Quadrupole time-of-flight mass spectrometer

描述(由申请人提供)：这项提案要求为高分辨率、高灵敏度和快速占空比的高精度四极飞行时间(QTOF)质谱仪提供资金，以帮助约翰霍普金斯大学医学院和公共卫生学院的一个富有成效的研究小组及其合作者进行蛋白质组学研究。该仪器将主要用于定性研究，如某些翻译后修饰的蛋白质鉴定和分析，以及用于测量蛋白质和翻译后修饰的相对丰度的定量研究。约翰霍普金斯大学现有的两台QTOF仪器(一台在蛋白质组学核心设施，一台在潘迪实验室)将于2009年淘汰，因为它们是老式仪器(分别使用了10年和7年)，缺乏较新仪器的许多先进特征，如快速占空比、高质量精度、大动态范围和准确的定量能力。由于缺乏用于蛋白质鉴定和定量的常规LC-MS/MS类型分析的高分辨率串联质谱仪，拟议的QTOF质谱仪有望成为这些项目的主要主力，在这些项目中，专家质谱学家和生物学家团队将受益匪浅。来自杰拉尔德·哈特、菲利普·科尔、罗伯特·科特、伯特·沃格尔斯坦、格雷格·塞门扎、科特·西文、哈罗德·瓦默斯和托马斯·肯斯勒实验室的研究小组将是主要用户。AkHilesh Pandey实验室的人员以及科特和HART实验室的人员将为那些没有质谱学专业知识的主要用户和其他用户提供质谱学实验设计和LC-MS/MS分析方面的专业知识。QTOF质谱仪的先进功能将用于iTRAQ和基于SILAC的质谱分析等用途。这些定量技术已经变得非常流行，并增加了我们对蛋白质组动态变化的了解。利用这种质谱学解决的生物学问题将从发现用于癌症早期检测的生物标记物和识别蛋白质复合体的成分到研究抗癌化合物的效果。最后，使用所建议的QTOF质谱计进行的一些实验的输出将有助于在其他类型的仪器(如三重四极杆质谱仪)上设计使用靶向蛋白质组学(例如多反应监测)的验证实验。 与公众健康相关：QTOF质谱仪是一种高分辨率质谱仪，具有高灵敏度，可以对蛋白质、多肽和翻译后修饰进行可靠的分析。它可以在多种操作模式下使用，包括数据依赖采集、母体离子和中性损耗扫描模式。该仪器将是发现阶段的关键，并为约翰·霍普金斯大学的一组富有成效的生物医学研究人员及其合作者提供关于多肽和蛋白质的基本信息。

DESCRIPTION (provided by applicant): This proposal requests funds for a high accuracy quadrupole time-of-flight (QTOF) mass spectrometer with high resolution, high sensitivity and fast duty cycle to assist a productive group of investigators at Johns Hopkins University School of Medicine and School of Public Health along with their collaborators in their proteomic studies. This instrument will mainly be used for qualitative studies such as protein identification and analysis of certain post-translational modifications (PTMs) and quantitative studies for measurement of relative abundance of proteins and PTMs. Two existing QTOF instruments (one in the proteomics core facility and one in the Pandey laboratory) are being retired at Johns Hopkins in 2009 as they are old instruments (10 and 7 years old, respectively) and lack many of the advanced features of newer instruments such as fast duty cycles, high mass accuracy, large dynamic range and accurate quantitation capabilities. Owing to this lack of availability of a high resolution tandem mass spectrometer for routine LC-MS/MS type analyses for protein identification and quantitation, the proposed QTOF mass spectrometer is expected to become a major workhorse for the projects in which a team of expert mass spectrometrists and biologists will be greatly benefited. The research groups from the laboratories of Gerald Hart, Philip Cole, Robert Cotter, Bert Vogelstein, Gregg Semenza, Curt Civin, Harold Varmus and Thomas Kensler will be among the major users. Personnel from Akhilesh Pandey's laboratory, along with personnel from Cotter and Hart laboratories, will provide expertise in design of mass spectrometry experiments and LC-MS/MS analysis for those major and other users who do not have mass spectrometry expertise. The advanced features of the QTOF mass spectrometer will be used for iTRAQ and SILAC-based mass spectrometric analysis, among other uses. These quantitative techniques have become very popular and have increased our knowledge about the dynamic changes in proteomes. The biological questions that will be addressed using this mass spectrometry will range from discovery of biomarkers for early detection of cancers and identification of components of protein complexes to studying effects of anti-cancer compounds. Finally, the output of some of the experiments carried out using the proposed QTOF mass spectrometer will be helpful for designing validation experiments using targeted proteomics (e.g. multiple reaction monitoring) on other types of instruments such as triple quadrupole mass spectrometers. PUBLIC HEALTH RELEVANCE: QTOF mass spectrometer is a high resolution mass spectrometer with high sensitivity that allows robust analysis of proteins, peptides and post-translational modifications. It can be used in several modes of operation including data dependent acquisition; parent ion and neutral loss scan modes. This instrument will be essential for the discovery phase and provide essential information about the peptide and proteins to a group of productive biomedical investigators at Johns Hopkins University and their collaborators.