From Multiplex Serology to Serolomics—A Novel Approach to the Antibody Response against the SARS-CoV-2 Proteome
Julia Butt,
Rajagopal Murugan,
Theresa Hippchen,
Sylvia Olberg,
Monique van Straaten,
Hedda Wardemann,
Erec Stebbins,
Hans-Georg Kräusslich,
Ralf Bartenschlager,
Hermann Brenner,
Vibor Laketa,
Ben Schöttker,
Barbara Müller,
Uta Merle,
Tim Waterboer
Affiliations
Julia Butt
Infections and Cancer Epidemiology, German Cancer Research Center (Deutsches Krebsforschungszentrum, DKFZ), 69120 Heidelberg, Germany
Rajagopal Murugan
B Cell Immunology, German Cancer Research Center (Deutsches Krebsforschungszentrum, DKFZ), 69120 Heidelberg, Germany
Theresa Hippchen
Department of Internal Medicine IV, University Hospital Heidelberg, 69120 Heidelberg, Germany
Sylvia Olberg
Department of Infectious Diseases, Virology, University Hospital Heidelberg, 69120 Heidelberg, Germany
Monique van Straaten
Division of Structural Biology of Infection and Immunity, German Cancer Research Center (Deutsches Krebsforschungszentrum, DKFZ), 69120 Heidelberg, Germany
Hedda Wardemann
B Cell Immunology, German Cancer Research Center (Deutsches Krebsforschungszentrum, DKFZ), 69120 Heidelberg, Germany
Erec Stebbins
Division of Structural Biology of Infection and Immunity, German Cancer Research Center (Deutsches Krebsforschungszentrum, DKFZ), 69120 Heidelberg, Germany
Hans-Georg Kräusslich
Department of Infectious Diseases, Virology, University Hospital Heidelberg, 69120 Heidelberg, Germany
Ralf Bartenschlager
German Center for Infection Research (DZIF), Heidelberg Partner Site, 69120 Heidelberg, Germany
Hermann Brenner
Division of Clinical Epidemiology and Aging Research, German Cancer Research Center (Deutsches Krebsforschungszentrum, DKFZ), 69120 Heidelberg, Germany
Vibor Laketa
Department of Infectious Diseases, Virology, University Hospital Heidelberg, 69120 Heidelberg, Germany
Ben Schöttker
Division of Clinical Epidemiology and Aging Research, German Cancer Research Center (Deutsches Krebsforschungszentrum, DKFZ), 69120 Heidelberg, Germany
Barbara Müller
Department of Infectious Diseases, Virology, University Hospital Heidelberg, 69120 Heidelberg, Germany
Uta Merle
Department of Internal Medicine IV, University Hospital Heidelberg, 69120 Heidelberg, Germany
Tim Waterboer
Infections and Cancer Epidemiology, German Cancer Research Center (Deutsches Krebsforschungszentrum, DKFZ), 69120 Heidelberg, Germany
The emerging SARS-CoV-2 pandemic entails an urgent need for specific and sensitive high-throughput serological assays to assess SARS-CoV-2 epidemiology. We, therefore, aimed at developing a fluorescent-bead based SARS-CoV-2 multiplex serology assay for detection of antibody responses to the SARS-CoV-2 proteome. Proteins of the SARS-CoV-2 proteome and protein N of SARS-CoV-1 and common cold Coronaviruses (ccCoVs) were recombinantly expressed in E. coli or HEK293 cells. Assay performance was assessed in a COVID-19 case cohort (n = 48 hospitalized patients from Heidelberg) as well as n = 85 age- and sex-matched pre-pandemic controls from the ESTHER study. Assay validation included comparison with home-made immunofluorescence and commercial enzyme-linked immunosorbent (ELISA) assays. A sensitivity of 100% (95% CI: 86–100%) was achieved in COVID-19 patients 14 days post symptom onset with dual sero-positivity to SARS-CoV-2 N and the receptor-binding domain of the spike protein. The specificity obtained with this algorithm was 100% (95% CI: 96–100%). Antibody responses to ccCoVs N were abundantly high and did not correlate with those to SARS-CoV-2 N. Inclusion of additional SARS-CoV-2 proteins as well as separate assessment of immunoglobulin (Ig) classes M, A, and G allowed for explorative analyses regarding disease progression and course of antibody response. This newly developed SARS-CoV-2 multiplex serology assay achieved high sensitivity and specificity to determine SARS-CoV-2 sero-positivity. Its high throughput ability allows epidemiologic SARS-CoV-2 research in large population-based studies. Inclusion of additional pathogens into the panel as well as separate assessment of Ig isotypes will furthermore allow addressing research questions beyond SARS-CoV-2 sero-prevalence.
