An AP-MALDI-FTICR Mass Spectrometer for Proteomics Research

该合同支持开发一种新型高灵敏度、常压、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），以推进蛋白质组学研究能力。本提案中针对离子源开发的特定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。大气压操作提供了机会来研究MALDI基质材料的功能要求，并仔细表征每个组件。由此开发的高灵敏度AP-MALDI离子源，结合FTICR-MS的精确质量、串联MS/MS能力，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能够同时使用APMALDI和电喷雾注射的仪器，为最佳肽质量指纹图谱提供准确的质量测量，并允许与在线色谱法耦合分析蛋白质混合物的消化。以下提案的知识价值集中在三个方面：1)通过实施多电离源和矩阵优化，结合离子光学的进步，提高AP-MALDI源的灵敏度；2)新设计的FTICR仪器与低成本的宽口径核磁共振磁体兼容；3)利用APMALDI- FTICR-MS技术开发精确质量的新方法，串联质谱能力用于蛋白质组学研究。这种仪器，当与自动化和在线色谱相结合时，将产生比目前实现的更高的通量蛋白质鉴定能力。按计划，该仪器可以利用目前用于核磁共振的相对便宜的宽孔磁铁，从而大大降低FTICR-MS仪器的成本。这将导致研究生研究人员在FTICR-MS相关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增加，研究生教育在工业、国家实验室和学术研究项目中的合作和参与增加。

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.