免疫学杂志2024,Vol.40Issue(3) :309-314.DOI:10.13431/j.cnki.immunol.j.20240040

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

蒋丹 舒纬童 李光琪 蓝华滔 黄倩莹 徐广贤
免疫学杂志2024,Vol.40Issue(3) :309-314.DOI:10.13431/j.cnki.immunol.j.20240040

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

蒋丹 1舒纬童 1李光琪 2蓝华滔 1黄倩莹 1徐广贤1
扫码查看

作者信息

  • 1. 523808 东莞,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
  • 2. 750000 银川,宁夏医科大学总医院医学实验中心
  • 折叠

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.

Abstract

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,Westem 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

Key words

Nanobody/SARS-CoV-2 S Protein/purification and expression

引用本文复制引用

基金项目

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)

出版年

2024
免疫学杂志
第三军医大学,中国免疫学会

免疫学杂志

CSTPCD
影响因子:0.704
ISSN:1000-8861
段落导航相关论文