Comparison of label-free biosensing in microplate, microfluidic, and spot-based affinity capture assays
Comparison of label-free biosensing in microplate, microfluidic, and spot-based affinity capture assays
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DOI：
10.1016/j.ab.2010.06.009
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发表时间：
2010-10-01
影响因子：
2.9
通讯作者：
Cunningham, Brian T.
中科院分区：
文献类型：
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作者：
Choi, Charles J.;Belobraydich, Alysia R.;Cunningham, Brian T.
使用实验测定和流体动力学有限元模拟模型，我们直接比较了四种不同的测定配置的生物传感器表面上的测试样品的分析物的无标记检测的可实现的性能限制。在这项工作中研究的测定配置包括生物传感器纳入微板孔和微流体通道的底部表面。对于每种配置，我们比较了流体处理容器的整个底表面涂覆有捕获配体的情况下的测定性能与以局部斑点的形式施加捕获配体的情况下的测定性能。作为一个模型系统，我们使用的蛋白质生物标志物肿瘤坏死因子-α（TNF-α）的检测使用固定的TNF-α捕获抗体。结果表明，微流控分析格式大大减少了建立稳定平衡所需的时间。基于斑点的测定对于基于微孔板的检测是有利的，以减少平衡传感器响应所需的时间。得出的结果通常适用于任何无标记的生物传感器技术和任何配体分析物系统与可调变量，包括传感器质量密度灵敏度，分析物-配体吸附/解吸速率常数，固定化配体密度，流动通道的几何形状，流速，和斑点大小。(C)2010年爱思唯尔公司All rights reserved.
Using both experimental assays and fluid-dynamic finite element simulation models, we directly compared the achievable performance limits of four distinct assay configurations for label-free detection of an analyte from a test sample on a biosensor surface. The assay configurations studied in this work included a biosensor incorporated into the bottom surface of a microplate well and a microfluidic channel. For each configuration, we compared assay performance for the scenario in which the entire bottom surface of the fluid-handling vessel is coated with capture ligands with assay performance for the scenario in which the capture ligands are applied in the form of localized spots. As a model system, we used detection of the protein biomarker tumor necrosis factor-alpha (TNF-alpha) using immobilized TNIF-alpha capture antibody. Results show that the microfluidic assay format dramatically reduces the time required to establish a stable equilibrium. Spot-based assays are advantageous for microplate-based detection for reducing the time required for equilibrium sensor response. The results derived are generally applicable to any label-free biosensor technology and any ligand-analyte system with adjustable variables that include sensor mass density sensitivity, analyte-ligand adsorption/desorption rate constants, immobilized ligand density, flow channel geometry, flow rate, and spot size. (C) 2010 Elsevier Inc. All rights reserved.