[Prokaryotic expression and polyclonal antibody preparation of truncated duck hepatitis B virus core protein].
OBJECTIVE
To construct a prokaryotic plasmid expressing truncated duck hepatitis B virus core protein (DHBc(1-214)), purify the recombinant protein, and to develop polyclonal antibodies against DHBc.


METHODS
DHBc(1-214) was cloned into vector pRSET-B, then expressed in E.coli Rosetta(DE3) pLacI induced by IPTG. The recombinant protein was purified using Ni-NTA spin column. Polyclonal antibody was developed by immunizing BALB/c mice with the purified recombinant protein, and their sensitivity and specificity were tested using enzyme-linked immunosorbent assay, immunohistochemical staining and Western blot analysis.


RESULTS
Recombinant plasmid expressing truncated DHBc(1-214) was successfully constructed. A protein of 28,000 was expressed and purified. Polyclonal serum antibody with a high specificity was obtained by immunizing BALB/c mice with the purified recombinant protein.


CONCLUSIONS
The truncated recombinant DHBc(1-214) developed in this study is purified and shown strong antigenecity. The polyclonal antibody against DHBc protein is generated by regular immunization method, demonstrating both high sensitivity and specificity. The protein and the antibody can be used for further clinical examination and research of DHBV.
Wang
Chao
C
Division of Clinical Immunology, Tongji Hospital of Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China. celful60@yahoo.com.cn
Tian
Yong-jun
YJ
Zhang
E-juan
EJ
Wu
Hong-kun
HK
Liu
Jia
J
Wang
Bao-ju
BJ
Zhao
Xi-ping
XP
Yang
Dong-liang
DL
D000818:Animals; D000917:Antibody Formation / Q000276:immunology*; D015153:Blotting, Western; D004372:Ducks / Q000821:virology; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D004926:Escherichia coli / Q000378:metabolism; D005822:Genetic Vectors; D006510:Hepatitis B Antibodies / Q000276:immunology* / Q000302:isolation & purification; D015611:Hepatitis B Virus, Duck / Q000276:immunology*; D051379:Mice; D008807:Mice, Inbred BALB C; D012680:Sensitivity and Specificity; D014758:Viral Core Proteins / Q000276:immunology* / Q000378:metabolism; D014764:Viral Proteins / Q000276:immunology
D006510:Hepatitis B Antibodies; D014758:Viral Core Proteins; D014764:Viral Proteins