Construction and expression of humanized anti-HBsAg scFv targeting interferon-alpha in escherichia coli / 中华肝脏病杂志

ABSTRACT

<p><b>OBJECTIVE</b>To develop a bacteria expression system to produce the fusion protein of humanized anti-HBsAg scFV and interferon-alpha.</p><p><b>METHODS</b>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.</p><p><b>RESULTS</b>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.</p><p><b>CONCLUSIONS</b>The construction and expression of the fusion gene of humanized anti-HBsAg scFv and interferon in E.coli are successful.</p>