(1089-B) NOMAD® biosensors for multiplexing Ca2+ and b-Arrestin functional assays of Proteinase-activated receptor 2 (PAR2)

Innoprot has developed a technology for multiplexing GPCR functional assays using fluorescent biosensors. This approach allows the measurement of the different signaling pathways involved in drug target activation in one single assay. It is the simplest way to identify GPCR biased ligands. Here we show a multiplexed assay using a stable cell line expressing green Ca2+ and red beta-arrestin Nomad biosensors and PAR2 protease-activated receptor to screen a library of 480 compounds. Both signals, calcium concentration and β-arrestin recruitment, were measured simultaneously by fluorescence intensity changes in living cells. After the screening campaign, positive compounds were chosen for further testing, based on the strength of the initial response and the lack of cytotoxicity. Our results indicate that Nomad Biosensors are effective and efficient tools for discovering differentiated GPCR biased ligands
We have performed a HTS campaign on a chemical and pharmacological diverse library (Prestwick Chemical Library: 1,200 FDA-approved & EMA-approved drugs) using Nomad biosensors. Ca2+ concentration changes and b-arrestin recruitment signals were measured simultaneously by fluorescence intensity changes in living cells. After the screening campaign, positive compounds were chosen for further testing, based on the strength of the initial response and the lack of cytotoxicity. Our results indicate that Nomad Biosensors are effective and efficient tools for discovering unknown GPCR ligands.