Date of Award
Master of Science (MS)
Mary J. Wirth
Committee Member 1
Committee Member 2
Protein drugs are increasingly developed in the pharmaceutical company. Under the regulation of FDA, high purity of therapeutic proteins needs to be maintained. Before putting those drugs in the market, fast and efficient method is in need to achieve homogeneity. Traditionally, polyacrylamide gel electrophoresis (PAGE), capillary electrophoresis (CE), and size-exclusion chromatography (SEC) are used for the purification process. These methods have the disadvantages of low time and cost efficiency, and this quality assurance process has become the bottleneck of production. In our group, sub-micron silica colloidal particles with polyacrylamide layer on the surface are packed inside capillaries to increase the separation efficiency. In this particular project, NanoOrange dye is non-covalently associated with the protein sample to best reserve their native conformation. Proteins were separated at a distance as short as 8.2 mm in 85 seconds with extremely low plate height. The efficiency can be improved by decrease the silica particle size. This high throughput separation has a potential to be adopted in industries.
Part II of this thesis describes the cost-effective DNA microarray project. Microarray is a technology evolved from southern blotting, and it is a common tool to measure the expression levels of the DNA samples. To quantify the DNA samples, fluorescently labeled target strands hybridize with the DNA probe which binds to a solid surface. DNA microarray is frequently utilized in clinical studies, and the efficiency is highly desired to be improved. When attach the probes on a smooth surface, the amount of DNA captured will be limited. In this research, the microarray sensitivity is increased by layering silica colloidal particles on the solid surface. Silica particles obtain face-centered cubic packing which increases the surface area to bind to the DNA probe, thus improve the microarray sensitivity. To reduce the cost of the microarray and to make it more point-of-care feasible, transparent plastic sheets is going to be researched to replace quartz silica plates. This new sensitive and cost effective microarray has a great possibility to be developed into point-of-care which detects a variety of diseases.
Hao, Yingxu, "Separation of On-Column Labeled Model Proteins with Packed Capillary Electrophoresis" (2014). Open Access Theses. 433.