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Label-Free Functional Selectivity Assays

  • Ann M. Ferrie
  • Vasiliy Goral
  • Chaoming Wang
  • Ye FangEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1272)

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.

Key words

β2-adrenergic receptor Biased agonism Biased antagonism Dynamic mass redistribution G protein-coupled receptor Microfluidics Resonant waveguide grating biosensor 

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Copyright information

© Springer Science+Business Media New York 2015

Authors and Affiliations

  • Ann M. Ferrie
    • 1
  • Vasiliy Goral
    • 1
  • Chaoming Wang
    • 1
    • 2
  • Ye Fang
    • 1
    Email author
  1. 1.Biochemical Technologies, Science and Technology DivisionCorning IncorporatedCorningUSA
  2. 2.Department of Mechanical, Materials and Aerospace Engineering, NanoScience Technology CenterUniversity of Central FloridaOrlandoUSA

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