Frost S J, Chakraborty J, Firth G B
Department of Clinical Biochemistry, Princess Royal Hospital, West Sussex, UK.
J Immunol Methods. 1996 Aug 14;194(2):105-11. doi: 10.1016/0022-1759(96)00057-9.
A homogeneous colorimetric immunoassay which has been developed for urinary microalbumin utilizes complement-mediated immunolysis of liposomes containing the dye, sulphorhodamine B. Unlike a previously described model complement-mediated liposomal assay for serum albumin (Frost et al., 1994) which was competitive, this assay uses a sandwich-type format and Fab' (antialbumin)-coated liposomes to increase the assay sensitivity. The liposomal assay, performed using a Cobas Bio analyser (Roche, Welwyn Garden City, UK), gave an acceptable correlation with a radioimmunoassay (NETRIA, London, UK): r = 0.94; y (liposomal assay) = 1.09 x (radioimmunoassay) - 1.54 mg/1. The imprecisions of the assays were similar and matrix effects due to the use of urine samples were determined to be acceptably small. The assay demonstrates the advantage of using Fab'-coated liposomes in sandwich-type liposomal immunoassays over liposomes coated with intact antibody, which failed to elicit complement-mediated immunolysis.
一种已开发用于检测尿微量白蛋白的均相比色免疫测定法,利用补体介导的对含有染料磺罗丹明B的脂质体的免疫溶解作用。与先前描述的用于血清白蛋白的补体介导脂质体测定模型(Frost等人,1994年)不同,该模型是竞争性的,此测定法采用夹心型模式和Fab'(抗白蛋白)包被的脂质体以提高测定灵敏度。使用Cobas Bio分析仪(罗氏公司,英国韦林花园城)进行的脂质体测定法与放射免疫测定法(英国伦敦NETRIA)具有可接受的相关性:r = 0.94;y(脂质体测定法)= 1.09x(放射免疫测定法)- 1.54mg/1。两种测定法的不精密度相似,并且确定由于使用尿液样本导致的基质效应小到可接受。该测定法证明了在夹心型脂质体免疫测定中使用Fab'包被的脂质体相对于完整抗体包被的脂质体的优势,完整抗体包被的脂质体无法引发补体介导的免疫溶解。