Urea gradient SEC in renaturation with simultaneous purification of rhSCF

    WANG Liª²Li£¬ GENG Xinª²Du

    Institute of Modern Separation Science, Shaanxi Provincial Key Lab of Modern Separation Science, Northwest University, Xi¬ðan 710069, China

    ¡¾Abstract¡¿ AIM: To study the application of urea gradient size exclusion chromatography (SEC) in the renaturation with simultaneous purification of recombinant human stem cell factor (rhSCF) expressed in E. coli. METHODS: SEC was performed by the property of different migration speed of protein and small molecules in gel. The urea was eluted by linear elution, while the protein was eluted by urea gradient, thus resulting in the renaturation and purification of the denatured protein. RESULTS: The human stem cell factor was renatured and purified successfully using SEC. The efficiency was improved significantly. It was also found that the elution time, speed, as well as protein concentration had evident influence on the efficiency of protein renaturation. CONCLUSION: The human stem cell factor expressed in E. coli can be renatured and purified using SEC. With gradient elution, the efficiency of renaturation is improved and the recovery of the recombinant protein increases from 35.2% to 51.4%. ......