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[Advances in Drug Discovery 2017] A Duplexed Calcium and β-Arrestin Assay Protocol for Human Histamine H1 Receptor PathHunter CHO-K1 HRH1 β-Arrestin Cell Line

[Advances in Drug Discovery 2017] A Duplexed Calcium and β-Arrestin Assay Protocol for Human Histamine H1 Receptor PathHunter CHO-K1 HRH1 β-Arrestin Cell Line
Version:
030617

File Name/Number:
2017 Advances in Drug Discovery

Year:
2017

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The screening and functional characterization of novel ligands for G-protein-coupled receptors (GPCRs) remains a key activity in drug discovery process. Furthermore, an increasing awareness of the subtleties of receptor signaling and potential for ligand bias requires multiple endpoints to be monitored. Efforts to reduce costs and improve efficiency also remain a key consideration. In this poster, we describe efforts to develop a duplexed assay for the Histamine H1 receptor employing the DiscoverX PathHunter® β-Arrestin clonal cell line. We have adapted existing protocols such that from a single plate we can monitor calcium  signaling with fluorescent dyes sensitive to intracellular Ca2+ ion flux in real time, followed by the recruitment of β-arrestin as an endpoint assay. Protocol development will be discussed and successful implementation of the final assay conditions will be exemplified with a characterization of H1 agonists and antagonist pharmacology.