Multiplexed G-Protein–Coupled Receptor Ca2+ Flux Assays for High-Throughput Screening
Pfizer Research Technology Center, 620 Memorial Drive, Cambridge, MA 02139
Pfizer Research Technology Center, 620 Memorial Drive, Cambridge, MA 02139; Pharmaceutical Sales and Marketing Department, Boehringer Ingelheim, Burlington, Ontario, Canada
Pfizer Research Technology Center, 620 Memorial Drive, Cambridge, MA 02139 An early drug discovery approach focusing on gene families can benefit fromstrategies that exploit common signalingmechanisms to more effectively identify and characterize novel chemical lead structures. Multiplexing, defined as the screening of multiple targets within the same experiment, is an example of this strategy. Here, the authors describe a technique that allows multiplexing of a common assay type used to study G-protein–coupled receptors: changes in intracellular Ca2+ levels as measured by Molecular Device's fluorometric imaging plate reader (FLIPR®). The multiplexed FLIPR assays showed the expected pharmacological properties of single assays, with good reproducibility and Z• factors. The authors used them to screen large compound libraries in 2 multiplexed assay designs. The 1st used a single-cell line expressing 2 different receptors and the 2nd amixture of 2 cell lines of the same type each expressing distinct receptors. Screening using thesemultiplexed assays produced significant savings in reagents, time, and human resources and allowed the authors to quickly identify specific and selective hits.
Key Words: GPCR • Ca 2+flux • FLIPR • multiplex • HTS
This version was published on December
1, 2005 Journal of Biomolecular Screening, Vol. 10, No. 8,
780-787 (2005) This article has been cited by other articles:
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