摘要
按katendel氏法用伊氏锥虫JG株制备全虫抗原,以其免疫BALB/C小鼠。以50％PEG(MW4000,pH8.0)作融合剂。使免疫鼠脾细胞与S P2/0 Ag14(简称S P2/0)骨髓瘤细胞融合。经系统筛选和克隆化,建立了9个分泌抗伊氏锥虫抗体的杂交瘤细胞株。他们分泌的McAb,4株为IgG1、1株为IgG3、另4株为IgM。9株McAb在琼脂免疫双扩散试验中均不形成沉淀线。用ELISA检测,这些细胞系培养上清和腹水与同源伊氏锥虫虫株抗原的滴度分别为1:64-1:1024和1:6.4×103-1:6.4×105;与泰氏锥虫和弓形虫抗原则全为阴性。用2个异源伊氏傩虫虫株抗原检测时,8株无任何交叉反应,8株与2个异源株均有交叉反应,另3株则只与1个异源株有交叉反应。用增值反应检查了5株IgG类McAb识别抗原决定簇的特异性,除2株共同识别一种抗原决定簇外,另8株各自识别不同的抗原决定簇。
SP2/0 myeloma cells were fused with spleen cells taken from BALB/C mice inmunized with intact trypanosoma antigen of Trypanosoma evansi JG strain. Nine hybridoma cell strains stably secreting specific an tibodies to T.evansi were obtained after systematically screening and cloning. Of these McAbs, 4 belonged to IgG1, 1 was IgG3 and others, IgM. No pre cipitatory line was formed in double immunodiffusion test with any of these McAbs. All the McAbs reacted with antigen of homologous strain of T. evansi and did not react with Trypanosoma theileri and Toxoplasma gondii antigens in ELISA. Titers of the 9 McAbs from culture supernatantand ascites fluid were 1 :64-1 :1024 and 1 :6.4×103-1 :6.4×106respectively. Of the 9 McAbs, 3 did not react and 3 reacted with antigens of 2 heterologous strains of T. evansi, and others reacted with antigen of 1 heterologous strain of T. evansi only. Antigenic determinant specificity of 5 McAbs of IgG class was determined by aiditivity test, 2 McAbs recognized same antigenic determinant and 3 recognized other 3 different, antige nic determinants.
关键词
伊氏锥虫
单克隆抗体
杂交瘤细胞株
Trypanosorna evansi
monoclonal antibody, hybridoma cell line
additivity testj additivity index
