抗大片吸虫ES抗原单克隆抗体的制备及特异性鉴定  

Production and characterization of monoclonal antibodies against excretory secretary antigen of Fasciola Gigantica

作　　者：沈涛[1] 沈永林[1] 何国声[2] 徐梅倩[2] 顾越星[2] 

机构地区：[1]南京农业大学动物医学院,南京210095 [2]中国农业科学院上海家畜寄生虫病研究所,上海200232

出　　处：《南京农业大学学报》2001年第3期81-84,共4页Journal of Nanjing Agricultural University

基　　金：欧洲联盟资助国际合作项目 (INCO -DC ;IC18-CT95 0 0 0 1)

摘　　要：用大片吸虫分泌排泄 (ES)抗原免疫BALB/c小鼠 ,取其脾细胞与SP2 / 0骨髓瘤细胞融合 ,经筛选和 3次克隆化培养后获得 2株分泌抗大片吸虫ES抗原单抗的杂交瘤细胞 (F5,F9)。经传代、冻存、复苏培养检测和染色体检查 ,证实其分泌单抗的性质稳定。通过间接ELISA试验证明 ,F5和F9单抗能与大片吸虫、肝片吸虫的ES抗原发生特异性反应 ,而不与它们的虫体抗原反应。SDS PAGE电泳和Western blot显示 ,F5McAb能与ES抗原的相对分子质量为 2 6 0 0 0～ 2 80 0 0的蛋白条带反应 ,而不与其可溶性虫体抗原反应。表明F5McAb有ES抗原特异性 ,是一株具有潜在诊断价值的McAb。Two hybridoma strains secreted monoclonal antibodies(F 5,F 9)against Fasciola gigantica excretory secretary antigen(ESAg)were screened.Specificity of the McAbs was evaluated by indirect ELISA antigens of F.gigantica and F.hepatica,body extract antigen(BA) of them and Schistosoma japonicum body antigen.McAbs(F 5,F 9)had reacted with the ES products from F.gigantica and F.hepatica,but did not with BA of F.gigantica,F.hepatica and S.japonicum.The specificity of the McAb was evaluated by Western blot.F 5 McAb had reacted with 26 000?28 000 protein bands of the ES products from F.gigantica,as well as the ES products from F.hepatica,but did not with BA of F.gigantica and F.hepatica.It demonstrated that the F 5 McAb could be as diagnostic reagent for detecting fasciolosis.

关 键 词：大片吸虫 分泌排泄抗原 单克隆抗体 ES抗原 

分 类 号：S852.735[农业科学—基础兽医学]