Toward Single-Molecule Nanomechanical Mass Spectrometry

描述(申请人提供)：质谱学(MS)正在成为蛋白质组学最重要的工具之一。最近开发的用于在非常大的背景下识别低浓度分析物的现代MS协议，已经成为系统生物学中许多重要进展的核心。然而，这些议定书伴随着重要的局限性，特别是在它们的敏感性和方法学简易性方面。纳米科学的进步可能会为MS提供新的方法。最近，利用真空中的纳米机电系统(NEMS)，提出者已经展示了7 Zeptogram水平的质量传感--一个4千道尔顿分子的质量。这比最近使用微尺度设备进行的最先进的实验室演示提高了100万倍，比商业设备提高了10亿倍。然而，这项工作仍处于初级阶段；NEMS的重大进一步改进迫在眉睫。 在最初的努力(R21)中，提出者将采取关键的初步步骤，将纳米科学的这一进步转化为直接适用于生物质谱学的真正技术。现在需要进行大量研究，以将这种前所未有的群体敏感性转变为一种可用和有效的新形式的多发性硬化症，认识到这一转变是拟议工作的主要重点和意图。 NEMS-MS提供了低至单分子水平的灵敏度承诺。一种新的单分子、基于NEMS的质谱学(NEMS-MS)将使利用极其微量的贵重/稀有分析物进行强大的新分析成为可能。与基因组学研究不同的是，在基因组学研究中，基因扩增可以产生大量相同的分析物，从而实现更“传统的”MS协议--极端敏感性对蛋白质组学至关重要。如果成功，这一探索性计划将提供单分子质量传感的基准演示，绘制实现蛋白质组单分子质谱学的方法论进程，并为充分实现这一愿景组装下一阶段的努力(R01)。合作团队包括一些基于蛋白质组的质谱学、纳米设备物理学和微流体、以及用于MS的新的表面化学方法的前沿实践者。

DESCRIPTION (provided by applicant): Mass spectrometry (MS) is emerging as one of the most important tools for proteomics. Modern MS protocols, recently developed for identification of low concentration analytes in the presence of very large backgrounds, have been central to many important advances in systems biology. However, these protocols are accompanied with important limitations, especially in terms of their sensitivity and ease of methodology. Advances emerging from nanoscience may offer new approaches to MS. Recently, using nanoelectromechanical systems (NEMS) in vacuo, the proposers have demonstrated mass sensing at the 7 zeptogram level -- the mass of an individual 4 kilodalton molecule. This represents a million-fold improvement over the most recent state-of-the-art laboratory demonstrations using microscale devices, and a billion-fold improvement over commercial devices. Yet the work is still in its infancy; significant further improvements with NEMS are imminent. In this initial (R21) effort, the proposers will make the crucial initial steps to transform this advance in nanoscience into a real technology that is directly applicable to biological mass spectrometry. A significant amount of research will now be required to transform this unprecedented mass sensitivity into a usable and efficient new form of MS. Realizing this transformation is the principal focus and intent of the proposed work. NEMS-MS offers the promise of sensitivity down to the single-molecule level. A new single-molecule, NEMS-based mass spectrometry (NEMS-MS) would enable powerful new assays utilizing extremely minute amounts of precious/rare analyte. In contrast to genomics research -- where gene amplification can produce large amounts of identical analytes enabling more "conventional" MS protocols -- extreme sensitivity is crucial for proteomics. If successful, this exploratory program will provide benchmark demonstrations of single-molecule mass sensing, chart a methodical course toward the realization of single-molecule mass spectrometry for proteomics, and assemble a next-phase effort (R01) for the full realization of this vision. The collaborative team includes some of the forefront practitioners of proteomic-based mass spectrometry, of nanodevice physics and microfluidics, and of novel surface chemistry approaches to MS.