Zhonghua shi yan he lin chuang bing du xue za zhi = Zhonghua shiyan he linchuang bingduxue zazhi = Chinese journal of experimental and clinical virology · 2008
[Prokaryotic expression, purification of prM of JEV and preparation of monoclonal antibody].
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Ning Bei-fang, Zhu Huai-min, Zhou Xiao-jun, Cao Yi, Zhou Ai-guo
Аннотация
OBJECTIVE: To prepare monoclonal antibody (mAb) against prM epitope.
METHODS: The gene encoding prM was isolated using RT-PCR from brain of JEV infected mouse and cloned into prokaryotic expression vector pET-32a. Recombinant plasmid was transformed into E.coli BL21/DE3/LysS, then the transformed cells were expressed with the induction of IPTG. The expression and purification of the prM protein was analyzed by SDS-PAGE. The BALB/c mice were immunized with purified prM protein. Hybridoma cell lines secreting monoclonal antibodies against prM were established after cell fusion of mouse splenic cell and P3-X63-Ag8.653 cells. The specificity of mAb was identified by ELISA, Western Blot and Immunohistochemistry assay.
RESULTS: mAb against prM epitope of JEV was prepared successfully.
CONCLUSION: The obtained prM specific mAb was valuable for the prevention and dignosis of Japanese encephalitis.
METHODS: The gene encoding prM was isolated using RT-PCR from brain of JEV infected mouse and cloned into prokaryotic expression vector pET-32a. Recombinant plasmid was transformed into E.coli BL21/DE3/LysS, then the transformed cells were expressed with the induction of IPTG. The expression and purification of the prM protein was analyzed by SDS-PAGE. The BALB/c mice were immunized with purified prM protein. Hybridoma cell lines secreting monoclonal antibodies against prM were established after cell fusion of mouse splenic cell and P3-X63-Ag8.653 cells. The specificity of mAb was identified by ELISA, Western Blot and Immunohistochemistry assay.
RESULTS: mAb against prM epitope of JEV was prepared successfully.
CONCLUSION: The obtained prM specific mAb was valuable for the prevention and dignosis of Japanese encephalitis.