链球菌蛋白G免疫球蛋白结合域C3区串联体的表达、筛选与初步应用  

Expression, Screening and Preliminary Application of the Concatenation C3 Region of the Streptococcal Protein G Immunoglobulin Binding Domain

作　　者：冯利利[1] 杜夜星 张强[1] 李娇娇 夏海锋[1,2] FENG Li-li;DU Ye-xing;ZHANG Qiang;LI Jiao-jiao;XIA Hai-feng(The Key Laboratory of Industrial Biotechnology,Ministry of Education,School of Biotechnology,Jiangnan University,Wuxi 214122,China;National Engineering Laboratory for Cereal Fermentation Technology,Jiangnan University,Wuxi 214122,China)

机构地区：[1]江南大学生物工程学院工业生物技术教育部重点实验室,江苏无锡214122 [2]江南大学粮食发酵工艺与技术国家工程实验室,江苏无锡214122

出　　处：《高校化学工程学报》2018年第4期885-892,共8页Journal of Chemical Engineering of Chinese Universities

基　　金：江苏省自然科学基金(BK20151123)

摘　　要：筛选并制备高效结合抗体的重组蛋白G亲和层析介质。以链球菌蛋白G C3区为结构单元,拼接形成3、4、5、6个重复C3片段的串联体,分别克隆至PET21,并在E.coli BL21(DE3)中表达。经Ni^(2+)纯化的重组蛋白分别偶联至Purose 4 Fast Flow制备亲和层析介质,比较介质的吸附情况,发现重组四串体对h IgG的吸附量最高,其静态饱和吸附量为66.79 mg×mL^(-1),动态吸附载量为35 mg×mL^(-1)介质,优于同类型商品介质。利用该介质分离纯化小鼠腹水和人血清中的IgG,纯度在95%左右,回收率不低于80%。In order to screen and prepare recombinant protein G affinity resins with high efficient antibody binding, the C3 region of Streptococcus protein G was used as the structural unit to prepare different tandem repeats（3, 4, 5 and 6） of C3 fragments, which were respectively cloned into PET21 and expressed in E. coli BL21（DE3）. The recombinant proteins purified by Ni^（2＋） affinity chromatography were coupled onto Purose 4 Fast Flow to prepare affinity chromatography resins. The 4-tandem repeats resin exhibits the highest adsorption capacity to h IgG with static adsorption capacity of 66.79 mg×mL^（-1） and dynamic adsorption capacity of 35 mg×mL^（-1）, which is better than same type commercial resins. Purity of 95% and recovery over 80% can be obtained for mouse ascites and human serum purification using this resin.

关 键 词：抗体 链球菌蛋白G 层析介质 饱和吸附量 

分 类 号：Q819[生物学—生物工程]