Botulinum type E toxin is a well recognized causative agent of seafood botulism poisoning. Underprocessing or postretort recontamination of preserved seafoods has resulted in sporadic cases of botulism. Currently, laboratory mice are being used to detect this toxin. However, it requires three to six days to obtain final results. A rapid method using monoclonal antibody (Mab) enzyme immunoassay was therefore developed. Hybridomas secreting specific Mab against the type E epitope were generated by fusion of SP/20‐Ag 14 myeloma cells with spleen cells from BALB/c mice immunized with botulinum type E neurotoxoid. Five potent, stable hybridomas were selected, cloned, propagated, and preserved in liquid nitrogen as cell lines. Immunoglobulin subisotyping showed these Mabs belonged to the IgG subclasses. No cross‐reaction was observed with culture supernatants of C. botulinum types A, B, and F or with crude toxins extracts of type C and D. Large quantities of Mabs were produced in ascites fluids, harvested, and affinity purified. A Mab‐based biotin‐avidin amplified double sandwich enzyme‐linked immunosorbent assay allowed detection of type E toxin in inoculated seafoods at levels equivalent to 1–10 MLDs/ml (
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