人CPNE5多克隆抗体的制备  

Preparation of Polyclonal Antibody Against Human CPNE5

作　　者：靳雁斌[1] 丁学锋[2] 司慧远[1] 吴彦瑞 王晓文[2] 范文红[2] 范明[2] 

机构地区：[1]空军总医院干部病房,北京100142 [2]军事医学科学院基础医学研究所脑保护与可塑性研究室,北京100850 [3]解放军第一八八医院感染内科,广东潮州521000

出　　处：《标记免疫分析与临床》2011年第6期411-415,共5页Labeled Immunoassays and Clinical Medicine

摘　　要：制备人CPNE5的多克隆抗体。将CPNE5基因中423bp(626-1048bp)的片段构建到原核表达载体pGEX-5X-1,将重组质粒转化大肠杆菌BL21,在IPTG诱导下表达蛋白,经SDS-PAGE分离纯化得到抗原肽段,质谱法鉴定后常规免疫家兔。用CL-4B柱纯化抗体。结果显示,成功表达并纯化了人CPNE5抗原肽段;ELISA显示抗血清具有高亲合性,Western blot检测表明抗体能特异性识别CPNE5。本文制备的抗体经检验证实为高亲合性和特异性的兔抗人CPNE5多克隆抗体。To prepare polyclonal antibody against human CPNE5,the 423bp specific sequence of CPNE5（626~1048bp） was cloned into the vector pGEX-5X-1.The recombinant plasmid was transformed into host strain E.coli BL21 and induced by IPTG.Recombinant protein was purified by SDS-PAGE and detected by mass spectrum.The protein was then used as antigen to immunize rabbit by the conventional method.The polyclonal antibody was purified by protein A sepharose CL-4B.The results showed that the antigen was successfully expressed and purified.The high affinity of the antiserum was confirmed by ELISA.The results of Western blot showed this antibody could specifically recognize CPNE5 protein.The CPNE5 antibody with high affinity and specificity was successfully prepared.

关 键 词：人CPNE5 pGEX-5X-1 原核表达 多克隆抗体 酶联免疫吸附实验 免疫印迹 

分 类 号：Q511[生物学—生物化学] R34[医药卫生—基础医学]