IDBR: A New Mass Spectrometer for the Detection and collection of Biomolecules Of High Molecular Mass

质谱（MS）是现代生物学家不可缺少的工具。质谱技术在生物科学中的应用提供了重要的新能力和分析，具有以前未知的灵敏度、特异性和选择性水平，对复杂的生物学问题产生了新的见解。本研究项目的目的是开发一种用于生物研究的新型质谱仪，能够分析和收集非常高分子质量的离子。具体来说，将设计和建造一个飞行距离质谱仪（DOFMS），根据每个离子在给定时间段内移动的距离分离不同质量电荷比（m/z）的离子，最终沿表面在空间上分离离子。根据m/z的这种离子的物理分布，然后使用一种新颖的位置选择和电荷敏感离子探测器来测量，这种探测器已经在pi ？实验室。与现有方法相比，该策略在质量分离手段和离子检测机制上都没有质量上限，使整个质量范围都可用于分析。离子的物理分离也允许离子在一个专门的二次表面上收集，这种技术被称为软着陆质谱。有些令人惊讶的是，当收集的中和离子随后从真空中取出时，它们通常保持其生物功效，并可用于其他生化或分析技术的分析。DOFMS将通过为生物学家提供新的和显著改进的MS功能来推进生物科学。该仪器的无限质量范围将被用于研究大量生物聚合物和生物聚集体，如病毒、酶-底物复合物、噬菌体、蛋白质复合物和表位复合物。从分子质量的分布中，生物学家将直接了解组成所研究生物群体的个体分布，以及对结合结合和诱变变异影响的重要见解。此外，这些物种将以质谱的灵敏度、准确性和特异性进行检测，然后通过生化或其他方法收集用于后续分析或扩增。这样，MS研究的结果可以直接与其他分析的结果进行比较，进一步提高了信息量。该项目满足了生物界对高分子量和复杂性生物物种的快速、敏感分析的新兴需求。拟议计划的更广泛影响有几个形式。研究生和本科生都参与了最初的研究，并将继续参与仪器的开发和应用。作出特别努力，纳入代表性不足的群体；事实上，一名女研究生和一名非洲裔研究生已经完成了部分初步工作。由于拟议的研究本质上是多学科的，因此与生物科学研究人员的合作对成功至关重要。这种关系是围绕重要的生物学问题形成的，他们的合作参与将解决重要的生物学问题并改进新仪器。此外，已经制定了一项计划，使这一新工具能够更广泛地为生物界所用，部分通过上述合作，但也通过商业开发和现有的联邦用户设施。总的来说，这种新的分析工具的成功部署将对其自身产生更广泛的影响，并且利用质谱的显著优势来研究复杂和大型生物实体将产生无法通过其他方式获得的新知识。

Mass spectrometry (MS) is an indispensible tool of the modern biologist. The application of MS techniques in the biological sciences has provided significant new capabilities and analyses with previously unknown levels of sensitivity, specificity, and selectivity, yielding new insight into intricate biological problems. The aim of this research project is the development a new type of mass spectrometer for biological research that is capable of analyzing and collecting ions of very high molecular mass. Specifically, a distance-of-flight mass spectrometer (DOFMS) will be designed and constructed that separates ions of differing mass-to-charge (m/z) ratio based upon the distance each travels during a given time period, ultimately spatially segregating the ions along a surface. This physical distribution of ions according to m/z is then measured by using a novel position-selective and charge-sensitive ion detector that has already been developed in the PIs? laboratory. In contrast to extant methods, this strategy exhibits no upper mass limit either in the means of mass separation or in the mechanism of the ion detection, making the entire mass range available for analysis. The physical separation of the ions also permits the ions to be collected upon a specialized secondary surface in a technique known as soft-landing mass spectrometry. Somewhat surprisingly, when the collected, neutralized ions are subsequently removed from vacuum they often retain their biological efficacy and are available for analysis by other biochemical or analytical techniques. The DOFMS will advance the biological sciences by providing new and significantly improved MS capabilities to the biologist. The unlimited mass range of the instrument will be exploited by studying massive biopolymers and aggregates of biological interest such as viruses, enzyme-substrate complexes, phages, protein complexes, and epitope complexes. From the distribution of molecular masses, biologists will learn directly about the distribution of individuals that comprises the biological population under study, as well as important insights about associative binding and the effects of mutagenic variation. Moreover, these species will be detected with the sensitivity, accuracy, and specificity of MS, then collected for subsequent analysis or amplification via biochemical or alternative means. In this way the results of MS study can be directly compared to results of other analyses, further improving information content. This project meets an emerging need within the biological community for rapid, sensitive analysis of biological species of high molecular weight and complexity. The broader impacts of the proposed program come in several forms. Both graduate and undergraduate students have been involved in the initial studies and will continue to participate in both instrument development and application. Special efforts are made to include underrepresented groups; indeed, portions of the preliminary work have been performed by a female graduate student and a graduate student of African-American descent. Because the proposed studies are inherently multidisciplinary, collaboration with investigators in the biological sciences is important to success. Such relationships have been formed around biological questions of significance, and their collaborative involvement will both address important biological questions and refine the new instrument. Also, a plan has already been established to make this new tool available to the broader biology community, in part through the collaborations mentioned above but also through commercial development and through an existing Federal user facility. Overall, the successful deployment of this new analytical tool will have a broader impact of its own, and leveraging the significant strengths of MS to the study of complex and large biological entities will yield new knowledge that cannot be gained by other means.