Putting the pieces together: contribution of fluorescence polarization assays to small-molecule lead optimization

Susan M. Keating; Jim Marsters; Maureen Beresini; Carmen Ladner; Kim Zioncheck; Kevin Clark; Fred Arellano; Sarah Bodary

doi:10.1117/12.382026

11 April 2000 Putting the pieces together: contribution of fluorescence polarization assays to small-molecule lead optimization

Susan M. Keating, Jim Marsters, Maureen Beresini, Carmen Ladner, Kim Zioncheck, Kevin Clark, Fred Arellano, Sarah Bodary

Author Affiliations +

Proceedings Volume 3913, In-Vitro Diagnostic Instrumentation; (2000) https://doi.org/10.1117/12.382026
Event: BiOS 2000 The International Symposium on Biomedical Optics, 2000, San Jose, CA, United States

Abstract

Fluorescence polarization assays with both purified receptor and intact cells have been developed to assess potency and selectivity of antagonists of the interaction of the lymphocyte receptor, LFA-1, and its endothelial ligand, ICAM-1. Fluorescein isothiocyanate conjugated small molecule probes were optimized for use in binding assay with LFA-1 and a closely related receptor, MAC-1. In the assays, the antagonists compete with the fluorescent probe for binding to the receptor. This enables the determination of IC₅₀ and consequently K_i values of the antagonists for each of the receptors. Routine use of polarization assay with tranfected cells, in addition to purified receptors, has become feasible with the availability of sensitive plate readers that are able to detect 1 nM fluorescent probe in 15 (mu) l sample volumes with good signal to noise. These measurements aid in the iterative synthesis of more potent and selective compounds.

Citation Download Citation

Susan M. Keating, Jim Marsters, Maureen Beresini, Carmen Ladner, Kim Zioncheck, Kevin Clark, Fred Arellano, and Sarah Bodary "Putting the pieces together: contribution of fluorescence polarization assays to small-molecule lead optimization", Proc. SPIE 3913, In-Vitro Diagnostic Instrumentation, (11 April 2000); https://doi.org/10.1117/12.382026

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