Novel Mass Spectrometer for Comprhensive No-Loss MS/MS of All Stored Ions

用于所有存储离子的全面无损 MS/MS 的新型质谱仪

• 批准号: 7405872
• 负责人: SUNNIE MYUNG
• 金额: $ 4.68万
• 依托单位: ROCKEFELLER UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-01-01 至 2010-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7405872
• 关键词: Argon; Beds; Biological; Canis familiaris; Cells; Classification; Complex; Condition; Cyclin-Dependent Kinases; Detection; Development; Devices; Diabetes Mellitus; Disease; Dissociation; Electronics; Electrospray Ionization; Future; Gases; Gene Expression; Genomics; Goals; Helium; Human Genome Project; Injection of therapeutic agent; Ions; Laboratories; Malignant Neoplasms; Mass Fragmentography; Mass Spectrum Analysis; Measurement; Messenger RNA; Methods; Microscope; Modification; Monitor; Nature; Optics; Peptides; Performance; Phase; Phosphoproteins; Phosphorylated Peptide; Phosphorylation; Physiologic pulse; Process; Property; Proteins; Proteome; Proteomics; Pulse taking; Range; Research; Resolution; Rheumatoid Arthritis; Ribosomes; Sampling; Spectrometry; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; Speed; Standards of Weights and Measures; Structure; System; Techniques; Testing; Time; Universities; Yeasts; data acquisition; detector; high throughput technology; instrument; instrumentation; interest; ion source; mass analyzer; mass spectrometer; novel; pressure; professor; prototype; research study; retinal rods; tool

DESCRIPTION (provided by applicant): The human genome project has inspired significant advancements in high-throughput technologies for the study of function and activation in gene expression. However, in order to truly tackle disease, the changes associated with gene expression at the mRNA and proteome levels has to be understood. Since the post-genomic era, the field of proteomics-the systematic profiling of proteins under a given set of conditions such as diseased versus healthy state- has grown tremendously. One of the major problems of proteomics is the low abundance of post-trans la tiona I ly modified proteins in complex biological mixtures such as phosphoproteins. This mandates a need for robust and highly sensitive detection system in which mass spectrometry (MS) has evolved as the method of choice for these analyses due to its fast experimental timescales and sensitivity. The long term goal of this proposal is to develop an ultra-sensitive ion trap mass spectrometry technique to routinely carry out comprehensive MS/MS analysis of complex biological mixtures. The key to this development is the ability to monitor all ions that are stored in an ion trap without discarding any, which is typically done in a standard proteomics experiment. The specific aims of this proposal are to: (1) Optimize the recently constructed high-capacity ion trap instrument by isolating the pressure within the ion trap to allow us to use optimal gases in the various regions of the instrument: helium in the high-capacity ion trap, N2 in the ion source cooling region, and N2 or argon in the collision cell. In addition, we will modify the geometry of the high-capacity ion trap to increase the resolution of the instrument. These straightforward improvements are expected to further enhance the performance of the high-capacity ion trap instrument. (2) The next phase of the research will couple an orthogonal injection reflectron TOP mass analyzer to the high-capacity ion trap. (3) Finally, future experiments will be geared towards the comprehensve sample analysis using this new high-capacity ion trap TOP MS instrument. Abnormal levels of protein phosphorylation are known to be associated with major diseases such as cancer, diabetes, and rheumatoid arthritis. The ability to identify and study these phosphorylation processes will ultimately bring us a step closer to finding a cure.

描述（由申请人提供）：人类基因组计划在基因表达功能和激活研究的高通量技术方面取得了重大进展。然而，为了真正治疗疾病，必须了解mRNA和蛋白质组水平上与基因表达相关的变化。自后基因组时代以来，蛋白质组学领域——在特定条件下（如患病与健康状态）对蛋白质进行系统分析——得到了极大的发展。蛋白质组学的主要问题之一是在复杂的生物混合物中，如磷蛋白，翻译后的I修饰蛋白丰度低。这就要求对强大和高灵敏度的检测系统的需求，其中质谱（MS）由于其快速的实验时间尺度和灵敏度，已经发展成为这些分析的首选方法。本提案的长期目标是开发一种超灵敏离子阱质谱技术，用于常规进行复杂生物混合物的综合MS/MS分析。这一发展的关键是能够监测存储在离子阱中的所有离子而不丢弃任何离子，这通常是在标准蛋白质组学实验中完成的。本提案的具体目标是：(1)通过隔离离子阱内的压力来优化最近建成的高容量离子阱仪器，使我们能够在仪器的各个区域使用最佳气体：高容量离子阱中的氦气，离子源冷却区中的氮气，碰撞池中的氮气或氩气。此外，我们将修改高容量离子阱的几何形状，以增加仪器的分辨率。这些直接的改进有望进一步提高高容量离子阱仪器的性能。(2)下一阶段的研究将把一个正交注入反射式TOP质谱仪耦合到高容量离子阱上。(3)最后，未来的实验将面向使用这种新型高容量离子阱TOP MS仪器进行全面的样品分析。蛋白质磷酸化水平异常与癌症、糖尿病和类风湿关节炎等重大疾病有关。识别和研究这些磷酸化过程的能力最终将使我们离找到治愈方法更近一步。