Development of high-throughput, high-sensitivity EPR sample handling capabilities for biomedical research

开发用于生物医学研究的高通量、高灵敏度 EPR 样品处理能力

• 批准号: 10323039
• 负责人: CANDICE S KLUG
• 金额: $ 37.19万
• 依托单位: MEDICAL COLLEGE OF WISCONSIN
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-01-01 至 2024-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10323039
• 关键词: Area; Automobile Driving; Biological; Biological Models; Biology; Biomedical Research; Biomedical Technology; Collection; Communities; Complex; Custom; Data Collection; Development; Disease; Drug Design; Electromagnetic Fields; Electron Spin Resonance Spectroscopy; Electrons; Engineering; Environment; Enzymes; Equipment; Frequencies; Geometry; Goals; Intervention; Kinetics; Magnetism; Measurement; Membrane; Metalloproteins; Muramidase; Oxidation-Reduction; Performance; Positioning Attribute; Protein Conformation; Protein Dynamics; Reaction; Research; Sample Size; Sampling; Science; Signal Transduction; Structure; System; Techniques; Technology; Temperature; Testing; Time; Tube; biophysical techniques; clinical diagnostics; data acquisition; design; drug discovery; innovation; instrument; instrumentation; magnetic field; millisecond; nano; new technology; novel; protein structure; prototype; simulation; structural biology; theories

Project Summary/Abstract Electron paramagnetic resonance (EPR) spectroscopy is a critically important technique in biomedical research with a unique ability to detect naturally occurring or engineered unpaired electrons in complex biological environments. EPR has wide-ranging applicability to structural biology, metalloprotein research, redox biology, rational drug design, and clinical diagnostics. Groundbreaking advancements in sample volume requirements for biomedical EPR spectroscopy applications were made nearly four decades ago with the development of the loop-gap resonator (LGR), which represented breakthrough benefits such as 10-fold lower sample volume requirements and higher resonator efficiencies to increase signals over the commonly used cavity resonator. To accommodate the biomedical needs of even further increased signal intensity and to provide the broader scientific community with accessible technology, we propose to capitalize on our recent development of the dielectric LGR (dLGR) concept and optimize this resonator technology to increase sensitivity beyond that achieved upon introduction of the LGR. A dLGR is effectively a small dielectric resonator placed inside the inner loop of an LGR where the return flux of the dielectric flows through the outer loops of the LGR. Analytic theory and our high-frequency structure simulations indicate that the dLGR enables an order-of-magnitude improvement in sensitivity over the LGR. To take full advantage of the extremely low volume requirements for the dLGR, we propose to develop efficient sample handling technologies that couple the dLGR to high- throughput sample handling instrumentation. We aim to develop two transformative technologies for biomedical EPR applications: i) a nano-dLGR with a 10-fold increase in sensitivity for ~0.2 µL sample volumes integrated with a customized autosampler and ii) a micro-dLGR for a dramatic increase in sensitivity for ~2 µL sample volumes with an optimized stopped-flow system for millisecond time scale kinetics measurements. These transformative and innovative prototypes with outstanding sensitivity will be easy to use and ultimately widely available to the scientific community. Where the LGR was a transformative advance compared with cavity resonators, the dLGR is expected to be another transformative leap from an LGR.

项目总结/文摘