Longitudinal Development of Antibody Responses in COVID-19 Patients of Different Severity with ELISA, Peptide, and Glycan Arrays: An Immunological Case Series

  1. Jasmin Heidepriem
  2. Christine Dahlke
  3. Robin Kobbe
  4. René Santer
  5. Till Koch
  6. Anahita Fathi
  7. Bruna Seco
  8. My Ly
  9. Stefan Schmiedel
  10. Dorothee Schwinge
  11. Sonia Serna
  12. Katrin Sellrie
  13. Niels-Christian Reichardt
  14. Peter Seeberger
  15. Marylyn Addo
  16. Felix Loeffler
  17. on behalf of the ID-UKE COVID-19 Study Group

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Abstract

The current COVID-19 pandemic is caused by the severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2). A better understanding of its immunogenicity can be important for the development of improved diagnostics, therapeutics, and vaccines. Here, we report the longitudinal analysis of three COVID-19 patients with moderate (#1) and mild disease (#2 and #3). Antibody serum responses were analyzed using spike glycoprotein enzyme linked immunosorbent assay (ELISA), full-proteome peptide, and glycan microarrays. ELISA immunoglobulin A, G, and M (IgA, IgG, and IgM) signals increased over time for individuals #1 and #2, whereas #3 only showed no clear positive IgG and IgM result. In contrast, peptide microarrays showed increasing IgA/G signal intensity and epitope spread only in the moderate patient #1 over time, whereas early but transient IgA and stable IgG responses were observed in the two mild cases #2 and #3. Glycan arrays showed an interaction of antibodies to fragments of high-mannose and core N-glycans, present on the viral shield. In contrast to protein ELISA, microarrays allow for a deeper understanding of IgA, IgG, and IgM antibody responses to specific epitopes of the whole proteome and glycans of SARS-CoV-2 in parallel. In the future, this may help to better understand and to monitor vaccination programs and monoclonal antibodies as therapeutics.

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  1. Version published to 10.3390/pathogens10040438
  2. ScreenIT

    SciScore for 10.1101/2020.04.14.20059733: (What is this?)

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

    Table 1: Rigor

    Institutional Review Board StatementConsent: Donors provided written informed consent.
    IRB: The protocol was approved by the Ethics Committee of the Hamburg Medical Association, Germany.
    Randomizationnot detected.
    Blindingnot detected.
    Power Analysisnot detected.
    Sex as a biological variablenot detected.

    Table 2: Resources

    No key resources detected.

    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:
    Key differences are the limitation of only using S1-proteins for the ELISA (vs. whole proteome on the microarray) and a higher sensitivity of peptide arrays. We detected an early and defined IgA response in patient #2 who had a very mild course of disease. In addition, our patient #4 (mild/moderate course of disease, no hospitalization, see Supporting Information Figures S1 & S2) also showed an early IgA response. IgA is considered a major effector molecule involved in defense mechanisms against viruses and infections with respiratory viruses can induce efficient IgA responses in secretions as well as in sera. Our data suggest that IgA antibodies are valuable diagnostic markers that show strong signals early after onset of mild COVID-19-associated symptoms and that IgA antibodies may be crucial for efficient clearance of SARS-CoV-2. We identified many spike protein epitopes that are bound by IgA antibodies, which might be crucial for neutralizing viral particles. Early and efficient binding of the spike protein by antibodies can limit viral spread following infection. We also identified several IgA epitopes in the spike protein, such as in the receptor binding domain (AA437–465) and in the fusion peptide domain (AA816–831). In addition, we also confirm parts of a SARS-CoV-2 and SARS-CoV cross-reactive IgG epitope (AA369–383) in the receptor binding domain of spike.12 In general, microarrays are valuable tools to identify crucial antibodies and antigens. Recently, COVID-19 pat...

    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 10.1101/2020.04.14.20059733v1 on medRxiv