Abstract Volume:6 Issue-5 Year-2018 Original Research Articles
|Online ISSN : 2347 - 3215
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2Department of Biotechnology, Acharya Nagarjuna University, Nagarjuna Nagar, Guntur, AP– 522 510, India
Human Granulocyte-Macrophage Colony-Stimulating Factor (hGM-CSF) is being produced by recombinant DNA technology and has been used as a therapeutic protein for different conditions. Different strategies have been reported to express and purify therapeutic grade hGM-CSF in different host systems including Escherichia coli. However, the reported yields are comparatively low and involve multiple steps of purification. In this present paper, we report a novel method of periplasmic secretion system for high expression, with relatively simple purification method using reversed phase chromatography and refolding technique. The final purity of the rhGM-CSF was more than 98 % with increased yield of ~300 mg per 35 g of wet weight cell pellet per litre of culture, which is relatively higher than the previous reports. Moreover, the final purified rhGM-CS proteinF has exhibited specific biological activity (1.1×107 IU/mg) in TF-1 cell proliferation assay similar to standard protein.
How to cite this article:Chitra Bajji, Ravi Kanth Reddy Kosana, Radha Madhavi Kanumuri and Murali Krishna Reddy Tummuru. 2018. Increased yield of high purity biologically active recombinant human granulocyte-macrophage colony-stimulating factor from Escherichia coli.Int.J.Curr.Res.Aca.Rev. 6(5): 50-65