Multiplexed, microscale, microarray-based serological assay for antibodies against all human-relevant coronaviruses

•Quantitative assay with LOD from 0.32 to 2.0 ng/mL (determined with mAbs).•Linear dynamic ranges were 76- to 911-fold, depending on antigen.•Accuracy (% recovery) ranged from 88 % to 97 % of expected result.•Precision across 3 days, 3 lots, 72 per day (n = 216) resulted in 11 % CV.•98.5 % PPA and 100 % NPA for SARS-CoV-2 (n = 263, human serum specimens). Rapid, sensitive, and precise multiplexed assays for serological analysis during candidate COVID-19 vaccine development would streamline clinical trials. The VaxArray Coronavirus (CoV) SeroAssay quantifies IgG antibody binding to 9 pandemic, potentially pandemic, and endemic human CoV spike antigens in 2 h with automated results analysis. IgG antibodies in serum bind to the CoV spike protein capture antigens printed in a microarray format and are labeled with a fluorescent anti-species IgG secondary label. The assay demonstrated excellent lower limits of quantification ranging from 0.3 to 2.0 ng/mL and linear dynamic ranges of 76 to 911-fold. Average precision of 11 % CV and accuracy (% recovery) of 92.5 % over all capture antigens were achieved over 216 replicates representing 3 days and 3 microarray lots. Clinical performance on 263 human serum samples (132 SARS-CoV-2 negatives and 131 positives based on donor-matched RT-PCR and/or date of collection) produced 98.5 % PPA and 100 % NPA.