畜牧兽医学报2012，Vol.43Issue(4)：588-595,8.
小反刍兽疫病毒核蛋白单克隆抗体的制备与初步应用
Preparation and Preliminary Application of Monoclonal Antibodies against Nucleoprotein of Peste des Petits Ruminants Virus
摘要
Abstract
The objective of this study was to prepare specific monoclonal antibodies (mAbs) against nucleoprotein of peste des petits ruminants virus (PPRV) , characterize its properties, and use it in ELISA. After immunization of BALB/c mice with purified recombinant protein Bac-mid-PPRV-N, two monoclonal antibodies against PPRV nucleoprotein were successfully prepared by polyethyleneglycol (PEG)-mediated fusion of sensitized lymphocytes and myeloma cells and named 5B11 and 3H10-3B8, respectively. The specificity and biological characterization of the mAbs were identified. A competitive ELISA using PPRV recombinant nucleoprotein as coating antigen and mAb 5B11 as competitive antibody was established. Our results about isotypes and subclasses indicate that 5B11 and 3H10-3B8 belong to IgG2b, the titres of ascitic fluids of 5B11 was up to 1:819 200 and 3H10-3B8 was 1 : 12 800. These mAbs could specifically recognize recombinant protein Bacmid-PPRV-N antigens in a serologic test. Antibodies additivity assay demonstrated that 5B11 and 3H10-3B8 recognized the different epitopes of PPRV nucleoprotein. The number of chromosome of the hybridoma cell lines was 99 to 104. A total of 222 serum samples were detected in parallel by c-ELISA and reference c-ELISA kit. The coincidence rate of the two assaya was 98.15%. We successfully prepared two specific monoclonal antibodies against PPRV nucleoprotein and established the competitive immunofluorescent method for detecting PPRV.关键词
小反刍兽疫/单克隆抗体/生物学特性,竞争ELISAKey words
peste des petits ruminants virus/ monoclonal antibody/ biological characteristic/ competitive ELISA分类
农业科技引用本文复制引用
龙云凤,陈朝银,花群义,杨仕标,尹尚莲,徐维佳,艾军,周晓黎,杨俊兴,王琼,祝贺,叶玲玲,董俊,吕建强,王金萍..小反刍兽疫病毒核蛋白单克隆抗体的制备与初步应用[J].畜牧兽医学报,2012,43(4):588-595,8.基金项目
国家高新技术研究发展计划(863计划)项目(2012AA100501) （863计划）
国家质检总局科技计划项目(2007IK025) （2007IK025）
