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Development and use of new affinity ligands for pharmaceutical analysis
Abstract
This work is a combination of three projects involving the study of a binding probe used in affinity chromatography for the study of drug-protein interaction, the use of affinity chromatography to measure binding parameters of an affinity ligand, and the development of affinity chromatographic theory. In the first project, a common binding probe for human serum albumin (HSA) was examined for its stability in aqueous solution. Using nuclear magnetic resonance (NMR), the change in warfarin structure was identified as a slow first-order conversion between two epimers. The subsequent effects on warfarin binding to HSA was then monitored using zonal elution chromatography on an immobilized HSA column. From these studies, a small, but measurable effect was seen on the binding of R- and S-warfarin to HSA. This change in structure may account for some of the variation that has appeared in literature values for the binding of warfarin to HSA. In the second project, an aptamer against thyroxine was studied for its potential use in a chromatographic immunoassay using a series of frontal analysis experiments. From these, the binding parameters of the affinity ligand were measured and compared to those of an anti-thyroxine antibody. The binding parameters suggested that aptamers could be utilized as antibody mimics in such a binding assay. The third project introduced a new linear transform which could be used with sequential binding assay data. Using sequential binding theory, equations for the slope and intercept were derived. This equation was then applied to data from previously reported sequential binding assays. Of these, only sequential binding assays with similarly sized analyte and label compounds resulted in linear calibration curves.
Subject Area
Analytical chemistry|Pharmacology
Recommended Citation
Moser, Annette C, "Development and use of new affinity ligands for pharmaceutical analysis" (2006). ETD collection for University of Nebraska-Lincoln. AAI3203636.
https://digitalcommons.unl.edu/dissertations/AAI3203636