Microfluidics-enabled Immunoassay Array for Multianalyte Detection

描述（由申请人提供）：拟议研究的目标是开发一种平台技术，以更高的效率和更低的成本同时检测多种分析物。目前，免疫分析广泛应用于生物医学研究、临床诊断、食品/水安全检测和生物战防御。虽然它们可以在微孔板（例如96孔）中进行以增加样品通量，但每孔中的免疫分析通常限于每次测试一种分析物。对于检测样品中的多种分析物的要求，这种格式是低效的，并且不具有成本效益。例如，应同时监测食品和饮用水中的一系列有毒物质。类似地，应该检测一组生物标志物，而不是单个生物标志物，以提高特定疾病的准确性。因此，为了提高效率和降低成本，需要一个同时检测多种分析物的平台。拟议研究的具体目标是：(1)设计和优化一系列微流体阀，这些阀用于容纳试剂，引导和调节流动；(2)以夹心检测的形式演示了微流控复用免疫检测阵列，并研究了表面修饰、流速、试剂浓度对检测的影响；(3)通过检测包括毒素和生物标志物在内的多种分析物来验证免疫分析阵列。本研究的意义在于以下几个方面。首先，提出的研究将导致一个同时检测多种分析物的平台。这种能力将导致高效率和成本效益的方法。其次，微流控免疫分析阵列将有更短的分析时间和减少所需的样品和试剂的消耗。由于免疫测定法广泛应用于化学、生物和临床实验室，这些优势可能会产生重大影响。第三，拟议的研究将导致一个可制造的过程，允许大规模生产低成本，一次性设备。一次性使用后的一次性能力可能会对顺序样品交叉污染的应用产生巨大影响。

DESCRIPTION (provided by applicant): The goal of the proposed research is to develop a platform technology for simultaneous detection of multiple analytes with better efficiency and lower cost. Currently, immunoassays are extensively used in biomedical research, clinical diagnostics, food/water safety testing, and biological warfare defense. Although they can be performed in a microplate (e.g. 96 wells) to increase sample throughput, the immunoassay in each well is generally limited to one analyte per test. Such a format is inefficient and not cost-effective for the requirement to detect multiple analytes in a sample. For instance, a spectrum of toxic agents should be simultaneously monitored in foods and drinking water. Similarly, a set of biomarkers, rather than an individual biomarker, should be detected for a particular disease to enhance the accuracy. Therefore, a platform to detect multiple analytes simultaneously is desirable for better efficiency and lower cost. The specific aims of the proposed research are to (1) design and optimize an array of microfluidic valves that are required for containing reagents, directing and regulating flows; (2) demonstrate the microfluidics-enabled multiplexed immunoassay array in the format of sandwich assays and study the effects of surface modification, flow rate, reagent concentration on the assay; and (3) validate the immunoassay array by detecting multiple analytes including toxins and biomarkers. The significance of the research lies in the following aspects. First, the proposed research will lead to a platform for simultaneous detection of multiple analytes. Such a capability will lead to efficient and cost- effective methods. Secondly, the microfluidic-enabled immunoassay array will have shorter analysis time and a reduction in the consumption of required sample and reagents. These advantages could have a significant impact since immunoassays are widely used in chemical, biological and clinical laboratories. Thirdly, the proposed research will result in a manufacturable process that allows mass production of low-cost, disposable devices. The ability to be disposable after a single use could have a tremendous impact to applications where cross-contamination of sequential samples is of concern.