Prokaryotic expression, purification of prM of JEV and preparation of monoclonal antibody / 中华实验和临床病毒学杂志

ABSTRACT

<p><b>OBJECTIVE</b>To prepare monoclonal antibody (mAb) against prM epitope.</p><p><b>METHODS</b>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.</p><p><b>RESULTS</b>mAb against prM epitope of JEV was prepared successfully.</p><p><b>CONCLUSION</b>The obtained prM specific mAb was valuable for the prevention and dignosis of Japanese encephalitis.</p>