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United States Department of Agriculture

Agricultural Research Service

Research Project: Technologies for Detecting and Determining the Bioavailability of Bacterial Toxins

Location: Foodborne Toxin Detection and Prevention

Title: Development and characterization of six monoclonal antibodies to hemagglutinin-70 (HA70) of Clostridium botulinum and their application in a sandwich ELISA

Authors
item Scotcher, Miles -
item Cheng, Luisa Wai Wai
item Ching, Kathryn
item McGarvey, Jeffery
item Hnasko, Robert
item Stanker, Larry

Submitted to: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: October 18, 2012
Publication Date: January 10, 2013
Citation: Scotcher, M.C., Cheng, L.W., Ching, K.H., Mcgarvey, J.A., Hnasko, R.M., Stanker, L.H. 2013. Development and characterization of six monoclonal antibodies to hemagglutinin-70 (HA70) of Clostridium botulinum and their application in a sandwich ELISA. Monoclonal Antibodies in Immunodiagnosis and Immunotherapy. 32(1):6-15. doi: 10.1089/mab.2012.0071.

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. Because of its high toxicity, the need for a long recovery period requiring extensive treatment, and the ease of producing BoNT, it is considered a class A bioterrorism agent. BoNT is secreted as a protein complex consisting of a toxin molecule surrounded by non-toxin associated proteins (NAPs). The exact role of the NAPs in disease is not known, but it is thought that they protect the toxin as it passes through the harsh environment of the stomach. We describe a set of monoclonal antibodies that bind to one of the NAP proteins , HA70, and using these antibodies we have developed a rapid assay for HA70. The ability to easily detect HA70, a protein associated with botulism, coupled with a previously developed assay for the toxin itself improves our ability to detect this lethal foodborne agent ultimatley resulting in a safer food supply.

Technical Abstract: Botulinum neurotoxins (BoNT),produced by Clostridium botulinum, cause severe neuroparalytic disease that if not treated quickly is often fatal. The toxin is synthesized as a 150 kDa precursor protein (holotoxin), which is then enzymatically cleaved to form two subunit chains linked by a single disulfide bond. The toxin serotypes A1 and B1 are secreted as precursor toxic complexes consisting of toxin, non-toxic heamagglutinins (HA), designated HA17, HA34 and HA70, and a 120 kDa non-toxin non-hemagglutinin (NTNH) protein. The exact structure of the complex is poorly understood although recent models suggest that for each molecule of toxin the PTC contains one molecule of the HTNH and multiple copies of the HA. In this paper we describe isolation of six monoclonal antibodies that specifically bind the HA70 protein from BoNT/A1 and /B1. Using these antibodies we demonstrate a rapid sandwich ELISA assay or detecting HA70.

Last Modified: 4/16/2014
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