大肠杆菌中表达的重组人粒细胞-巨噬细胞集落刺激因子/白细胞介素-3(rhGM-CSF/IL-3)融合蛋白的纯化  

Purification of Recombinant Human Granulocyte Macrophage Colony Stimulating Factor/Interleukin-3 Fusion Protein Expressed in E. coli

作　　者：戴盛明 肖桂元 周少雄 郑兴武 饶颖竹 周甘平 吴茂莲 

机构地区：[1]广东湛江市临床医学研究所,湛江524037

出　　处：《药物生物技术》1998年第2期70-74,共5页Pharmaceutical Biotechnology

基　　金：卫生部科研基金课题;广东省重点科技开发项目资助

摘　　要：采用 SP- Sepharose FF及 DEAE- Sepharose FF离子交换层析对大肠杆菌中表达的重组人粒细胞 -巨噬细胞集落刺激因子 /白细胞介素 - 3(rh GM- CSF/ IL- 3)融合蛋白进行了纯化 ,进而用 SDS- PAGE、RP- HPLC、内毒素测定等方法对该蛋白进行了鉴定 ,表明经两步离子交换层析的 rh GM- CSF/ IL- 3达到电泳纯度为 96.6%、HPLC纯度为 97.2 % ,比活性为 8.1× 1 0 8U/ mg,回收率为 1 4 .4%。内毒素含量极低 ,为大批量生产 rh GM- CSF/ IL-Recombinant human granulocyte macrophage colony stimulating factor/interleukin 3 fusion protein expressed in E. coli was isolated and purified to more than 96. 6% homogeneity through ion exchange chromatography on SP Sepharose FF and DEAE Sepharose FF, respectively. The purified protein with a specific activity of 8.10×10 8U/mg protein and the recovery rate of 14.4% appeared as a single band on the SDS PAGE and a single peak on the RP HPLC. This study constructed the basis for scale preparation and clinical use of rhGM CSF/IL 3 fusion protein with very low endotoxin.

关 键 词：GM-CSF 白细胞介素-3 融合蛋白 蛋白纯化 

分 类 号：R392.11[医药卫生—免疫学] Q784[医药卫生—基础医学]