噬菌体单链抗体导向溶栓剂的构建  

Construction of Phage Single-chain Antibody-targeted Thrombolytics

作　　者：孙雷[1] 黄仪秀[1] 朱圣庚[1] 

机构地区：[1]北京大学生命科学学院,北京100871

出　　处：《北京大学学报（自然科学版）》2000年第6期881-884,共4页Acta Scientiarum Naturalium Universitatis Pekinensis

基　　金：国家自然科学基金资助项目!(396 70 170 );高等学校博士学科点专项科研基金资助项目!(95 0 0 12 8)

摘　　要：Fibrin\|specific antibodies were isolated from a phage\|displayed single\|chain antibody(ScAb) library using affinity selection or panning.DNA shuffling was introduced to realign the specific antibody genes in order to mimic the antibody maturation in vivo. After cloning of shuffling products,a secondary library was constructed,from which the specific antibody against fibrin with higher activity was selected through panning and screening.The fibrin\|specific antibody gene and uPA functional domain gene fragment were joined together,then inserted into expression plasmid pET\|21a.The ScAb/uPA fusion protein was expressed with the induction of IPTG.In substrate S 2444 assay,the uPA activity of the chimera was about 3.1×10 3?IU/mg periplasmic protein of host E.coli .It also kept up the affinity to fibrin.Fibrin\|specific antibodies were isolated from a phage\|displayed single\|chain antibody(ScAb) library using affinity selection or panning.DNA shuffling was introduced to realign the specific antibody genes in order to mimic the antibody maturation in vivo. After cloning of shuffling products,a secondary library was constructed,from which the specific antibody against fibrin with higher activity was selected through panning and screening.The fibrin\|specific antibody gene and uPA functional domain gene fragment were joined together,then inserted into expression plasmid pET\|21a.The ScAb/uPA fusion protein was expressed with the induction of IPTG.In substrate S 2444 assay,the uPA activity of the chimera was about 3.1×10 3?IU/mg periplasmic protein of host E.coli .It also kept up the affinity to fibrin.

关 键 词：噬菌体 单链抗体 DNA改组 导向溶栓剂 尿激酶 

分 类 号：R944[医药卫生—药剂学]