Monoclonal antibody-based immunoassay for type A Clostridium botulinum toxin is comparable to the mouse bioassay.
A monoclonal antibody (BA11) has been produced against Clostridium botulinum type A neurotoxin by the fusion of myeloma cells (P3 NS1/1-Ag4-1) with spleen cells from BALB/c mice immunized with botulinum type A neurotoxoid. The antibody bound specifically to botulinum type A neurotoxin, showing no cross-reactivity with types B and E botulinum toxins or with any of several other bacterial toxins tested. The monoclonal antibody did not bind to botulinum type A neurotoxin which had been denatured with sodium dodecyl sulfate and bound only weakly to each of the separated heavy and light subunits of the neurotoxin, suggesting a conformational requirement for the antigenic determinant of the antibody. A sensitive immunoassay for C. botulinum type A toxin with monoclonal antibody BA11 in conjunction with an enzyme amplication system has been developed which allows detection of 5 to 10 mouse 50% lethal doses ml-1 of purified neurotoxin. The assay was equally sensitive when applied to the detection of crude toxin in food stuffs; the average value for the minimum level of detectable toxin in extracts of tinned salmon or corned beef was 9 +/- 3.1 mouse 50% lethal doses ml-1.
D000818:Animals; D000911:Antibodies, Monoclonal; D000918:Antibody Specificity; D001681:Biological Assay; D001905:Botulinum Toxins / Q000032:analysis* / Q000276:immunology / Q000302:isolation & purification; D003429:Cross Reactions; D004168:Diphtheria Toxoid; D004768:Enterotoxins; D004797:Enzyme-Linked Immunosorbent Assay; D005516:Food Microbiology; D006825:Hybridomas; D005779:Immunodiffusion; D007124:Immunoenzyme Techniques; D007928:Lethal Dose 50; D051379:Mice; D008807:Mice, Inbred BALB C; D009500:Neutralization Tests
D000911:Antibodies, Monoclonal; D004168:Diphtheria Toxoid; D004768:Enterotoxins; D001905:Botulinum Toxins