<li>We will determine enteric dose-response relationships for crude preparations of botulinum neurotoxin (BoNT)in food using rodent models.</li>
<li>We will optimize sample preparation and develop rapid immunological and biochemical tests for BoNT.</li>
Approach: 1. We will administer BoNT in buffer and BoNT spiked food to mice, testing various time and temperature storage conditions (including heating) for effects on toxicity. We will compare commercially available purified BoNT to crude toxin (sterile culture fractions). 2. We will evaluate BoNT assays currently available commercially or through collaborators. We will develop new monoclonal antibodies to BoNT toxin and toxoid. Initial evaluations will be made in ELISA and other standard formats, and selected assays will be ported onto new assay platforms. Examples of new platforms include microbead assays read via microfluorimetry, paramagnetic fluorescent nanosphere assays, microarray analyses employing glass or nitrocellulose surfaces, "dip-stick" format rapid ELISAs, and "black-box" turnkey assay systems. Biochemical assays will include measurements of protease activity using fluorescence polarization and/or fluorescence resonance energy transfer. Documents SCA with the University of Wisconsin-Madison.