Kakabakos S E, Christopoulos T K, Diamandis E P
Department of Clinical Biochemistry, Toronto Western Hospital, Ontario, Canada.
Clin Chem. 1992 Mar;38(3):338-42.
We describe a new multianalyte immunoassay principle and apply it to the simultaneous immunoassay of lutropin, follitropin, choriogonadotropin, and prolactin in serum. The method is based on the coating of distinct areas of polystyrene with analyte-specific antibodies. These antibodies react with the analyte and immobilize it in a specific area while another biotinylated antibody also reacts with the analyte to form a sandwich. After addition of streptavidin labeled with the fluorescent europium chelate of 4,7-bis(chlorosulfophenyl)-1,10-phenanthroline-2,9-dicarboxylic acid, fluorescent areas are formed, the intensity of which is related to the amount of each analyte present in the sample. The fluorescent areas are quantified on the dry solid phase with laser-excited time-resolved fluorometric measurements. The assays developed are highly sensitive, precise, and accurate. We believe that this system shows potential for multianalyte immunoassay of diverse groups of compounds in disciplines such as endocrinology, infectious disease, hematology, and oncology.
我们描述了一种新的多分析物免疫测定原理，并将其应用于血清中促黄体生成素、促卵泡生成素、绒毛膜促性腺激素和催乳素的同时免疫测定。该方法基于用分析物特异性抗体包被聚苯乙烯的不同区域。这些抗体与分析物反应并将其固定在特定区域，而另一种生物素化抗体也与分析物反应形成夹心结构。加入用4,7-双（氯磺苯基）-1,10-菲咯啉-2,9-二羧酸的荧光铕螯合物标记的链霉亲和素后，形成荧光区域，其强度与样品中每种分析物的量相关。通过激光激发时间分辨荧光测量对干燥固相上的荧光区域进行定量。所开发的测定方法具有高灵敏度、精密度和准确性。我们认为，该系统在内分泌学、传染病学、血液学和肿瘤学等学科中对多种化合物进行多分析物免疫测定显示出潜力。
