Expression, Purification, and Characterization of a Functional Mutant Recombinant Human Interleukin-2

被引：8

作者：

Liu, Mingjun ^{[1
]}

Wang, Bin ^{[2
]}

Sun, Guirong ^{[1
]}

Qian, Dongmeng ^{[2
]}

Yan, Zhiyong ^{[2
]}

Song, Xuxia ^{[2
]}

Ding, Shouyi ^{[2
]}

机构：

[1] Med Coll Qingdao Univ, Affiliated Hosp, Clin Immunol Lab, Qingdao 266003, Peoples R China

[2] Qingdao Univ Med Coll, Key Lab Med & Biotechnol, Qingdao 266021, Peoples R China

来源：

PROTEIN AND PEPTIDE LETTERS | 2010年 / 17卷 / 10期

关键词：

Biological activity; chromatography; expression; interleukin-2; mutant; purification; soluble form; ESCHERICHIA-COLI; CELLS; IDENTIFICATION; DOMAIN; IL-2; CDNA;

D O I：

10.2174/092986610792231474

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

In the current study, a mutant recombinant human interleukin-2 (MhIL-2) was generated using site-directed mutagenesis. The bacteria transformed with plasmid pET15b-MhIL-2 were cultured in LB medium containing 0.6mM IPTG for 8 hours at 27 degrees C. Approximately 90% of His-MhIL-2 was efficiently expressed in soluble form. Purification efficiency was optimized using a number of strategies, including nickel ion chelating chromatography, desalting chromatography, thrombin cleavage and Superdex 75 gel filtration chromatography. The final product had >95% purity. PBMCs, CD4+ and CD8+ T cell proliferation assays revealed that one such mutant has identical functional property to the wildtype hIL-2. In summary, we generated a mutant hIL-2 that is functionally identical to wild-type hIL-2.

引用

页码：1280 / 1284

页数：5

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