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Application of Micro Arrayed Compound Screening (pcARCS) to Identify Inhibitors of Caspase-3

Abbott Laboratories, Pharmaceutical Products Division, Abbott Park, IL

Jarkko Karvinen

PerkinElmer Life Sciences, Wallac OY, Turku, Finland

James L. Kofron

Abbott Laboratories, Pharmaceutical Products Division, Abbott Park, IL

David J. Burns

Abbott Laboratories, Pharmaceutical Products Division, Abbott Park, IL

Usha Warrior

Abbott Laboratories, Pharmaceutical Products Division, Abbott Park, IL

Micro Arrayed Compound Screening (pARCS) is a miniaturized ultra-high-throughput screening platform developed at Abbott Laboratories. In this format, 8640 discrete compounds are spotted and dried onto a polystyrene sheet, which has the same footprint as a 96-well plate. A homogeneous time-resolved fluorescence assay format (LANCE) was applied to identify the inhibitors of caspase-3 using a peptide substrate labeled with a fluorescent europium chelate and a dabcyl quencher. The caspase-3 enzyme was cast into a thin agarose gel, which was placed on a sheet containing test compounds. A second gel containing caspase substrate was then laid above the enzyme gel to initiate the reaction. Caspase-3 cleaves the substrate and separates the europium from the quencher, giving rise to a time-resolved fluorescent signal, which was detected using a ViewLux charge-coupled device imaging system. Potential inhibitors of caspase-3 appeared as dark spots on a bright fluorescent background. Results from the pARCS assay format were compared to those from a conventional 96-well plate-screening format.

Journal of Biomolecular Screening, Vol. 7, No. 4, 317-323 (2002)
DOI: 10.1177/108705710200700403


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