[Construction and expression of humanized anti-HBsAg scFv targeting interferon-alpha in escherichia coli].
PMID 11856497 · Zhonghua gan zang bing za zhi = Zhonghua ganzangbing zazhi = Chinese journal of hepatology · 2002
Authors: Xiaobing Xia; Jun Cheng; Jizhen Yang; Yanwei Zhong; Gang Wang; Hongqing Fang; Yan Liu; Ke Li; Jing Dong
Abstract
To develop a bacteria expression system to produce the fusion protein of humanized anti-HBsAg scFV and interferon-alpha. The expression vector was constructed after cleaving the plasmids harboring the humanized anti-HBsAg scFv and interferon alpha respectively and ligating to linearized pET22b subsequence. The expression of fusion protein in E.coli was analyzed by SDS-PAGE. The binding activity and antiviral activity of the fusion protein was characterized by competing inhibition test and cytopathic effect reduction. The plasmid harboring the in frame arranged fusion gene was constructed and identified. After induction for 12h, a new band close to 4.5 10(4) was observed using SDS-PAGE. Results of competing ELISA and cytopathic effect reduction showed the fusion protein retained its specific binding activity and antiviral activities. The construction and expression of the fusion gene of humanized anti-HBsAg scFv and interferon in E.coli are successful.
Electrophoresis, Polyacrylamide Gel; Escherichia coli; Gene Expression; Hepatitis B Antibodies; Hepatitis B Surface Antigens; Hepatitis B virus; Humans; Immunoglobulin Fragments; Interferon-alpha; Recombinant Fusion Proteins
Cite
Xiaobing Xia; Jun Cheng; Jizhen Yang; Yanwei Zhong; Gang Wang; Hongqing Fang; Yan Liu; Ke Li; Jing Dong. [Construction and expression of humanized anti-HBsAg scFv targeting interferon-alpha in escherichia coli]. Zhonghua gan zang bing za zhi = Zhonghua ganzangbing zazhi = Chinese journal of hepatology (2002). PMID 11856497.