Stable and efficient expression of hepatitis B virus S antigen and preS1 epitope fusion protein (S/preS1) in CHO cells.
- Author:
Zhenxi YANG
;
Shichong LI
;
Hong LIU
;
Miao ZHANG
;
Lingling YE
;
Yanzhuo WU
;
Mingbo XU
;
Zhaolie CHEN
- Publication Type:Journal Article
- MeSH:
Animals;
CHO Cells;
Cricetulus;
Epitopes;
biosynthesis;
genetics;
Hepatitis B Surface Antigens;
biosynthesis;
genetics;
immunology;
Hepatitis B Vaccines;
biosynthesis;
genetics;
Hepatitis B virus;
Protein Precursors;
biosynthesis;
genetics;
immunology;
Recombinant Fusion Proteins;
biosynthesis;
genetics;
Transfection
- From:
Chinese Journal of Biotechnology
2013;29(12):1808-1816
- CountryChina
- Language:Chinese
-
Abstract:
Hepatitis B surface antigen (HBsAg) carrying preS sequences could be an ideal candidate for a new hepatitis B virus (HBV) vaccine with higher efficacy. Here we report the success in achieving efficient and stable expression of hepatitis B virus S antigen and preS1 epitope fusion protein (S/preS1) in CHO cells. The HMRCHEF53u/Neo-S/preS1 expression vector carrying S/preS1 gene was constructed and transfected into CHO-S cells. A stable and high-expression CHO cell line, named 10G6, was selected by ELISA and limiting dilution analysis. Western blotting analysis showed S/preS1 expressed from 10G6 cells possessed both S and preS1 antigenicity. 10G6 cells displayed characters of favorable growth and stable S/preS1 expression in repeated batch cultures as evaluated by viable cell density, viability and S/preS1 concentration. And cultivation of 10G6 cells in fed-batch mode resulted in S/preS1 production at 17-20 mg/L with viable cell density at 7 x 10(6)-10 x 10(6) cells/mL.
