A downstream purification procedure for recombinant human interferon-γ(rhIFN-γ) expressed in E. coli was described. An essentially two-step chromatographic purification procedure, i. e., size exclusion chromatograph...A downstream purification procedure for recombinant human interferon-γ(rhIFN-γ) expressed in E. coli was described. An essentially two-step chromatographic purification procedure, i. e., size exclusion chromatography(SEC) and high-performance hydrophobic in-teraction chromatography (HPHIC), was used for purification of homogeneity of rhIFN-γ from the inclusion body. The specific activity of purified rhIFN-Y was 1.0×108 IU/mg pro-tein. The product of purification rhIFN-γ was analyzed by an analytical high-per formance SEC and a significant sigle symmetrical peak had been found. The purity of purified rhIFN-Y was greater than 95% from analysis, determination by analytical HPSEC and SDS-PAGE with Coomssie Blue. The molecular weight was ca. 15 000. It was shown that this procedure was an effective method for purifying rhIFN-γ.展开更多
SP Sepharose Fast Flow离子交换层析柱作为蛋白质复性系统,采用尿素梯度法进行重组人γ-干扰素包涵体蛋白质的复性实验.结果表明,尿素梯度离子交换层析法能有效地复性重组人γ-干扰素包涵体,复性后的重组人γ-干扰素的纯度达95%,蛋白...SP Sepharose Fast Flow离子交换层析柱作为蛋白质复性系统,采用尿素梯度法进行重组人γ-干扰素包涵体蛋白质的复性实验.结果表明,尿素梯度离子交换层析法能有效地复性重组人γ-干扰素包涵体,复性后的重组人γ-干扰素的纯度达95%,蛋白收率达54%,比活为7.5×105IU?mg-1.以Superdex 75凝胶作为体积排阻层析介质,对离子交换层析复性的样品进行分析,表明离子交换层析复性的样品中没有重组人γ-干扰素聚集体存在.荧光光谱分析表明重折叠的重组人γ-干扰素的构象接近于其天然二聚体构象.展开更多
文摘A downstream purification procedure for recombinant human interferon-γ(rhIFN-γ) expressed in E. coli was described. An essentially two-step chromatographic purification procedure, i. e., size exclusion chromatography(SEC) and high-performance hydrophobic in-teraction chromatography (HPHIC), was used for purification of homogeneity of rhIFN-γ from the inclusion body. The specific activity of purified rhIFN-Y was 1.0×108 IU/mg pro-tein. The product of purification rhIFN-γ was analyzed by an analytical high-per formance SEC and a significant sigle symmetrical peak had been found. The purity of purified rhIFN-Y was greater than 95% from analysis, determination by analytical HPSEC and SDS-PAGE with Coomssie Blue. The molecular weight was ca. 15 000. It was shown that this procedure was an effective method for purifying rhIFN-γ.
文摘SP Sepharose Fast Flow离子交换层析柱作为蛋白质复性系统,采用尿素梯度法进行重组人γ-干扰素包涵体蛋白质的复性实验.结果表明,尿素梯度离子交换层析法能有效地复性重组人γ-干扰素包涵体,复性后的重组人γ-干扰素的纯度达95%,蛋白收率达54%,比活为7.5×105IU?mg-1.以Superdex 75凝胶作为体积排阻层析介质,对离子交换层析复性的样品进行分析,表明离子交换层析复性的样品中没有重组人γ-干扰素聚集体存在.荧光光谱分析表明重折叠的重组人γ-干扰素的构象接近于其天然二聚体构象.
