New microsystems for protein and peptide separations
Abstract
This dissertation consists of five chapters and two appendices where chapter I serves as general introduction. Chapter II was to miniaturize HPLC system on a chip by fabricating individual support structures equivalent to particles and form the entire column in situ through photolithography. Channels between the support structure were 1.5 μm in width. Various monolith shapes were fabricated in a quartz wafer. The chip columns were 4096 particles wide and 1, 3, or 5 cm in length. Of monolith support structures were coated with C 18 reversed phase for small molecule and peptide separations. Additionally, a crosslinked PEI anion exchange stationary phase for protein separations was tested. New types of on-chip injectors with hydrophobic coated monolith structure for HPLC-on-a-Chip has been proposed in the appendix II. Chapter III investigates new types reversed phase stationary phase and restricted access reversed phase media based on poly(succinimide) for analyzing small molecule in protein media. Poly(succinimide) reacted with aminated silica particles. Excess succinimide would react with hydrophobic amine to form a reversed phase for protein and small molecule separations. Hydrophilic Jeffamine was grafted to the poly(succinimide) based reversed phase packing to form restricted access media which excluded proteins and peptides while separating small molecules. Chapter IV discusses hydrophobic and positively charged coating for small neutral molecule separations in open tabular CEC. PEI was absorbed in a capillary and crosslinked with hydrophobic EPON. Chapter V describes several hydrophilic coatings for protein separations under neutral conditions. A dynamic coating was developed for fast protein separations by applying surfactants, either octanoyl-n-methylglucamine or n-dodecyl-β-D-maltoside to a C 18 coated capillary. Appendix I discusses that chitosan was absorbed and crosslinked with EDGE to silica gels to form an anion exchange chromatographic column for protein separation. The surface charges were increased by addition of N, N-dimethylethylenediamine.
Degree
Ph.D.
Advisors
Regnier, Purdue University.
Subject Area
Analytical chemistry
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