中文摘要：

采用体积色谱法对由E．coli表达的重组人粒细胞一巨噬细胞集落刺激因子（rhGM—CSF）复性与同时纯化工艺进行了研究。分别讨论了不同浓度的脲、流动相pH以及在流动相中添加不同比例的GSSG／GSH对rhGM—CSF复性与同时纯化的影响。结果表明：三者对其复性效率和质量回收率均有较大的影响，GSSG／GSH可有效提高rhGM—CSF二硫键的正确对接和复性效率，而低浓度的脲则可有效抑制蛋白沉淀，可大大提高质量回收率。在最优化条件下，SEC可在30min内使rhGM—CSF在分离纯化的同时进行复性，其比活为1．31×10^7IU／mg，质量回收率为42．3％，纯度可达到86．4％。

英文摘要：

The renaturation and simultaneous purification of recombinant human granulocyte macrophage colonly stimulation factor （rhGM-CSF） expressed in E.coli with size exclusion chromatography （SEC） was studied. The effects of the different pH values, the ratios of concentrations of GSH/GSSG and urea concentrations in the mobile phase on the renaturation and simultaneous purification of rhGM-CSF with SEC were investigated respectively. The results show the three factors have the remarkable effects on the renaturation efficiency and mass recovery ofrhGM-CSF. GSH/GSSG can improve to form the correct disulfide bonds as a result that the renaturation efficiency ofrhGM-CSF can be increased. In addition, in order to enhance the mass recovery ofrhGM-CSF with SEC, the low concentration urea can be used in the mobile phase to prevent denatured protein aggregation. Under the optimal condition, rhGM-CSF can be renatured with simultaneous purification with SEC in 30 rain only by one step, and its specific activity, mass recovery and purity can be obtained to be 1.31× 10^7IU/mg,42.3% and 86.4%, respectively.