抗华支睾吸虫代谢抗原单克隆抗体的制备及鉴定研究

Preparation and characterization of monoclonal antibodies against the metabolic antigen of Clonorchis sinensis

目的建立分泌抗华支睾吸虫代谢抗原的单克隆抗体杂交瘤细胞株。方法用华支睾吸虫成虫代谢抗原免疫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全虫可溶性抗原、卫氏并殖吸虫成虫抗原和猪囊尾蚴抗原的交叉反应。结果获得3株分泌高滴度抗华支睾吸虫代谢抗原的杂交瘤细胞株,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,3株单抗均属IgG。结论制备的抗华支睾吸虫代谢抗原的杂交瘤细胞株能分泌高滴度和高特异性的单抗,为制备免疫诊断试剂盒奠定了基础。

To develop and identify monoclonal antibody(McAb) against the metabolic antigen of Clonorchis sinensis, BALB/c mice were immunized with metabolic antigen of Clonorchis inensis, and 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. And the antigens recognized by the McAbs were identified with immunoblot analysis. The cross reactions between the McAbs and antigens of Schistosoma japonicum, Paragonimus westermani and Cysticercus cellulosae were detected. Three hybridoma cell lines secreting high titer of McAbs agains metabolic antigen of Clonorchis sinensis were established. All three clones of McAbs belonged to IgG subtype. The McAbs had no cross reactions with antigens of Schistosoma japonicum, Paragonimus westermani and Cysticercus cellulosae. It is concluded that 3 hybridoma cell lines against the metabolic antigen of Clonorchis sinensis secreting high titer of McAbs with high specificity were established.