Document Detail

Botulinum neurotoxin serotype A specific cell-based potency assay to replace the mouse bioassay.
MedLine Citation:
PMID:  23185348     Owner:  NLM     Status:  MEDLINE    
Botulinum neurotoxin serotype A (BoNT/A), a potent therapeutic used to treat various disorders, inhibits vesicular neurotransmitter exocytosis by cleaving SNAP25. Development of cell-based potency assays (CBPAs) to assess the biological function of BoNT/A have been challenging because of its potency. CBPAs can evaluate the key steps of BoNT action: receptor binding, internalization-translocation, and catalytic activity; and therefore could replace the current mouse bioassay. Primary neurons possess appropriate sensitivity to develop potential replacement assays but those potency assays are difficult to perform and validate. This report describes a CBPA utilizing differentiated human neuroblastoma SiMa cells and a sandwich ELISA that measures BoNT/A-dependent intracellular increase of cleaved SNAP25. Assay sensitivity is similar to the mouse bioassay and measures neurotoxin biological activity in bulk drug substance and BOTOX® product (onabotulinumtoxinA). Validation of a version of this CBPA in a Quality Control laboratory has led to FDA, Health Canada, and European Union approval for potency testing of BOTOX®, BOTOX® Cosmetic, and Vistabel®. Moreover, we also developed and optimized a BoNT/A CBPA screening assay that can be used for the discovery of novel BoNT/A inhibitors to treat human disease.
Ester Fernández-Salas; Joanne Wang; Yanira Molina; Jeremy B Nelson; Birgitte P S Jacky; K Roger Aoki
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't     Date:  2012-11-21
Journal Detail:
Title:  PloS one     Volume:  7     ISSN:  1932-6203     ISO Abbreviation:  PLoS ONE     Publication Date:  2012  
Date Detail:
Created Date:  2012-11-27     Completed Date:  2013-05-22     Revised Date:  2013-07-11    
Medline Journal Info:
Nlm Unique ID:  101285081     Medline TA:  PLoS One     Country:  United States    
Other Details:
Languages:  eng     Pagination:  e49516     Citation Subset:  IM    
Department of Biological Sciences, Allergan Inc., Irvine, California, United States of America.
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MeSH Terms
Antibodies, Monoclonal / chemistry
Biological Assay / methods*
Botulinum Toxins, Type A / chemistry,  physiology*
Cell Line, Tumor
Enzyme-Linked Immunosorbent Assay / methods
Neurons / metabolism
Neurotoxins / chemistry
PC12 Cells
Quality Control
Sensitivity and Specificity
Surface Plasmon Resonance / methods
Synaptosomal-Associated Protein 25 / genetics*,  metabolism
Reg. No./Substance:
0/Antibodies, Monoclonal; 0/Neurotoxins; 0/Snap25 protein, mouse; 0/Synaptosomal-Associated Protein 25; 0/onabotulinumtoxinA; EC Toxins, Type A

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