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重组人尿激酶原在CHO细胞中的高效表达及其纯化

Efficient expression of recombinant human pro-urokinase in CHO cells and its purification
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摘要 用鸡β-globin的MAR序列和人看家基因延伸因子1α(hEF-1α)的调控序列以及旱獭RNA稳定与输出序列,构建了重组人尿激酶原(recombinant human pro-urokinase,rhPro-UK)的高效表达载体,在CHO细胞中获得了rhPro-UK的高效稳定表达,rhPro-UK的表达水平达到1 299 IU(以百万细胞1 d的表达量计)。采用阳离子交换层析、疏水层析和凝胶排阻层析的三步工艺纯化表达rhPro-UK的CHO细胞培养上清液,rhPro-UK的纯度达到98%、回收率为60%～70%。 The efficient expression vector for recombinant human pro-urokinase （rhPro-UK） was construc- ted with the combination of chicken β-globin MAR sequences, human elongation factor alpha! （hEF-lα） housekeeping gene regulatory sequences, and RNA stability and the output sequence. The rhPro-UK ex pression level reached 1 299 IU/cells6/d in CHO （Chinese hamster ovary） cell lines. The rhPro-UK containing supernatant purified with cation exchange chromatography, hydrophobic interaction chromatog- raphy, gel exclusion chromatography process resulted in rhPro-UK purity over 95 % with recovery rate of 60% - 70%.
作者 李世崇 叶玲玲 刘红 张正光 高丽华 胡显文 陈昭烈
机构地区 军事医学科学院生物工程研究所
出处 《生物加工过程》 CAS CSCD 2012年第5期50-54,共5页 Chinese Journal of Bioprocess Engineering
基金 "重大新药创制"科技重大专项资助项目(2009ZX09503-011)
关键词 CHO细胞 重组人尿激酶原 表达 纯化 CHO cells rhupro-UK expression purification
分类号 R392 [医药卫生—免疫学]
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参考文献17

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生物加工过程
2012年 第5期

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