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IJBB Vol.45 [2008] >
IJBB Vol.45(2) [April 2008] >


Title: Optimized conditions for high-level expression and purification of recombinant human interleukin-2 in E. coli
Authors: Sengupta, Paromita
Meena, Kalpana
Mukherjee, Rama
Jain, S K
Maithal, Kapil
Keywords: Interleukin-2
Inclusion bodies
Cytokines
Refolding
Issue Date: Apr-2008
Publisher: CSIR
Abstract: Interleukin-2 (IL-2), a potent cytokine has been used in anti-cancer therapy for over a decade now. IL-2, originally identified as a growth factor for T lymphocytes is a 15 kDa hydrophobic glycoprotein that induces the activation, clonal proliferation and differentiation of T and B-lymphocytes and enhances the cytotoxicity of monocytes and natural killer (NK) cells. Here, we report a simple method for the cloning, high-level expression and purification of IL-2 protein, which can be easily extended to other bioactive therapeutic proteins. The IL-2 gene was amplified from human spleen cDNA and cloned in a prokaryotic (E. coli) expression system. An optimal expression of the IL-2 protein was determined by varying the expression conditions like temperature, inducer concentration and duration of induction. The protein was expressed as inclusion bodies and a panel of reagents including detergents, urea and guanidine hydrochloride were used to solubilize it. After solubilization, the protein was renatured and subjected to a single step gel-filtration chromatography to yield immunobioactive IL-2 protein with >99% purity.
Page(s): 91-97
ISSN: 0301-1208
Source:IJBB Vol.45(2) [April 2008]

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