Development of a highly sensitive cell-based assay for detecting botulinum neurotoxin type a through neural culture media optimization

Won S. Hong, Hannah M. Pezzi, Andrea R. Schuster, Scott M. Berry, Kyung E. Sung, David J. Beebe

Research output: Contribution to journalArticlepeer-review

4 Scopus citations

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 ECof 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.

Original languageEnglish
Pages (from-to)65-73
Number of pages9
JournalJournal of Biomolecular Screening
Volume21
Issue number1
DOIs
StatePublished - Jan 1 2016

Bibliographical note

Publisher Copyright:
© 2015 Society for Laboratory Automation and Screening.

Keywords

  • botulinum neurotoxin
  • cell-based assays
  • media optimization
  • toxicology

ASJC Scopus subject areas

  • Analytical Chemistry
  • Biotechnology
  • Biochemistry
  • Molecular Medicine
  • Pharmacology
  • Drug Discovery

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