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Microfabricated Amperometric Cells for Multicomponent Analysis

机译：用于多组分分析的超细安培池

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Towards the development of multianalyte electrochemical immunoassays three individually addressable micro-electrode array (MEA) type working electrodes and a reference electrode were integrated into a 4 mu L volume, planar electrochemical cell. To model the simultaneous determination of multiple antigens in the cell with enzyme linked immunosorbent assays (ELISAs) glucose oxidase (GOx), alkaline phosphatase (ALP), and beta-galactosidase (beta-GAL) were immobilized site specifically onto the individual MEA surfaces and the biocatalitic activity of these surface confined enzymes were evaluated by measuring the products of the enzyme catalyzed reactions directly on the gold MEA surfaces by chronoamperometry or by imaging the enzyme patterned microelectrode array surfaces by Scanning Electrochemical Microscopy (SECM). ALP and beta-GAL were selected as model enzymes because they are the most commonly used enzymes labels in ELISAs. In these measurements glucose, ascorbic acid phosphate (AAP), and p-aminophenyl-beta-D-galactopyranoside (PAPG) served as enzyme substrates, respectively. The electrochemical surface area of the gold MEAs did not change during the multistep immobilization process. All enzyme modified MEAs presented selective and proportional responses to their substrates and the response characteristics of the enzyme modified sensors were identical in separate and simultaneous calibration protocols, i.e., there was no cross-contamination between the closely placed MEAs. The SECM images of the enzyme patterned MEA surfaces suggest that nonspecific adsorption is negligible on the insulating polyimide surface of the MEA separating the individual microelectrode sites.
机译：为了发展多分析物电化学免疫测定法，将三个可单独寻址的微电极阵列（MEA）型工作电极和参比电极集成到一个4μL体积的平面电化学池中。为了模拟使用酶联免疫吸附测定（ELISA）同时测定细胞中多种抗原的方法，将葡萄糖氧化酶（GOx），碱性磷酸酶（ALP）和β-半乳糖苷酶（β-GAL）固定在特定的MEA表面上，并通过计时安培法直接在金MEA表面上测量酶催化反应的产物或通过扫描电化学显微镜（SECM）对酶构图的微电极阵列表面成像，可以评估这些表面受限酶的生物催化活性。选择ALP和β-GAL作为模型酶，因为它们是ELISA中最常用的酶标记。在这些测量中，葡萄糖，抗坏血酸磷酸酯（AAP）和对氨基苯基-β-D-吡喃半乳糖苷（PAPG）分别用作酶底物。在多步固定过程中，金MEA的电化学表面积没有变化。所有酶修饰的MEA均表现出对其底物的选择性和比例响应，并且酶修饰的传感器的响应特性在单独和同时的校准方案中是相同的，即在紧密放置的MEA之间没有交叉污染。酶构图的MEA表面的SECM图像表明，在分隔各个微电极部位的MEA绝缘聚酰亚胺表面上，非特异性吸附可忽略不计。

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