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.
出处
《药物生物技术》
CAS
CSCD
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
关键词
GM-CSF
白细胞介素-3
融合蛋白
蛋白纯化
Granulocyte macrophage colony stimulating factor, Interleukin 3, Fusion protein, Protein purification
