重组人甲状旁腺激素的生产工艺研究  

Production Technology of Recombinant Human Parathyroid Hormone

作　　者：李珊珊 黄宗庆 马洁 陆伟根[1] 路建光[1,2] LI Shanshan;HUANG Zongqing;MA Jie;LU Weigen;LU Jianguang(State Key Lab.of New Drug and Pharmaceutical Process,Shanghai Institute of Pharmaceutical Industry,China Institute of Pharmaceutical Industry,Shanghai 201203;Shanghai Duomirui Bio-Technology Co.,Ltd.,Shanghai 201203)

机构地区：[1]中国医药工业研究总院上海医药工业研究院,创新药物与制药工艺国家重点实验室,上海201203 [2]上海多米瑞生物技术有限公司,上海201203

出　　处：《中国医药工业杂志》2020年第7期843-848,共6页Chinese Journal of Pharmaceuticals

基　　金：国家十三五“重大新药创制”科技重大专项“儿童用药品种及关键技术研究”(2018ZX09721003-008-010)。

摘　　要：本研究采用基因重组大肠埃希菌发酵的方法,以可溶形式表达人甲状旁腺激素融合蛋白。菌体经高压破壁和加热沉淀后获得融合蛋白,经Ulp1酶切去除泛素标签后获得目的蛋白粗品,经阳离子交换色谱法和反相色谱法获得纯度为99.2%的目的蛋白。产品经相对分子质量测定、肽图分析和生物活性测定,证明自制甲状旁腺激素的结构和功能与理论值一致。本研究为工业化制备重组人甲状旁腺激素提供了参考。In this study,genetically recombinant E.coli fermentation was used to express the human parathyroid hormone fusion protein in soluble form.The fusion protein was obtained with high pressure wall breaking and heating precipitation.The crude protein was obtained after removing the ubiquitin tag by Ulp1 digestion.The purity of the target protein was 99.2%by cation exchange and reversed phase chromatography.According to the studies of relative molecular weight determination,peptide map analysis and biological activity determination,it was proved that the structure and function of the self-made parathyroid hormone were consistent with the theory value.This study provided a guide for the preparation of recombinant human parathyroid hormone in industry.

关 键 词：生物重组 甲状旁腺激素 泛素标签 离子交换 

分 类 号：R945[医药卫生—微生物与生化药学] Q78[医药卫生—药剂学]