A throughput serological Western blot system using whole virus lysate for the concomitant detection of antibodies against SARS-CoV-2 and human endemic Coronaviridae

  1. Simon Fink
  2. Felix Ruoff
  3. Aaron Stahl
  4. Matthias Becker
  5. Philipp Kaiser
  6. Bjoern Traenkle
  7. Daniel Junker
  8. Frank Weise
  9. Natalia Ruetalo
  10. Sebastian Hörber
  11. Andreas Peter
  12. Annika Nelde
  13. Juliane Walz
  14. Gérard Krause
  15. Katja Schenke-Layland
  16. Thomas Joos
  17. Ulrich Rothbauer
  18. Nicole Schneiderhan-Marra
  19. Michael Schindler
  20. Markus F. Templin
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Abstract

B <sc>ackground</sc>

Seroreactivity against human endemic coronaviruses has been linked to disease severity after SARS-CoV-2 infection. Assays that are capable of concomitantly detecting antibodies against endemic coronaviridae such as OC43, 229E, NL63, and SARS-CoV-2 may help to elucidate this question. We set up a platform for serum-screening and developed a bead-based Western blot system, namely DigiWest, capable of running hundreds of assays using microgram amounts of protein prepared directly from different viruses.

M <sc>ethods</sc>

The parallelized and miniaturised DigiWest assay was adapted for detecting antibodies using whole protein extract prepared from isolated SARS-CoV-2 virus particles. After characterisation and optimization of the newly established test, whole virus lysates of OC43, 229E, and NL63 were integrated into the system.

R <sc>esults</sc>

The DigiWest-based immunoassay system for detection of SARS-CoV-2 specific antibodies shows a sensitivity of 87.2 % and diagnostic specificity of 100 %. Concordance analysis with the SARS-CoV-2 immunoassays available by Roche, Siemens, and Euroimmun indicates a comparable assay performance (Cohen’s Kappa ranging from 0.8799-0.9429). In the multiplexed assay, antibodies against the endemic coronaviruses OC43, 229E, and NL63 were detected, displaying a high incidence of seroreactivity against these coronaviruses.

C <sc>onclusion</sc>

The DigiWest-based immunoassay, which uses authentic antigens from isolated virus particles, is capable of detecting individual serum responses against SARS-CoV-2 with high specificity and sensitivity in one multiplexed assay. It shows high concordance with other commercially available serologic assays. The DigiWest approach enables a concomitant detection of antibodies against different endemic coronaviruses and will help to elucidate the role of these possibly cross-reactive antibodies.

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