Synthetic substrate for application in both high and low throughput assays for botulinum neurotoxin B protease inhibitors

A FRET peptide substrate was synthesized and evaluated for enzymatic cleavage by the BoNT/B light chain protease. The FRET substrate was found to be useful in both a high throughput assay to uncover initial 'hits' and a low throughput HPLC assay to determine kinetic parameters and modes of inhibition.