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中国生物工程杂志

China Biotechnology
China Biotechnology
China Biotechnology  2015, Vol. 35 Issue (4): 42-47    DOI: 10.13523/j.cb.20150406
    
Expression and Purification of Biological-active Recombinant HBV Precore Protein-Mouse IgG Fc Based on Baculovirus Expression Vector System
WANG Xiao-yan, CHEN Na-zi, AI Jun, ZHAO Yang, WU Mei-yu, HUANG Jin-zhi, JIANG Chao, LI Xiao-kun
Wenzhou Medical College, Key Laboratory of Biotechnology and Pharmaceutical Engineering of Zhejiang Province, Wenzhou 325035, China
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Abstract  

Objective: The fusion protein HBV pre-C protein - mouse IgG Fc (HBV pre-c-Fc) protein was expressed based on the baculovirus expression vector system, and identify its immunogenicity. Methods: target gene HBV pre-c-Fc connected to pFastBac1 vector, pFastBac1-HBV pre-c-Fc plasmid was transformed DH10Bac competent, transposase Bacmid through Th7 transposon into the target gene, got Bacmid-HBV pre -c-Fc shuttle vector, Cellfectin ⅡReagent transfected Sf9 insect cells to obtain P1 generation virus, transfected Sf9 repeated to obtain high titer virus. Cell supernatants were collected after ultrafiltration to obtain the target protein HBV pre-c-Fc purified by Protein G affinity chromatography. The purified protein thigh intramuscularly immunized BALB/c mice and detected the concentration of serum hepatitis B virus core protein antibody. Results: HBV pre-c-Fc successfully expressed in insect cells, the purity of purified protein was over 90%, the production of protein is about 3.03mg/L, purified protein can effectively immunize the BALB/c mice to produce specific antibodies. Conclusion: The expression of immunogenic HBV pre-c-Fc protein based on the baculovirus expression vector system can be the production of hepatitis B therapeutic vaccine foundation.

Key words： Hepatitis B virus core protein      Baculovirus expression vector system      Sf9 cells     
Received: 22 January 2015      Published: 25 April 2015
ZTFLH:  Q789  
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Cite this article:

WANG Xiao-yan, CHEN Na-zi, AI Jun, ZHAO Yang, WU Mei-yu, HUANG Jin-zhi, JIANG Chao, LI Xiao-kun. Expression and Purification of Biological-active Recombinant HBV Precore Protein-Mouse IgG Fc Based on Baculovirus Expression Vector System. China Biotechnology, 2015, 35(4): 42-47.

URL:

https://manu60.magtech.com.cn/biotech/10.13523/j.cb.20150406     OR     https://manu60.magtech.com.cn/biotech/Y2015/V35/I4/42


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