Lutz, Barry BLHsu, Yu-Hsuan2025-08-012025-08-012025-08-012025Hsu_washington_0250O_28548.pdfhttps://hdl.handle.net/1773/53396Thesis (Master's)--University of Washington, 2025Continuous flow paper electrophoresis (paper CFE) is a field separation technique utilized for the separation of various charged molecules. It is capable of processing any volume and serving as both an analytical and preparative tool. However, traditional paper CFE systems are limited by their large size, high power requirements, and long operation times, primarily suited for well-resourced laboratory settings. This project aims to address these limitations by miniaturizing paper CFE to a portable scale and reduce dependency on high voltage sources to expand the accessibility of electrophoresis separation. By changing the driven force from gravity in the traditional paper CFE to a capillary-driven system, and manipulating buffer composition, the project seeks to design a platform to achieve the separation of analytes. The project investigated the utility of the platform on detecting sickle cell hemoglobin as an analytical tool. Sickle cell hemoglobin results from a point mutation in the wild-type hemoglobin, altering a single amino acid in its structure, leading to different hydrophilicity and isoelectric point. The separation principle in this platform mainly depends on the difference in isoelectric point of the analytes. The successful development of this platform will provide a new tool contributing to improved disease management and patient outcomes. As the result, the designed prototype, with dimensions of approximately 4 cm*10 cm and operating at 30 V to 50 V, offers portability and reduced power requirements, and the separation of low molecular weight analytes and proteins was observed. Further work is needed to optimize the separation between proteins. The platform also shows potential for hemoglobin quantification. Overall, this project is a proof of feasibility and an advancement toward creating an analytic or preparative device that can be used in both basic lab settings and point-of-care tests in the field, making it more accessible outside of traditional labs.application/pdfen-USnoneHemoglobin electrophoresisIsoelectric focusingPaper-based microfluidicsPoint-of-care diagnosticsProtein separationSickle cell diseaseBioengineeringBioengineeringThesis