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Indian Journal of Biochemistry and Biophysics (IJBB) >
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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