抗鸡传染性贫血病毒VP2 蛋白单克隆抗体的制备及其应用研究
Development and Application of Monoclonal Antibody against VP2 Protein of Chicken Infectious Anemia Virus
侯力丹 1薛麒 1王嘉 1孔冬妮 1吴宏举 1翟天舒 1邓永 1毛娅卿 1吴华伟 1孙瑶 2刘丹1
作者信息
- 1. 中国兽医药品监察所(农业农村部兽药评审中心),北京 100081
- 2. 中牧实业股份有限公司,北京 100070
- 折叠
摘要
为制备针对鸡传染性贫血病毒(Chicken infectious anemia virus,CIAV)的单克隆抗体,以CIAV的VP2 原核表达蛋白作为抗原免疫小鼠,经杂交瘤技术获得3 株能稳定分泌抗VP2 蛋白特异性抗体的细胞株,进一步经IFA筛选出特异性和灵敏度俱佳的杂交瘤细胞Mab-VP2-4 株,制备的小鼠腹水效价达到 1∶2000.结果显示,制备的抗 VP2 蛋白单克隆抗体可与多株 CIAV 感染的MDCC-MSB1 细胞发生特异性反应,而对EDSV、ALV、ILTV、ARV 以及空白MDCC-MSB1 细胞、CEF细胞等均无交叉反应,特异性良好,且应用于 IFA 方法时对 CIAV 感染的最低检出限为5 TCID50.以该单克隆抗体对市场上10 种不同来源的禽活疫苗进行病毒分离结合单克隆抗体IFA检测,同时与《中国兽药典》规定的鸡检查法进行对比检测,结果一致,均未检出CIAV污染.本研究制备了针对CIAV VP2 蛋白的特异性单克隆抗体,为该病特异性诊断方法的建立奠定了基础.
Abstract
To prepare chicken infectious anemia virus(CIAV)specific monoclonal antibody,codon-optimized VP2 protein was used as antigen and hybridoma technology to obtain three positive cell lines which could secrete specific antibody stably,the MAB-VP2-4 strain with high specificity and sensitivity was screened by IFA,and the ascites of mice was prepared with the titer of 1∶2000.The prepared monoclonal antibody could react specifically with CIAV infected MDCC-MSB1 cells,but had no cross-reaction with EDSV,AlV,ILTV,ARV,MDCC-MSB1 cells and CEF cells,and the minimum detection limit is 5TCID50.The monoclonal antibody was used to detect 10 kinds of live poultry vaccines from different sources on the market,and the SPF chickens inoculated with the vaccine were tested by the method stipulated according to the Chinese Veterinary Pharmacopoeia,no CIAV contamination was detected.In this study,the monoclonal antibody against VP2 protein of CIAV was prepared,which laid a foundation for the establishment of the specific diagnostic method and the related research of CIAV.
关键词
鸡传染性贫血病毒/VP2蛋白/单克隆抗体/间接免疫荧光Key words
chicken infectious anemia virus/VP2 protein/monoclonal antibody/indirect immunoinfluscent assay引用本文复制引用
基金项目
国家重点研发计划(2022YFD1800602)
中国兽医药品监察所公益性重点专项(GY202104)
中国兽医药品监察所公益性重点专项(GY202105)
出版年
2024
国家科技期刊平台
NSTL
万方数据