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[Construction and expression of humanized anti-HBsAg scFv targeting interferon-alpha in escherichia coli]
Xiaobing Xia1, Jun Cheng, Jizhen Yang
1Gene Therapy Research Centre, 302nd Hospital of PLA, Beijing 100039, China.
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.