ReScan, a Multiplex Diagnostic Pipeline, Pans Human Sera for SARS-CoV-2 Antigens.

Zamecnik, Colin R; Rajan, Jayant V; Yamauchi, Kevin A; Mann, Sabrina A; Loudermilk, Rita P; Sowa, Gavin M; Zorn, Kelsey C; Alvarenga, Bonny D; Gaebler, Christian; Caskey, Marina; Stone, Mars; Norris, Philip J; Gu, Wei; Chiu, Charles Y; Ng, Dianna; Byrnes, James R; Zhou, Xin X; Wells, James A; Robbiani, Davide F; Nussenzweig, Michel C; DeRisi, Joseph L; Wilson, Michael R

Comprehensive understanding of the serological response to SARS-CoV-2 infection is important for both pathophysiologic insight and diagnostic development. Here, we generate a pan-human coronavirus programmable phage display assay to perform proteome-wide profiling of coronavirus antigens enriched by 98 COVID-19 patient sera. Next, we employ ReScan, a method to efficiently sequester phage expressing the most immunogenic peptides and print them onto paper-based microarrays using acoustic liquid handling, which isolates and identifies 9 candidate antigens, 8 of which are derived from the 2 proteins used for SARS-CoV-2 serologic assays: spike and nucleocapsid proteins. After deployment in a high-throughput assay amenable to clinical lab settings, these antigens show improved specificity over a whole protein panel. This proof-of-concept study demonstrates that ReScan will have broad applicability for other emerging infectious diseases or autoimmune diseases that lack a valid biomarker, enabling a seamless pipeline from antigen discovery to diagnostic using one recombinant protein source.