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低温共烧陶瓷微通道,四面具有可单独寻址的丝网印刷金电极,用于自成体系的电化学免疫分析。

Low-temperature co-fired ceramic microchannels with individually addressable screen-printed gold electrodes on four walls for self-contained electrochemical immunoassays.

机构信息

Department of Chemistry and Biochemistry, University of Arkansas, Fayetteville, AR 72701, USA.

出版信息

Anal Bioanal Chem. 2010 Nov;398(6):2605-15. doi: 10.1007/s00216-010-4098-5. Epub 2010 Aug 28.

Abstract

Microchannel devices were constructed from low-temperature co-fired ceramic (LTCC) materials with screen-printed gold (SPG) electrodes in three dimensions--on all four walls--for self-contained enzyme-linked immunosorbant assays with electrochemical detection. The microchannel confines the solution to a small volume, allowing concentration of electroactive enzymatically generated product and nearby electrodes provide high-speed and high-sensitivity detection: it also facilitates future integration with microfluidics. LTCC materials allow easy construction of three-dimensional structures compared with more traditional materials such as glass and polymer materials. Parallel processing of LTCC layers is more amenable to mass production and fast prototyping, compared with sequential processing for integrating multiple features into a single device. LTCC and SPG have not been reported previously as the basis for microchannel immunoassays, nor with integrated, individually addressable electrodes in three dimensions. A demonstration assay for mouse IgG at 5.0 ng/mL (3.3 × 10(-11) M) with electrochemical detection was achieved within a 1.8 cm long × 290 μm high × 130 μm wide microchannel (approximately 680 nL). Two of four SPG electrodes span the top and bottom walls and serve as the auxiliary electrode and the assay site, respectively. The other two (0.7 cm long × 97 μm wide) are centered lengthwise on the sidewalls of the channel. One serves as the working and the other as the pseudoreference electrode. The immunoassay components were immobilized at the bottom SPG region. Enzymatically generated p-aminophenol was detected at the internal working electrode within 15 s of introducing the enzyme substrate p-aminophenyl phosphate. A series of buffer rinses avoided nonspecific adsorption and false-positive signals.

摘要

微通道器件采用低温共烧陶瓷(LTCC)材料构建,在三维空间中(四面均有)印刷有金(SPG)电极,用于进行内置酶联免疫吸附分析和电化学检测。微通道将溶液限制在小体积内,允许电活性酶促生成产物浓缩,并且附近的电极提供高速和高灵敏度检测:它还便于与微流控技术集成。与更传统的材料(如玻璃和聚合物材料)相比,LTCC 材料允许更容易地构建三维结构。与为将多个特征集成到单个器件中而进行的顺序处理相比,LTCC 层的并行处理更适合大规模生产和快速原型制作。LTCC 和 SPG 以前并未被报道为微通道免疫分析的基础,也未与集成的、可单独寻址的三维电极一起使用。在 1.8 厘米长×290 微米高×130 微米宽的微通道(约 680 nL)中,实现了电化学检测的 5.0 ng/mL(3.3×10(-11) M)的小鼠 IgG 检测演示。四个 SPG 电极中的两个横跨顶壁和底壁,分别用作辅助电极和检测部位。另外两个(0.7 厘米长×97 微米宽)沿通道的侧壁居中。一个用作工作电极,另一个用作伪参比电极。免疫分析组件固定在底 SPG 区域。在引入酶底物对氨基苯磷酸酯 15 秒内,在内部工作电极处检测到酶促生成的对氨基酚。一系列缓冲冲洗避免了非特异性吸附和假阳性信号。

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