Quantifying Fluorescent Ligand Binding to GPCRs in Live Cells—A New HTS Format Replaces Radioactive Ligand Studies: Unlabeled Competitor Displacement IC_(50) Curves in < 3 Minutes in 96-Well Format Using PHERAstar FS

摘要

Ligand binding affinities at G-protein coupled receptors (GPCRs) have historically been determined using a radioligand that competes for receptor binding sites against an unlabeled drug-like compound. However, the potential hazards of open-source radioisotope handling, and the environmental impact of radioisotope disposal, make this a less desirable, costly technology. Development of fluorescent li-gands for GPCRs provides a safer method for determining ligand binding affinities. However, quantification of fluorescent ligand binding tends to rely on high-resolution fluorescence image capture. This approach can be lengthy and tedious, resulting in 96-well plate read times of approximately 30 minutes or more.