Enzymatically-generated fluorescent detection in micro-channels with internal magnetic mixing for the development of parallel microfluidic ELISA
Enzymatically-generated fluorescent detection in micro-channels with internal magnetic mixing for the development of parallel microfluidic ELISA
复制标题
DOI：
10.1039/b516031f
复制
发表时间：
2006-01-01
期刊：
影响因子：
6.1
通讯作者：
Tabrizian, M
中科院分区：
文献类型：
--
作者：
Herrmann, M;Veres, T;Tabrizian, M
酶联免疫吸附试验（ELISA）通常用于定量小浓度的特定蛋白质，用于各种各样的目的，从医学诊断到环境分析和食品安全。然而，这种技术需要大量昂贵的试剂和较长的潜伏期。微流体的使用允许人们通过减少微通道内的体积和扩散距离来专门解决这些缺点。现有的微流体系统受到极低流速的必要控制的限制，以提供足够的时间使分子仅通过扩散相互作用。在本文中，我们描述了一种新的微流体设计，用于在停止流动条件下实现平行ELISA。磁珠被用作固体相，以支持反应性免疫复合物的形成，并在通道内实现磁混合。为了测试检测程序，在将反应珠注入反应室之前，在芯片外进行免疫复合物的形成。抗链霉亲和素抗体的定量具有低皮摩尔灵敏度（0.1 - 6.7 pM），线性范围为2个数量级，重复性好。这项工作代表了一个简单、高效、平行的微流体ELISA新平台的第一步。
The Enzyme-Linked Immuno-Sorbent Assay, or ELISA, is commonly utilized to quantify small concentrations of specific proteins for a large variety of purposes, ranging from medical diagnosis to environmental analysis and food safety. However, this technique requires large volumes of costly reagents and long incubation periods. The use of microfluidics permits one to specifically address these drawbacks by decreasing both the volume and the distance of diffusion inside the micro-channels. Existing microfluidic systems are limited by the necessary control of extremely low flow rates to provide sufficient time for the molecules to interact with each other by diffusion only. In this paper, we describe a new microfluidic design for the realization of parallel ELISA in stop-flow conditions. Magnetic beads were used both as a solid phase to support the formation of the reactive immune complex and to achieve a magnetic mixing inside the channels. In order to test the detection procedure, the formation of the immune complex was performed off-chip before the reactive beads were injected into the reaction chamber. Anti-streptavidin antibodies were quantified with low picomolar sensitivity (0.1 - 6.7 pM), a linear range of 2 orders of magnitude and good reproducibility. This work represents the first step toward a new platform for simple, highly effective and parallel microfluidic ELISA.