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Chinese Journal of Biotechnology ; (12): 1808-1816, 2013.
Article in Chinese | WPRIM | ID: wpr-242451

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.


Subject(s)
Animals , CHO Cells , Cricetulus , Epitopes , Genetics , Hepatitis B Surface Antigens , Genetics , Allergy and Immunology , Hepatitis B Vaccines , Genetics , Hepatitis B virus , Protein Precursors , Genetics , Allergy and Immunology , Recombinant Fusion Proteins , Genetics , Transfection
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