Hybridization detection of enzyme-labeled DNA at electrically heated electrodes
Hybridization detection of enzyme-labeled DNA at electrically heated electrodes
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DOI：
10.1007/s00216-013-6815-3
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发表时间：
2013-02
影响因子：
4.3
通讯作者：
A. Walter;Annette‐Enrica Surkus;G. Flechsig
中科院分区：
文献类型：
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作者：
A. Walter;Annette‐Enrica Surkus;G. Flechsig
在这份报告中，我们描述了一种电化学DNA杂交传感器的方法，其中使用加热电极与作为DNA标记的酶一起实现信号放大。在可加热低温共烧陶瓷（LTCC）金电极表面，以碱性磷酸酶（SA-ALP）为报告分子，将巯基化的捕获探针与生物素标记的靶分子杂交。该酶标记转化成氧化还原活性的1-萘酚氧化还原活性的1-磷酸萘酯（NAP）在改性金LTCC电极伏安测定。在测量期间，仅加热电极，将本体溶液留在环境温度下。检测过程中温度升高导致酶活性增加，DNA杂交检测的分析信号增强。在53 °C电极温度下的检测限为1.2 nmol/L。
In this report we describe an electrochemical DNA hybridization sensor approach, in which signal amplification is achieved using heated electrodes together with an enzyme as DNA-label. On the surface of the heatable low temperature co-fired ceramic (LTCC) gold electrode, an immobilized thiolated capture probe was hybridized with a biotinylated target using alkaline phosphatase (SA-ALP) as reporter molecule. The enzyme label converted the redox-inactive substrate 1-naphthyl phosphate (NAP) into the redox-active 1-naphthol voltammetrically determined at the modified gold LTCC electrode. During the measurement only the electrode was heated leaving the bulk solution at ambient temperature. Elevated temperature during detection led to increased enzyme activity and enhanced analytical signals for DNA hybridization detection. The limit of detection at 53 °C electrode temperature was 1.2 nmol/L.