猪德尔塔冠状病毒S蛋白纳米颗粒的构建及免疫原性评价
Construction and Immunogenicity Evaluation of Porcine deltacoronavirus Spike Protein Nanoparticles
张晓蓉 1吴建晓 2高銘泽 2高娜 1陈建飞 3冯力2
作者信息
- 1. 黑龙江八一农垦大学,黑龙江 大庆 163319;中国农业科学院哈尔滨兽医研究所,黑龙江 哈尔滨 150069
- 2. 中国农业科学院哈尔滨兽医研究所,黑龙江 哈尔滨 150069
- 3. 中国农业科学院哈尔滨兽医研究所,黑龙江 哈尔滨 150069;黑龙江省生猪现代农业产业技术协同创新推广体系,黑龙江 哈尔滨 150006
- 折叠
摘要
为评价猪德尔塔冠状病毒(PDCoV)纤突(S)蛋白纳米颗粒的免疫原性,首先利用pcDNA3.1(+)-S1-CTD-Spa-His、pET28a-SpyCatcher-mi3分别表达了重组蛋白S1-CTD-Spy-His和SpyCatcher-mi3,纯化后,经SDS-PAGE鉴定表明在55 ku和37 ku处有目的条带.通过SpyTag-SpyCatcher蛋白连接酶系统,获得了S1-CTD-SpyCatcher-mi3纳米颗粒,在电镜下能够观察到直径35～40 nm的颗粒.血清抗体检测结果表明,S1-CTD-SpyCatcher-mi3纳米颗粒可诱导小鼠产生较强的体液免疫反应,S1-CTD-SpyCatcher-mi3组的特异性IgG抗体水平显著高于S1-CTD组(P＜0.05),S1-CTD-SpyCatcher-mi3组和S1-CTD组与PBS对照组相比差异均极显著(P＜0.01).该研究表明,S1-CTD-SpyCatcher-mi3纳米颗粒具有良好的免疫原性,可诱导小鼠产生免疫保护力,为新型猪德尔塔冠状病毒感染疫苗的研制奠定了基础.
Abstract
To evaluate the immunogenicity of Porcine deltacoronaviurs(PDCoV)spike(S)proteinnanoparticles,the recombinant proteins S1-CTD-Spy-His and SpyCatcher-mi3 were firstly expressed by using pcDNA3.1(+)-S1-CTD-Spa-His and pET28a-SpyCatcher-mi3,After purification,SDS-PAGE identification showed that there were target bands at 55 ku and 37 ku.Utilizing the SpyTag-SpyCatcher protein ligase system,S1-CTD-Spycatcher-mi3 nanoparticles were obtained,and the diameter of 35～40 nm nanoparticles were observed under electron microscopy.The results of serum antibody assay showed that S1-CTD-SpyCatcher-mi3 nanoparticles induced a strong humoral immune response in mice,and the specific IgG antibody levels of the S1-CTD-SpyCatcher-mi3 group were significantly higher than those of the S1-CTD group(P＜0.05),and the differences between the S1-CTD-SpyCatcher-mi3 group and the S1-CTD group compared with the PBS control group were both highly significant(P＜0.01).This study shows S1-CTD-SpyCatcher-mi3 nanoparticles have good immunogenicity and can induce immune protection in mice,which lays a foundation for the development of novel Porcine deltacoronavirus infection vaccine.
关键词
冠状病毒/连接酶/纳米颗粒/小鼠/免疫原性Key words
coronavirus/ligase/nanoparticle/mouse/immunogenicity引用本文复制引用
基金项目
国家重点研发计划项目(2021YFD1801105)
中国农业科学院科技创新工程科学中心重点任务(CAAS-CSLPDCP-202301)
出版年
2024
国家科技期刊平台
NSTL
万方数据