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Cross-Linking an Unsaturated Phospholipid Bilayer for a Coating in Capillary Electrophoresis
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Cross-Linking an Unsaturated Phospholipid Bilayer for a Coating in Capillary Electrophoresis
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Dissertation
Cross-Linking an Unsaturated Phospholipid Bilayer for a Coating in Capillary Electrophoresis
2023
Overview
Semi-permanent capillary coatings are an effective tool in capillary electrophoresis (CE), allowing an analyst to control the conditions inside the capillary such as the electroosmotic flow (EOF) for the purposes of separating analytes. However, they pose a dilemma in CE due to their ability to be easily degraded through the separation process if not constantly being refreshed. Phospholipid bilayers, a type of semi-permanent coating, are diverse option for an assortment of separations but prove difficult in maintaining a consistent EOF and are easily removed by mechanical means over consecutive rinses. This work looks to utilize two unsaturated phospholipid bilayers, DPPC and DOPC, and cross-link their individual monomers with the binding agent osmium tetroxide. To determine the efficacy of this coating, the integrity of the coating and the consistency of the EOF over consecutive runs were observed after various modifications to the phospholipid preparation process were made. Both DPPC and DOPC showed significant masking of the capillary’s negative charge as indicated by a decreased EOF magnitude, but still showed signs of gradually increasing over time as portions of the coating were rinsed away. Charged proteins often provide a challenge in CE due to their tendency to adsorb to the capillary surface and was a way to further test the coatings’ integrity. While some proteins did experience band broadening issues, the cross-linked coating maintained its integrity through multiple runs with anionic proteins, indicating how effective the coating became when cross-linked. Finally, crosslinking the phospholipids made a more robust coating which showed a lowered EOF magnitude when left exposed to the air and when rinsed with the organic solvent methanol (MeOH). This work provides a greater insight into the efficacy of cross-linking phospholipid bilayers as a capillary coating in CE, which are less expensive than permanent coatings and respect projects that are time sensitive.
Publisher
ProQuest Dissertations & Theses
Subject
ISBN
9798380614429
