<p></p><p>The global COVID-19 pandemic has created an urgent
demand for large numbers of inexpensive, accurate, rapid, point-of-care
diagnostic tests. Analyte-based assays are suitably inexpensive and can be rapidly
mass-produced, but for sufficiently accurate performance they require highly
optimized antibodies and assay conditions. We used an automated liquid handling
system, customized to handle arrays of lateral flow immunoassay (LFA) tests in
a high-throughput screen, to identify anti-nucleocapsid antibodies that will
perform optimally in an LFA. We tested 1021 anti-nucleocapsid antibody pairs as
LFA capture and detection reagents with the goal of highlighting pairs that
have the greatest affinity for unique epitopes of the nucleocapsid protein of
SARS-CoV-2 within the LFA format. In contrast to traditional antibody screening
methods (e.g., ELISA, bio-layer interferometry), the method described here
integrates real-time reaction kinetics with transport in, and immobilization
directly onto, nitrocellulose. We have identified several candidate antibody
pairs that are suitable for further development of an LFA for SARS-CoV-2.</p><p></p>
Antigen production and development of an indirect ELISA based on the nucleocapsid protein to detect human SARS-CoV-2 seroconversion
