Early Kinetic Characterization of SARS-CoV-2 Main Protease Inhibitors: A Review and Guidance for Biochemical Assessments

Optical, microplate reader-based assays are a standard tool for the initial biochemical analysis of SARS-CoV-2 main protease (M^pro) inhibitor candidates. Such assays monitor M^pro-catalyzed substrate proteolysis in order to investigate the kinetic impact of inhibitors under examination on the catalytic reaction. This review outlines the numerous intricate considerations involved in establishing suitable assay protocols. Commonly employed M^pro substrates that explo