SBIR Phase I:De novo assays for detection of the proteolytic activity in botulinum neurotoxin-based pharmaceuticals

This Small Business Innovation Research (SBIR) Phase I project proposes to develop a universal, in vitro assay for determining the enzymatic activity of botulinum neurotoxin (BoNT)-based pharmaceuticals. BoNTs are used in many cosmetic and pharmaceutical applications due to their long lasting effects and specificity for motor neurons. Currently, the strength and quality of BoNT-based pharmaceuticals is assessed using a mouse bioassay where BoNT is injected into mice and a unit of activity depends on the rate or proportion of deaths. Beyond animal testing concerns, the mouse bioassay's inaccuracy and the lack of standardized protocols among manufacturers put patient safety and clinical outcomes at risk. The proposed technology will reduce animal use and provide a method to uniformly assess the activity of BoNT-based pharmaceuticals regardless of manufacturer, thus increasing patient safety and the likelihood of successful treatment. The broader/commercial impacts of this research are increased patient safety, reduced animal testing, and the potential to enable point-of-use/point-of-care testing. The proposed technology could be used for decontamination testing at manufacturing sites, authenticity (against counterfeit drugs) testing at border control points, and in-office testing for high-dose procedures where toxin concentration is critical for clinical outcomes. The proposed technology would be the first of its kind on the commercial market. Other commercially available assays are not suitable for assessing BoNT-based pharmaceuticals because formulation stabilizers interfere with the assays. The proposed technology could be further adapted for testing BoNT activity in highly complex matrices extending the technology's utility into food, water, biodefense, and environmental testing.