Fu Zhifeng, Hao Chen, Fei Xiaoqing, Ju Huangxian
Key Laboratory of Analytical Chemistry for Life Science (Ministry of Education of China), Department of Chemistry, Nanjing University, Nanjing 210093, PR China.
J Immunol Methods. 2006 May 30;312(1-2):61-7. doi: 10.1016/j.jim.2006.02.006. Epub 2006 Mar 6.
A flow-injection chemiluminescent immunoassay system based on a novel transparent immunoaffinity reactor is proposed for the quantitation of alpha-fetoprotein. The reactor prepared with alpha-fetoprotein immobilized epoxysilane modified glass microbeads was used as an immunosensor for chemiluminescent detection. With a non-competitive immunoassay format, the proposed immunosensor system is a low cost, flexible and rapid assay for alpha-fetoprotein. After an off-line incubation of the analyte alpha-fetoprotein with horseradish peroxidase-labeled alpha-fetoprotein antibody as enzyme tracer, the mixture was injected into the reactor, which led to trapping of the free enzyme tracer by the reactor. The trapped enzyme tracer was detected by the p-iodophenol-luminol-H2O2 chemiluminescence system. Under optimal conditions, the decrease in chemiluminescence intensity was proportional to the alpha-fetoprotein concentration in the range of 5.0-100 ng/ml with a detection limit of 2.7 ng/ml at a signal/noise ratio of 3. The immunosensor system showed an acceptable reproducibility and stability. Clinical serum samples were assayed with this method and the results were in acceptable agreement with those obtained from immunoradiometric assay.
提出了一种基于新型透明免疫亲和反应器的流动注射化学发光免疫分析系统，用于定量检测甲胎蛋白。用固定有甲胎蛋白的环氧硅烷修饰玻璃微珠制备的反应器用作化学发光检测的免疫传感器。所提出的免疫传感器系统采用非竞争性免疫分析形式，是一种用于甲胎蛋白的低成本、灵活且快速的检测方法。将分析物甲胎蛋白与辣根过氧化物酶标记的甲胎蛋白抗体作为酶标记物进行离线孵育后，将混合物注入反应器，这导致反应器捕获游离的酶标记物。通过对碘苯酚 - 鲁米诺 - H₂O₂ 化学发光系统检测捕获的酶标记物。在最佳条件下，化学发光强度的降低与甲胎蛋白浓度在 5.0 - 100 ng/ml 范围内成正比，在信噪比为 3 时检测限为 2.7 ng/ml。该免疫传感器系统具有可接受的重现性和稳定性。用该方法对临床血清样本进行检测，结果与免疫放射分析获得的结果具有可接受的一致性。
