Instrumentation for Analysis of High Mass Biomolecules

这项工作的目的是推进最先进的全蛋白混合物分析，重点是电动力离子阱作为单分子和离子/离子反应化学的反应容器。全蛋白串联质谱法在很大程度上仅限于几种质谱技术，并且在大多数情况下是基于电喷雾电离的。该建议将强调线性离子阱的使用，它可以作为独立的质量分析仪，也可以很容易地与其他形式的质量分析接口。此外，该提案的一个方面致力于将单电荷离子气相转化为多电荷离子，这将有助于使用解吸电离方法进行全蛋白串联质谱分析，而解吸电离方法往往会形成单电荷离子。这样的发展将使非常适合表面空间分析的技术能够享受到现在主要限于通过电喷雾电离产生的带电离子的结构工具。该提案的其他方面有望扩大离子/离子反应在混合物分析和整个蛋白质离子（包括翻译后修饰的物种）结构信息的推导中的应用。

The objective of this work is to advance the state-of-the-art in whole protein mixture analysis with heavy emphasis on electrodynamic ion traps as reaction vessels for unimolecular and ion/ion reaction chemistries. Whole protein tandem mass spectrometry has been largely restricted to only a few types of mass spectrometry technologies and has, for the most part, been based on electrospray ionization. This proposal will emphasize the use of linear ion traps, which can serve as stand-alone mass analyzers or can readily be interfaced with other forms of mass analysis. Furthermore, one aspect of the proposal is devoted to the gasphase conversion of singly charged ions to multiply charged ions, which will facilitate whole protein tandem mass spectrometry using desorption ionization methods, which tend to form singly charged ions. Such a development would enable technologies that are well-suited to the spatial interrogation of surfaces to enjoy the structural tools now largely restricted to multiply charged ions derived via electrospray ionization. Other aspects of the proposal promise to expand the use of ion/ion reactions for mixture analysis and for the derivation of structrual information from whole protein ions, including post-translationally modified species. Furthermore, this work promises to make much more widely accessible means for studying whole proteins and protein complexes without prior recourse to enzymatic digestion. Particular emphasis will be placed on improved understanding and performance of electrospray ionization and matrix assisted laser desorption ionization applied to protein mixtures, charge state manipulation reactions, including charge inversion and, in some cases, sequential charge inversion steps, ion/ion reactions that provide structural information that is complementary to collisional activation of protonated species, and instrumentation development focused on stand-alone linear ion trap instrumentation and linear ion traps coupled with orthogonal acceleration time-of-flight mass analysis.