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Cloning and Expression of Human Interleukin 2 (IL-2) in E. coli and its Antitumor Activity

Cloning and Expression of Human Interleukin 2 (IL-2) in E. coli and its Antitumor Activity

Qurrat ul Ain Shafique1, Sana Batool2, Hanfa Ashfaq2, Asima Tayyab2, Roquyya Gul3 and Mahjabeen Saleem1*

1School of Biochemistry and Biotechnology, University of the Punjab, Quaid-Azam Campus, Lahore, 54590, Pakistan 
2School of Biological Sciences, University of the Punjab, Quaid-Azam Campus, Lahore, 54590, Pakistan
3Faculty of Life Sciences, Gulab Devi Educational Complex, Lahore, Ferozpur Road, Lahore, Pakistan
 
* Corresponding author: mahjabeen.ibb@pu.edu.pk

ABSTRACT

Interleukin 2, also known as T cell growth factor, brought a great revolution in the field of immunology. IL-2 is a pleiotropic cytokine and one of the cytokines that is approved by FDA for cancer immunotherapy particularly kidney and skin cancer. Interleukin-2 (IL-2) is a 4-helix-bundle type I cytokine possessing a cytokine receptor chain essential for the immune response. The current study presents a vector/host combination (pET28a(+)/IL-2) and optimization of expression in LB,TB and M9 media with varying concentrations of inducers (IPTG and lactose) and post induction time. Maximum expression of IL-2 was achieved in LB with 0.5 mM IPTG for 6 hours post induction time. A denaturing purification scheme (Immobilized metal affinity chromatography) was employed for the purification of IL-2. The refolding of the purified IL-2 was achieved by stepwise dialysis method using urea gradient (8M-0M). Human IL-2 was recovered from 1 litre culture to a purity level of ~95%.  In vitro cytotoxic potential of IL-2 on HepG-2 and MCF-7 cell lines revealed that it possess sufficient cytotoxic potential and can inhibit the growth of these cell lines directly. Thus, refolded IL-2 had activity identical to that of authentic IL-2 and enhanced the anti-tumor activity of HepG-2 as compared to MCF-7 cells. These conclusions suggest the potential use of the refolded cytokine as immunotherapeutic agent for treatment of hepatocellular carcinoma.

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Pakistan Journal of Zoology

June

Pakistan J. Zool., Vol. 56, Iss. 3, pp. 1001-1500

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