白细胞介素—2—绿脓杆菌外毒素融合蛋白的分高纯化及其复性研究

表达的白细胞介素－２－绿脓杆菌外毒素（ＩＬ－２－ＰＥ）融合蛋白以包含体形式存在于宿主菌中，为分离纯化表达产物提供了方便，但因需进行复性，也增加了后处理的难度．我们采用４ｍｏｌ／Ｌ尿素、０．５％ＴｒｉｔｏｎＸ－１００的１×ＰＢＳ洗涤包含体两遍，再经ＳｅｐｈａｃｒｙｌＳ－３００分子筛及ＤＥＡＥ－ＳｅｐｈａｒｏｓｅＦＦ阴离子交换柱层析后，获得的融合蛋白纯度可达９０％～９５％。此外，我们从ＧＳＳＧ浓度、Ｌ－精氨酸浓度、复性蛋白质的起始浓度、复性液的ｐＨ值、复性温度及复性时间等参数入手，系统地研究了融合蛋白的复性条件，探索到了ＩＬ－２－ＭＥ４０和ＩＬ－２－ＰＥ６６４Ｇｌｕ融合蛋白复性的最适条件。

The Purification and Renaturation of the Interleukin-2-Pseudomonas Exotoxin Fusion Protein (IL-2-PE)

The IL-2-PE fusion protein expressed in E. coli forms the inclusion body which can be isolated easily. The purity of the fusion protein was 90%-95% after washing the inclusion body with PBS containing 4M urea and 0.5% Triton X-100followed by chromatography on Sephacryl S-300 and DEAE-Sepharose FF. Then,parameters relative to the renaturation, including the concentrations of GSSG and L-Arg, the initiation concentrations of the protein, pH, temperature andreaction time etc, were systematically analysed. An optimun condition for IL-2-PE renaturation was proposed.