寨卡病毒NS2B蛋白的表达纯化及单克隆抗体制备  

Prokaryotic Expression and Monoclonal Antibody Preparation of Zika Virus NS2B Protein

作　　者：吴钰彬 李姝璇 侯汪衡 杨宏伟 赵欢[2] 吴林丽 吴文伟 朱峻毅 张军[2] 程通[1] WU Yu-Bin;LI Shu-Xuan;HOU Wang-Heng;YANG Hong-Wei;ZHAO Huan;WU Lin-Li;WU Wen-Wei;ZHU Jun-Yi;ZHANG Jun;CHENG Tong(National Institute of Diagnostics and Vaccine Development of Infectious Disease,School of Life Science,Xiamen University,Xiamen 361102;School of Public Health,Xiamen University,Xiamen 361102,China)

机构地区：[1]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,福建厦门361102 [2]厦门大学公共卫生学院,福建厦门361102

出　　处：《生物技术通讯》2020年第1期12-18,共7页Letters in Biotechnology

基　　金：国家自然科学基金(81701999)。

摘　　要：目的:应用大肠杆菌重组表达并纯化寨卡病毒NS2B蛋白,制备抗NS2B蛋白的单克隆抗体。方法:构建携带NS2B基因的重组原核表达质粒,采用大肠杆菌ER2566作为表达菌株,以IPTG诱导表达,通过亲和层析纯化获得NS2B目的蛋白。用纯化后的NS2B蛋白免疫BALB/c小鼠,筛选制备抗NS2B蛋白的单克隆抗体,并初步分析抗体的反应活性和特异性。结果:表达并纯化获得寨卡病毒NS2B蛋白,筛选获得可与NS2B蛋白结合并具有较好反应活性的单克隆抗体2H11,该单抗可识别寨卡病毒感染细胞后表达的NS2B蛋白。结论:筛选获得靶向寨卡病毒NS2B蛋白的单克隆抗体,可为后期深入开展寨卡病毒NS2B蛋白的功能和相关抗病毒药物研究提供支持。Objective:To express the NS2B protein of Zika virus and prepare its monoclonal antibodies with good reactivity.Methods:The recombinant prokaryotic expression plasmid containing NS2B gene of Zika virus was constructed,then the NS2B protein was soluble expressed based on E.coli ER2566 strain and purified by affinity chromatography.BALB/c mice were immunized subcutaneously with purified NS2B protein suspended in Freund's adjuvant,and monoclonal antibodies against NS2B protein were screened by ELISA.The reactivity and specificity of the antibody were preliminarily analyzed by Western blot and immunofluorescence assay.Results:The NS2B protein of Zika virus was successfully expressed,and a mouse monoclonal antibody,named 2H11,which has good activity with NS2B protein was obtained.The monoclonal antibody could recognize the NS2B protein expressed in cells infected with Zika virus.Conclusion:A monoclonal antibody against NS2B of Zika virus was obtained,which will provide support for the further function study of NS2B protein and anti-viral drug research.

关 键 词：寨卡病毒 NS2B蛋白 原核表达 单克隆抗体 

分 类 号：Q78[生物学—分子生物学] R392.1[医药卫生—免疫学]