Immunoassays based on electrochemical detection using microelectrode arrays
Immunoassays based on electrochemical detection using microelectrode arrays
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DOI：
10.1016/j.bios.2004.06.049
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发表时间：
2004-11-01
影响因子：
12.6
通讯作者：
Oleinikov, AV
中科院分区：
文献类型：
--
作者：
Dill, K;Montgomery, DD;Oleinikov, AV
我们表明，CombiMatrix的VLSI阵列的可单独寻址的电极，使用传统的CMOS集成电路，可用于检测各种分析物，通过免疫分析协议。这些微阵列每平方厘米提供超过1000个电极。芯片上涂有一层多孔材料，在其上合成特定的亲和标签，靠近选定的电极位点。CombiMatrix微阵列用于开发从小分子到细胞的各种大小的生物实体的空间多路复用检测格式。用编码的亲和标记物标记抗体，然后使其在适当的电极测定位点上自组装。每种分析物特异性抗体通过自组装过程被陪伴到单独的预定位置。由此产生的芯片可以执行许多不同的分析物特异性免疫测定，同时，我们提出了新的检测技术的基础上使用的芯片上的活性可单独寻址的微电极：氧化还原酶放大电化学检测。对人α 1酸性糖蛋白、蓖麻毒素、M13噬菌体、球形芽孢杆菌孢子和荧光素的结果表明，该方法是最灵敏的方法之一，检测限在阿托摩尔范围内。检测范围为4-5个对数的分析物浓度，测定体积为50穆尔或更少。我们展示了如何在CombiMatrix微阵列平台上优化检测以获得最佳性能。(C)2004 Elsevier B. V.保留所有权利。
We show that CombiMatrix's VLSI arrays of individually addressable electrodes, using conventional CMOS integrated circuitry, can be used in detecting various analytes via immunoassay protocols. These microarrays provide over 1000 electrodes per square centimeter. The chips are coated with a porous material on which specific affinity tags are synthesized proximate to selected electrode sites.CombiMatrix microarrays are used to develop spatially multiplexed assay formats for biological entities over a wide range of sizes, from small molecules to cells. Antibodies are tagged with coded affinity labels and then allowed to self-assemble on the appropriate electrode assay sites. Each analyte-specific antibody is chaperoned to individual, predetermined locations by the self-assembly process. The resulting chip can perform numerous different analyte-specific immunoassays, simultaneously.We present new detection technologies based upon the use of the active individually addressable microelectrodes on the chip: redox enzyme amplified electrochemical detection. The results for human alpha1 acid glycoprotein, ricin, M13 phage, Bacillus globigii spores, and fluorescein indicate that this method is one of the most sensitive available, with limits of detection in the attomole range. The detection range is 4-5 logs of analyte concentration, with an assay volume of 50 mul or less. The system provides for a host of multiplexed immunoassays because of the large number of electrodes available. We show how the assays can be optimized for maximum performance on the CombiMatrix microarray platform. (C) 2004 Elsevier B.V. All rights reserved.