detect antibodies, we used the Euroimmun Anti-SARS-CoV-2 ELISA (IgG), which was manufactured by Euroimmun Medizinische Labordiagnostika AG, Lübeck, Germany.
    Anti-SARS-CoV-2 ELISA (IgG)
    suggested: None
    First, the diluted patient samples were incubated in the wells which lead to specific IgG antibodies binding to the antigens.
    specific IgG
    suggested: None
    In order to detect the bound antibodies, an enzyme-labelled anti-human IgG antibody detected antigen-antibody complexes and catalysed a colour reaction.
    anti-human IgG
    suggested: None
    Experimental Models: Cell Lines
    SentencesResources
    The break-off microplate wells were covered with the antigen, a recombinant structural spike 1 (S1) protein of SARS-CoV-2 expressed in HEK293 [FDA, 2020].
    HEK293
    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

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  3. Version published to 10.1101/2020.11.24.20216663v1 on medRxiv