Sensitive and quantitative detection of botulinum neurotoxin in neurons derived from mouse embryonic stem cells

• Published in: Biochemical and biophysical research communications Vol. 404; no. 1; pp. 388 - 392
• Main Authors: Pellett, Sabine, Du, Zhong-wei, Pier, Christina L., Tepp, William H., Zhang, Su-chun, Johnson, Eric A.
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 07.01.2011
• Subjects: Animals; bioassays; Biological Assay; biopharmaceuticals; Bioterrorism; BoNT; Botulinum toxin; Botulinum Toxins, Type A - analysis; Botulinum Toxins, Type A - toxicity; Cell assay; Cell Count; cell culture; Cell Culture Techniques; Cells, Cultured; embryonic stem cells; Embryonic Stem Cells - cytology; mES; Mice; Mouse embryonic stem cells; Neurogenesis; neurons; Neurons - chemistry; Neurons - cytology; Neurons - drug effects; neurotoxicity; Sensitivity and Specificity; mES; Cell assay; BoNT; MBA; Botulinum toxin; Mouse embryonic stem cells
• ISSN: 0006-291X; 1090-2104
• DOI: 10.1016/j.bbrc.2010.11.128

► Botulinum toxins are both a serious bioterrorism threat and an important bio-pharmaceutical. ► A major emphasis by NIH and FDA is to replace the current mouse bioassay. ► Cell-based assays are an excellent platform to quantitatively detect botulinum toxins. ► Currently primary cells are required to reach the necessary sensitivity. ► Neuron cultures derived from mouse stem cells provide a novel assay platform. Botulinum neurotoxins (BoNTs), the most poisonous protein toxins known, represent a serious bioterrorism threat but are also used as a unique and important bio-pharmaceutical to treat an increasing myriad of neurological disorders. The only currently accepted detection method by the United States Food and Drug Administration for biological activity of BoNTs and for potency determination of pharmaceutical preparations is the mouse bioassay (MBA). Recent advances have indicated that cell-based assays using primary neuronal cells can provide an equally sensitive and robust detection platform as the MBA to reliably and quantitatively detect biologically active BoNTs. This study reports for the first time a BoNT detection assay using mouse embryonic stem cells to produce a neuronal cell culture. The data presented indicate that this assay can reliably detect BoNT/A with a similar sensitivity as the MBA.