肠道病毒71型2C蛋白多克隆抗体的制备及鉴定

Preparation and characterization of polyclonal antibody against enterovirus A712C protein

目的制备肠道病毒71型(enterovirus 71,EV-A71)2C蛋白的多克隆抗体并测试抗体的应用效果。方法分析EV-A712C的抗原性,设计合成1条EV-A712C蛋白多肽,偶联钥孔血蓝蛋白KLH作为抗原,免疫新西兰大白兔。ELISA测定免疫效价,达到要求后,收集血清并纯化。利用ELISA、免疫印迹、免疫沉淀和免疫荧光法对纯化后的2C蛋白抗体进行鉴定。结果设计的2C蛋白肽段刺激新西兰大白兔产生了特异性抗体,免疫印迹、免疫沉淀和免疫荧光法证明制备的抗体可特异性识别并结合2C蛋白。结论成功获得了EV-A71的2C蛋白抗体,为进一步研究2C蛋白功能提供了有效工具。

Objective To prepare polyclonal antibody against the 2 C protein of enterovirus 71(EV-A71).Methods A polypeptide of EV-A712 C protein was designed after analyzing the antigenicity of 2 C protein.The 2 C polypeptide coupled with KLH was used as the antigen to immunize New Zealand white rabbits.After the immunization,the immune-potency was determined by ELISA.The 2 C antibody purified using protein A beads was characterized by ELISA,Western blotting,immunoprecipitation and immunofluorescence assays.Results The 2 C antibody could recognize the 2 C antigen specifically in Western blot,immunoprecipitation,and immunofluorescence.Conclusions The EV-A712 C antibody is successfully generated.