抗隐孢子虫子孢子表膜单克隆抗体的制备及鉴定

PREPARATION AND CHARCTERIZATION OF MONOCLONAL ANTIBODIES AGAINST SURFACE ANTIGENS OF CRYPTOSPORIDIUM PARVUM SPOROZOITES

利用牛源微小隐孢子虫 (Cryptosporidiumparvum)超声粉碎物免疫BALB C小鼠 ,取脾细胞与SP2 0细胞融合 ,经 3～ 5次有限稀释克隆化 ,获得 4株持续分泌抗体的杂交瘤细胞株 ,分别命名为 1D5 ,2E9,3D6和 4G4。经检测 ,其分泌的抗体亚类 3株为IgG1,1株为IgM。通过免疫荧光鉴定 ,4株单抗均作用于子孢子抗原 ,其中 2株针对子孢子表膜。对杂交瘤细胞株的染色体计数 ,结果染色体均数为 90～ 98条。ELISA检测细胞培养上清效价分别为 1∶16 0 ,1∶32 0 ,1∶32 0和 1∶6 4 0 ,诱生小鼠腹水的抗体效价分别为 1∶4 0 9,6 0 0 ,1∶819,2 0 0 ,1∶819,2 0 0和 1∶1,6 38,4 0 0 ,特异性试验结果显示 ,4株单抗均不与贾第虫滋养体和结肠小袋纤毛虫发生交叉反应。

Four hybirdoma (1D5,2E9,3D6 and 4G4) producing McAbs against occyst antigens of Cryptosporidium parvum were developed by fusion of SP2/0 mouse myeloma cells and spleen cells from BALB/C mice immunized with sonicated oocysts.The immunoglobulin subclasses of these four McAbs were identified as IgG1(1D5,3D6,4G4) and IgM(2E9). All McAbs were reacted with the sporozoites of C parvum by indirect immunofluorescent assay,two of which were reacted with surface of sporozoites.The number of chromosome of the hybirdoma cell line was 90 to 98.The ELISA titers of acites were 1∶409,600,1∶819,200,1∶819,200 and 1∶1,638,400,and the ELISA titers of cell supernatant were 1∶160 ,1∶320,1∶320 and 1∶640 respectively.Four McAbs did not react at all with Giardia lambila and Balantidium sp.