Refolding and purification of interferon-gamma in industry by hydrophobic interaction chromatography

Journal of Biotechnology
Xindu GengDan Wu

Abstract

A new technology for renaturation with simultaneous purification of the recombinant human interferon-gamma (rhIFN-gamma) in downstream of biotechnology is presented. The strategies to develop the new technology in industry scale were suggested. Based on chemical equilibrium and molecular interactions, the principle of rhIFN-gamma refolding by HPHIC was described. The kind of stationary and mobile phases were evaluated and found the former to contribute to the rhIFN-gamma refolding more than the latter. The extract containing the rhIFN-gamma in gram scale in 7.0 mol L(-1) guanidine hydrochloride solution of 700 mL was directly pumped into a unit of simultaneous renaturation and purification of proteins (USRPP, 10 x 300 mm i.d.) packed by small particle packings of hydrophobic interaction chromatography and a satisfactory recovery of bioactivity and mass of the rhIFN-gamma was obtained. With flow rate 100 mL min(-1) and a gradient elution by only one step in 4h, the purity and specific bioactivity approach to 95% and 8.7 x 10(7) IU(-1) mg, respectively. To evaluate the goodness of the presented new technology in this study, a usual method with the renaturation by dilution method firstly and then purification with a series of LC i...Continue Reading

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Citations

Jul 2, 2010·Journal of Industrial Microbiology & Biotechnology·Surianarayanan BhuvaneshSubramanian Ramalingam
Apr 7, 2010·Protein Expression and Purification·Stefan PetrovIvan Ivanov
Mar 28, 2008·Protein Expression and Purification·Zhan-Xia ShiXin-Du Geng
Oct 4, 2012·Biomedical Chromatography : BMC·Ke CongyuGeng Xindu
Oct 20, 2006·Protein Expression and Purification·Praveen K ReddyRaju C Reddy
Dec 3, 2014·Protein Expression and Purification·Chunfang LingAilin Tao
Dec 3, 2014·Journal of Chromatography. a·Adrian M GospodarekErik J Fernandez
Feb 26, 2008·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·Xindu Geng, Lili Wang
Nov 1, 2012·Journal of Separation Science·Fei WangXindu Geng
Aug 8, 2006·Biotechnology Journal·Klaus Graumann, Andreas Premstaller
Feb 10, 2018·Preparative Biochemistry & Biotechnology·Rajat Pandey, Venkata Dasu Veeranki
Feb 10, 2018·Preparative Biochemistry & Biotechnology·Rajat PandeyVenkata Dasu Veeranki

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