DEVELOPMENT OF AN ELECTROCHEMILUMINESCENT ASSAY TO SUPPORT HIGH THROUGHPUT SCREENING FOR BOTULINUM NEUROTOXIN SEROTYPE A INHIBITORS

Abstract

Botulinum neurotoxin (BoNT) is the causative agent for the disease Botulism and is secreted by the bacterium, Clostridium botulinum. Botulinum neurotoxins gain entry into motor neurons and selectively cleave a component of the SNARE protein complex, inhibiting the complex from binding and releasing the neurotransmitter, acetylcholine, resulting in flaccid paralysis and ultimately death. Botulinum neurotoxin serotype A (BoNT/A) is the most potent known toxin in nature and has the potential to be used as a bioweapon. The medical usage of BoNT has increased and the potential for illicit medical procedures could lead to an increase in accidental BoNT intoxications. Both the health and bioterror threats necessitate the development of improved therapeutic options. To identify potential BoNT/A inhibitors, this study focuses on the development of a Meso Scale Discovery Electrochemiluenescence (MSD-ECL) assay for high throughput screening. This assay was found to be sensitive, reproducible and reliable. As an initial proof of concept study, a 1200-compound library was screened with this assay and 21 hits were identified as potential BoNT/A inhibitors.