Label-Free Profiling of Ligands for Endogenous GPCRs Using a Cell-Based High-Throughput Screening Technology

This article reports the use of Corning Epic system— a label-free and noninvasive optical system that is centered on resonant waveguide grating biosensors—to profile endogenous G protein-coupled receptors (GPCRs) in living cells under physiologically relevant conditions. The endogenous GPCRs examined were bradykinin B2 receptor in A431 cells and protease-activated receptor subtype 1 (PAR1) in Chinese hamster ovary (CHO) cells. The activation of either receptor led to Gq-mediated signaling in the respective cells, as confirmed by Fluo-3 assays. Stimulation of CHO cells with thrombin, a PAR1 natural agonist, resulted in an optical response relating to dynamic mass redistribution that is similar to that induced by bradykinin in A431 cells. Based on the kinetics of agonist-mediated optical signatures, two time points, one before and another 5 min after the stimulation, were chosen to develop high-throughput (HT) screening assays. Results showed that such endpoint measurements enable not only HT screening of c...