Volume 25, Issue 125 (6-2015)                   J Mazandaran Univ Med Sci 2015, 25(125): 10-20 | Back to browse issues page

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Pouranvari S, Ebrahimi F, Javadi G, Maddah B. Production of Recombinant Human Epidermal Growth Factor and Assessment of its Activity in Cell Viability. J Mazandaran Univ Med Sci 2015; 25 (125) :10-20
URL: http://jmums.mazums.ac.ir/article-1-5716-en.html
Abstract:   (8235 Views)
Background and purpose: Human epidermal growth factor (hEGF) is a polypeptide of 53 amino acids with various medical application such as wound healing. The purpose of this study was cloning, expression, and purification of recombinant human EGF (rhEGF) and assessment of its mitogenic effect on NIH 3T3 cells. Materials and methods: Subcloninig of hEGF was performed in to pET24a (+). Protein expression was done under standard conditions. According to the protein expression as inclusion body, mild solubilization using alkaline pH buffer was utilized for protein solubilization. Ultimately, after refolding of solubilized proteins, MTT assay was performed to assess the mitogenic effect of rhEGF in NIH 3T3 cells treated with various concentrations of rhEGF. Results: Mild solubilization of inclusion bodies with alkaline buffer and subsequent refolding had a very high efficiency. MTT assay showed that cells treated with our rhEGF exhibited significantly higher proliferation compared to control after 72 h (P < 0.0001). Conclusion: It seems cytoplasmic expression system is an efficient system for production of recombinant hEGF. The method presented in this study is a simple, accessible, affordable and of high efficiency for solubilization of inclusion bodies which is also helpful in achieving bioactive form of human epidermal growth factor.
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Type of Study: Research(Original) | Subject: genetic

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