Glycerol-Assisted Hydrophobic Interaction Chromatography Improving Refolding of Recombinant Human Granulocyte Colony-Stimulating Factor |
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Authors: | Fangwei Wang Yongdong Liu Guanghui Ma Zhiguo Su |
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Institution: | (1) Second Clinical College, Dalian Medical University, Dalian, 116027, China;(2) State Key Laboratory of Biochemical Engineering, Institute of Process Engineering, Chinese Academy of Sciences, Beijing, 100190, China; |
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Abstract: | Efficient refolding of recombinant proteins in the forms of inclusion bodies at higher concentration remains challenging.
Here, we report a strategy of a dual-gradient hydrophobic interaction chromatography (HIC) mode to refold recombinant human
granulocyte colony-stimulating factor from its inclusion bodies at high protein concentration. The strategy was taken to meet
the demand of dynamic refolding proceeding by gradually decrease the denaturant (guanidine-HCl) concentration and gradually
increase the hydrophilicity of media (column of Poros PE 20) with glycerol as additive to provide a mild refolding surroundings.
Compared with dilution method, this dual-gradient HIC process gave about 8.5-fold of increase in specific activity and 30%
increase in soluble protein recovery. Furthermore, much higher protein concentration could be obtained at the same time. |
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