Instrumentation for Analysis of High Mass Biomolecules

这项工作的目的是推进全蛋白质混合物分析的最新进展，重点是电动离子陷阱作为单分子和离子/离子反应化学的反应容器。整个蛋白质串联质谱学在很大程度上仅限于几种类型的质谱学技术，并且在很大程度上是基于电喷雾电离。这项建议将强调使用线性离子陷阱，它可以作为独立的质量分析器，也可以很容易地与其他形式的质量分析相结合。此外，该提案的一个方面是将单电荷离子的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整个蛋白质串联质谱分析，因为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于表面空间询问的技术能够享受目前在很大程度上限于通过电喷雾电离获得的多电荷离子的结构工具。该提案的其他方面承诺扩大离子/离子反应的使用，用于混合物分析和从整个蛋白质离子，包括翻译后修饰物种中获得结构信息。 此外，这项工作有望为研究整个蛋白质和蛋白质复合体提供更广泛的手段，而不需要事先求助于酶消化。 将特别强调提高对应用于蛋白质混合物的电喷雾电离和基质辅助激光解吸电离的理解和性能，电荷状态操纵反应，包括电荷反转以及在一些情况下的顺序电荷反转步骤，提供补充质子化物种碰撞激活的结构信息的离子/离子反应，以及侧重于独立的线性离子陷阱仪器和线性离子陷阱与正交加速飞行时间质量分析相结合的仪器开发。

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.