Fakunle Eyitayo S, Aguilar Zoraida P, Shultz John L, Toland Alan D, Fritsch Ingrid
Department of Chemistry and Biochemistry and Arkansas Analytical Laboratories, University of Arkansas, Fayetteville, Arkansas 72701, USA.
Langmuir. 2006 Dec 5;22(25):10844-53. doi: 10.1021/la061304n.
Screen-printed gold (SPG, Dupont gold conductor 5734) on low-temperature co-fired ceramic (LTCC) materials (Dupont dielectric tape 951, mostly composed of alumina and silica) has been demonstrated to be a substrate for electrochemical enzyme-linked immunosorbant assays. The effect of two different cleaning treatments and the extent of nonspecific adsorption on the SPG/LTCC and plain LTCC surfaces were also evaluated. LTCC materials hold promise for constructing a new generation of devices for microelectrochemical sensing and assays. Facile fabrication in three dimensions with integrated conducting elements makes them attractive. A standard sandwich immunoassay for a model analyte, mouse IgG, was used to evaluate the LTCC materials. After the assembly of components onto chips of SPG/LTCC and plain LTCC, p-aminophenol that was generated enzymatically by the enzyme label was detected electrochemically with a separate glassy carbon electrode. Cleaning SPG/LTCC with a piranha solution (7:1 vol/vol of concentrated H2SO4/30% H2O2), traditionally used for other gold surfaces prior to SAM assembly, resulted in a notable decrease in assay signal and an increase in nonspecific adsorption when compared to cleaning with water alone. Assay components assemble specifically on plain LTCC, with only a small percent attributed to NSA. Environmental scanning electron microscopy, energy-dispersive X-ray spectroscopy, and X-ray photoelectron spectroscopy reveal the tremendous chemical heterogeneity and complexity of both SPG/LTCC and plain LTCC surfaces and aid in the explanation of assay results. A 10% acetate Tween bovine serum albumin solution and an ethanolic solution of 4 mM 1-butanol eliminate assay signals originating from plain LTCC. The outcomes of these studies can now be used to achieve miniaturized electrochemical immunoassays on LTCC materials where both plain and SPG surfaces are present.
低温共烧陶瓷（LTCC）材料（杜邦介电胶带951，主要由氧化铝和二氧化硅组成）上的丝网印刷金（SPG，杜邦金导体5734）已被证明是用于电化学酶联免疫吸附测定的一种基质。还评估了两种不同清洗处理的效果以及SPG/LTCC和平坦LTCC表面上非特异性吸附的程度。LTCC材料有望构建新一代用于微电化学传感和测定的器件。其具有集成导电元件的三维简易制造使其具有吸引力。使用针对模型分析物小鼠IgG的标准夹心免疫测定法来评估LTCC材料。在将组件组装到SPG/LTCC和平坦LTCC芯片上之后，通过单独的玻碳电极对酶标记物酶促产生的对氨基苯酚进行电化学检测。与仅用水清洗相比，用传统上在自组装单分子层之前用于其他金表面的王水（浓硫酸/30%过氧化氢体积比7:1）清洗SPG/LTCC会导致测定信号显著降低以及非特异性吸附增加。测定组件特异性地组装在平坦的LTCC上，只有一小部分归因于非特异性吸附。环境扫描电子显微镜、能量色散X射线光谱和X射线光电子能谱揭示了SPG/LTCC和平坦LTCC表面巨大的化学异质性和复杂性，并有助于解释测定结果。10%乙酸吐温牛血清白蛋白溶液和4 mM 1-丁醇的乙醇溶液可消除源自平坦LTCC的测定信号。这些研究的结果现在可用于在同时存在平坦表面和SPG表面的LTCC材料上实现小型化电化学免疫测定。
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