A glucose dehydrogenase biosensor as an additional signal amplification step in an enzyme-flow immunoassay.
A glucose dehydrogenase biosensor as an additional signal amplification step in an enzyme-flow immunoassay.
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葡萄糖脱氢酶生物传感器作为酶流免疫测定中的附加信号放大步骤。
DOI：
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发表时间：
2002
期刊：
影响因子：
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通讯作者：
J. Emnéus
中科院分区：
文献类型：
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作者：
C. Nistor;A. Rose;U. Wollenberger;D. Pfeiffer;J. Emnéus
抗体亲和力和标记物的可检测性在决定非均相免疫测定的最终特征中是必不可少的。本文描述了一种获得通过使用酶标记产生的信号的补充增强的方法，例如，通过在用基于葡萄糖脱氢酶（GDH）的电流型生物传感器进行的检测步骤期间将酶促反应的产物包括在附加的扩增循环中。采用具有标记的分析物衍生物的免疫测定形式，所述标记的分析物衍生物与样品中存在的分析物竞争有限量的抗体结合位点。β-半乳糖苷酶标记物水解底物氨基苯基-β-吡喃半乳糖苷，然后产生的氨基苯酚进入GDH生物传感器的生物电催化放大循环。该原理适用于测定4-硝基苯酚，最佳最小浓度为1.5 μ M，校准中点为24 μ M。这种系统的潜力和局限性进行了讨论。
Both the antibody affinity and the detectability of the label are essential in deciding the final characteristics of a heterogeneous immunoassay. This paper describes an approach to obtain a supplementary enhancement of the signal generated by using an enzyme label, e.g., by including the product of the enzymatic reaction in an additional amplification cycle during the detection step performed with an amperometric biosensor based on glucose dehydrogenase (GDH). An immunoassay format with a labelled analyte derivative that competes with the analyte present in the sample for a limited amount of antibody binding sites was employed. The beta-galactosidase label hydrolyses the substrate aminophenyl-beta-galactopyranoside, and the generated aminophenol enters then into a bioelectrocatalytic amplification cycle at the GDH biosensor. The principle was applied for determination of 4-nitrophenol, with the best minimal concentration of 1.5 microM and a midpoint of the calibration of 24 microM. The potentials and limitations of such a system are discussed.