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A 384-HTS for Human Factor VIIa: Comparison With 96- and 864-Well Formats

Lead Discovery Unit, Glaxo Wellcome Research and Development, Medicines Research Centre, Gunnels Wood Road, Stevenage, Hertordshire, SGl 2NY, UK

Alastair Binnie

Lead Discovery Unit, Glaxo Wellcome Research and Development, Medicines Research Centre, Gunnels Wood Road, Stevenage, Hertordshire, SGl 2NY, UK

Caroline Bonk

Lead Discovery Unit, Glaxo Wellcome Research and Development, Medicines Research Centre, Gunnels Wood Road, Stevenage, Hertordshire, SGl 2NY, UK

John G. Houston

Lead Discovery Unit, Glaxo Wellcome Research and Development, Medicines Research Centre, Gunnels Wood Road, Stevenage, Hertordshire, SGl 2NY, UK

A homogenous fluorescent HTS for recombinant human factor VIIa (FVIIa) using soluble tissue factor has been developed in 384-well microplates. In this report we discuss our experiences with assay development, liquid handling using a Tomtec Quadra and Matrix PlateMate, fluorescent detection and screening of -200,000 compounds against FVIIa in 384-well plate format. Assays using the entire Helix 864-well plate were prototyped using contact dispensing with a modified Hamilton Microlab 2200. FVIIa was used as a model assay to compare between 96-, 384-, and 864-plate formats in a total assay volume of 100, 25, and 10 µl, respectively. FVIIa was assayed in 864 to the same degree of sensitivity as 384- and 96-well assays and dose-response curves for a standard inhibitor (benzamidine) in the FVIIa assay were identical in all plate formats. Finally, we review the prospects for HTS in 864-well microplates.

Journal of Biomolecular Screening, Vol. 2, No. 3, 171-178 (1997)
DOI: 10.1177/108705719700200307


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