Microfluidic Devices for Enhancing the Sensitivity of ELISA methods

用于增强 ELISA 方法灵敏度的微流体装置

• 批准号: 0964211
• 负责人: Debashis Dutta
• 金额: $ 48.93万
• 依托单位: University of Wyoming
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-07-01 至 2015-06-30
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=0964211&HistoricalAwards=false
• 关键词: Microfluidic; Devices; Enhancing; Sensitivity; ELISA

Enzyme-linked immunosorbent assay (ELISA) is a method used to detect the presence of an antibody or an antigen.This project outlines a research plan to develop microfluidic devices that can significantly increase the sensitivity of ELISA methods when compared to currently available systems. This will be achieved by selecting the concentration of ELISA reaction products in a trapping zone. Because the trapping zone in the microfluidic devices has a volume of the order of nanoliters or less,entrapment of a small number of product molecules in this region results in a relatively large concentration within it, thereby significantly enhancing the sensitivity of the assay.In addition to characterizing and optimizing the performance of the devices described above, we will also extend their applicability to simultaneous detection of two analytes. Moreover, we will explore new protocols that could significantly simplify the fabrication and operation of the outlined microfluidic systems, thereby making them available to researchers across several disciplines and leading to a broad impact of this research onthe scientific community. The performance of the ELISA devices described above will be evaluated by side-by-side comparison with current ELISA methods in use by collaborators on this project (Dr. James Mecham from the USDA-ARS-ABADRL, Prof. Derek Smith from the Department of Kinesiology and Prof. Mark Stayton from the Department of Molecular Biology at the University of Wyoming). We anticipate determining concentrations of protein biomarkers for acute myocardial infarction and oxidative stress in serum samples, and monitoring of virus-cell receptor interactions for blue tongue virus.Besides training graduate and undergraduate students, educational activities aimed at disseminating the results of our research to a broad range of audiences, as well as directly involving high school students in research activities will be pursued as part of this project. This will include incorporating ELISA methods into a graduate level chemistry course at the University of Wyoming, and initiating a summer research apprentice program for high school students. At the same time, the PI will also develop an outreach program with the Central Wyoming College that has about 20% Native American and 80% first-generation college student population to broaden the participation of minorities and underprivileged communities.The research findings from this work will be published in scientific journals and presented at national/international conferences. In addition, the PI will also post some of the project outcomes, including protocols for carrying out the new assay methods at his research group website http://www.uwyo.edu/ddutta

酶联免疫吸附测定（ELISA）是用于检测抗体或抗原存在的方法。本项目概述了开发微流控装置的研究计划，与现有系统相比，该装置可以显着提高ELISA方法的灵敏度。这将通过选择捕获区中ELISA反应产物的浓度来实现。由于在微流控装置中的捕获区具有纳升或更小的数量级的体积，在该区域中的少量产品分子的捕获导致在其内的相对大的浓度，从而显著地增强了测定的灵敏度。除了表征和优化上述装置的性能之外，我们还将其适用性扩展到同时检测两种分析物。此外，我们将探索新的协议，可以显着简化所概述的微流体系统的制造和操作，从而使它们可用于跨多个学科的研究人员，并导致这项研究对科学界的广泛影响。将通过与本项目合作者（USDA-ARS-ABADRL的James Mecham博士、怀俄明州大学运动机能学系的Derek Smith教授和分子生物学系的Mark Stayton教授）使用的当前ELISA方法进行并行比较，评价上述ELISA装置的性能。我们预计确定血清样本中急性心肌梗死和氧化应激的蛋白质生物标志物的浓度，并监测蓝舌病毒的病毒-细胞受体相互作用。除了培训研究生和本科生，旨在向广泛的受众传播我们的研究结果的教育活动，以及让高中生直接参与研究活动，将作为该项目的一部分。这将包括将ELISA方法纳入怀俄明州大学的研究生水平化学课程，并为高中生启动暑期研究学徒计划。与此同时，PI还将与中央怀俄明州学院（约有20%的美国原住民和80%的第一代大学生）一起制定一项外展计划，以扩大少数民族和贫困社区的参与。这项工作的研究成果将发表在科学期刊上，并在国家/国际会议上发表。此外，PI还将在其研究小组网站http://www.uwyo.edu/ddutta上发布一些项目成果，包括执行新测定方法的方案