肠道病毒71型3A蛋白肽段抗体的制备  

Preparation of polyclonal antibody against polypeptide of enterovirus 71 3Aprotein

作　　者：邹欣然 吴静[1,2] 郑蔚[2] 顾佳奇 茅凌翔 ZOU Xin-ran;WU Jing;ZHENG Wei;GU Jia-qi;MAO Ling-xiang(Department of Laboratory Medicine,The Affiliated People's Hospital,Zhenjiang,Jiangsu 212000,China;School of Medicine,Jiangsu University,Zhenjiang,Jiangsu 212000,China)

机构地区：[1]江苏大学附属人民医院检验科,江苏镇江212002 [2]江苏大学医学院,江苏镇江212000

出　　处：《中国病毒病杂志》2019年第4期263-267,共5页Chinese Journal of Viral Diseases

基　　金：国家自然科学基金(81601751);江苏省青年医学人才计划(QNRC2016453);江苏省“333”工程科研项目(BRA2017144)

摘　　要：目的制备肠道病毒71型(EV-A71)3A蛋白的肽段抗体。方法利用ExPasy和DNAstar软件对EV-A71 3A蛋白的跨膜结构域、二级结构、亲水性及疏水性等生物学特性进行分析,设计并合成2条14个氨基酸的多肽。用钥孔戚血蓝蛋白(keyhole limpet hemocyanin,KLH)偶联肽段作为抗原分别免疫Balb/c小鼠,免疫结束后取小鼠全血分离血清,用免疫印迹实验(Western blot)对血清中的3A蛋白抗体进行鉴定。结果Western blot结果显示,第一条3A蛋白的多肽不能刺激小鼠产生特异性抗体,而第二条多肽能够刺激小鼠产生特异性抗体。结论成功制备特异性较好的3A肽段抗体,为进一步研究3A蛋白提供了工具。Objective To prepare and characterize antibody against polypeptide of enterovirus 71(EV-A71)3 A protein.Methods Two different polypeptide fragments of EV-A71 3 Aprotein were designed and synthesized to immunize mice,and the antiserum was collected respectively.The anti-3 Aantibody in antiserum was detected by Western blot.The localization of 3 Aprotein was observed in the infected RD cells by using this polyclonal antibody.Results Western blot analysis showed that the first polypeptide fragment failed to stimulate the mice to produce antibody of 3 Aprotein,whereas the second polypeptide fragment was able to stimulate the mice to produce antibody of 3 Aprotein.Conclusions The 3 Aprotein antibody was prepared successfully,which can be used for further research on the funcion of 3 Aprotein.

关 键 词：人肠道病毒71型 3A蛋白 多克隆抗体 

分 类 号：R373.2[医药卫生—病原生物学]