Two simple and generic antibody-independent kinase assays: Comparison of a bioluminescent and a microfluidic assay format
Two simple and generic antibody-independent kinase assays: Comparison of a bioluminescent and a microfluidic assay format
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DOI：
10.1177/1087057104264175
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发表时间：
2004-08-01
影响因子：
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通讯作者：
Kannt, A
中科院分区：
文献类型：
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作者：
Pommereau, A;Pap, E;Kannt, A
In this study, the authors have compared the performance of 2 high-throughput screening assays for a serin/threonine kinase: a microplate-based, bioluminescent assay that uses the luciferin/luciferase system to monitor ATP consumption, and a microfluidic assay that measures the change in mobility in an electric field of a fluorescently labeled peptide upon phosphorylation. Both assays are homogeneous, nonradioactive, antibody independent and could be miniaturized to a reaction volume of 4 mul. The robustness of both formats was demonstrated by Z' values > 0.8. Screening of a small library (2133 compounds) showed that the results obtained with both technologies correlate very well. Although the threshold for hits was set to a comparably low value-22.2% and 13.7% inhibition for the ATP consumption and microfluidic assay, respectively, corresponding to mean plus 3 standard deviations-the overlap of active compounds identified with the 2 assay formats was greater than 94%. Thus, both assays allow the identification of even low potency inhibitors with a high level of confidence.
