Bead-assisted displacement immunoassay for staphylococcal enterotoxin B on a microchip
Bead-assisted displacement immunoassay for staphylococcal enterotoxin B on a microchip
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DOI：
10.1021/ac061057s
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发表时间：
2006-12-15
影响因子：
7.4
通讯作者：
Collins, Greg E.
中科院分区：
文献类型：
--
作者：
Haes, Amanda J.;Terray, Alex;Collins, Greg E.
提出了一种基于微芯片的位移免疫分析法，用于敏感的激光诱导荧光检测葡萄球菌肠毒素B。该玻璃微芯片装置由包含用于支持抗体功能化微珠的双堰结构的微通道组成。在30分钟的样品制备步骤后，在没有用户干预的情况下进行位移测定，并在额外的20分钟内产生定量结果。线性检测响应超过6个数量级，检测限低至1 fM（28.5 fg/mL）。令人惊讶的低检测限被假设是由类似于场放大堆叠的场基富集、色谱效应和微珠床中有限的扩散长度引起的。该试验用牛血清白蛋白、酪蛋白和牛奶样品基质进行挑战。该系统具有为目标生物分子提供高灵敏度检测能力的潜力。
A microchip-based, displacement immunoassay for the sensitive laser-induced fluorescence detection of staphylococcal enterotoxin B is presented. The glass microchip device consists of a microchannel that contains a double weir structure for supporting antibody-functionalized microbeads. After a 30-min sample preparation step, the displacement assay was performed without user intervention and produced quantitative results in an additional 20 min. Linear detection responses were observed over 6 orders of magnitude and provided detection limits down to 1 fM(28.5 fg/mL). The surprisingly low detection limits are hypothesized to arise from field-based enrichment analogous to field-amplified stacking, chromatographic effects, and limited diffusion lengths in the microbead bed. The assay was challenged with bovine serum albumin, casein, and milk sample matrixes. This system has the potential to provide highly sensitive detection capabilities for target biomolecules.