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Journal of Biomolecular Screening
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Miniaturization and Validation of a High-Throughput Serine Kinase Assay Using the Alpha Screen Platform

Achim Von Leoprechting

Perkin Elmer LAS (Germany) GmbH, Rodgau-Jügesheim, Germany.

Renate Kumpf

Susanne Menzel

Dominique Reulle

Department of Integrated Lead Discovery, Boehringer Ingelheim Pharma GmbH & Co. KG, Biberach an der Riss, Germany.

Ralf Griebel

Perkin Elmer LAS (Germany) GmbH, Rodgau-Jügesheim, Germany.

Martin J. Valler

Frank H. Büttner

Department of Integrated Lead Discovery, Boehringer Ingelheim Pharma GmbH & Co. KG, Biberach an der Riss, Germany.

Reducing costs while maintaining the highest readout quality is a precept of modern high-throughput screening. Given the trend toward nonradiometric screening platforms, this has been a big challenge for some kinase target classes. Common issues include lowsensitivity, susceptibility to nonspecific interference, or the need for costly reagents. In this study, the authors describe the feasibility ofminiaturization of a serine kinase assay using generic reagents in the Alpha Screen format. They have validated the robustness of this assay in the course of miniaturization from a 35-to 4.375-µ L final assay volume in 384-and 1536-well formats. Within this volume range, they consistently obtained Z• values above 0.5 and have investigated the suitability of these assay formats for measuring compound effects by testing a set of 25 previously identified active compounds. These active compounds were also reliably identified in the miniaturized assay formats. The results presented here show that the Alpha Screen technology permits robust and cost-efficient miniaturization of serine/threonine kinase assays.

Key Words: serine kinase • Alpha Screen • homogeneous assay • miniaturization • HTS

Journal of Biomolecular Screening, Vol. 9, No. 8, 719-725 (2004)
DOI: 10.1177/1087057104268805
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