Matriptase-2多克隆抗体的制备及鉴定  

Generation and Identifi cation of a Polyclonal Antibody to Matripase-2

作　　者：杨剑峰[1] 赵赟霄[2] 左斌[1] 何杨[1,2] 

机构地区：[1]苏州大学唐仲英血液学研究中心,江苏苏州215123 [2]苏州大学附属第一医院江苏省血液研究所,江苏苏州215006

出　　处：《中国血液流变学杂志》2014年第4期472-475,共4页Chinese Journal of Hemorheology

基　　金：江苏省高校优势学科建设工程资助项目;江苏省临床医学中心项目（ZX201102）

摘　　要：目的：探讨Matriptase-2重组蛋白的表达及多克隆抗体制备。方法 Matriptase-2胞外区近跨膜段（Glu81-Gly228）克隆入pMAL-C2X，重组质粒转化BL21（DE3）大肠杆菌，经IPTG诱导表达，提取细菌蛋白后用直链淀粉树脂纯化柱进行纯化。将纯化后的融合蛋白免疫新西兰大白兔，制备Matriptase-2抗血清；免疫血清用protein G亲和层析柱进行纯化，获得多克隆抗体。结果成功表达了Matriptase-2重组融合蛋白，获得了高效价的抗血清，Western blot结果显示具有抗原特异性识别。结论成功制备了高特异性、高效价的抗人Matriptase-2多克隆抗体，为今后深入研究Matriptase-2的生物学功能提供了有用的实验工具。Objective To investigate the expression of Matriptase-2 recombinant protein inE.coli and the generation of its polyclonal antibody.Methods Extracellular region （Glu81-Gly228） proximal to transmembrane domain of Matriptase-2 was cloned into pMAL-C2X. The recombinant plasmid was transformed to BL21 （DE3） bacteria. After IPTG induction, the total protein was extracted and the recombinant Matriptase-2 protein was purifi ed with amylose resin. The Matriptase-2 recombinant protein was used to immunize New Zealand Rabbits to prepare serum with anti-Matriptase-2 polyclonal antibody. The antibody was purifi ed from serum by protein G sepharose 4B.Results Recombinant Matriptase-2 protein was successfully expressed inE.coliand anti-serum with high titer was obtained. Western blotting result showed that the antibody specifi cally recognized Matriptase-2. Conclusion It is concluded that the anti-Matriptase-2 polyclonal antibody with high titer and specificity is successfully generated. The antibody provides an useful experimental tool to investigate the biological function of Matriptase-2.

关 键 词：Matriptase-2 原核表达 多克隆抗体 

分 类 号：R55[医药卫生—血液循环系统疾病]