Date of Award

5-2016

Document Type

Thesis

Degree Name

Master of Science (MS)

Legacy Department

Chemistry

Committee Member

Dr. R. Kenneth Marcus, Committee

Committee Member

Chair Dr. Jeffry Anker

Committee Member

Dr. Carlos Garcia

Abstract

Developing new stationary phases for liquid chromatography is continuing to drive high performance liquid chromatography (HPLC) into the future. In this regard the Marcus’ group has been leveraging the continued advances of Capillary-Channeled Polymer (C-CP) fibers in an attempt to meet the demand of high throughput biomarcomolecule chromatography. Separation mechanisms studied include: ion-exchange (IC), reversed phase (RP), affinity, hydrophobic interaction chromatography (HIC). In this work, the next generation of C-CP fiber stationary phases was thoroughly evaluated with respect to hydrodynamic concerns relating to protein chromatography. Traditionally C-CP fibers have eight channels that run co-linearly along the length of the fiber. Packed C-CP fibers form a network of pseudo-open capillary structures through channels interdigitating. The fibers studied have a much higher surface area to volume ratio compared to circular fibers with similar diameters. The open tubular network has an added bonus of operating at low back pressures. C-CP fibers are non-porous with regards to biomarcomolecules, resulting in fast mass transfer kinetics causing no significant C-term band broadening. The next generation of C-CP fiber has been developed with three larger more ridged channels. This design allows for tighter packing densities without compromising channel integrity. This advancement allows the fibers to operate at higher linear velocities leading to a separation of a six-protein suit (ribonuclease A, cytochrome C, lysozyme, transferrin, bovine serum albumin, and α-chimotrypsinogen) under reversed phase conditions. Surface modification of the C-CP fibers has been accomplished with a variety of techniques, both through covalent and physical adsorption modification. Of particular interest to this work is the Lipid Tethered Ligand (LTL) surface modification modality, which has seen excellent success when employed on polypropylene C-CP fibers. LTLs functionalize a surface with ion-exchange or affinity ligands through hydrophobic physical adsorption to augment the available surface chemistry in a quick and simple flow-through system. In the work presented here, the LTL system was applied to the most commonly used polymer resin, polystyrene-divinylbenzene. The effectiveness of LTL loading, stability, and kinetics on PS-DVB was evaluated. Ligand availability was evaluated with both biotin-LTL for the extraction of streptavidin and iminodiacetic acid-LTL for the extraction of methylene blue.

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