Nanoliter-Scale NMR of Biomolecule Solutions using Dynamic Nuclear Polarization

Objective

Dynamic nuclear polarization (DNP) is an exciting and rapidly developing field in magnetic resonance, with solid-state and dissolution DNP techniques emerging as powerful tools across various applications, ranging from materials science to medicine. However, direct DNP in solutions still suffers from a lack of general applicability to macromolecules. The LiquidStateDNP project aims to establish DNP spectroscopy in the liquid state as an innovative tool for chemistry and biology, enabling the analysis of biomacromolecules with small sample volumes as low as a few hundred nanoliters – an achievement beyond the capabilities of traditional NMR techniques. This research will focus on using the solid-effect mechanism at high magnetic fields and room temperature, which offers significant potential for investigating a wide range of molecules. Notably, the solid-effect mechanism exhibits a favorable dependence on the molecular weight of the analyte, becoming increasingly effective for larger macromolecules. To achieve the project’s goals, I will pursue the following objectives: (i) develop a DNP probehead with improved signal sensitivity and resolution; (ii) establish methodologies for various nuclei, including ¹H, ¹³C, ¹⁹F, and ³¹P; (iii) optimize the structure of polarizing agents to maximize their effectiveness in solid-effect DNP; and (iv) demonstrate the application of liquid-state DNP to biomolecules for the first time. This will involve monitoring ligand binding to nucleic acids or proteins, investigating the dynamics of native membranes, and conducting in-cell NMR studies on living cells to detect drug binding and penetration. The LiquidStateDNP project aims to develop an innovative tool for liquid-state NMR, unlocking fundamentally new opportunities for a variety of biological applications at the nanoliter scale under physiological conditions.