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Proceedings Paper

Putting the pieces together: contribution of fluorescence polarization assays to small-molecule lead optimization
Author(s): Susan M. Keating; Jim Marsters; Maureen Beresini; Carmen Ladner; Kim Zioncheck; Kevin Clark; Fred Arellano; Sarah Bodary
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Paper 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 Ki 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.

Paper Details

Date Published: 11 April 2000
PDF: 10 pages
Proc. SPIE 3913, In-Vitro Diagnostic Instrumentation, (11 April 2000); doi: 10.1117/12.382026
Show Author Affiliations
Susan M. Keating, Genentech, Inc. (United States)
Jim Marsters, Genentech, Inc. (United States)
Maureen Beresini, Genentech, Inc. (United States)
Carmen Ladner, Genentech, Inc. (United States)
Kim Zioncheck, Genentech, Inc. (United States)
Kevin Clark, Genentech, Inc. (United States)
Fred Arellano, Genentech, Inc. (United States)
Sarah Bodary, Genentech, Inc. (United States)

Published in SPIE Proceedings Vol. 3913:
In-Vitro Diagnostic Instrumentation
Gerald E. Cohn, Editor(s)

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