Netter H J, Guldner H H, Szostecki C, Lakomek H J, Will H
Max Planck Institut für Biochemie, Martinsried, Federal Republic of Germany.
Arthritis Rheum. 1988 May;31(5):616-22. doi: 10.1002/art.1780310506.
A human liver complementary DNA expression library was screened using sera from patients with high titers of autoantibodies, to search for clones expressing major autoantigens that are relevant in connective tissue diseases. One of the clones isolated expressed a major epitope(s) that was immunoreactive with anti-U1 RNP sera, as shown by several techniques. Affinity-purified autoantibodies from the cloned RNP protein specifically recognized the 68-kd U1 RNP protein of HeLa cell nuclear extracts. All sera containing anti-U1 RNP antibodies detected by immunodiffusion, counterimmunoelectrophoresis, or immunoblotting also recognized the cloned RNP protein. The RNP antigen-expressing bacterial colonies and the partially purified cloned RNP fusion protein have been applied to fast and sensitive immunologic assays for the detection and quantification of anti-U1 RNP antibodies.
利用高滴度自身抗体患者的血清筛选人肝脏互补DNA表达文库,以寻找表达与结缔组织病相关的主要自身抗原的克隆。通过多种技术表明,分离出的一个克隆表达了一个主要表位,该表位与抗U1 RNP血清发生免疫反应。从克隆的RNP蛋白中亲和纯化的自身抗体特异性识别HeLa细胞核提取物中的68-kd U1 RNP蛋白。所有通过免疫扩散、对流免疫电泳或免疫印迹检测到的含有抗U1 RNP抗体的血清也识别克隆的RNP蛋白。表达RNP抗原的细菌菌落和部分纯化的克隆RNP融合蛋白已应用于快速灵敏的免疫测定,用于检测和定量抗U1 RNP抗体。