Ohe Y, Nishino T, Shin S, Hirano H, Higashino K, Amuro Y, Tamura S, Hada T, Fujioka H
Otsuka Assay Laboratories, Otsuka Pharmaceutical Co. Ltd., Tokushima, Japan.
J Immunoassay. 1990;11(4):459-75. doi: 10.1080/01971529008055045.
Pseudouridine, a component of tRNA, was modified to yield the derivatives: succinyl, palmitoyl pseudouridine, and protein conjugates. These derivatives were used in preparation of monoclonal antibodies specifically directed to pseudouridine. MAbs from three hybridomas (OAL 881, 812 and 814) were established and shown to be directed to pseudouridine, uridine and uracil. OAL 881 was equally reactive with the three substrates, OAL 812 showed the same reactivity for pseudouridine and uracil, while OAL 814 showed a reactivity mainly for pseudouridine. MAb OAL 814 was used in a radioimmunoassay (RIA) system unique for pseudouridine. A good dose-response curve was observed in the range between 31.3 and 2000 nmol/ml. Intra- and inter-assay CV values were below 4.1% and 7.4% respectively, and good results were obtained from recovery of added material and dilution tests. The ratio of pseudouridine/creatinin in urine was significantly higher in patients with cancer than in normal subjects.
假尿苷是转运核糖核酸（tRNA）的一个组成部分，经过修饰后产生了以下衍生物：琥珀酰化假尿苷、棕榈酰化假尿苷以及蛋白质缀合物。这些衍生物被用于制备特异性针对假尿苷的单克隆抗体。建立了来自三种杂交瘤（OAL 881、812和814）的单克隆抗体，并证明它们针对假尿苷、尿苷和尿嘧啶。OAL 881与这三种底物的反应性相同，OAL 812对假尿苷和尿嘧啶表现出相同的反应性，而OAL 814主要对假尿苷有反应性。单克隆抗体OAL 814被用于一种针对假尿苷的独特放射免疫分析（RIA）系统。在31.3至2000 nmol/ml的范围内观察到了良好的剂量反应曲线。批内和批间变异系数（CV）值分别低于4.1%和7.4%，并且从添加物质的回收率和稀释试验中获得了良好的结果。癌症患者尿液中假尿苷/肌酐的比值显著高于正常受试者。
