Further comparison of primary hit identification by different assay technologies and effects of assay measurement variability.
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Wu, Xiang, Sills, Matthew and Zhang, Ji (2005) Further comparison of primary hit identification by different assay technologies and effects of assay measurement variability. Journal of Biomolecular Screening : the official journal of the Society for Biomolecular Screening, 10 (6). pp. 581-589. ISSN 1087-0571
Abstract
High-throughput screening (HTS) has grown rapidly in the past decade, with many advances in new assay formats, detection technologies, and laboratory automation. Recently, several studies have shown that the choice of assay technology used for the screening process is particularly important and can yield quite different primary screening outcomes. However, because the screening assays in these previous studies were performed in a single-point determination, it is not clear to what extent the difference observed in the screening results between different assay technologies is attributable to inherent assay variability and day-to-day measurement variation. To address this question, a nuclear receptor coactivator recruitment assay was carried out in 2 different assay formats, namely, AlphaScreen and time-resolved fluorescence resonance energy transfer, which probed the same biochemical binding events but with different detection technologies. For each assay format, 4 independent screening runs in a typical HTS setting were completed to evaluate the run-to-run screening variability. These multiple tests with 2 assay formats allow an unambiguous comparison between the discrepancies of different assay formats and the effects of the variability of assay and screening measurements on the screening outcomes. The results provide further support that the choice of assay format or technology is a critical factor in HTS assay development.
| Item Type: | Article |
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| Additional Information: | Publisher's version/PDF cannot be used |
| Keywords: | high-throughput screening; assay comparison, AlphaScreen™;TR-FRET; assay variability; assay concordance |
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| Date Deposited: | 14 Dec 2009 14:07 |
| Last Modified: | 31 Jan 2013 01:32 |
| URI: | https://oak.novartis.com/id/eprint/28 |