Dual Spray Reactor for Biological Mass Spectrometry

描述（由申请人提供）：质谱仪已经变得越来越复杂，分析功能强大的仪器，为肽测序和蛋白质组学应用提供了完善的MS/MS方法。然而，尽管有一系列令人印象深刻的高性能仪器，但仍然需要为目标应用扩展质谱分析策略的灵活性，以及提高选择性和可调性。本提案的目的是开发一种新的前端方法，用于在引入质谱仪之前操作离子，从而增加串联质谱工作流程的通用性。该策略基于双喷雾反应器，允许分析物离子与第二批试剂离子或分子相交，促进气态离子-分子和离子-离子在大气压下的反应。喷雾反应器将有助于分析物离子的在线衍生化，基于酸碱化学的电荷状态操作，以及用于定量和校准目的的内部标准的添加。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作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，促进选择性反应的能力，以添加可切割的报告标签，附加发色团，或方便地添加内部标准以提高精度，包括离子化学前端控制的一些方式，为实验灵活性提供了额外的维度。一系列离子-离子和离子-分子反应将通过双喷雾反应器实现，包括：(i)通过酸碱化学和可电离基团的添加来操纵电荷状态，（ii）基于官能团的选择性反应性，（iii）添加发色团以增强光解作用，（iv）通过SRM扫描（数据依赖模式）附着报告标签以方便跟踪离子，以及(v)注入参考标准物以提高运行到运行的精度。自底向上定向蛋白质组学应用的适应性为扩展现有高性能质谱仪的功能提供了新的范例，并为扩展用于蛋白质组学应用的标准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