[Expression of hepatitis C virus subunit fusion protein and analysis of its immunogenicity].
OBJECTIVE
Obtain the hepatitis C virus high purified subunit fusion protein and detect its immunogenicity.


METHODS
With the vector of pET-11d, fusion protein was expressed in Escherichia coli BL21 (DE3) after induced by IPTG. The protein was then purified by DEAE negative ion exchange chromatography and Ni2+ affinity chromatography. Western Blot analysis was used to detect the antigenicity of the fusion protein. At the same time, the sera were collected and prepared from the immunized experimental animals in order to investigate the immunogenicity of the protein by EIA.


RESULTS
High purified hepatitis C virus subunit fusion protein was obtained successfully. The EIA indicated that the fusion protein could elicit specific antibodies in the animals with very high titers.


CONCLUSIONS
The hepatitis C virus subunit fusion protein expressed in prokaryotic system was proved to have strong immunogenicity. It could provide some helpful and useful information to the hepatitis C virus prophylactic and therapeutic vaccine development.
State Key Laboratory for Viral Genetic Engineering, Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing, China.
Feng
F
Qiu
Zhi-Yuan
ZY
Jia
Min-Zhuo
MZ
Guo
Si-Yong
SY
Chen
Yao
Y
Yi
Li-Ping
LP
Shen
Tao
T
Yu
Yong-Liang
YL
Fei
Yu
Y
Guo
Sheng-Li
SL
Bi
D000818:Animals; D004797:Enzyme-Linked Immunosorbent Assay; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D016174:Hepacivirus / Q000235:genetics / Q000378:metabolism*; D007118:Immunoassay; D011817:Rabbits; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology* / Q000378:metabolism*; D014764:Viral Proteins / Q000235:genetics / Q000276:immunology* / Q000378:metabolism*
D011993:Recombinant Fusion Proteins; D014764:Viral Proteins