An AP-MALDI-FTICR Mass Spectrometer for Proteomics Research

该奖项支持开发一种新型的高灵敏度、大气压、基质辅助激光解吸/电离(AP-MALDI)傅立叶变换离子回旋共振质谱仪(FTICR-MS)，以提高蛋白质组学研究能力。本提案针对的离子源开发的具体领域将包括基质效应和反应性，以及改进的离子捕获和传输。常压操作提供了调查MALDI基质材料的功能要求并仔细地单独表征每个组分的机会。由此开发的高灵敏度AP-MALDI离子源，与FTICR-MS的准确质量、串联MS/MS能力相结合，将为蛋白质组学研究和蛋白质鉴定提供理想的平台。这些努力将产生一种既能进行APMALDI，又能进行电喷雾注射的仪器，为最佳的多肽质量指纹图谱提供准确的质量测量，并允许与在线色谱联用，以分析蛋白质混合物的消化。以下建议的智力优势集中在三个方面：1)通过实施多个电离源和矩阵优化以及离子光学的进步来提高AP-MALDI源的灵敏度，2)与低成本大口径核磁共振磁体兼容的FTICR仪器的新设计，以及3)利用APMALDI-FTICR-MS技术进行蛋白质组研究的精确质量、串联MS能力的新方法。这样的仪器，当与自动化和在线层析相结合时，将产生比目前实现的更高的蛋白质识别能力。按照计划，该仪器可以利用目前用于核磁共振的相对便宜的大口径磁铁，从而显著降低FTICR-MS仪器的成本。这将增加对研究生研究人员在FTICR-MS所涉及技能方面的培训，增加下一代蛋白质组学研究的传播和适应，并加强与工业、国家实验室和学术研究项目的研究生教育的合作和参与。

This award supports development of a novel high sensitivity, atmospheric pressure, matrix-assisted laser desorption/ionization (AP-MALDI) Fourier transform Ion Cyclotron Resonance mass spectrometer (FTICR-MS) to advance proteomics research capabilities. Specific areas of ion source development targeted in this proposal will include matrix effects and reactivity, and improved ion capture and transmission. Atmospheric pressure operation presents the opportunity to investigate the functional requirements of MALDI matrix materials and carefully characterize each component individually. The resultant development of a high sensitivity AP-MALDI ion source, combined with the accurate mass, tandem MS/MS capabilities of FTICR-MS will provide an ideal platform for proteomics research and protein identification. These efforts will result in an instrument that is capable of both APMALDI and ElectroSpray Injection, giving accurate mass measurements for optimal peptide mass fingerprinting and allowing coupling with on-line chromatography for analysis of protein mixture digests. The intellectual merit of the following proposal centers on three aspects 1) the development of improved sensitivity of AP-MALDI sources through implementation of multiple ionization sources and matrix optimization combined with advances in ion optics, 2) a new design for an FTICR instrument compatible with lower cost wide-bore NMR magnets, and 3) new methods to exploit accurate mass, tandem MS capabilities with APMALDI- FTICR-MS technology for proteomics research. Such an instrument, when coupled with automation and on-line chromatography will result in much higher throughput protein identification capabilities than is currently achieved. As planned, the instrument can make use of relatively inexpensive wide bore magnets currently used for NMR, leading to significantly lower cost FTICR-MS instruments. This will lead to increased training of graduate student researchers in the skills involved in FTICR-MS, increased dissemination and adaptation of next-generation proteomics research and increased collaboration with, and involvement of graduate student education in industry, national laboratory and academic research programs.