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《China Biotechnology》 2009-01
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Efficient Expression and Purification Technique of a Mutant of Human Interleukin 2

LIU Ming-jun1 WANG Bin2 SONG Xu-xia2 QIAN Dong-meng2(1 Department of Clinical Immunologic Laboratory,Affiliated Hospital of Qingdao University Medical College,Qingdao 266003,China)(2 Key Laboratory of Medicine and Biotechnology,Qingdao University Medical College,Qingdao 266021,China)  
Mutants of recombinant hIL-2(rhIL-2),generated by using site-directed mutagenesis strategy,can increase anti-tumor activity and decrease toxicity.A mutant of hIL-2(MhIL-2) in which Asn88 was substituted by Arg88 was generated by using the strategy.To reduce the undesirable formation of inclusion body and maximize the yield of soluble MhIL-2 fusion protein,a protocol for the expression of soluble MhIL-2 fusion protein was adapted.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.
【Fund】: 山东省自然科学基金资助项目(E99C02)
【CateGory Index】: Q786
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【Citations】
Chinese Journal Full-text Database 2 Hits
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【Co-citations】
Chinese Journal Full-text Database 10 Hits
1 REN Jin_rong, ZHENG Zhen_hai, WANG Zeng_lin, LIU Lu_ning, YU Feng_ling(1. Hebei Cancer Institute The Fourth Hospital of Hebei Medical University, Shijiazhuang 050011 , China; 2. Hebei lingshou Zheng's Bialogcal Cancer Institute, Linshou County 050500 , China);ESTABLISHMENT OF IMMUNE TOLERANCE MODEL IN BALB/C MOUSE AND PRELIMINARY STUDY ON THE TRANSPLANTING HUMAN LUNG CANCER[J];Carcinogenesis,Teratogenesis and Mutagenesis;2003-01
2 DAI Zhong-hua,SHI Nian ,LIU Gong-ping,WEI Yan-hong LI Huang-yuan,CHEN Dan,WANG Bin (Department of Health Toxicology,Tongji Medical College,Huazhong University of Science and Technology,Wuhan Hubei430030,China);The Effects of Deltamethrin on Fas,FasL,TNFR1 Protein Expre s sion and Apoptosis in PC12 Cells[J];Carcinogenesis,Teratogenesis and Mutagenesis;2005-02
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1 ZHAO Dan-ping1,TAO Bin1,CHEN Li-gong,LI Yan-qin,SONG Qin-ye2,LI Tan-qing,YANG Fang,LI Hong-yuan,YUAN Hong-xin(Experimental Station of Animal Pathogen Biology,Ministry of Agriculture /Agricultural University of Hebei,Baoding Hebei 071000,China);INFLUENCE OF PORCINE CIRCOVIRUS TYPE 2 INFECTION ON IMMUNE RESPONSE OF PIGS TO CLASSICAL SWINE FEVER VACCINE[A];[C];2012
2 XU Li, WANG Ming - yan, XU Dong - qing, ZHOU Chun - xiang (Nanjing University of Traditional Chinese Medicine,Nanjing ,Jiangsu Province ,China,210029);Experimental Study on Effect of Modified Sanwubai Powder in Affecting MDR Gene Expression of Tumor[A];[C];2005
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