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目的 制备具有广谱种属反应性的PrP单克隆抗体 (McAb) ,用于可传播性海绵样脑病(TSE)的诊断及致病机制研究。方法 分别将对应于牛PrP2 9～ 4 8(BoP1 )、PrP89～ 10 8(BoP2 )的两种多肽与匙孔血蓝蛋白 (KLH)偶联 ,免疫BALB c小鼠 ,经细胞融合后获得分泌针对上述两种多肽的杂交瘤细胞株 ,用Western blot方法检测这些细胞株分泌的McAb与牛 (Bo)、人 (Hu)和仓鼠 (Ha)PrP蛋白的反应性。结果 通过细胞融合和 2～ 3轮克隆化 ,用ELISA筛选出分泌抗BoP1 和BoP2 抗体的杂交瘤细胞株D1 1 和D8。Westernblot显示 ,获得的McAb均能分别与重组BoPrP(2 5～ 2 4 2 )、重组HuPrP(2 3～ 2 31)和HaPrP(2 3～ 2 31)反应。结论 获得了可与牛、人和仓鼠PrP反应的两种McAb ,可用于哺乳类PrP检测及TSE致病机制研究。
Objective To prepare a monoclonal antibody (McAb) against PrP with broad spectrum species-reactivity for the diagnosis and pathogenesis of transmissible spongiform encephalopathy (TSE). Methods The two polypeptides corresponding to the PrP2 9 ~ 48 (BoP1) and PrP89 ~ 108 (BoP2) were conjugated to the keyhole limpet haemocyanin (KLH) respectively and immunized BALB mice. After fusion, The hybridoma cell lines secreting these two polypeptides were secreted and the reactivity of McAbs secreted by these cell lines with the Bo, Hu and PrP proteins was examined by Western blot. Results The hybridoma cell lines D1 1 and D8 secreting anti-BoP1 and BoP2 antibodies were screened by ELISA by cell fusion and 2 to 3 rounds of cloning. Westernblot showed that McAbs were able to react with recombinant BoPrP (25 ~ 24 2), recombinant HuPrP (23 ~ 231) and HaPrP (23 ~ 231) respectively. Conclusions Two McAbs that can react with PrP in cattle, humans and hamsters were obtained and could be used for the detection of mammalian PrP and the pathogenesis of TSE.