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Journal of Biomolecular Screening
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Miniaturization of Whole Live Cell-Based GPCR Assays Using Microdispensing and Detection Systems

Oleg Kornienko

Merck Research Laboratories, Department of Automated Biotechnology, North Wales, PA.

Raul Lacson

Merck Research Laboratories, Department of Automated Biotechnology, North Wales, PA.

Priya Kunapuli

Merck Research Laboratories, Department of Automated Biotechnology, North Wales, PA.

Jonathan Schneeweis

Merck Research Laboratories, Department of Automated Biotechnology, North Wales, PA.

Ira Hoffman

Merck Research Laboratories, Department of Automated Biotechnology, North Wales, PA.

Todd Smith

Merck Research Laboratories, Department of Automated Biotechnology, North Wales, PA.

Melissa Alberts

Merck Research Laboratories, Department of Automated Biotechnology, North Wales, PA.

James Inglese

Merck Research Laboratories, Department of Automated Biotechnology, North Wales, PA.

Berta Strulovici

Merck Research Laboratories, Department of Automated Biotechnology, North Wales, PA.

Cell-based ß-lactamase reporter gene assays designed to measure the functional responses of G-protein-coupled receptors (GPCRs) were miniaturized to less than 2 µL total assay volume in a 3456-well microplate. Studies were done to evaluate both receptor agonists and antagonists. The pharmacology of agonists and antagonists for target GPCRs originally developed in a 96-well format was recapitulated in a 3456-well microplate format without compromising data quality or EC50/IC50 precision. These assays were employed in high-throughput screening campaigns, allowing the testing of more than 150,000 compounds in 8 h. The instrumentation used and practical aspects of the assay development are discussed.

Key Words: miniaturization • ultra-high throughput • screening • live cell-based assay • nanoplates • piezo crystal dispensers

Journal of Biomolecular Screening, Vol. 9, No. 3, 186-195 (2004)
DOI: 10.1177/1087057103260070
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