Dual Spray Reactor for Biological Mass Spectrometry

描述(申请人提供)：质谱仪已经变得越来越复杂，分析功能强大，为肽序列和蛋白质组学应用提供了成熟的MS/MS方法。然而，尽管有一系列令人印象深刻的高性能仪器，仍然需要扩大针对目标应用的质谱学策略的灵活性，以及提高选择性和可调性。这项提议的目标是开发一种新的前端方法，用于在引入质谱仪之前操纵离子，从而增加串联质谱仪工作流程的多功能性。该策略基于双喷雾反应器，允许分析物离子与第二组试剂离子或分子相交，促进常压下的气态离子-分子和离子-离子反应。喷雾反应器将促进分析物离子的在线衍生化，基于酸碱化学操纵电荷状态，并增加用于定量和校准目的的内标。对于自下而上的蛋白质组学应用，双喷雾反应器将通过允许操纵电荷状态和化学衍生化的方便手段来扩大激活方法的灵活性。此外，促进选择性反应以添加可切割的报告标签、附着发色团或方便地添加内部标准以提高精密度的能力，包括前端控制离子化学提供额外的实验灵活性的一些方法。通过双喷雾反应器将实现一系列离子-离子和离子-分子反应，包括：(I)通过酸/碱化学和添加可电离基团来操纵电荷状态，(Ii)基于官能团的选择性反应，(Iii)添加发色团以增强光解离，(Iv)附着报告标签以便于通过SRM扫描来跟踪离子(数据依赖模式)，以及(V)输入参考标准以提高跑到跑的精确度。针对目标自下而上的蛋白质组学应用的适应为扩展现有高性能质谱仪的能力提供了新的范例，并为扩展用于蛋白质组应用的标准ESI-MS平台的可调性、灵活性和多功能性提供了巨大的潜力。1

DESCRIPTION (provided by applicant): Mass spectrometers have become increasingly sophisticated, analytically powerful instruments offering well-established MS/MS methods for peptide sequencing and proteomics applications. However, despite the impressive array of high performance instruments, there remains a need for extending the flexibility of mass spectrometric strategies for targeted applications as well as enhancing both the selectivity and tunability. The objective of this proposal is the development of a new front-end method for manipulation of ions prior to introduction into the mass spectrometer, thus adding versatility to tandem mass spectrometry workflows. The strategy is based on a dual spray reactor that allows intersection of analyte ions with a second population of reagent ions or molecules, promoting gaseous ion-molecule and ion-ion reactions at atmospheric pressure. The spray reactor will facilitate on-line derivatization of analyte ions, manipulation of charge states based on acid-base chemistry, and addition of internal standards for quantitative and calibration purposes. For bottom-up proteomics applications, the dual spray reactor will expand the flexibility of activation methods by allowing a convenient means for manipulation of charge states and chemical derivatization. Morever, the capabilities of promoting selective reactions to add cleavable reporter tags, to attach chromophores, or to conveniently add internal standards to improve precision, comprise some of the ways that front-end control over ion chemistry offers an added dimension of experimental flexibility. An array of ion-ion and ion-molecule reactions will be implemented via the dual spray reactor including: (i) manipulation of charge states via acid/base chemistry and addition of ionizable groups, (ii) selective reactivity based on functional group, (iii) addition of chromophores to enhance photodissociation, (iv) attachment of reporter tags to facilitate tracking ions via SRM scans (data-dependent mode), and (v) infusion of reference standard to improve run-to-run precision. Adaptation for targeted bottom-up proteomics applications offers a new paradigm for extending the capabilities of existing high performance mass spectrometers and affords significant potential for extending the tunability, flexibility, and versatility of the standard ESI-MS platform for proteomics applications. 1