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Duplexed Label-Free G Protein—Coupled Receptor Assays for High-Throughput Screening

Biochemical Technologies, Science and Technology Division, Corning Incorporated, Sullivan Park, Corning, New York

Ye Fang

Biochemical Technologies, Science and Technology Division, Corning Incorporated, Sullivan Park, Corning, New York, fangy2{at}corning.com

This article describes duplexed label-free optical biosensor cellular assays for simultaneously assaying 2 endogenous receptors, the G_q-coupled histamine receptor (H ₁) and the G_s-coupled β₂-adrenergic receptor (β₂AR), in A431 cells. The biosensor cellular assays consist of 2 sequential steps—an initial agonist screening using Sigma LOPAC (Library of Pharmaceutically Active Compounds) and a subsequent antagonist screening using a solution mixture containing the H₁ agonist histamine and the β₂AR agonist epinephrine. Results showed that costimulating A431 cells with histamine and epinephrine led to an optical response additive to individual responses. The agonist screening not only identified all full agonists for both the H₁ and β₂ receptors, but also detected pathway-biased ligands for the β₂AR. Furthermore, the succeeding antagonist screening documented all known antagonists in the library for either the H₁ or β₂ receptors. This is the 1st demonstration of a single cellular assay that is capable of screening ligands against 2 GPCRs coupled to distinct G proteins, and highlights the power of pathway-unbiased and label-free biosensor cellular assays for GPCR screens. (Journal of Biomolecular Screening 2008:975-985)

Key Words: optical biosensor • G protein—coupled receptor • histamine H1 receptor • β2 adrenoceptor • high-throughput screening • multiplexing • ligand-directed functional selectivity • compartmentalization

This version was published on December 1, 2008

Journal of Biomolecular Screening, Vol. 13, No. 10, 975-985 (2008)
DOI: 10.1177/1087057108326141


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