分泌抗包虫囊液抗原McAb杂交瘤细胞株的建立

Establishment of Hybridoma Cell Lines Secreting McAbs Against Echinococcus Granulosus Cyst Fluid Antigens

应用Oriel法制备的包虫囊液抗原经腹腔注入供实验的小鼠,作为脾细胞供体。此脾细胞与不能分泌的骨髓瘤细胞株SP2/0和P3u_1相融合,以生成杂交瘤细胞株。用ELISA法从这些混合细胞培养基中筛选出上清液,用稀释法将上清液克隆,再克隆化之后,生成能分泌单克隆抗体、抗包虫囊液活性的杂交瘤,其染色体数目在80～100范围之内。单克隆抗体类及亚类13Fs和31G_(12)分别是IgG_1和IgA。DD和ELISA用于鉴定它们的敏感性和特异性。应用ELISA法比较杂交瘤和绦虫囊尾蚴液等其它寄生虫抗原制备物,包虫囊液和Pf抗原,表明单克隆抗体对包虫囊寄生虫有很高的特异性。

Mice experimentally immunized by intraperitoneal injection of Echinococcus granulosus cyst fluid antigens (E.g CF) prepared by the method described by Oriel were used as spleen cell donors,the spleen cells were fused to the non-secreting myeloma cell lines SP 2/0 and P3U1 for hybridoma production.Su-pernants from those hybrid cell cultures were screend by ELISA and those hybrids of which supernants were positive in this assay were cloned and recloned by the technique of limited dilution,after that,two hybridomas (13F5,31G12) were produced which secreted mcAb anti-E.gCF activity and their chromosome numbers were demonstrated in the range of 80-100.The class and subclass of McAb 13F5 and 31G12 are IgG1 and IgM,respectively.DD and ELISA were used for identifying their sensitivity and specificity.ELISA comparing the binding of these hybridoma derived antibodies versus several other parasite antigen preparations from Taenia solium cysticerci fluid,Echinococcus multilocularis cyst fluid and P.f antigen showed that the McAbs had a high degree of specificity for the hydatid parasite.