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 on the 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 www.uwyo.edu/ddutta
