Electrokinetically driven microfluidic chips with surface-modified chambers for heterogeneous immunoassays
Electrokinetically driven microfluidic chips with surface-modified chambers for heterogeneous immunoassays
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DOI：
10.1021/ac0015366
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发表时间：
2001-07-15
影响因子：
7.4
通讯作者：
de Rooij, NF
中科院分区：
文献类型：
--
作者：
Dodge, A;Fluri, K;de Rooij, NF
本文介绍了微流控芯片异质生物分析的第一个例子，使用局部固定的生物特异性层和电动操作。反应室具有皮升尺寸，并集成到蚀刻在玻璃上的微通道网络中。利用蛋白A （PA）对兔免疫球蛋白G （rIgG）的高亲和力进行芯片检测，将PA固定在微通道壁上，荧光标记（Cy5） rIgG缝制为样品。可以使用电动泵送溶液以免疫亲和层析的方式操作芯片。证明了抗体从稀溶液到固相的浓度，信号增益可能接近30。在浓度低于50 nM、孵育时间为200 s的条件下，获得了Cy5-rIgG的剂量响应曲线。使用样品/示踪剂联合孵育和这些溶液的顺序添加，证明了芯片布局的灵活性，用于竞争性免疫分析rIgG。对于这种未优化的设备，分析时间通常低于5分钟，所描述的微流控方法在许多高通量筛选应用中显示出巨大的潜力。
This article presents the first example of a microfluidic chip for heterogeneous bioassays using a locally immobilized biospecific layer and operated electrokinetically. The reaction chamber has picoliter dimensions and is integrated into a network of microchannels etched in glass. The high affinity of protein A (PA) for rabbit immunoglobulin G (rIgG) was exploited for chip testing, with PA being immobilized on microchannel walls and fluorescently labeled (Cy5) rIgG sewing as sample. It was possible to operate the chip in an immunoaffinity chromatographic manner, using electrokinetically pumped solutions. Concentration of antibody from dilute solution onto the solid phase was demonstrated, with signal gains of similar to 30 possible. A dose-response curve for Cy5-rIgG was obtained for concentrations down to 50 nM, for an incubation time of 200 s. The flexibility of chip layout was demonstrated for competitive immunoassay of rIgG using both a combined sample/tracer incubation and sequential addition of these solutions, With assay times generally below 5 min for this unoptimized device, the microfluidic approach described shows great potential for many high-throughput screening applications,