Abstract

Polymeric microchips have received increasing attention in chemical analysis because polymers have attractive properties, such as low cost, ease of fabrication, biocompatibility and high flexibility. However, commercial polymers usually exhibit analyte adsorption on their surfaces, which can interfere with microfluidic transport in, for example, chemical separations such as chromatography or electrophoresis. Usually, surface modification is required to eliminate this problem. To perform stable and durable surface modification, a new polymer, poly(methyl methacrylate-co-glycidyl methacrylate) (PGMAMMA) was prepared for microchip fabrication, which provides epoxy groups on the surface. Whole surface atom transfer radical polymerization (ATRP) and in-channel ATRP approaches were employed to create uniform and dense poly(ethylene glycol) (PEG)-functionalized polymer brush channel surfaces for capillary electrophoresis (CE) separation of biomolecules, such as peptides and proteins. In addition, a novel microchip material was developed for bioanalysis, which does not require surface modification, made from a PEG-functionalized copolymer. The fabrication is easy and fast, and the bonding is strong. Microchips fabricated from this material have been applied for CE separation of small molecules, peptides, proteins and enantiomers. Electric field gradient focusing (EFGF) is an attractive technique, which depends on an electric field gradient and a counter-flow to focus, concentrate and separate charged analytes, such as peptides and proteins. I used the PEG-functionalized copolymer to fabricate EFGF substrates. The separation channel was formed in an ionically conductive and protein resistant PEG-functionalized hydrogel, which was cast in a changing cross-sectional cavity in the plastic substrate. The hydrogel shape was designed to create linear or non-linear gradients. These EFGF devices were successfully used for protein focusing, and their performance was optimized. Use of buffers containing small electrolyte ions promoted rapid ion transport in the hydrogel for achieving the designed gradients. A PEG-functionalized monolith was incorporated in the EFGF separation channel to reduce dispersion and improve focusing performance. Improvement in peak capacity was proposed using a bilinear EFGF device. Protein concentration exceeding 10,000-fold was demonstrated using such devices.

Degree

PhD

College and Department

Physical and Mathematical Sciences; Chemistry and Biochemistry

Rights

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