Multiplexed protein detection using antibody-conjugated microbead arrays in a microfabricated electrophoretic device.
Multiplexed protein detection using antibody-conjugated microbead arrays in a microfabricated electrophoretic device.
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DOI：
10.1039/c0lc00044b
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发表时间：
2010-11-21
期刊：
影响因子：
6.1
通讯作者：
Huang X
中科院分区：
文献类型：
--
作者：
Barbee KD;Hsiao AP;Roller EE;Huang X
我们报道了一种用于组装高密度抗体偶联微珠阵列的微加工电泳装置的发展，该装置用于基于芯片的蛋白质检测。该装置由在具有蚀刻在二氧化硅膜中的微孔阵列的金涂覆的硅芯片和具有一系列薄的金对电极线的玻璃盖玻片之间形成的流动池组成。我们已经证明，通过在腔室中施加脉冲电场，可以将与抗体缀合的0.4和1 μ m珠快速组装到微孔中。通过逐步组装荧光标记的抗体缀合的微珠的混合物，我们将空间和荧光编码策略结合以展示显著的多路复用能力。我们已经证明，这些抗体偶联的微珠阵列可用于进行芯片上夹心免疫测定，以检测浓度低至40 pM（6 ng/mL）的测试抗原。还开发了一个有限元模型，以检查不同的反电极配置的线间距和腔室高度的范围内的设备内的电场分布。该装置将用于组装高密度编码抗体阵列，用于蛋白质和其他类型的蛋白质缀合微珠的多重检测，用于诸如蛋白质-蛋白质相互作用的分析的应用。
We report the development of a microfabricated electrophoretic device for assembling high-density arrays of antibody-conjugated microbeads for chip-based protein detection. The device consists of a flow cell formed between a gold-coated silicon chip with an array of microwells etched in a silicon dioxide film and a glass coverslip with a series of thin gold counter electrode lines. We have demonstrated that 0.4 and 1 μm beads conjugated with antibodies can be rapidly assembled into the microwells by applying a pulsed electric field across the chamber. By assembling step-wise a mixture of fluorescently labeled antibody-conjugated microbeads, we incorporated both spatial and fluorescence encoding strategies to demonstrate significant multiplexing capabilities. We have shown that these antibody-conjugated microbead arrays can be used to perform on-chip sandwich immunoassays to detect test antigens at concentrations as low as 40 pM (6 ng/mL). A finite element model was also developed to examine the electric field distribution within the device for different counter electrode configurations over a range of line pitches and chamber heights. This device will be useful for assembling high-density, encoded antibody arrays for multiplexed detection of proteins and other types of protein-conjugated microbeads for applications such as the analysis of protein-protein interactions.
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影响因子：
2.9
作者：
Diercks AH;Ozinsky A;Hansen CL;Spotts JM;Rodriguez DJ;Aderem A
通讯作者：
Aderem A
影响因子：
46.9
作者：
de Wildt, RMT;Mundy, CR;Tomlinson, IM
通讯作者：
Tomlinson, IM
影响因子：
4
作者：
Cordovez, Bernardo;Psaltis, Demetri;Erickson, David
通讯作者：
Erickson, David
影响因子：
7.4
作者：
Blicharz, Timothy M.;Siqueira, Walter L.;Helmerhorst, Eva J.;Oppenheim, Frank G.;Wexler, Philip J.;Little, Frederic F.;Walt, David R.
通讯作者：
Walt, David R.
影响因子：
46.9
作者：
Han, MY;Gao, XH;Nie, S
通讯作者：
Nie, S