Development of a Highly Sensitive Cell-Based Assay for Detecting Botulinum Neurotoxin Type A through Neural Culture Media Optimization. (January 2016)

• Record Type: Journal Article
• Authors: Hong, Won S.; Pezzi, Hannah M.; Schuster, Andrea R.; Berry, Scott M.; Sung, Kyung E.; Beebe, David J.
• Is Part Of: Journal of biomolecular screening. Volume 21:Number 1(2016)
• Page Start: 65
• Page End: 73
• Publication Date: 2016-01
• Subjects: botulinum neurotoxin -- cell-based assays -- media optimization -- toxicology; Drugs -- Analysis -- Periodicals; Drugs -- Testing -- Periodicals; Biomolecules -- Analysis -- Periodicals; 572.36
• DOI: 10.1177/1087057115608103
• Languages: English
• ISSNs: 1087-0571

Abstract

Botulinum neurotoxin (BoNT) is the most lethal naturally produced neurotoxin. Due to the extreme toxicity, BoNTs are implicated in bioterrorism, while the specific mechanism of action and long-lasting effect was found to be medically applicable in treating various neurological disorders. Therefore, for both public and patient safety, a highly sensitive, physiologic, and specific assay is needed. In this paper, we show a method for achieving a highly sensitive cell-based assay for BoNT/A detection using the motor neuron–like continuous cell line NG108-15. To achieve high sensitivity, we performed a media optimization study evaluating three commercially available neural supplements in combination with retinoic acid, purmorphamine, transforming growth factor β1 (TGFβ1), and ganglioside GT1b. We found nonlinear combinatorial effects on BoNT/A detection sensitivity, achieving an EC50 of 7.4 U ± 1.5 SD (or ~7.9 pM). The achieved detection sensitivity is comparable to that of assays that used primary and stem cell–derived neurons as well as the mouse lethality assay.