%0 Journal Article
%T Chromatography-assisted refolding of a fusion protein containing multiple disulfide bonds<br>层析辅助体外复性含有多对二硫键的融合蛋白质
%A Weiquan Xie
%A Guifeng Zhang
%A Ling Gao
%A Yongdong Liu
%A Rong Yu
%A Zhiguo Su
%A <br>谢伟全
%A 张贵锋
%A 高玲
%A 刘永东
%A 余蓉
%A 苏志国
%J 生物工程学报
%D 2010
%I 
%X To establish a refolding process for the protein fused with 12-peptide of hirudin and reteplase (HV12p-rPA), we developed an anion-exchange chromatography assisted method to form correct disulfide bonds. After evaluating various parameters by orthogonal experiments with Q Sepharose XL as refolding medium, we found that urea gradient, sample loading size and L-Arg concentration were three major factors to affect the refolding outcomes, and urea gradient was critical to the recovery yield. Meanwhile, enzymatic activity of the refolded protein was decreased by the increase of sample loading size, and the optimal concentration of L-Arg in the eluting buffer was 1 mol/L. Thus, a dual-gradient of urea and pH on the anion-exchange chromatography resulted in remarkable increase of specific fibrinolytic and anti-coagulative activities of the refolded protein. Compared with the dilution method for refolding HV12p-rPA, the present approach was more effective and advantageous.
%K fusion protein
%K anion exchange chromatography
%K refolding
%K disulfide bond<br>融合蛋白质，阴离子交换层析，复性，二硫键
%U http://www.alljournals.cn/get_abstract_url.aspx?pcid=90BA3D13E7F3BC869AC96FB3DA594E3FE34FBF7B8BC0E591&jid=A66E90C274451689E69F6F0291467824&aid=14F727E8B18FE2CE981748DE1FED9ECE&yid=140ECF96957D60B2&vid=96C778EE049EE47D&iid=5D311CA918CA9A03&sid=CD1AB74EAF0667FF&eid=4E85BC78FC25985C&journal_id=1000-3061&journal_name=生物工程学报&referenced_num=0&reference_num=0