Detection of stable epitopes on formaldehyde-detoxified Pasteurella multocida toxin by monoclonal antibodies.
Progressive atrophic rhinitis in pigs can be prevented by vaccination of pregnant sows with formaldehyde-detoxified preparations of either crude extract of toxigenic Pasteurella multocida or purified P. multocida toxin (PMT). The protective value of a vaccine is expected to be related to its content of detoxified immunogenic PMT, but previously described methods for detection of PMT cannot be used for the quantification of formaldehyde-detoxified PMT. In contrast to the epitopes on PMT recognized by previously produced anti-PMT monoclonal antibodies (mAbs), two of the epitopes recognized by mAbs developed in this study showed a significant stability after treatment with formaldehyde under conditions relevant for production of vaccines. When analysed in a sandwich ELISA based on these two mAbs, the titre of a preparation of PMT, detoxified by treatment with 1% (w/v) formaldehyde for 48 h at 20 degrees C, decreased by less than 30% when compared to the titre of native PMT. A close relationship between the amount of formaldehyde-treated PMT in a vaccine determined by the sandwich ELISA and its immunogenic properties in mice was observed.
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National Veterinary Laboratory, Animal Biotechnology Research Centre, Copenhagen, Denmark.
D000818:Animals; D000907:Antibodies, Bacterial / Q000276:immunology*; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000942:Antigens, Bacterial / Q000187:drug effects / Q000276:immunology*; D001426:Bacterial Proteins; D001427:Bacterial Toxins / Q000276:immunology* / Q000633:toxicity; D001428:Bacterial Vaccines; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000276:immunology*; D005557:Formaldehyde / Q000494:pharmacology; D051379:Mice; D008807:Mice, Inbred BALB C / Q000276:immunology; D016979:Pasteurella multocida / Q000276:immunology*; D011489:Protein Denaturation
D000907:Antibodies, Bacterial; D000911:Antibodies, Monoclonal; D000942:Antigens, Bacterial; D001426:Bacterial Proteins; D001427:Bacterial Toxins; D001428:Bacterial Vaccines; D000939:Epitopes; C062258:Pasteurella multocida toxin; D005557:Formaldehyde