水稻条纹叶枯病毒单克隆抗体的制备及其特性研究  

Production of a monoclonal antibody to rice stripe virus and research of its characteristics

作　　者：戈林泉[1,2] 赵伟春[3] 祝树德[2] 娄永根[1] 

机构地区：[1]浙江大学昆虫科学研究所,浙江杭州310029 [2]扬州大学园艺与植物保护学院,江苏扬州225009 [3]浙江中医药大学生物工程学院,浙江杭州310053

出　　处：《扬州大学学报（农业与生命科学版）》2010年第1期76-80,共5页Journal of Yangzhou University：Agricultural and Life Science Edition

基　　金：浙江省自然科学基金资助项目(2006C22003;2006C30040);杭州市科技发展计划项目(20051332B31)

摘　　要：以提纯的水稻条纹叶枯病毒(RSV)粒子作为抗原免疫BALB/c小鼠脾细胞,并与SP2/0骨髓瘤细胞融合,经克隆筛选,获得1株能稳定分泌针对RSV蛋白的单克隆抗体的杂交瘤细胞株1H2。利用该细胞株制备了腹水型单克隆抗体,经检测,该单克隆抗体的间接ELISA效价达1∶1.024×106,且与水稻、灰飞虱、褐飞虱和白背飞虱的蛋白均不发生交叉反应,检测的灵敏度为抗原蛋白稀释1∶2.14×104倍,对应的抗原浓度为25.89μg.L-1。按常规饱和硫酸铵沉淀法纯化了单克隆抗体1H2,测定其IgG含量为6.05 g.L-1。该单克隆抗体可用于监测田间RSV的发生动态。One cell strain by hybridoma bridoma cell linesecreting monoclonal antibodies （McAbs） against RSV was produced by fusing mouse myeloma cells （SP2/0） with spleen cells from BALB/c immunized by the purification RSV particles, which was named 1 HZ. We prepared ascite fluids types of McAbs with 1HZ. Based on 1 HZ, a McAb against RSV was developed, and the titer of the McAb was 1 ： 1. 024×10^6 , when tested by indirect ELISA. The McAbs didn＇t cross- react with proteins extracted from Oryza sativa , Niaparvata lugens （Still）, Laodelphax stratells and sogatella furcifera （Horvath） virus. Dectection of sensitivity was antigen protein 1 ： 2.14× 10^4 dilution, and corresponding antigen chroma was 25.89 μg· L^- 1. The monoclonal antibody 1H2 was purified by saturated （NH4）2SO4. The content of IgG was 6.05 g · L^- 1. The results suggest that the McAb antibody could be used to detect the dynamic of RSV in the field.

关 键 词：灰飞虱 水稻条纹叶枯病毒 单克隆抗体 ELISA 

分 类 号：S432.41[农业科学—植物病理学]