Cui J, Bystryn J C
Department of Dermatology, New York University School of Medicine, NY 10016.
J Immunol Methods. 1992 Feb 14;147(1):13-9. doi: 10.1016/s0022-1759(12)80023-8.
An improved assay for complement-mediated cytolysis is described. The target cells are labeled with europium complexed to diethylenetriaminopentaacetate (Eu-DTPA). Cytolysis caused by antibody plus complement leads to the release of the Eu-DTPA complex which is then formed into a highly fluorescent chelate by the addition of 2-naphthoyltrifluoroacetone (2-NTA). The amount of europium chelate formed--a measurement of cell death--is then quantified with a time-resolved fluorometer. The results of the assay are reproducible. Complement-mediated cytolysis when measured by europium release was five times more sensitive than when measured by conventional 51Cr release and three times than when measured by trypan blue exclusion. Because europium does not decay, target cells can be labelled in batches and stored frozen until use, which speeds and simplifies the assay. Thus, europium release assay is a simple and quantitative method to measure complement-mediated cytolysis which is sensitive and more rapid than conventional assays.
本文描述了一种改进的补体介导的细胞溶解检测方法。靶细胞用与二乙烯三胺五乙酸络合的铕（Eu-DTPA）进行标记。抗体加补体引起的细胞溶解导致Eu-DTPA复合物的释放，然后通过加入2-萘甲酰三氟丙酮（2-NTA）形成高荧光螯合物。然后用时间分辨荧光计对形成的铕螯合物量（细胞死亡的一种测量方法）进行定量。该检测结果具有可重复性。通过铕释放测量的补体介导的细胞溶解比通过传统的51Cr释放测量时敏感五倍，比通过台盼蓝排斥测量时敏感三倍。由于铕不会衰变，靶细胞可以批量标记并冷冻保存直至使用，这加快并简化了检测过程。因此，铕释放检测是一种简单且定量的方法，用于测量补体介导的细胞溶解，它比传统检测方法更敏感、更快速。
