ANALYSIS OF PROTEASE ACTIVITY USING MEMBRANE- COATED BIOMIMETIC MICROSPHERES

Poisoning by Botulinum Neurotoxins (BoNTs) leads to the severe neuroparalytic illness known as botulism, which is characterized by flaccid paralysis, respiratory failure, and death. BoNTs are the most acutely toxic substances on the planet, so detection at physiologically relevant concentrations is difficult. Because current treatments for botulism cannot reverse existing paralysis, rapid detection is key, and current diagnostic techniques are not meeting this need. We have developed a flow-based assay platform that allows for detection of the Light Chain of BoNT/A as well as a means to study the action of the toxin on its protein substrate in a system that mimics many aspects of a physiologically relevant environment. The total assay time has been reduced to approximately 20 hours, and this platform has multiplex and high throughput screening capabilities for drug discovery.NIH 1R21EB016339-01A