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门冬胰岛素原的酶切及产物分离纯化方法
周孟能; 陈文琼; 陈颖; 刘永东
2018-10-20
Source Publication西部医学
Volume30Issue:10Pages:1434-1437+1442
Abstract

目的建立门冬胰岛素原的酶切及纯化方法,获得高纯度的活性门冬胰岛素。方法以大肠杆菌制备含有前导肽和折叠伴侣C肽的门冬胰岛素原为底物,采用胰蛋白酶和羧肽酶B对门冬胰岛素原进行酶切同时去除前导肽及C肽,酶切条件:酶与门冬胰岛素原的质量比为1:1000,室温反应30min。利用SPFF层析柱纯化酶切产物,获得高纯度的门冬胰岛素。结果电泳结果显示酶切产物与门冬胰岛素分子量相似,离子交换纯化可以分开门冬胰岛素及酶切的多余肽段。高效凝胶过滤层析分析洗脱峰纯度大于95%,蛋白收率为30%。MALDI-TOF检测纯化产物的分子量与门冬胰岛素的理论分子量完全一致。结论建立门冬胰岛素原酶切及分离纯化工艺可用于指导门冬胰岛素的规模制备。

Keyword门冬胰岛素原 门冬胰岛素 酶切 分离纯化
Language中文
Document Type期刊论文
Identifierhttp://ir.ipe.ac.cn/handle/122111/28865
Collection中国科学院过程工程研究所
Recommended Citation
GB/T 7714
周孟能,陈文琼,陈颖,等. 门冬胰岛素原的酶切及产物分离纯化方法[J]. 西部医学,2018,30(10):1434-1437+1442.
APA 周孟能,陈文琼,陈颖,&刘永东.(2018).门冬胰岛素原的酶切及产物分离纯化方法.西部医学,30(10),1434-1437+1442.
MLA 周孟能,et al."门冬胰岛素原的酶切及产物分离纯化方法".西部医学 30.10(2018):1434-1437+1442.
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