两种分泌抗PrP单克隆抗体杂交瘤细胞株的建立及其初步鉴定  

Establishment and preliminary characterization of hybridoma cell lines secreting monoclonal antibodies against Prion Proteins

作　　者：赵丽[1] 计融[2] 王健伟[1] 韩春卉[2] 于修平[3] 董小平[1] 洪涛[1] 

机构地区：[1]中国疾病预防控制中心病毒病预防控制所洪涛院士实验室 [2]中国疾病预防控制中心病毒病预防控制所营养与食品安全所生物污染监控室,北京100052 [3]山东大学医学院微生物教研室

出　　处：《中华实验和临床病毒学杂志》2003年第2期133-136,共4页Chinese Journal of Experimental and Clinical Virology

基　　金：国家高技术应用发展项目资助 ( 2 0 0 1)

摘　　要：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 obtain monoclonal antibodies (McAbs) which can be widely used to detect mammalian prions (PrP) and to develop diagnostic tests for screening transmissile spongiform encephalopathies (TSE) as well as for studying pathogenesis of prion related diseases Methods BALB/c mice were immunized separately with bovine PrP peptide 29 48 (BoP 1) and 89 108 (BoP 2) coupled to keyhole limpt hemocyan Two hybridoma cell lines secreting monoclonal antibodies against these peptides were established by cell fusion and 2 to 3 rounds of cell cloning The reactions of the McAbs to the recombinant bovine (Bo)PrP(25 242), human (Hu)PrP(23 231) and hamster (Ha) PrP (23 231) were tested separately by Western bloting Results Through cell fusion, two hybridoma cell lines secreting McAbs against BoP 1 and BoP 2, designated D 11 and D 8 accordingly, were identified by ELISA and cell cloning The McAbs produced by these cell lines reacted well with the recombinant PrP proteins; (Bo) PrP (25 242), (Hu) PrP (23 231), and (Ha) PrP (23 231), respectively Conclusion Two McAbs reacting with bovine, human and hamster PrPs were successfully generated, they are potential to be used to detect PrPs in mammals and to study the mechanism of pathogenesis of TSE

关 键 词：PRP 单克隆抗体 杂交瘤 细胞株 McAb 可传播性海绵样脑病 诊断 免疫印迹法 朊病毒 

分 类 号：R392[医药卫生—免疫学]