抗SARS-CoV-2 S蛋白纳米抗体的筛选、表达和鉴定  

Screening,expression and characterization of nanobody targeting SARS-CoV-2 S proteins

作　　者：蒋丹 舒纬童 李光琪 蓝华滔 黄倩莹 徐广贤 JIANG Dan;SHU Weitong;LI Guangqi;LAN Huatao;HUANG Qianying;XU Guangxian(Dongguan Key Laboratory of Molecular Immunology and Cell Therapy,First Dongguan Affiliated Hospital,School of Medical Technology,Guangdong Medical University,Dongguan 523808,China;Medical Laboratory Center,General Hospital of Ningxia Medical University,Yinchuan 750000,China)

机构地区：[1]广东医科大学医学技术学院附属东莞第一医院,东莞市分子免疫与细胞治疗重点实验室,东莞523808 [2]宁夏医科大学总医院医学实验中心,银川750000

出　　处：《免疫学杂志》2024年第3期309-314,共6页Immunological Journal

基　　金：广东医科大学高层次人才科研资助计划(4SG23190G);广东医科大学博士科研启动基金(GDMU2022030);宁夏回族自治区重点研发计划(2022CMG03123)。

摘　　要：目的筛选特异性抗SARS-CoV-2 S蛋白纳米抗体序列,表达纯化重组抗SARS-CoV-2纳米抗体,评估其特性及应用潜能。方法基于前期构建的天然噬菌体纳米抗体展示库,以生物素化SARS-CoV-2 S蛋白受体结合结构域抗原为靶点,利用生物素-链霉亲和素液相筛选法、噬菌体-ELISA和测序筛选抗SARS-CoV-2 S蛋白纳米抗体序列;构建重组抗SARS-CoV-2 S蛋白纳米抗体表达载体,IPTG诱导表达并纯化,采用SDS-PAGE、Western blot和ELISA分析其特征及应用潜能。结果成功获得2条抗SARS-CoV-2 S蛋白纳米抗体序列,并构建了相应原核表达菌;成功表达纯化重组抗SARS-CoV-2 S蛋白纳米抗体,其与SARS-CoV-2 S蛋白可特异性结合。结论筛选及表达的重组抗SARS-CoV-2 S蛋白纳米抗体为其应用于SARS-CoV-2检测和治疗及相关研究提供基础。This study was performed to screen the sequence of specific nanobody targeting SARS-CoV-2 S protein,and to express and purify recombinant anti-SARS-CoV-2 nanobodies,and evaluate their characteristics and application potential.Based on the previously constructed natural bacteriophage nanobody display library,the anti-SARS-CoV-2 S protein nanobody sequences were screened,with biotinylated SARS-CoV-2 S protein receptor-binding domain antigen as target,by biotin-streptavidin liquid phase screening method,phage-ELISA and sequencing.Recombinant anti-SARS-CoV-2 S protein nanobody expression vectors were constructed,the protein was induced by IPTG and then purified,and their characteristics and application potential were analyzed by SDS-PAGE,Western blot and ELISA methods.Two anti-SARS-CoV-2 S protein nanobody sequences were successfully obtained,and the corresponding prokaryotic expression bacteria were constructed.The recombinant anti-SARS-CoV-2 S protein nanobody was expressed and purified successfully,which can specifically bind to SARS-CoV-2 S protein.In conclusion,the recombinant anti-SARS-CoV-2 S protein nanobodies have screened and expressed successfully,which provides a basis for its application in SARS-CoV-2 detection and treatment,and facilitates related researches.

关 键 词：纳米抗体 新型冠状病毒(SARS-CoV-2)S蛋白 纯化表达 

分 类 号：Q939.48[生物学—微生物学] R392-33[医药卫生—免疫学]