Accelerating the Throughput of Affinity Mass Spectrometry-Based Ligand Screening toward a G Protein-Coupled Receptor

摘要: Affinity mass spectrometry (MS) enables rapid screening of compound mixtures for ligands bound to a specific protein target, yet its current throughput is limited individually assay pools 400-2000 compounds. Typical affinity MS screens implemented in pharmaceutical industry laboratories identify putative based on qualitative analysis binding the target whereas no quantitative information acquired discriminate high- and low-affinity phase. Furthermore, these require purification stabilized form which poses great challenge membrane receptor targets. Here, we describe new, potentially general strategy that allows 20,000 compounds one pool highly efficient ligand discovery toward G protein-coupled (GPCR) target. Quantitative measurement high-affinity selection using both purified receptor-embedded cell membranes. This high-throughput, label-free screen resulted three new antagonists A2A adenosine receptor.