公开(公告)号：EP1179181A1

公开(公告)日：2002-02-13

申请号：EP00932644.8

申请日：2000-05-19

申请人： University of Washington

发明人： WEIGL, Bernhard, H. ,  YAGER, Paul ,  KAMHOLZ, Andrew ,  HATCH, Anson

IPC分类号： G01N33/558 ,  G01N33/557 ,  G01N33/543 ,  G01N15/06 ,  G01N33/00 ,  G01N33/48 ,  G01N21/00 ,  G01N21/29 ,  G01N21/41 ,  G01N21/47 ,  B01D11/00 ,  B01D11/04

CPC分类号： G01N33/558 ,  G01N30/0005

摘要： Methods and apparatuses are provided for determining presence and concentration of analytes by exploiting molecular binding reactions and differential diffusion rates. Analyte particles and binding particles are allowed to diffuse toward each other, and slowing of the diffusion front is detected when they meet. From the position of the diffusion front, presence and concentration of analyte particles can be determined. One embodiment provides a competitive immunoassay in a microfluidic format. This diffusion immunoassay (DIA) relies on measuring the concentration of labeled antigen along one dimension of a microchannel after allowing it to diffuse for a short time into a region containing specific antibodies. A simple microfluidic device, the T-Sensor, was used to implement a DIA to measure the concentration of phenytoin, a small drug molecule. Concentrations of analyte over the range of 50 to 1600 nM can be measured in less than a minute. The assay is homogenous, rapid, requires only microliter volumes of reagents and sample, and is applicable to a wide range of analytes, including therapeutic drugs, molecular biological markers, and environmental contaminants. Methods for separating particles of similar size in a diffusion separator are also provided.

摘要翻译： 提供了通过利用分子结合反应和差异扩散速率来确定分析物的存在和浓度的方法和设备。 允许分析物颗粒和结合颗粒朝向彼此扩散，并且当它们相遇时检测到扩散前沿的减慢。 从扩散前沿的位置可以确定分析物颗粒的存在和浓度。 一个实施方案提供微流体形式的竞争性免疫测定。 该扩散免疫测定法（DIA）依赖于在允许其短时间扩散到含有特异性抗体的区域中后，沿着微通道的一维测量标记抗原的浓度。 使用简单的微流体装置T传感器来实施DIA以测量苯妥英（一种小分子药物）的浓度。 在不到一分钟的时间内可以测量50至1600 nM范围内的分析物浓度。 该检测方法均匀，快速，只需要微升体积的试剂和样品，适用于各种分析物，包括治疗药物，分子生物标志物和环境污染物。 还提供了用于在扩散分离器中分离相似尺寸的颗粒的方法。