%0 Journal Article
%T Efficient Expression and Purification Technique of a Mutant of Human Interleukin 2<br>一种白细胞介素2基因变构体的高效表达与纯化工艺
%A Ming-jun Liu
%A <br>刘明军
%A 王斌
%A 宋旭霞
%A 钱冬萌
%J 中国生物工程杂志
%D 2009
%I 
%X Mutants of recombinant hIL-2 (rhIL-2), generated by using site-directed mutagenesis strategy, can increase anti-tumor activity and decrease toxicity. We have used site-directed mutagenesis of hIL-2 to generate a mutant of hIL-2(MhIL-2) in which Asn88 was substituted by Arg88. To reduce the undesirable formation of inclusion body and maximize the yield of soluble MhIL-2 fusion protein, we adapted a protocol for the expression of soluble MhIL-2 fusion protein. Our results have indicated that soluble form of the MhIL-2 fusion protein is expressed from E. coli.. Moreover, it also has facilitated purification of the MhIL-2. SDS-PAGE analysis revealed that the MhIL-2 protein was efficiently purified to 95% purity by the combination of nickel ion Chelating column chromatography, Desalting column chromatography, thrombin cleavage and Superdex 75 gel filtration column chromatography. Proliferation assay of T lymphocyte of purified MhIL-2 showed that the biological activity of MhIL-2 was higher than that of standard hIL-2 in vitro. This work not only describes an efficient preparation strategy of MhIL-2, but also introduces a highly active MhIL-2 that may have important clinical applicability.
%K 白细胞介素-2
%K 基因变构体
%K 表达
%K 纯化
%U http://www.alljournals.cn/get_abstract_url.aspx?pcid=90BA3D13E7F3BC869AC96FB3DA594E3FE34FBF7B8BC0E591&jid=951380788B355DEA7D75520FA3B199C9&aid=405DAAFE969FE7FB2C00754607E465FD&yid=DE12191FBD62783C&vid=771469D9D58C34FF&iid=CA4FD0336C81A37A&sid=014B591DF029732F&eid=BFE7933E5EEA150D&journal_id=1671-8135&journal_name=中国生物工程杂志&referenced_num=0&reference_num=9