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Journal of Biomolecular Screening
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What's this?

A Novel Method for Analyzing [Ca2+] Flux Kinetics in High-Throughput Screening

Philip Gribbon

Chris Chambers

Pfizer HTS Centre of Emphasis, Sandwich, UK.

Kaupo Palo

Juergen Kupper

Juergen Mueller

Evotec Technologies, Hamburg, Germany.

Andreas Sewing

Primary Pharmacology Group, IPC654, Ramsgate Road, Sandwich CT13 9NJ, United Kingdom andreas.sewing{at}pfizer.com

Driven by multiparameter fluorescence readouts and the analysis of kinetic responses from biological assay systems, the amount and complexity of high-throughput screening data are constantly increasing. As a consequence, the reduction of data to a simple number, reflecting a percentage activity/inhibition, is no longer an adequate approach because valuable additional information, for example, about compound-or process-induced artifacts, is lost. Time series data such as the transient calcium flux observed after activation of Gq-coupled G protein-coupled receptors (GPCRs), are especially challenging with respect to quantity of data; typically, responses are followed for several minutes. Based on measurements taken on the fluorometric imaging plate reader, the authors have introduced a mathematical model to describe the time traces of cellular calcium fluxes mediated by the activation of GPCRs. The model describes the time series using 13 parameters, reducing the amount of data by 90% while guiding the detection of compound-induced artifacts as well as the selection of compounds for further characterization.

Key Words: calcium • GPCR • HTS • FLIPR

This version was published on August 1, 2006

Journal of Biomolecular Screening, Vol. 11, No. 5, 511-518 (2006)
DOI: 10.1177/1087057106287929
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 J Biomol ScreenHome page
K. J. Cassutt, M. J. Orsini, M. Abousleiman, D. Colone, and W. Tang
Identifying Nonselective Hits from a Homogeneous Calcium Assay Screen
J Biomol Screen, March 1, 2007; 12(2): 285 - 287.
[Abstract] [PDF]