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Date of Award
Doctor of Philosophy (PhD)
Nuclear magnetic resonance (NMR) is a nondestructive experimental technique that often relies on radio frequency and large magnetic fields to investigate molecular structure and dynamics. The technique is well suited to study complex biological systems; however, universal application, to all samples, is hindered by low sensitivity. While several methods to increase NMR sensitivity currently exist, dynamic nuclear polarization (DNP) is one method gaining increased attention in the solid-state NMR community. This dissertation describes the development of DNP instrumentation for magic-angle spinning (MAS) NMR, including an efficient counterflow liquid nitrogen heat exchanger and novel spherical rotors. Furthermore, contained herein is the development of in-cell DNP. For the first time, DNP enhanced NMR signals from the intracellular regions of intact human cells are confirmed through the application of new fluorescent trimodal polarizing agents. The combination of fluorescent microscopy and fluorescence activated cell sorting with MAS DNP NMR provides a unique analytical platform to investigate intracellular molecular structures and dynamics. The in-cell DNP techniques developed in this research are expanded to investigate activation of HIV virus from latently infected human cell lines by NMR. Lastly, novel combinations of latency reversing agents, typically utilized in the “shock and kill” approach to HIV eradication to potently activate HIV from its latent state, are presented.
Chair and Committee
Alexander B. Barnes
George B. Kyei, Sophia E. Hayes, Jacob Schaefer, Timothy A. Wencewicz,
Albert, Brice Johnathan, "Instrumentation and Methods for In-cell Dynamic Nuclear Polarization HIV Cure Research" (2019). Arts & Sciences Electronic Theses and Dissertations. 1879.
Available for download on Tuesday, August 15, 2119