Open AccessDevelopment of a Highly Sensitive Cell-Based Assay for Detecting Botulinum Neurotoxin Type A through Neural Culture Media Optimization
Author(s) -
Won S. Hong,
Hannah M. Pezzi,
Andrea R. Schuster,
Scott M. Berry,
Kyung Eun Sung,
David J. Beebe
Publication year - 2016
Publication title -
slas discovery
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.907
H-Index - 75
eISSN - 2472-5560
pISSN - 2472-5552
DOI - 10.1177/1087057115608103
Subject(s) - neurotoxin , clostridium botulinum , cell culture , botulinum neurotoxin , retinoic acid , toxin , recombinant dna , cell , neural stem cell , chemistry , pharmacology , biology , biochemistry , microbiology and biotechnology , stem cell , gene , genetics
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.