Chan M A, Bellem A C, Diamandis E P
CyberFluor Inc., Toronto, Ontario, Canada.
Clin Chem. 1987 Nov;33(11):2000-3.
We describe a new "sandwich"-type nonisotopic immunoassay of alpha-fetoprotein (AFP) in serum and amniotic fluid. In the assay, AFP binds to a monoclonal antibody immobilized in a microtiter well and to a polyclonal soluble biotinylated antibody. A fluorescent product is created on the solid-phase after adding streptavidin labeled with a new Eu3+ chelate, 4,7-bis(chlorosulfophenyl)-1,10-phenanthroline-2,9-dicarboxylic acid (BCPDA), and excess Eu3+. The fluorescent complex, monoclonal antibody-AFP-polyclonal antibody-biotin-streptavidin-BCPDA-Eu3+, is quantified by nitrogen laser excitation at 337.1 nm, the emission at 615 nm being monitored in an especially designed gated fluorometer working in a time-resolved mode. This assay of AFP has a broad dynamic range (up to 1 mg/L), and is precise and accurate. The detection limit is approximately 0.1 microgram/L. Results agree well with those obtained by established techniques.
我们描述了一种用于血清和羊水甲胎蛋白（AFP）检测的新型“夹心”式非同位素免疫分析方法。在该分析中，AFP与固定于微量滴定孔中的单克隆抗体以及多克隆可溶性生物素化抗体相结合。加入用新型铕（Eu3+）螯合物4,7-双（氯磺苯基）-1,10-菲咯啉-2,9-二羧酸（BCPDA）标记的链霉亲和素和过量的Eu3+后，在固相中产生荧光产物。通过在337.1nm处用氮激光激发对荧光复合物（单克隆抗体-AFP-多克隆抗体-生物素-链霉亲和素-BCPDA-Eu3+）进行定量，在特别设计的以时间分辨模式工作的门控荧光计中监测615nm处的发射光。这种AFP分析方法具有较宽的动态范围（高达1mg/L），且精确、准确。检测限约为0.1μg/L。结果与通过既定技术获得的结果非常吻合。
