Lieu T S, Jiang M, Steigerwald J C, Tan E M
J Immunol Methods. 1984 Jul 6;71(2):217-28. doi: 10.1016/0022-1759(84)90068-1.
Autoantibodies to SS-A/Ro antigen have been described in the sera of certain patients with Sjögren's syndrome, systemic lupus erythematosus and in neonates with lupus and/or congenital heart block. SS-A/Ro is a trypsin-sensitive intracellular antigen which appears to be associated with another intracellular antigen SS-B/La in tissue extracts. This study describes a simple method of separation of SS-A/Ro and SS-B/La antigenic activity by differential salt elution in polybuffer ion exchange chromatography. The partially purified SS-A/Ro antigen was separated by polyacrylamide gel electrophoresis under non-denaturing conditions and identified by Western blot analysis as a single polypeptide of 61,000 Da. The pI of SS-A/Ro antigen was 4.67. The partially purified SS-A/Ro antigen derived from gel electrophoresis could be adapted to an enzyme linked immunosorbent assay (ELISA) for the detection of antibody.
在某些干燥综合征、系统性红斑狼疮患者以及患有狼疮和/或先天性心脏传导阻滞的新生儿血清中,已发现针对SS - A/Ro抗原的自身抗体。SS - A/Ro是一种对胰蛋白酶敏感的细胞内抗原,在组织提取物中似乎与另一种细胞内抗原SS - B/La相关。本研究描述了一种通过在多缓冲离子交换色谱中进行差异盐洗脱来分离SS - A/Ro和SS - B/La抗原活性的简单方法。部分纯化的SS - A/Ro抗原在非变性条件下通过聚丙烯酰胺凝胶电泳分离,并通过蛋白质印迹分析鉴定为一条61,000道尔顿的单一多肽。SS - A/Ro抗原的等电点为4.67。源自凝胶电泳的部分纯化的SS - A/Ro抗原可用于酶联免疫吸附测定(ELISA)以检测抗体。