戊型肝炎病毒衣壳蛋白特异性人源单克隆抗体的筛选与鉴定  

Screening and identification of human monoclonal antibodies to hepatitis E virus capsid protein

作　　者：张旭 温桂平[1] 唐自闽 王思令 陈佳昕 应东 刘畅 郑子峥(指导)[1] 夏宁邵[1] ZHANG Xu;WEN Gui-Ping;TANG Zi-Min;WANG Si-Ling;CHEN Jia-Xin;YING Dong;LIU Chang;ZHENG Zi- Zheng;XIA Ning-Shao.(State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, National Institute of Diagnostics and Vaccine Development of Infectious Diseases, School of Public Health, Xiamen University, Xiamen 361102, China)

机构地区：[1]厦门大学分子疫苗学与分子诊断学国家重点实验室,厦门大学国家传染病诊断试剂与疫苗工程技术研究中心,厦门大学公共卫生学院,厦门361102

出　　处：《中国免疫学杂志》2018年第5期703-707,共5页Chinese Journal of Immunology

基　　金：国家自然科学基金(No.81571996); 福建省自然科学基金(No.2016J05201)资助

摘　　要：目的:建立从外周血快速筛选戊型肝炎病毒(Hepatitis E virus,HEV)衣壳蛋白特异性人源抗体的方法,从疫苗免疫者外周血中筛选出相应抗体并进行鉴定。方法:采用分选型流式细胞仪获得外周血中HEV衣壳蛋白特异性的记忆B细胞,通过单细胞RT-PCR的方法获得抗体序列,并进行重组表达,最后对获得的人源单克隆抗体进行初步性质鉴定。结果:成功筛选到识别HEV衣壳蛋白的6株人源单克隆抗体,6株抗体均具有抗原结合活性,4株抗体具有中和活性。结论:成功获得HEV衣壳蛋白特异性人源单克隆抗体序列,并进行真核表达,对抗体的性质进行初步鉴定,为后期研究疫苗免疫的人体内的抗体演化打下基础。Objective： To establish a method for rapidly screening human antibodies recognizing HEV capsids proteins from peripheral blood. The antibodies recognizing HEV capsids proteins were screened from the peripheral blood of vaccinator and the properties of the antibodies were analyzed. Methods： The HEV capsids proteins specific memory B cells in peripheral blood were obtained by flow cytometry sorting. Then antibody variable genes were acquired through single-cell RT-PCR and recombined to express in eukaryocyte. Finally,the properties analysis of recombinant expressed human monoclonal antibodies were carried out. Results： Six humanized monoclonal antibodies recognizing HEV capsids proteins were successfully obtained,and most of them had binding activity and neutralizing activity. Conclusion： The sequence of human monoclonal antibodies recognizing HEV capsid proteins is successfully screened and successfully expressed in the eukaryocyte. The properties of the antibodies are identified,which lay the foundation for studying antibody evolution in the human body after vaccination.

关 键 词：戊型肝炎病毒 衣壳 人源单克隆抗体 流式细胞分选 单细胞RT-PCR 

分 类 号：R373.2[医药卫生—病原生物学]