One important consideration in any assay that uses an enzymatic reporter is inhibition of that reporter enzyme by compounds from the library (1). High-throughput, luciferase-coupled assays do not require expensive antibodies or fluorescent labels and therefore provide a platform for screening that reduces assay development time and reagent costs. Firefly luciferase can be used to measure ATP concentrations, as a reporter of cell-based gene expression and to assay the turnover of proluciferin substrates in the presence of target enzymes. High-throughput screening (HTS) is widely used by the pharmaceutical industry for drug discovery. It leverages automation to quickly assay the biological or biochemical activity of a large number of drug-like compounds and is useful for discovering ligands for receptors, enzymes, ion-channels or other pharmacological targets, or for profiling cellular or biochemical pathways. Firefly luciferase is a common reporter used in HTS because of its sensitivity, ease of use, and low expense (1).
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