Extraction and Detect on Immunization of Outer Membrane Protein of Bordetella bronchiseptica

Abstract:

In order to evaluate the immunogenicity and their protection of outer membrane protein of Bordetella bronchiseptica and effective antibody (OMP68, we established a mice model. The research improved on the method of Wooldridge in order to extract OMP of Bordetella bronchiseptica (P13, P11), and the differences of OMP were analyzed by SDS-PAGE. The P13-OMP was analyzed with Western blotting and OMP68 was gained by the way of electrophoretic elution. Then we maked the effective antibodies of P13-OMP. Then P13-OMP, OMP68 and whole-cell bacterin (WCB)immunizing antigen was prepared 70 mice were equally assigned to 7 groups at random. They were immunized by P13-OMP, OMP68 and P13-WCB of different dose respectively. The indirect ELISA was used to detect antibody responses. 80 mice were equally assigned to 8 groups at random. Group a and b were immunized by P13-OMP (25 μg) respectively. Groups c and d were immunized by OMP68(25 μg); Groups e and f were immunized by WCB(P13). Ten days after the immunizations, group a, c, e and f were challenged with P13. The other groups were challenged with P11. Then we registered the rate of protective potency and evaluated protection given by P13-OMP, OMP68 and P13-WCB. Results showed that at 42th day, the titer of serum reach the top: P13-OMP(215.3)> OMP68(214.8). Then, the titer of serum started the slow-moving descent At 70th day, the titer of serum reach the OMP68 (210.2)> P13-OMP (29.1). P13-OMP and OMP68 group challenged with P13 and P11 can be efectivly protected; P13-WCB group challenged with P13 and P11 can not be efectivly protected; the control group were died out. The P13-OMP and OMP68 of Bordetella bronchiseptica has good immunogenicity and protection, so the results of this study lay good theoretical foundation for OMP subunit vaccine.