Submitted to: The Botulinum Journal
Publication Type: Proceedings
Publication Acceptance Date: 8/29/2012
Publication Date: 10/22/2012
Citation: Stanker, L.H., Cheng, L.W. 2012. Monoclonal antibodies and reagents for botulinum research. The Botulinum Journal. 2(2):150-155 doi: 10.1504/tbj2012.050197. Interpretive Summary: Botulism is a serious, often fatal neuroparalytic disease in humans and animals caused by a protein toxin (botulinum toxin, BoNT) produced by the bacterium Clostridium botulinum. BoNT is considered the most toxic biological toxin known. It is produced as a protein complex containing the toxin as well as nontoxic associated proteins. In this report we report development of monoclonal antibodies to different forms of the toxin and antibodies to the nontoxic proteins associated with the toxin complex found in nature, i.e., the nontoxic associated proteins (NAPs). Antibodies to the NAPs together with our previously generated anti-toxin monoclonal antibodies will allow us to develop sensitive immunoassays capable of detecting the toxin complex. The development of this immunoassay will facilitate rapid detection of botulinum contamination in food.
Technical Abstract: Botulinum neurotoxin (BoNT) is produced by Clostridium botulinum as a dichain protein of ~ 150 kDa that blocks acetylcholine release resulting in muscular paralysis. Diagnosis of BoNT often relies on the mouse bioassay that has a detection limit of 10–20 pg/mL and can take up to four days to complete. Rapid in vitro methods for toxin detection are needed and to date, most rely on either immunoassay or endopeptidase activity. In the latter, many also use specific antibodies to concentrate the toxin. We have developed panels of monoclonal antibodies (mAbs) to purified toxin, serotypes A, B and E, as well as mAbs to the non-toxic associated proteins. Application of these mAbs in sandwich ELISAs and assay performance in milk and other foods is discussed. The assays described here are able to detect toxin in a few hours, at levels lower than the mouse bioassay.