摘要
目的:建立分泌抗华支睾吸虫成虫的单克隆抗体杂交瘤细胞株并进行鉴定。方法:用华支睾吸虫成虫可溶性抗原免疫BALB/c小鼠,取其脾细胞与小鼠骨髓瘤细胞SP2/0融合,筛选分泌高滴度单克隆抗体的杂交瘤细胞株,测定单抗免疫球蛋白亚类和单抗效价,检测单抗与日本血吸虫虫卵抗原、卫氏并殖吸虫成虫抗原和猪囊尾蚴抗原的交叉反应,用IFAT进行单抗识别的抗原定位。结果:获得5株分泌高滴度抗华支睾吸虫成虫单抗的杂交瘤细胞株,其分泌的抗体与日本血吸虫、卫氏并殖吸虫和猪囊尾蚴抗原均不发生交叉反应;5株单抗均属IgM;单抗所识别的抗原定位于华支睾吸虫肠管壁。结论:制备的抗华支睾吸虫成虫的杂交瘤细胞株能分泌高滴度和高特异性的单抗。
AIM:To develope and identify monoclonal antibody(McAb) against adult Clonorchis sinensis. METHODS:BALB/c mice were immunized with soluble antigen of adult Clonorchis sinensis ,the spleen cells of immunized mice were fused with myeloma cell line SP2/0.Then the hybridoma cell lines secreting high titer of McAbs were screened.The Ig subtypes and titer of the McAbs were assayed with ELISA.The antigens recognized by the McAbs were examined with IFAT and identified by immunoblot analysis.The cross reactions between the McAbs and antigens of Schistosoma japonicum,Paragonimus westermani and Cysticercus cellulosae were detected.RESULTS:5 hybridoma cell lines secreting high titer of McAbs against adult Clonorchis sinensis were established.All five clones of McAbs belonged to IgM subtype.The antigens recognized by the McAbs were located at the intestinal tract wall of adult Clonorchis sinensis. The McAbs had no cross reactions with antigens of Schistosoma japonicum,Paragonimus and Cysticercus cellulosae .CONCLUSION:5 hybridoma cell lines against adult Clonorchis sinensis secreting high titer of McAbs with high specificity were established.
出处
《中国寄生虫学与寄生虫病杂志》
CAS
CSCD
北大核心
1998年第1期42-44,共3页
Chinese Journal of Parasitology and Parasitic Diseases