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过程工程学报 2005
Study on Dilution and Ion Exchange Chromatographic Refolding of Recombinant Human Granulocyte Colony-stimulating Factor by Reversed Phase Chromatography
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Abstract:
A novel way to refold rhG-CSF by combining ion exchange chromatography (IEC) and L-arginine was developed: denatured rhG-CSF from inclusion body was attached to a DEAE column and eluted by a gradient from 6 mol/L urea to 0.5 mol/L L-arginine. Refolding of rhG-CSF was also done by dilution and by passing the rhG-CSF inclusion body through the DEAE column perfused with 0.5 mol/L arginine without adsorption. RPC analysis and bioactivity assay showed that rhG-CSF refolded by IEC had different pattern from those of the rhG-CSFs refolded by dilution and flowing-through, representing fast but asynchronous kinetics, due to the adsorption of the DEAE medium to the rhG-CSF and the gradient of denaturant. It was also found that subsequent dilution and incubation of the IEC-refolded rhG-CSF furthered its refolding and enhanced its bioactivity up to 10 folds.