Characterization of Immune Responses to SARS-CoV-2 and Other Human Pathogenic Coronaviruses Using a Multiplex Bead-Based Immunoassay

Abstract

Serological assays that simultaneously detect antibodies to multiple targets of SARS-CoV-2 and to other structurally related coronaviruses provide a holistic picture of antibody response patterns. Well-validated multiplex immunoassays are scarce. Here, we evaluated the performance of an 11-plex serological assay capable of detecting antibodies directed to four antigenic targets of SARS-CoV-2 and to S1 proteins of other human pathogenic coronaviruses. We used 620 well-characterized sera (n = 458 seropositive and n = 110 seronegative for SARS-CoV-2 in the pre-SARS-CoV-2 era and n = 52 seronegative for SARS-CoV-2 in the era of SARS-CoV-2) as positive and negative standards. We calculated the sensitivity, specificity, as well as positive and negative predictive values, including a 95% confidence interval. The difference in mean fluorescence intensity (95% CI) was used to assess a potential cross-reaction between antibodies to SARS-CoV-2 and the other coronaviruses. The sensitivity (95% CI) of detecting anti-SARS-CoV-2 antibodies to four antigenic targets ranged from 83.4% (76.7–86.7) to 93.7% (91.0–95.7) and the specificity from 98.2% (93.6–99.8) to 100% (96.7–100). We observed no obvious cross-reaction between anti-SARS-CoV-2 antibodies and antibodies to the other coronaviruses except for SARS-CoV-1. The high sensitivity and specificity warrant a reliable utilization of the assay in population-based seroprevalence surveys or vaccine efficacy studies.

Immune response after two doses of the BNT162b2 COVID-19 vaccine and risk of SARS-CoV-2 breakthrough infection in Tyrol, Austria: an open-label, observational phase 4 trial

“…This notion is supported by previous data from before COVID-19 vaccines were available, which showed a considerable degree of correlation between anti-N and anti-S IgG antibodies generated after infection. 23 Our finding of poor correlations between antibodies and T-cell responses is consistent with some other studies showing no correlation to only moderate correlation between humoral and cellular immune parameters in vaccinated 24 , 25 , 26 and convalescent 27 , 28 , 29 individuals.…”

Immune response to 2-dose BNT162b2 vaccination and risk of SARS-CoV-2 breakthrough infection: The Shieldvacc-2 study

“…This notion is supported by previous data from pre-SARS-CoV-2 vaccine era that showed a considerable degree of correlation between anti-N and anti-S IgG antibodies generated post-infection. 24 Our finding of poor correlations between antibodies and T-cell responses is consistent with some other studies showing no or low correlations between humoral and cellular immune parameters in vaccinated 25–30 and convalescent 31–33 individuals.…”

Development of a Rapid Live SARS-CoV-2 Neutralization Assay Based on a qPCR Readout

“…Twenty non-COVID-19 sera, collected prior to the emergence of COVID-19, were used as negative controls. In order to test for cross-reactivity, a total of 20 prepandemic sera with antibodies against endemic coronaviruses (229E, HKU1, OC43, NL-63), as detected by ProcartaPlex Human Coronavirus Ig Total 11-Plex Panel (Invitrogen, Thermo Fisher Scientific, MA, USA) ( 38 ), were tested in the RT-VNA. All sera were heat-inactivated at 56°C for 30 min prior to use.…”