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A novel in-cell ELISA assay allows rapid and automated quantification of SARS-CoV-2 to analyse neutralizing antibodies and antiviral compounds

View ORCID ProfileLara Schöler, View ORCID ProfileVu Thuy Khanh Le-Trilling, View ORCID ProfileMareike Eilbrecht, View ORCID ProfileDenise Mennerich, Olympia E. Anastasiou, View ORCID ProfileAdalbert Krawczyk, Anke Herrmann, View ORCID ProfileUlf Dittmer, View ORCID ProfileMirko Trilling
doi: https://doi.org/10.1101/2020.06.05.135806
Lara Schöler
1Institute for Virology, University Hospital Essen, University of Duisburg-Essen, Essen, Germany
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Vu Thuy Khanh Le-Trilling
1Institute for Virology, University Hospital Essen, University of Duisburg-Essen, Essen, Germany
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Mareike Eilbrecht
1Institute for Virology, University Hospital Essen, University of Duisburg-Essen, Essen, Germany
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Denise Mennerich
1Institute for Virology, University Hospital Essen, University of Duisburg-Essen, Essen, Germany
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Olympia E. Anastasiou
1Institute for Virology, University Hospital Essen, University of Duisburg-Essen, Essen, Germany
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Adalbert Krawczyk
1Institute for Virology, University Hospital Essen, University of Duisburg-Essen, Essen, Germany
2Department of Infectious Diseases, West German Centre of Infectious Diseases, University Hospital Essen, University of Duisburg-Essen, Essen, Germany
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Anke Herrmann
1Institute for Virology, University Hospital Essen, University of Duisburg-Essen, Essen, Germany
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Ulf Dittmer
1Institute for Virology, University Hospital Essen, University of Duisburg-Essen, Essen, Germany
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Mirko Trilling
1Institute for Virology, University Hospital Essen, University of Duisburg-Essen, Essen, Germany
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  • For correspondence: Mirko.Trilling@uk-essen.de
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Abstract

The coronavirus disease 2019 (COVID-19) caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is currently the most pressing medical and socioeconomic challenge. Constituting important correlates of protection, determination of virus-neutralizing antibodies (NAbs) is indispensable for convalescent plasma selection, vaccine candidate evaluation, and immunity certificates. In contrast to standard serology ELISAs, plaque reduction neutralization tests (PRNTs) are laborious, time-consuming, expensive, and restricted to specialized laboratories. To replace microscopic counting-based SARS-CoV-2 PRNTs by a novel assay exempt from genetically modified viruses, which are inapplicable in most diagnostics departments, we established a simple, rapid, and automated SARS-CoV-2 neutralization assay employing an in-cell ELISA (icELISA) approach.

After optimization of various parameters such as virus-specific antibodies, cell lines, virus doses, and duration of infection, SARS-CoV-2-infected cells became amenable as direct antigen source for quantitative icELISA. Using commercially available nucleocapsid protein-specific antibodies, viral infection could easily be quantified in human and highly permissive Vero E6 cells by icELISA. Antiviral agents such as human sera containing NAbs or antiviral interferons dose-dependently reduced the SARS-CoV-2-specific signal. Applying increased infectious doses, the icNT was superior to PRNT in discriminating convalescent sera with high from those with intermediate neutralizing capacities.

The SARS-CoV-2 icELISA test allows rapid (<48h in total, read-out in seconds) and automated quantification of virus infection in cell culture to evaluate the efficacy of NAbs as well as antiviral drugs, using reagents and equipment present in most routine diagnostics departments. We propose the icELISA and the icNT for COVID-19 research and diagnostics.

Competing Interest Statement

The authors have declared no competing interest.

Footnotes

  • Brief summary: Knowledge concerning SARS-CoV-2-neutralizing antibodies (NAbs) is indispensable for COVID-19 convalescent plasma selection, evaluation of vaccines, and immunity certificates. Our in-cell ELISA (icELISA) test allows rapid (<48h) and high-throughput detection and quantification of SARS-CoV-2-specific NAbs and antiviral activity of drug candidates.

Copyright 
The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. All rights reserved. No reuse allowed without permission.
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Posted June 05, 2020.
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A novel in-cell ELISA assay allows rapid and automated quantification of SARS-CoV-2 to analyse neutralizing antibodies and antiviral compounds
Lara Schöler, Vu Thuy Khanh Le-Trilling, Mareike Eilbrecht, Denise Mennerich, Olympia E. Anastasiou, Adalbert Krawczyk, Anke Herrmann, Ulf Dittmer, Mirko Trilling
bioRxiv 2020.06.05.135806; doi: https://doi.org/10.1101/2020.06.05.135806
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A novel in-cell ELISA assay allows rapid and automated quantification of SARS-CoV-2 to analyse neutralizing antibodies and antiviral compounds
Lara Schöler, Vu Thuy Khanh Le-Trilling, Mareike Eilbrecht, Denise Mennerich, Olympia E. Anastasiou, Adalbert Krawczyk, Anke Herrmann, Ulf Dittmer, Mirko Trilling
bioRxiv 2020.06.05.135806; doi: https://doi.org/10.1101/2020.06.05.135806

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