[Construction and expression of humanized anti-HBsAg scFv targeting interferon-alpha in escherichia coli].
作者信息
Xia Xiaobing, Cheng Jun, Yang Jizhen, Zhong Yanwei, Wang Gang, Fang Hongqing, Liu Yan, Li Ke, Dong Jing
机构信息
Gene Therapy Research Centre, 302nd Hospital of PLA, Beijing 100039, China.
出版信息
Zhonghua Gan Zang Bing Za Zhi. 2002 Feb;10(1):28-30.
OBJECTIVE
To develop a bacteria expression system to produce the fusion protein of humanized anti-HBsAg scFV and interferon-alpha.
METHODS
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.
RESULTS
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.
CONCLUSIONS
The construction and expression of the fusion gene of humanized anti-HBsAg scFv and interferon in E.coli are successful.
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