2012 Annual Report 1a.Objectives (from AD-416): The objective of this cooperative research project is to develop an electrochemiluminescence assay for BoNT in foods, using novel monoclonal antibodies developed by our ARS laboratory. Specifically, BoNT occurs in many serotype variations. This Agreement addresses BoNT serotypes A, B, and E. The foods covered under this Agreement include ground beef, liquid eggs, milk, and leafy greens, such as lettuce and spinach. 1b.Approach (from AD-416): We will test various monoclonal antibodies developed by ARS Scientists at Albany, CA as well as additional monoclonal and polyclonal antibodies available through commercial license. Antibodies will be employed in pairs, in sandwich format, using the Meso Scale platform for electrochemiluminescence. For validation, crude supernatant from cultures of each serotype will be spiked into foods, and the Lower Limit Of Detection (LLOD) determined. 3.Progress Report: In FY 12 monoclonal antibodies against BoNT serotypes A and B were optimized for the detection of toxin in sera using a multiwell plate-based electrochemiluminescence(ECL) assay system. This assay system was used to study the pharmacokinetics of BoNTs in sera and the effects of potentially therapeutic antibodies. Additional details for the research can be found in the report for the parent project 5325-42000-048-00D, TECHNOLOGIES FOR DETECTING AND DETERMINING THE BIOAVAILABILITY OF BACTERIAL TOXINS. Research progress reported, addresses objectives 1 and 2 of the parent project.