Toivonen E, Hemmilä I, Marniemi J, Jørgensen P N, Zeuthen J, Lövgren T
Clin Chem. 1986 Apr;32(4):637-40.
We describe a two-site "sandwich"-type time-resolved immunofluorometric assay for human insulin, based on use of two monoclonal antibodies with different specificities. The first antibody is immobilized on the surface of microtiter plate strip wells, the other is labeled with Eu3+. Serum samples can be assayed with one incubation step; two incubation steps are required when plasma samples are assayed. After the immunoreactions are complete, the bound fraction of Eu3+-label is quantified by dissociating it in a fluorescence-enhancement solution and measuring its fluorescence with a fluorometer with time-resolution. The sensitivity of the assay is 0.24 micro-int. units/mL. The standard curve is linear from 0.24 to 2400 micro-int. units/mL.
我们描述了一种用于人胰岛素的双位点“夹心”型时间分辨免疫荧光分析法，该方法基于使用两种具有不同特异性的单克隆抗体。第一种抗体固定在微量滴定板条孔的表面，另一种用Eu3+标记。血清样品可通过一步温育进行检测；检测血浆样品时需要两步温育。免疫反应完成后，通过在荧光增强溶液中解离Eu3+标记的结合部分并用具有时间分辨率的荧光计测量其荧光来定量。该分析方法的灵敏度为0.24微国际单位/毫升。标准曲线在0.24至2400微国际单位/毫升范围内呈线性。
