水痘带状疱疹病毒ORF41的单克隆抗体制备及其初步研究  

Preparation of monoclonal antibody to varicella-zoster virus ORF41 and its preliminary study

作　　者：蔡雨泽 潘德全 杨睿曦 袁成先 赵灿阳 赵欢[1] 程通[2] 王玮(指导) 夏宁邵[1] CAI Yuze;PAN Dequan;YANG Ruixi;YUAN Chengxian;ZHAO Canyang;ZHAO Huan;CHENG Tong;WANG Wei;XIA Ningshao(School of Public Health,Xiamen University,Xiamen 361102,China)

机构地区：[1]厦门大学公共卫生学院,厦门361102 [2]厦门大学生命科学学院,国家传染病诊断试剂与疫苗工程技术研究中心,厦门361102

出　　处：《中国免疫学杂志》2023年第5期1040-1045,共6页Chinese Journal of Immunology

基　　金：国家自然科学基金(81871648,82171833)资助。

摘　　要：目的:制备水痘-带状疱疹病毒(VZV)衣壳蛋白ORF41的单克隆抗体(mAbs),并对该蛋白性质进行初步研究。方法:截取VZV ORF41蛋白亲水性及免疫原性较优的片段,用大肠杆菌蛋白表达系统对相关区段进行表达并纯化,免疫BALB/c小鼠后,通过杂交瘤细胞技术制备并筛选出针对VZV ORF41的mAbs,并利用制备得到的mAbs进行ELISA、Western blot和免疫荧光实验,检测所得抗体的亲和性和特异性,同时分析VZV ORF41蛋白在细胞中的表达与分布特征。结果:本研究筛选到的VZV OR41 mAb 12C3具有较高的亲和性和特异性,可特异性识别在质粒转染及VZV感染细胞中表达的ORF41蛋白,且发现检测出的VZV ORF41蛋白在细胞质与细胞核中均有分布。结论:本研究获得针对VZV ORF41反应性、特异性良好的mAb,为进一步开展VZV ORF41蛋白功能和作用机制的研究提供基础。Objective:To prepare monoclonal antibodies(mAbs)target varicella-zoster virus(VZV)capsid protein ORF41 and preliminarily study its properties.Methods:Fragments with good hydrophilicity and immunogenicity of VZV ORF41 protein were selected for production using an E.coli protein expression system and purified.After immunization in BALB/c mice,mAbs against VZV ORF41 were screened and prepared by hybridoma cell technology.With the mAbs,expression and distribution of VZV ORF41 protein in cells were analyzed by Western blot and immunofluorescence.Results:VZV OR41 mAb 12C3 screened in this study has high affinity and specificity,which could specifically recognize the ORF41 protein expressed in either plasmid-transfected or VZV-infected cells,and it is found that the VZV ORF41 protein is distributed in both cytoplasm and nucleus with the help of 12C3.Conclusion:In this study,an mAb with high reactivity and specificity against VZV ORF41 was obtained,which provided a basis for further studies on function and mechanism of VZV ORF41 protein.

关 键 词：水痘-带状疱疹病毒 衣壳蛋白 ORF41 单克隆抗体 

分 类 号：R392.12[医药卫生—免疫学] R511.5[医药卫生—基础医学]