CAREER: Capillary Electrophoretic Mobility Spectrometry - A Solution Phase Analog to Ion Mobility Spectrometry

职业：毛细管电泳迁移谱分析 - 离子迁移谱分析的溶液相模拟

• 批准号: 1944902
• 负责人: Christopher Baker
• 金额: $ 62万
• 依托单位: University of Tennessee Knoxville
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-12-01 至 2020-11-30
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1944902&HistoricalAwards=false
• 关键词: CAREER; Capillary; Electrophoretic; Mobility; Spectrometry

The ability to measure the structure of molecules is key to understanding their behavior, such as how chemical species move and why they interact with other molecules and themselves. Creating new measurement methods that provide answers to these questions is especially important when studying complex molecules containing hundreds and thousands of atoms, such as biological molecules. Of great interest are proteins, because their behavior at the molecular level is strongly influenced by their environment, which often changes on a rapid time scale. From a long-term outlook, developing basic science tools that offer insight into protein behavior can lead to new perspectives on factors that impact the health of organisms. Dr. Christopher Baker at the University of Tennessee Knoxville develops technologies that afford unprecedented, high-speed measurements of the structures of proteins in their native environment. In this project, a new tool called capillary electrophoretic mobility spectrometry enables the simultaneous measurement of protein size and ionic charge in extremely tiny volumes of aqueous solution. This project is enabled by developments in 3-dimensional printing to create precision measurement tools. The educational outreach activities broaden the impact of the project by integrating research topics into curricula created in collaboration with middle school science-technology-engineering-mathematics (STEM) educators. This collaboration helps educators in resource-limited environments meet learning objectives outlined in Common Core Math and Next Generation Science standards. The educational outreach program focuses on the design and use of hands-on experiential learning projects and include 3-dimensional printing of practical scientific measurement devices and computer-aided simulation of difficult-to-observe chemical properties and events. With this award, the Chemical Measurement and Imaging Program is funding Dr. Christopher Baker at the University of Tennessee Knoxville to develop and integrate two multi-detector strategies in capillary electrophoresis (CE) to enable a new technology termed capillary electrophoretic mobility spectrometry (CEMS). Protein structure evaluation is a highly specific motivating force behind the development of methods for the selective detection of proteins in dynamic biological systems. Nuclear magnetic resonance spectroscopy and X-ray crystallography offer excellent structural resolution, but require relatively large quantities of highly purified proteins, which may be impossible for many analyses. Ion mobility spectrometry (IMS) is emerging as a tool for structural characterization and selective detection, but as a gas-phase ion analysis technique, IMS characterizes proteins under non-native conditions that influence protein conformation. Capillary electrophoresis (CE) is a separation technique highly analogous to IMS, however CE is performed in the solution phase. Unfortunately, the results of CE cannot be interpreted as straightforwardly as those of IMS, due to deleterious intermolecular interactions and dynamic properties of the solution environment. CEMS is based on the online integration of Fourier transform CE (FT-CE), which dramatically improves resolution over conventional CE, and Taylor dispersion analysis (TDA), which enables the direct and calibration-free measurement of hydrodynamic radii. Improved resolution and direct observation of structurally-descriptive properties are the key distinguishing factors between IMS and CE, and therefore, the integration of FT-CE with TDA may enable IMS-like analyses in the solution phase.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

测量分子结构的能力是理解其行为的关键，例如化学物质如何移动以及它们与其他分子和自身相互作用的原因。当研究包含成百上千个原子的复杂分子（如生物分子）时，创建新的测量方法来回答这些问题尤为重要。人们对蛋白质非常感兴趣，因为它们在分子水平上的行为受到环境的强烈影响，而环境往往在快速的时间尺度上发生变化。从长远来看，开发能够深入了解蛋白质行为的基础科学工具可以为影响生物体健康的因素带来新的视角。田纳西大学诺克斯维尔的克里斯托弗·贝克博士开发了一种技术，可以在蛋白质的自然环境中对蛋白质的结构进行前所未有的高速测量。在该项目中，一种名为毛细管电泳迁移率光谱法的新工具可以同时测量极小体积水溶液中的蛋白质大小和离子电荷。该项目得益于三维打印的发展，以创建精确的测量工具。教育推广活动通过将研究课题纳入与中学科学-技术-工程-数学（STEM）教育工作者合作创建的课程，扩大了该项目的影响。 这种合作有助于教育工作者在资源有限的环境中满足共同核心数学和下一代科学标准中概述的学习目标。教育推广计划侧重于设计和使用实践体验式学习项目，包括实用科学测量设备的三维打印和难以观察的化学性质和事件的计算机辅助模拟。凭借该奖项，化学测量和成像计划资助田纳西大学诺克斯维尔的Christopher Baker博士开发和整合毛细管电泳（CE）中的两种多检测器策略，以实现称为毛细管电泳迁移率光谱（CEMS）的新技术。蛋白质结构评估是动态生物系统中蛋白质选择性检测方法开发背后的高度特定的动力。核磁共振光谱和X射线晶体学提供了极好的结构分辨率，但需要相对大量的高度纯化的蛋白质，这对于许多分析来说可能是不可能的。离子迁移谱（IMS）是新兴的结构表征和选择性检测的工具，但作为一种气相离子分析技术，IMS表征蛋白质在非天然条件下，影响蛋白质构象。毛细管电泳（CE）是一种与IMS高度相似的分离技术，但CE是在溶液相中进行的。不幸的是，由于有害的分子间相互作用和溶液环境的动态特性，CE的结果不能像IMS的结果那样直接解释。CEMS基于傅里叶变换CE（FT-CE）和泰勒色散分析（TDA）的在线集成，前者大大提高了传统CE的分辨率，后者能够直接和免校准地测量流体动力学半径。改进的分辨率和直接观察结构描述属性是IMS和CE之间的关键区别因素，因此，FT-CE与TDA的集成可能使IMS类似的分析在solution phase.This award reflects NSF的法定使命，并已被认为是值得通过使用基金会的智力价值和更广泛的影响审查标准进行评估的支持。