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A Homogeneous Fluorescence Polarization Assay Adaptable for a Range of Protein Serine/Threonine and Tyrosine Kinases

Molecular Devices Corporation, 1311 Orleans Ave. Sunnydale, CA 94089. Liz_Gaudet@MolDev.com

Kuo-Sen Huang

Yan Zhang

Wei Huang

David Mark

J. Richard Sportsman

Recently, a new technology for high-throughput screening has been developed, called IMAP(patent pending). IMAP technology has previously been implemented in an assay for cyclic nucleotide phosphodiesterases (PDE). The authors describe the development of a homogeneous, non-antibody-based fluorescence polarization (FP) assay for a variety of protein kinases. In this assay, fluorescently labeled peptide substrate phosphorylated by the kinase is captured on modified nanoparticles through interactions with immobilized metal (M^III) coordination complexes, resulting in a change from low to high polarization values. This assay is applicable to protein kinases that phosphorylate serine, threonine, or tyrosine residues. The IMAP platform is very compatible with high-throughput robotics and can be applied to the 1536-well format. As there are hundreds of different kinases coded for in the human genome, the assay platform described in this report is a valuable new tool in drug discovery. (Journal of Biomolecular Screening 2003:164-175)

Key Words: high-throughput screening • fluorescence polarization • protein kinases • IMAP platform

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