Microfluidic Devices for Enhancing the Sensitivity of ELISA methods

酶联免疫吸附试验（ELISA）是一种用于检测抗体或抗原存在的方法。该项目概述了一项研究计划，开发微流体装置，与目前可用的系统相比，可以显着提高ELISA方法的灵敏度。这将通过在捕获区选择ELISA反应产物的浓度来实现。由于微流控装置的捕获区体积为纳升数量级或更小，因此在该区域捕获少量产物分子会导致该区域内相对较大的浓度，从而显着提高检测的灵敏度。除了表征和优化上述设备的性能外，我们还将扩展其同时检测两种分析物的适用性。此外，我们将探索新的方案，可以大大简化概述的微流体系统的制造和操作，从而使它们可供多个学科的研究人员使用，并导致这项研究在科学界产生广泛的影响。上述ELISA设备的性能将通过与该项目的合作者（来自USDA-ARS-ABADRL的James Mecham博士、运动机能系的Derek Smith教授和来自怀俄明大学分子生物学系的Mark Stayton教授）使用的当前ELISA方法进行并排比较来评估。我们期望在血清样本中确定急性心肌梗死和氧化应激的蛋白质生物标志物浓度，并监测蓝舌病毒的病毒-细胞受体相互作用。除了培养研究生和本科生外，旨在向广泛受众传播我们的研究成果以及直接让高中生参与研究活动的教育活动将作为该项目的一部分进行。这将包括将ELISA方法纳入怀俄明大学研究生水平的化学课程，并为高中生启动暑期研究学徒计划。与此同时，PI还将与中央怀俄明学院（Central Wyoming College）开展一项推广计划，以扩大少数民族和贫困社区的参与。该学院约有20%的美国原住民和80%的第一代大学生。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还将在他的研究小组网站http://www.uwyo.edu/ddutta上公布一些项目成果，包括实施新检测方法的协议

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