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Toxins 2017, 9(7), 230; doi:10.3390/toxins9070230

SiMa Cells for a Serotype Specific and Sensitive Cell-Based Neutralization Test for Botulinum Toxin A and E

Division of Bacteriology, National Institute for Biological Standards and Control (NIBSC), A Centre of the Medicines and Healthcare Products Regulatory Agency, Hertfordshire EN6 3QG, UK
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Author to whom correspondence should be addressed.
Academic Editors: Jianlong Lou and James D. Marks
Received: 11 May 2017 / Accepted: 18 July 2017 / Published: 20 July 2017
(This article belongs to the Special Issue Botulinum Neurotoxins Antibody and Vaccine)
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Abstract

Botulinum toxins (BoNTs), of which there are seven serotypes, are among the most potent neurotoxins, with serotypes A, B and E causing human botulism. Antitoxins form the first line of treatment for botulism, and functional, highly sensitive in vitro methods for toxin neutralization are needed to replace the current in vivo methods used for determination of antitoxin potency. In this preliminary proof of concept study, we report the development of a neutralization test using the neuroblastoma SiMa cell line. The assay is serotype specific for either BoNT/A or BoNT/E, which both cleave unique sequences on SNAP-25 within SiMa cells. The end point is simple immunodetection of cleaved SNAP-25 from cell lysates with antibodies detecting only the newly exposed sequence on SNAP-25. Neutralizing antibodies prevent the toxin-induced cleavage of SNAP-25. The toxin neutralization assay, with an EC50 of ~2 mIU/mL determined with a standardized reference antiserum, is more sensitive than the mouse bioassays. Relevance was demonstrated with commercial and experimental antitoxins targeting different functional domains, and of known in vivo neutralizing activities. This is the first report describing a simple, specific, in vitro cell-based assay for the detection of neutralizing antibodies against BoNT/A and BoNT/E with a sensitivity exceeding that of the mouse bioassay. View Full-Text
Keywords: SiMa cells; BoNT/A; BoNT/E; cell-based assay; toxin neutralization test (TNT); SNAP-25; capture ELISA; immunodetection SiMa cells; BoNT/A; BoNT/E; cell-based assay; toxin neutralization test (TNT); SNAP-25; capture ELISA; immunodetection
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This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. (CC BY 4.0).

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Bak, N.; Rajagopal, S.; Stickings, P.; Sesardic, D. SiMa Cells for a Serotype Specific and Sensitive Cell-Based Neutralization Test for Botulinum Toxin A and E. Toxins 2017, 9, 230.

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