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Journal of Biomolecular Screening
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A 1536 Colorimetric SPAP Reporter Assay: Comparison with 96- and 384-Well Formats

John C.W. Comley

Lead Discovery Unit, Glaxo Wellcome Research and Development, Stevenage, UK

Tony Reeves

Lead Discovery Unit, Glaxo Wellcome Research and Development, Stevenage, UK

Phil Robinson

Lead Discovery Unit, Glaxo Wellcome Research and Development, Stevenage, UK

We report the successful miniaturization of a functional cell-based reporter gene assay. Utilizing interleukin-1beta (IL-1/ß)-induced secreted placental alkaline phosphatase (SPAP)-catalyzed colorimetric readout, we reduced the assay volume to 10 µl using a Greiner 1536-well microplate. Our experiences of assay development, liquid handling (using a Hydra® 96; Robbins Scientific, Sunnyvale, CA), and detection (using the SpectraImage and SpectraFluor-Plus plate readers, Tecan Austria GmbH, Grodig, Austria) in 1536 wells are discussed. The effect of a set of 1,280 compounds in this SPAP reporter assay were compared between 96-, 384-, and 1536-well formats and were shown to be very similar. We conclude that cell-based reporter gene assays using SPAP-catalyzed color readouts are sensitive and highly reproducible in 1536-well plates and should be considered as a cost-effective alternative to luciferase reporters for miniaturized assay formats. Finally, we review the prospects for the implementation of routine HTS in 1536-well plates based around the instrumentation investigated.

Journal of Biomolecular Screening, Vol. 3, No. 3, 217-225 (1998)
DOI: 10.1177/108705719800300308
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