上海海洋大学学报2024，Vol.33Issue(1)：18-29,12.DOI:10.12024/jsou.20230104053
3型鲤疱疹病毒白介素10单克隆抗体制备与鉴定
Preparation and characterization of Cyprinid herpesvirus 3 interleukin 10 monoclonal antibody
摘要
Abstract
Cyprinid herpesvirus 3(CyHV3),also known as Koi herpesvirus(KHV),is a highly contagious pathogen that threatens the sustainability of cyprinid aquaculture.The ORF134 gene of CyHV3 encodes an interleukin(IL)10 like protein.In this study,we constructed a prokaryotic expression plasmid of ORF134 and transformed it into the E.coli Rosetta(DE3)cells for expression of recombinant protein.The recombinant CyHV3-IL10 protein was purified by size exclusion chromatography and used for immunization of mice to generate monoclonal antibodies.The antibodies were then characterized by ELISA,Western blotting and immunofluorescent assay.SDS-PAGE analysis showed that the recombinant CyHV3-IL10 protein was approximately 18 ku in size and was expressed mainly in the form of inclusion bodies.The inclusion bodies were denatured,refolded and subjected to size exclusion chromatography.The recombinant CyHV3-IL10 protein was of high purity.Western blotting and immunofluorescence analyses revealed that the CyHV3-IL10 monoclonal antibodies recognized the recombinant CyHV3-IL10 protein expressed in the E.coli and HEK293 cells and could specifically detect the HEK293 cells expressing the CyHV3-IL10 protein.In summary,the availability of CyHV3-IL10 monoclonal antibodies lays the foundation for the functional study of CyHV3-IL10 and the establishment of CyHV3 diagnostic tools,and provides novel strategies for the prevention and control of the diseases caused by CyHV3.关键词
鲤疱疹病毒3型/白介素10/单克隆抗体/Western blotting/免疫荧光分析Key words
CyHV3/IL10/monoclonal antibody/Western blotting/immunofluorescence assay分类
农业科技引用本文复制引用
陈静,邹钧,张艳维,刘琴,朱晓真,贾钊,王俊亚,高娃,贾智英,桂朗..3型鲤疱疹病毒白介素10单克隆抗体制备与鉴定[J].上海海洋大学学报,2024,33(1):18-29,12.基金项目
国家自然科学基金(32030112) （32030112）
