INSTRUMENTATION FOR ANALYSIS OF HIGH MASS BIOMOLECULES

这项工作的目的是通过扩展其混合物分析能力和结构分析的有用范围，从质量范围为2-4 kDa的分子扩展到质量范围为数万千道尔顿的分子，从而推动生物分子电喷雾/离子阱质谱技术的发展。这项工作的两个主要重点领域是提高对中等大小蛋白质（小于60 kDa）解离行为的因素的理解，以及开发基于离子/离子化学的复杂生物聚合物混合物分析的新实验。这项工作将允许考虑涉及直接审讯的应用，例如，整个蛋白质。这些应用可能包括快速蛋白质鉴定、快速鉴定翻译后修饰、快速分析蛋白质变异等。四极离子阱已经是一种非常强大的工具，用于生物衍生离子的操作和质量/电荷分析。这项工作将允许生物聚合物的尺寸急剧扩大，可以通过串联质谱与台式仪器进行研究。提出的努力旨在开发新的工具，以获得高质量离子的结构信息，特别强调蛋白质。将开发仪器以允许进行新的研究，旨在提高对蛋白质的离子/离子反应和蛋白质的单分子（即解离）反应的理解。具体地说，新的仪器发展将加强离子/离子化学的范围，可以研究和提高高质量离子的质量分析的优点数字。

The objective of this work is to advance the state-of-the-art in electrospray/ion trap mass spectrometry of biomolecules by extending its mixture analysis capabilities and its useful range of structural analysis from molecules ranging in mass up to about 2-4 kDa to molecules ranging in mass up to several tens of kiloDaltons. The two major emphasis areas in this work are to improve understanding of the factors that underlie the dissociation behavior of moderately sized proteins (less than 60 kDa) and to develop new experiments for complex bio-polymer mixture analysis based on ion/ion chemistry. This work will permit applications to be considered that involve the direct interrogation of, for example, whole proteins. Such applications might include rapid protein identification, rapid identification of post-translational modifications, rapid analysis of protein variants, etc. The quadrupole ion trap is already a remarkably powerful tool for the manipulation and mass/charge analysis of biologically-derived ions. This work will allow for a dramatic expansion in the size of bio-polymer that can be studied via tandem mass spectrometry with a bench-top instrument. The proposed effort seeks to develop new tools for deriving structural information from high mass ions, with particular emphasis on proteins. Instrumentation will be developed to allow for new studies designed to improve understanding of the ion/ion reactions of proteins and the unimolecular (i.e. dissociation) reactions of proteins. Specifically, new instrumental developments will be made to enhance both the range of ion/ion chemistry that can be studied and to improve the figures of merit of mass analysis of high mass ions.