Abstract
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°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 wild-type hIL-2. In summary, we generated a mutant hIL-2 that is functionally identical to wild-type hIL-2.
Keywords: Biological activity, chromatography, expression, interleukin-2, mutant, purification, soluble form
Protein & Peptide Letters
Title: Expression, Purification, and Characterization of a Functional Mutant Recombinant Human Interleukin-2
Volume: 17 Issue: 10
Author(s): Mingjun Liu, Bin Wang, Guirong Sun, Dongmeng Qian, Zhiyong Yan, Xuxia Song and Shouyi Ding
Affiliation:
Keywords: Biological activity, chromatography, expression, interleukin-2, mutant, purification, soluble form
Abstract: 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°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 wild-type hIL-2. In summary, we generated a mutant hIL-2 that is functionally identical to wild-type hIL-2.
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Cite this article as:
Liu Mingjun, Wang Bin, Sun Guirong, Qian Dongmeng, Yan Zhiyong, Song Xuxia and Ding Shouyi, Expression, Purification, and Characterization of a Functional Mutant Recombinant Human Interleukin-2, Protein & Peptide Letters 2010; 17 (10) . https://dx.doi.org/10.2174/092986610792231474
DOI https://dx.doi.org/10.2174/092986610792231474 |
Print ISSN 0929-8665 |
Publisher Name Bentham Science Publisher |
Online ISSN 1875-5305 |
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