Flow sandwich-type immunoassay in microfluidic devices based on negative dielectrophoresis
Flow sandwich-type immunoassay in microfluidic devices based on negative dielectrophoresis
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DOI：
10.1016/j.bios.2006.11.010
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发表时间：
2007-05-15
影响因子：
12.6
通讯作者：
Matsue, Tomokazu
中科院分区：
文献类型：
--
作者：
Yasukawa, Tomoyuki;Suzuki, Masato;Matsue, Tomokazu
利用负介电泳技术在微流控通道中操纵微粒，并将该方法应用于非均相免疫分析系统。一个微流控装置，与三维（3-D）微电极上制作的两个基板上，被用来操纵颗粒流的通道，并捕获在笼状区域，由集电极包围的颗粒。用抗小鼠免疫球蛋白G（IgG）修饰的聚苯乙烯微粒（直径6 μ m）被操纵，并在被强烈的n-DEP电场包围时被捕获在笼状区域中。具体地，当将振幅为6-15 V-峰值和频率超过500 kHz的AC电压施加到两个面对的微电极时，捕获颗粒。通过将含有小鼠抗原（IgG）的样品溶液和含有具有信号源的二次抗体（FITC标记的抗小鼠IgG抗体）的溶液相继注入通道中，实现异质夹心免疫测定。来自笼区中捕获颗粒的荧光强度随着小鼠IgG浓度的增加（10 ng/ml至10 μ g/ml）而增加。所描述的系统能够在40分钟内测定小鼠IgG。因此，使用基于n-DEP的微流体装置可以实现从所需分析物和标记抗体中自动分离游离级分。(c)2006 Elsevier B. V.保留所有权利。
Microparticles have been manipulated in a microfluidic channel by means of negative dielectrophoresis (n-DEP), and the approach applied to a heterogeneous immunoassay system. A microfluidic device, with three-dimensional (3-D) microelectrodes fabricated on two substrates, was used to manipulate particle flow in the channel and to capture the particles in the caged area that was enclosed by the collector electrodes. Polystyrene microparticles (6 mu m diameters) modified with anti-mouse immunoglobulin G (IgG) were manipulated and captured in the caged area when surrounded by intense n-DEP electric fields. Specifically, particles were trapped when AC voltages with amplitudes of 6-15 V-peak and frequencies over 500 kHz were applied to the two facing microelectrodes. A heterogeneous sandwich immunoassay was achieved by successively injecting a sample solution containing mouse antigen (IgG), and a solution containing a secondary antibody with a signal source (FITC-labeled anti-mouse IgG antibody), into the channel. The fluorescence intensity from captured particles in the caged area increased with increasing concentrations (10 ng/ml to 10 mu g/ml) of mouse IgG. The described system enables mouse IgG to be assayed in 40 min. Thus, the automatic separation of free fractions from desired analytes and labeled antibodies can be achieved using a microfluidic device based on n-DEP. (c) 2006 Elsevier B.V. All rights reserved.