中国兽医科学2024，Vol.54Issue(12)：1569-1576,8.DOI:10.16656/j.issn.1673-4696.2024.0203
新型冠状病毒受体结合域亚单位疫苗的制备及其免疫原性评价
Preparation and immunogenicity evaluation of SARS-CoV-2 receptor binding domain subunit vaccine
摘要
Abstract
The aim of this study was to prepare a receptor binding domain(RBD)subunit vaccine of SARS-CoV-2 and evaluate its immunogenicity.Firstly,recombinant plasmid pFastBacl-gp67-RBD was con-structed by PCR.Recombinant baculovirus P3-RBD was obtained using bac to bac technique.The expression of RBD was identified by indirect immunofluorescence assay.After purification of RBD by Ni2+column affinity chromatography,its immunoreactivity and angiotensin-converting enzyme 2(ACE2)binding affinity were identified.Finally,the immunogenicity of the subunit vaccine was evaluated by immuniz-ing BALB/c mice with a mixture of RBD and Salmonella typhimurium flagellin(FliC),and setting up the RBD group and the aluminum adjuvant(Alum)group as controls,respectively.The results showed that green fluorescence was observed in Sf9 cells infected with P3-RBD,and the RBD protein could react with anti-RBD polyclonal antibody and ACE2 protein.After 2 weeks of second immunization,the results showed that both FliC and Alum adjuvant groups could significantly increase IgG titers in serum com-pared to the RBD group.In addition,the IgG subtypes(IgG1 and IgG2a)titers in the FliC adjuvant group were significantly higher than that in the RBD group,which demonstrated that FliC promotes the mixed Th1/Th2 immune response in the RBD vaccine.In conclusion,Salmonella FliC enhances the immunogenicity of the SARS-CoV-2 RBD subunit vaccine,which shows good application prospects and lays the foundation for the further development of a safe and effective COVID-19 subunit vaccine.关键词
新型冠状病毒/杆状病毒表达载体系统/RBD蛋白/鞭毛蛋白/佐剂/免疫原性Key words
SARS-CoV-2/baculovirus expression vector system/RBD protein/flagellin/adjuvant/immunogenicity分类
农业科技引用本文复制引用
崔耀丹,宋丽,焦新安,潘志明..新型冠状病毒受体结合域亚单位疫苗的制备及其免疫原性评价[J].中国兽医科学,2024,54(12):1569-1576,8.基金项目
国家自然科学基金项目(32102679) （32102679）
江苏省高校优势学科建设工程项目(PAPD) （PAPD）
江苏省重点研发计划(现代农业)重点项目(BE2021331) （现代农业）
高等学校学科创新引智计划项目(D18007) （D18007）
江苏省第六期"333高层次人才培养工程"项目 （）
