重组人成纤维细胞生长因子-21的表达及纯化工艺的优化  

Optimization of Procedure for Expression and Purification of Recombinant Human Fibroblast Growth Factor-21

作　　者：赵宏鑫[1,2] 邵明龙[1] 杨苹[1,2] 张耀方[1] 万晓姗[1] 孔祥鑫[1] 王会岩[1] 李校堃[1] 

机构地区：[1]吉林农业大学生物反应器与药物开发教育部工程研究中心,长春130118 [2]吉林农业大学中药材学院,长春130118

出　　处：《中国生物制品学杂志》2009年第11期1113-1116,共4页Chinese Journal of Biologicals

基　　金：吉林省科技厅资助项目(20080712);长春净月开发区资助项目(2008B003)

摘　　要：目的优化重组人成纤维细胞生长因子-21(SUMO-rhFGF-21)融合表达工程菌的表达条件及目的蛋白纯化工艺。方法对工程菌的诱导温度、时间及诱导剂浓度进行优化,并进行罐发酵,对菌体裂解液进行离子交换层析、Ni离子亲和层析及分子筛层析等。纯化产物经SDS-PAGE和HPLC鉴定纯度。结果在诱导温度为37℃,诱导时间为4h,IPTG浓度为0.5mmol/L时,目的蛋白的表达量最高,达20.5%;罐发酵收菌量可达66g/L,目的蛋白的表达量达20.2%;纯化后的rhFGF-21纯度达96%以上。结论优化了rhFGF-21的表达条件及纯化工艺,为rhFGF-21用于新药开发奠定了基础。Objective To optimize the condition for fermentation of recombinant E. coli with fusion gene of SUMO and human fibroblast growth factor-2l （SUMO-rhFGF-21） as well as the procedure for purification of target protein. Methods The temperature and time for induction as well as concentration of inducer of recombinant E. coil was optimized. The recombinant E. coli was ferment- ed under the optimal condition, and the lysate of bactefa was purified by ion exchange, nickel ion affinity and molecular sieve chromatography. The purified target protein was determined for purity by SDS-PAGE and HPLC. Results The optimal temperature and time for induction of recombinant E. coil were 37℃ and 4 h respectively, while the optimal concentration of IPTG as an inducer was 0. 5 mmol/L. The expression level of target protein reached a peak value of 20. 5% under the optimal condition. The yield of bacteria after fermentation reached 66 g/L, and the expression level of target protein was 20. 2%. After purification, the purity of target protein was more than 96%. Conclusion The procedure for expression and purification of rhFGF-21 was optimized, which laid a foundation of developing rhFGF-21 as a novel drug.

关 键 词：重组人成纤维细胞生长因子-21 SUMO 融合表达 纯化 

分 类 号：Q78[生物学—分子生物学]