Single-Molecule Mass Spectrometry Enabled by Nanomechanical Systems (NEMS-MS)

描述（由申请人提供）：质谱是蛋白质组学最重要的工具之一。尤其重要的是最近实施的用于在非常大的背景存在下鉴定低浓度分析物的新方案。基于NEMS的质谱（NEMS-MS）可以为极其稀有或稀释的分析物提供互补和强大的新检测。如果成功的话，该计划将在生物质谱新方法的原型演示中达到高潮，提供前所未有的分辨率-与蛋白质组质谱领域的世界杰出领导者密切合作。下面的应用图表的实现单分子NEMS-MS的有条不紊的过程。我们已经从理论上表明，基于纳米机电系统（NEMS）的质量检测器的固有分辨率远低于1 Da。对于生物质谱（MS）领域的影响是深远的。为了继续我们成功的R21资助的试点计划-我们已经实现了真实的单分子NEMS质量检测的首次演示-我们提出了一个为期5年的研发计划（R 01），以开发具有单分子分辨率的紧凑型下一代高通量质谱仪。这些将基于微流体接口NEMS的大规模集成。 公共卫生相关性：蛋白质组学是研究所有生命过程的生物机制-它是生物医学和制药研究的关键。蛋白质鉴定的最重要工具是质谱（MS），但现有的工具通常只有在可以从细胞培养物中提取数亿个相同分子进行分析时才有效。该项目提出从纳米科学中开发一种新技术，已经以原型形式验证，允许在单分子水平上进行MS研究，这将明确阐明赋予细胞功能的生化网络的细节。

DESCRIPTION (provided by applicant): Mass spectrometry is one of the most important tools for proteomics. Especially important are new protocols recently implemented for the identification of low- concentration analytes in the presence of very large backgrounds. NEMS-based mass spectrometry (NEMS-MS) can provide complementary and powerful new assays for extremely rare or dilute analytes. If successful, the proposed program will culminate in prototype demonstrations of new methods for biological mass spectrometry providing unprecedented resolution - carried out in close collaboration with the world's preeminent leaders in the field of proteomic mass spectrometry. The application below charts a methodical course toward the realization of single-molecule NEMS-MS. We have shown theoretically that the intrinsic resolution of nanoelectromechanical systems (NEMS) -based mass detectors is well below 1 Da. For the field of biological mass spectrometry (MS) the implications of this are profound. To follow-on from our successful R21-funded pilot program - in which we have achieved the first demonstration of single-molecule NEMS mass detection in real time - we propose a 5- year research and development program (R01) to develop compact, next-generation, high-throughput mass spectrometers with single-molecule resolution. These will be based on the large-scale integration of microfluidic-interfaced NEMS. PUBLIC HEALTH RELEVANCE: Proteomics is the study of the biological machinery that underlies all life processes - and it is key to biomedical and pharmaceutical research. The paramount tool for protein identification is mass spectrometry (MS), but existing tools typically work only if hundreds of millions of identical molecules can be extracted from cell cultures for analysis. This project proposes to develop a new technique from nanoscience, already validated in prototype form, allowing MS studies at the single-molecule level that will unambiguously elucidate the details of biochemical networks that give cells their function.