Highly sensitive electrochemical detection of alkaline phosphatase

被引：71

作者：

Ito, S ^{[1
]}

Yamazaki, S ^{[1
]}

Kano, K ^{[1
]}

Ikeda, T ^{[1
]}

机构：

[1] Kyoto Univ, Grad Sch Agr, Div Appl Life Sci, Kyoto 6068502, Japan

来源：

ANALYTICA CHIMICA ACTA | 2000年 / 424卷 / 01期

关键词：

bioelectrocatalysis; alkaline phosphatase; enzyme immunoassay; tyrosinase; phenol;

D O I：

10.1016/S0003-2670(00)01149-1

中图分类号：

O65 [分析化学];

学科分类号：

070302 ; 081704 ;

摘要：

Alkaline phosphatase (ALP), a labeling enzyme frequently used for enzyme immunoassay, was determined at the attomole level using a mushroom tyrosinase (TN)-embedded carbon paste electrode. The detection scheme consists of two successive amplifications. The first reaction is the ALP-catalyzed hydrolysis of phenyl phosphate to accumulate phenol as a product. The second is the bioelectrocatalytic detection of phenol with the TN-embedded electrode at 0 V versus Ag/AgCl, which allowed sensitive detection of phenol down to 35 nM. The two-enzymatic reactions of ALP and TN were made to proceed successively in one electrochemical cell by adjusting pH to their optimum ones. This method provided a Linear relation between the ALP concentration and the steady-state current and allowed the ALP detection as low as 7 amol/10 mul sample. The ALP detection system was applied to a streptavidin-biotin binding assay. The lowest detection limit of biotin was 4 fmol/10 mul sample with an ALP reaction time of 50 min. (C) 2000 Elsevier Science B.V. All rights reserved.

引用

页码：57 / 63

页数：7

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