Dual Spray Reactor for Biological Mass Spectrometry

用于生物质谱分析的双喷雾反应器

• 批准号: 8164272
• 负责人: Jennifer S. Brodbelt
• 金额: $ 15.97万
• 依托单位: UNIVERSITY OF TEXAS AT AUSTIN
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-09-15 至 2014-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8164272
• 关键词: Acids; Address; Algorithms; Amines; Ammonium; Atmospheric Pressure; Biological; Calibration; Charge; Chemicals; Chemistry; Cleaved cell; Complex Mixtures; Data; Databases; Development; Dimensions; Dissociation; Evaluation; Infusion procedures; Ions; Label; Lysine; Mass Spectrum Analysis; Methods; Modification; Monitor; Pattern; Peptide Conformation; Peptides; Performance; Population; Proteomics; Protons; Reaction; Reagent; Reference Standards; Reporter; Research; Running; Scanning; Side; Signal Transduction; Site; Solutions; Source; Techniques; base; carboxylate; chemical reaction; chromophore; flexibility; functional group; human disease; improved; instrument; instrumentation; ionization; mass spectrometer; protein aminoacid sequence; tandem mass spectrometry

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 PUBLIC HEALTH RELEVANCE (provided by applicant): Advances in the analytical performance and sensitivity of tandem mass spectrometers have allowed increasingly challenging biological questions related to the origins and progression of human diseases to be addressed. The proposed research will extend the flexibility and versatility of high performance mass spectrometers for broad-scale and targeted proteomics applications by exploiting on-line chemical manipulation of ions.

描述（由申请人提供）：质谱仪已变得越来越复杂，具有分析功能强大的工具，为肽测序和蛋白质组学应用提供了良好的MS/MS方法。但是，尽管有许多令人印象深刻的高性能工具，但仍需要扩展针对目标应用的质谱策略的灵活性，并提高了选择性和可调性。该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对离子进行操纵，从而增加了串联质谱工作流的多功能性。该策略基于双重喷雾反应器，该反应器允许将分析物与第二个群体的试剂离子或分子相交，从而在大气压下促进气态离子 - 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，基于酸碱化学的电荷状态以及添加内部标准的电荷状态以进行定量和校准目的。对于自下而上的蛋白质组学应用，双重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展激活方法的灵活性。此外，促进选择性反应以添加可切换的记者标签，附加发色团或方便地添加内部标准以提高精度的能力，包括对离子化学的前端控制提供了实验灵活性的一些方法。 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模式）和（v）输注参考标准以提高运行到运行精度。针对目标自下而上的蛋白质组学应用程序的适应为扩展现有高性能质谱仪的功能提供了一种新的范式，并为扩展蛋白质组学应用标准ESI-MS平台的可调性，灵活性和多功能性提供了巨大潜力。 1 公共卫生相关性（由申请人提供）：串联质谱仪的分析性能和敏感性的进步允许越来越具有挑战性的生物学问题与人类疾病的起源和进展有关。拟议的研究将通过利用离线的在线化学操作来扩展高性能质谱仪的灵活性和多功能性，以实现广泛的蛋白质组学应用。