RIG: Metabolite Target Analysis using Microdevice Electrophoresis

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This project will develop new microdevice (microfluidic) electrophoresis methods to improve the separation and detection of metabolites unique to certain cellular processes. Electrophoresis, a technique used to separate molecules based on differential migration rates in an electric field, remains one of the most powerful and widely used analytical tools in biology and in chemistry. The modern capillary format, microdevice electrophoresis, offers numerous advantages over the traditional slab gel, such as sensitive on-line detection, speed, cost, and improved dissipation of joule heating. Microfluidic methods to analyze estrogen metabolites will be created using two different strategies, free solution and hydrogel immobilization. Both approaches will utilize competitive immunoassays. The free solution microfluidic method will employ conventional competitive immunoassays whereas the hydrogel immobilization strategy will rely on the physical entrapment of antibodies specific for estrogen metabolites within acrylamide gel plugs. Antigen specific hydrogels will be evaluated for their ability to capture and concentrate estrogen metabolites at various concentrations. Limits of detection and non-specific binding will be assessed to directly compare the utility and diverse applications of the two methods. The broader impacts of this research are linked to its potential to:1. train numerous minority graduate and postdoctoral students in interdisciplinary research, 2. enhance current outreach activities designed to give economically and socially disadvantaged high school students hands-on research experience, 3. pioneer knowledge relating to microfluidic electrophoresis, and4. revolutionize the speed and resolution with which scientists can analyze macromolecules.