A Novel In-Cell ELISA Assay Allows Rapid and Automated Quantification of SARS-CoV-2 to Analyze Neutralizing Antibodies and Antiviral Compounds
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, the determination of virus-neutralizing antibodies (NAbs) is indispe...
| Main Authors: | , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2020-10-01
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| Series: | Frontiers in Immunology |
| Subjects: | |
| Online Access: | https://www.frontiersin.org/article/10.3389/fimmu.2020.573526/full |
| _version_ | 1818335310422999040 |
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| author | Lara Schöler Vu Thuy Khanh Le-Trilling Mareike Eilbrecht Denise Mennerich Olympia E. Anastasiou Adalbert Krawczyk Adalbert Krawczyk Anke Herrmann Ulf Dittmer Mirko Trilling |
| author_facet | Lara Schöler Vu Thuy Khanh Le-Trilling Mareike Eilbrecht Denise Mennerich Olympia E. Anastasiou Adalbert Krawczyk Adalbert Krawczyk Anke Herrmann Ulf Dittmer Mirko Trilling |
| author_sort | Lara Schöler |
| collection | DOAJ |
| description | 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, the determination of virus-neutralizing antibodies (NAbs) is indispensable for convalescent plasma selection, vaccine candidate evaluation, and immunity certificates. In contrast to standard serological 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. 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 icELISA-based neutralization test (icNT) was superior to PRNT in discriminating convalescent sera with high from those with intermediate neutralizing capacities. In addition, the icNT was found to be specific, discriminating between SARS-CoV-2-specific NAbs and those raised against other coronaviruses. Altogether, the SARS-CoV-2 icELISA test allows rapid (<48 h in total, read-out in seconds) and automated quantification of virus infection in cell culture to evaluate the efficacy of NAbs and antiviral drugs using reagents and equipment present in most routine diagnostics departments. |
| first_indexed | 2024-12-13T14:21:24Z |
| format | Article |
| id | doaj.art-90489618534b4068ac4c2b8414530f1e |
| institution | Directory Open Access Journal |
| issn | 1664-3224 |
| language | English |
| last_indexed | 2024-12-13T14:21:24Z |
| publishDate | 2020-10-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Immunology |
| spelling | doaj.art-90489618534b4068ac4c2b8414530f1e2022-12-21T23:42:05ZengFrontiers Media S.A.Frontiers in Immunology1664-32242020-10-011110.3389/fimmu.2020.573526573526A Novel In-Cell ELISA Assay Allows Rapid and Automated Quantification of SARS-CoV-2 to Analyze Neutralizing Antibodies and Antiviral CompoundsLara Schöler0Vu Thuy Khanh Le-Trilling1Mareike Eilbrecht2Denise Mennerich3Olympia E. Anastasiou4Adalbert Krawczyk5Adalbert Krawczyk6Anke Herrmann7Ulf Dittmer8Mirko Trilling9Institute for Virology, University Hospital Essen, University of Duisburg-Essen, Essen, GermanyInstitute for Virology, University Hospital Essen, University of Duisburg-Essen, Essen, GermanyInstitute for Virology, University Hospital Essen, University of Duisburg-Essen, Essen, GermanyInstitute for Virology, University Hospital Essen, University of Duisburg-Essen, Essen, GermanyInstitute for Virology, University Hospital Essen, University of Duisburg-Essen, Essen, GermanyInstitute for Virology, University Hospital Essen, University of Duisburg-Essen, Essen, GermanyDepartment of Infectious Diseases, West German Centre of Infectious Diseases, University Hospital Essen, University of Duisburg-Essen, Essen, GermanyInstitute for Virology, University Hospital Essen, University of Duisburg-Essen, Essen, GermanyInstitute for Virology, University Hospital Essen, University of Duisburg-Essen, Essen, GermanyInstitute for Virology, University Hospital Essen, University of Duisburg-Essen, Essen, GermanyThe 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, the determination of virus-neutralizing antibodies (NAbs) is indispensable for convalescent plasma selection, vaccine candidate evaluation, and immunity certificates. In contrast to standard serological 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. 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 icELISA-based neutralization test (icNT) was superior to PRNT in discriminating convalescent sera with high from those with intermediate neutralizing capacities. In addition, the icNT was found to be specific, discriminating between SARS-CoV-2-specific NAbs and those raised against other coronaviruses. Altogether, the SARS-CoV-2 icELISA test allows rapid (<48 h in total, read-out in seconds) and automated quantification of virus infection in cell culture to evaluate the efficacy of NAbs and antiviral drugs using reagents and equipment present in most routine diagnostics departments.https://www.frontiersin.org/article/10.3389/fimmu.2020.573526/fullSARS-CoV-2COVID-19neutralizingantibodiesvaccineinterferon |
| spellingShingle | Lara Schöler Vu Thuy Khanh Le-Trilling Mareike Eilbrecht Denise Mennerich Olympia E. Anastasiou Adalbert Krawczyk Adalbert Krawczyk Anke Herrmann Ulf Dittmer Mirko Trilling A Novel In-Cell ELISA Assay Allows Rapid and Automated Quantification of SARS-CoV-2 to Analyze Neutralizing Antibodies and Antiviral Compounds Frontiers in Immunology SARS-CoV-2 COVID-19 neutralizing antibodies vaccine interferon |
| title | A Novel In-Cell ELISA Assay Allows Rapid and Automated Quantification of SARS-CoV-2 to Analyze Neutralizing Antibodies and Antiviral Compounds |
| title_full | A Novel In-Cell ELISA Assay Allows Rapid and Automated Quantification of SARS-CoV-2 to Analyze Neutralizing Antibodies and Antiviral Compounds |
| title_fullStr | A Novel In-Cell ELISA Assay Allows Rapid and Automated Quantification of SARS-CoV-2 to Analyze Neutralizing Antibodies and Antiviral Compounds |
| title_full_unstemmed | A Novel In-Cell ELISA Assay Allows Rapid and Automated Quantification of SARS-CoV-2 to Analyze Neutralizing Antibodies and Antiviral Compounds |
| title_short | A Novel In-Cell ELISA Assay Allows Rapid and Automated Quantification of SARS-CoV-2 to Analyze Neutralizing Antibodies and Antiviral Compounds |
| title_sort | novel in cell elisa assay allows rapid and automated quantification of sars cov 2 to analyze neutralizing antibodies and antiviral compounds |
| topic | SARS-CoV-2 COVID-19 neutralizing antibodies vaccine interferon |
| url | https://www.frontiersin.org/article/10.3389/fimmu.2020.573526/full |
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