Renaturation and Purification of rhGM-CSF with Ion-Exchange Chromatography

摘要

The renaturation and purification of recombinant human granulocyte macrophage colony stimulation factor(rhGM-CSF)expressed in Escherichia coli with strong anion-exchange chromatography(SAX)were studied.The effects of pH values,ratios of concentrations of GSH/ GSSG,and urea concentrations in the mobile phase on the renaturation and purification of rhGM-CSF with SAX were investigated,respectively.The results show that the above three factors have remarkable influences on the efficiency of renaturation and mass recovery of rhGM-CSF.The addition of GSH/GSSG in the mobile phase can improve the formation of correct disulfide bonds in rhGM-CSF so that its renaturation yield increases.In addition,to enhance the mass recovery of rhGM-CSF with SAX,the low concentration of urea was added in the mobile phase to prevent denatured protein aggregation.Under the optimal conditions,rhGM-CSF was renatured with simultaneous purification on SAX column within 30 min only by one step.After that its specific bioactivity,mass recovery,and purity reached 1.66 x 10~7 IU·mg,58.8%,and 96.2%,respectively.