tetano
Editor, Senior Moderator
Viruses
. 2021 Apr 24;13(5):749.
doi: 10.3390/v13050749.
From Multiplex Serology to Serolomics-A Novel Approach to the Antibody Response against the SARS-CoV-2 Proteome
Julia Butt[SUP] 1 [/SUP], Rajagopal Murugan[SUP] 2 [/SUP], Theresa Hippchen[SUP] 3 [/SUP], Sylvia Olberg[SUP] 4 5 [/SUP], Monique van Straaten[SUP] 6 [/SUP], Hedda Wardemann[SUP] 2 [/SUP], Erec Stebbins[SUP] 6 [/SUP], Hans-Georg Kr?usslich[SUP] 4 5 [/SUP], Ralf Bartenschlager[SUP] 5 7 8 [/SUP], Hermann Brenner[SUP] 9 10 [/SUP], Vibor Laketa[SUP] 4 5 [/SUP], Ben Sch?ttker[SUP] 9 10 [/SUP], Barbara M?ller[SUP] 4 [/SUP], Uta Merle[SUP] 3 [/SUP], Tim Waterboer[SUP] 1 [/SUP]
Affiliations
Abstract
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.
Keywords: SARS-CoV-2; multiplex serology.
. 2021 Apr 24;13(5):749.
doi: 10.3390/v13050749.
From Multiplex Serology to Serolomics-A Novel Approach to the Antibody Response against the SARS-CoV-2 Proteome
Julia Butt[SUP] 1 [/SUP], Rajagopal Murugan[SUP] 2 [/SUP], Theresa Hippchen[SUP] 3 [/SUP], Sylvia Olberg[SUP] 4 5 [/SUP], Monique van Straaten[SUP] 6 [/SUP], Hedda Wardemann[SUP] 2 [/SUP], Erec Stebbins[SUP] 6 [/SUP], Hans-Georg Kr?usslich[SUP] 4 5 [/SUP], Ralf Bartenschlager[SUP] 5 7 8 [/SUP], Hermann Brenner[SUP] 9 10 [/SUP], Vibor Laketa[SUP] 4 5 [/SUP], Ben Sch?ttker[SUP] 9 10 [/SUP], Barbara M?ller[SUP] 4 [/SUP], Uta Merle[SUP] 3 [/SUP], Tim Waterboer[SUP] 1 [/SUP]
Affiliations
- PMID: 33923338
- DOI: 10.3390/v13050749
Abstract
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.
Keywords: SARS-CoV-2; multiplex serology.