%0 Journal Article
%T Expression Purification and Verification of HBscFv-IFNγ in Pichia pastoris x33<br>HBscFv–IFNγ在毕赤酵母X33中的表达、纯化及鉴定
%A Shishui Zhou
%A Xiaoning Wang
%A <br>周世水
%A 王小宁
%J 生物工程学报
%D 2008
%I 
%X In order to effectively cure hepatitis B virus (HBV), we studied on fusion protein HBscFv-IFNgamma, which was connected with single-chain Fv against HBV surface antigen(HBscFv) and gamma-interferon(IFNgamma) of being used in clinic against HBV. Adopting overlap PCR, the hbscfv and the ifngamma were connected into hbscfv-ifngamma. Then the pPICZalphaA/(hbscfv-ifngamma)(1,2,4) of multi-copy recombinant plasmid were constructed and transformed into Pichia pastoris x33. The engineering strain x4 was screened from transformed x33 and could secretively express HBscFv-IFNgamma. The preliminary verification indicates that HBscFv-IFNgamma has the bioactivity of HBscFv and IFNgamma by SDS-PAGE, Western blotting and ELISA. The supernatant of culturing X4 was purified by 14F7 affinity chromatography to HBscFv-IFNgamma with purity of 95%-98%. The HBscFv-IFNgamma is able to bind 27.9% HBV surface antigen (HBsAg) in the serum of HBV transgenic mice, which shows the antibody of HBscFv-IFNgamma has bioactivity in vivo. Therefore HBscFv-IFNgamma can shed light on the development of a new promising HBV-targeted drug.
%K 重叠PCR
%K HBscFv-IFNγ
%K 巴斯德毕赤酵母X33
%K 14F7亲合层析
%U http://www.alljournals.cn/get_abstract_url.aspx?pcid=90BA3D13E7F3BC869AC96FB3DA594E3FE34FBF7B8BC0E591&jid=A66E90C274451689E69F6F0291467824&aid=A728D0E9E82D76CD82D27AB5713B521B&yid=67289AFF6305E306&vid=B91E8C6D6FE990DB&iid=38B194292C032A66&sid=F27A401E323B6FAD&eid=9BA67A0B76A3DBA8&journal_id=1000-3061&journal_name=生物工程学报&referenced_num=0&reference_num=15