[Prokaryotic expression, purification of prM of JEV and preparation of monoclonal antibody].
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.


CONCLUSIONS
The obtained prM specific mAb was valuable for the prevention and dignosis of Japanese encephalitis.
Ning
Bei-fang
BF
Department of Etiology of Second Military Medical University, Shang hai 200433, China.
Zhu
Huai-min
HM
Zhou
Xiao-jun
XJ
Cao
Yi
Y
Zhou
Ai-guo
AG
D000818:Animals; D000911:Antibodies, Monoclonal / Q000032:analysis / Q000276:immunology* / Q000302:isolation & purification*; D000918:Antibody Specificity; D041702:BALB 3T3 Cells; D002460:Cell Line; D003001:Cloning, Molecular; D004591:Electrophoresis, Polyacrylamide Gel; D004664:Encephalitis Virus, Japanese / Q000235:genetics / Q000276:immunology; D000939:Epitopes / Q000276:immunology; D004926:Escherichia coli / Q000235:genetics; D051379:Mice; D010957:Plasmids / Q000235:genetics / Q000378:metabolism; D011387:Prokaryotic Cells / Q000378:metabolism*; D017422:Sequence Analysis, DNA; D014764:Viral Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification*
D000911:Antibodies, Monoclonal; D000939:Epitopes; D014764:Viral Proteins