Homogeneous High Throughput Live Cell GPCR Functional and Surface Binding Assays

Oksana Sirenko, Estelle N'Garwate, Jean-Luc Tardieu, Marie-Laure Lebreton, Delphine Jaga, Michael Katzlinger, Laurence Monnet, Caroline Cardonnel, Cathy Olsen, Francois Degorce, Evan F. Cromwell


Molecular Devices, Inc. Sunnyvale, CA. Cisbio Bioassays, Codolet, France.

G-protein coupled receptors (GPCR) are the largest class of cell-surface receptors and are targets for almost 40% of existing drugs. Lead discovery, testing the efficacy of prospective drugs (in the area of cardiovascular diseases and other fields), and understanding of mechanism of action of drug candidates requires assays that can measure the binding of ligands to the receptors, receptor oligomerization, and/or internalization. Accordingly, there is a real need for robust and sensitive assays of this type that are suitable for high throughput screening. The Tag-lite® cellular screening platform was designed to increase the flexibility of cell-surface receptor research. This platform is ideal for primary and secondary screening and can be applied to a variety of assay formats for pharmacological characterization and development of therapeutic antibodies. Here we show results from use of this assay platform with SpectraMax® Paradigm plate reader for characterization of GPCR agonist and antagonists in Tag-lite receptor ligand binding assays including Dopamine D3, Glucagon GLP1, and Mu Opioid assays. Excellent performance was observed as measured by Z-prime and assay window values. We also demonstrated performance for cAMP detection, pERK and pAKT kinase assays.

HTRF microplate readers