%0 Journal Article
%T Expression of a Human Single-chain Fv Antibody Against HBsAg in Pichia pastoris<br>人源抗HBsAg单链抗体在巴氏毕赤酵母中的表达
%A XIONG Sheng
%A REN Xiang Rong
%A TANG Yong Hong
%A SU Kuan Yuan
%A YU Zhou Yao
%A LUO Yong
%A WANG Yi Fei
%A LI Jiu Xiang
%A <br>熊盛
%A 任向荣
%A 唐永红
%A 粟宽源
%A 余宙耀
%A 罗勇
%A 王一飞
%A 李久香
%J 生物工程学报
%D 2003
%I 
%X To express and secrete native HBscFv (anti-HBsAg single-chain Fv) in P. pastoris, HBscFv was amplified from plasmid pGEM-HBscFv, and then sub-cloned into expression vector pPICZalphaA. The resulting plasmid pPIC-HBscFv was linearized and transformed into P. pastoris GS115. The recombinant Pichia strains, identified by direct PCR and Zeocin-resistant screening of Pichia transformants, were cultured and induced with methanol. It was found that recombinant HBscFv, lead by alpha-factor, could be secreted into the culture supernatant to a level of 80mg/L. The bioactivity of Pichia produced HBscFv was confirmed by indirect ELISA, which also suggested that the bioactivity of HBscFv in the culture supernatant reached its peak in 72h and decreased in the late-stage of the induction. PAS staining suggests that HBscFv produced by yeast is poorly glycosylated or none-glycosylated protein.
%K hepatitis B
%K single
%K chain Fv
%K Pichia pastoris
%K expression<br>乙型肝炎，
%K 单链抗体，
%K 巴氏毕赤酵母，
%K 表达
%U http://www.alljournals.cn/get_abstract_url.aspx?pcid=90BA3D13E7F3BC869AC96FB3DA594E3FE34FBF7B8BC0E591&jid=A66E90C274451689E69F6F0291467824&aid=D11504079FF1446B&yid=D43C4A19B2EE3C0A&vid=2A8D03AD8076A2E3&iid=CA4FD0336C81A37A&sid=2A8D03AD8076A2E3&eid=EA389574707BDED3&journal_id=1000-3061&journal_name=生物工程学报&referenced_num=6&reference_num=16