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Abstract                 Volume:6  Issue-5  Year-2018          Original Research Articles


Online ISSN : 2347 - 3215
Issues : 12 per year
Publisher : Excellent Publishers
Email : editorijcret@gmail.com

Increased yield of high purity biologically active recombinant human granulocyte-macrophage colony-stimulating factor from Escherichia coli
Chitra Bajji1,2*, Ravi Kanth Reddy Kosana1, Radha Madhavi Kanumuri1 and Murali Krishna Reddy Tummuru1
1Virchow Research Centre, Hyderabad – 500 055, India
2Department of Biotechnology, Acharya  Nagarjuna University, Nagarjuna Nagar, Guntur, AP– 522 510, India

*Corresponding author
Abstract:

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.

Keywords: Escherichia coli, hGM-CSF, signal peptidase, pelB (pectate lyase B) signal sequence, reversed phase chromatography, Triton X 100
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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
doi: https://doi.org/10.20546/ijcrar.2018.605.009