title: Label-free functional selectivity assays.
abstract: G protein-coupled receptors (GPCRs) represent the largest class of drug targets. Ligand-directed functional selectivity or biased agonism opens new possibility for discovering GPCR drugs with better efficacy and safety profiles. However, quantification of ligand bias is challenging. Herein, we present five different label-free dynamic mass redistribution (DMR) approaches to assess ligand bias acting at the β2-adrenergic receptor (β2AR). Multiparametric analysis of the DMR agonist profiles reveals divergent pharmacology of a panel of β2AR agonists. DMR profiling using catechol as a conformational probe detects the presence of multiple conformations of the β2AR. DMR assays under microfluidics, together with chemical biology tools, discover ligand-directed desensitization of the receptor. DMR antagonist reverse assays manifest biased antagonism. DMR profiling using distinct probe-modulated cells detects the biased agonism in the context of self-referenced pharmacological activity map.
authors.affiliation: Biochemical Technologies, Science and Technology Division, Corning Incorporated, Corning, NY, 14831, USA.
authors.forename: Ann M
authors.initials: AM
authors.lastname: Ferrie
authors.forename: Vasiliy
authors.initials: V
authors.lastname: Goral
authors.forename: Chaoming
authors.initials: C
authors.lastname: Wang
authors.forename: Ye
authors.initials: Y
authors.lastname: Fang
mesh_terms: D000318:Adrenergic beta-Agonists / Q000494:pharmacology; D000319:Adrenergic beta-Antagonists / Q000494:pharmacology; D000526:Alprenolol / Q000494:pharmacology; D015374:Biosensing Techniques; D002396:Catechols / Q000494:pharmacology; D045744:Cell Line, Tumor; D004129:Dimethylpolysiloxanes; D004847:Epithelial Cells; D015870:Gene Expression; D057166:High-Throughput Screening Assays; D006801:Humans; D008024:Ligands; D044085:Microfluidics / Q000295:instrumentation*; D055616:Microtechnology; D055096:Optical Devices; D018343:Receptors, Adrenergic, beta-2 / Q000235:genetics / Q000378:metabolism*; D016008:Statistical Distributions
chemical_list: D000318:Adrenergic beta-Agonists; D000319:Adrenergic beta-Antagonists; D002396:Catechols; D004129:Dimethylpolysiloxanes; D008024:Ligands; D018343:Receptors, Adrenergic, beta-2; C013830:baysilon; D000526:Alprenolol; C034221:catechol