The broad goals of this research are to further develop capillary electrophoresis (CE) as a technique to study in-capillary microreactions and to study biomolecular non-covalent interactions. Its focus is on demonstrating principles and on developing useful bioanalytical procedures that can be applied to the analysis of enzyme kinetics and receptor-ligand interactions. Approaches will include in-capillary enzyme-catalyzed microreactions involving both single cell and non- single cell analysis using laser-induced fluorescence (LIF) and UV VIS detection, and affinity capillary electrophoresis (ACE). The research will examine enzymes and small biomolecules that are important in the development of disease and, hence, to the issue of public health. This understanding will strengthen the scientific base underlying the design, preparation, and application of studied. This research effort will provide the technician and graduate student technicians a solid background in chemistry by examining problems at the interface of chemistry, chemistry-related fields, and medicine. The research projects planned for the technicians are aimed at demonstrating the interdisciplinary nature of CE and its importance as a new and exciting instrument in solving broad scientific problems. The specific objectives of the research are to: * Expand the CE-microreactor concept in the analysis of enzyme- catalyzed reactions. * Examine receptor-ligand interactions using affinity CE (ACE). * Study the physical biochemistry of derivatized proteins using CE. This research will: * Extend the usefulness of CE using microscale quantities in bioanalytical applications. * Extend the utility of CE in examining biomolecular non-covalent interactions. * Examine the changes in protein-ligand interactions using covalently modified proteins.
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