IDBR: Development of an Advanced Quadrupole Ion Trap Mass Spectrometer for Proteomics

这一奖项授予一位为蛋白质组学开发改进的四极离子陷阱质谱仪的初学者。一种新的电动力学方法将用于在四极离子陷阱(QIT)中碎裂生物离子。与传统方法相比，这种新方法有四个潜在的优势：1)每秒获得比商业仪器更多的串联质谱图的能力，从而在与自下而上的分离系统相结合时能够实现更完整的蛋白质组覆盖；2)能够探测更高能量的裂解途径，这将为传统方法提供互补的序列覆盖；3)重新捕获和大量分析低质量产物离子的能力，这对于测序肽和执行定量蛋白质组学非常有利；4)能够增加定量蛋白质组中可以裂解的前体离子的尺寸，从而扩展了自上而下的蛋白质组学的能力。新仪器将利用一种名为动态碰撞诱导解离(DCID)的碰撞激活方法，从而在质量采集扫描期间选择性地将前体离子碎裂。这项工作的大部分将涉及软件开发，以便以完成DCID所需的非常规方法运行离子陷阱。还将编写程序来执行顺序循环实验，这些实验能够以自动方式检查或优化不同实验参数的影响。为了帮助简化这一发展在未来仪器中的实施，将作出相当大的努力来确定一套或多套普遍适用于各种前体离子的最佳实验参数。由于这种新的碎裂方法的应用不需要任何硬件修改，通过这项工作所取得的进展将很容易实现到现有的或新的四极或线性离子陷阱中，因此可以立即用于各种生物应用中。质谱学是蛋白质组学的核心技术。蛋白质组学是对蛋白质的大规模研究，对理解生物体至关重要。除了这一科学利益外，该奖项还将具有宝贵的教育利益。本科生和研究生将第一手学习生物研究中使用的仪器开发和质谱学的各种方法。该奖项将在暑期为两名女研究生和本科生提供资助。学生将学习与仪器仪表开发相关的技术和工程技能，这些技能在仪器仪表行业中并不常见，而且非常受欢迎。

This award is to a beginning investigator for developing an improved quadrupole ion trap mass spectrometer for proteomics. A new electrodynamic approach will be used for fragmenting biological ions in quadrupole ion traps (QIT). This new approach has four potential advantages over conventional methods: 1) the ability to acquire more tandem mass spectra per second than commercial instruments, thus enabling more complete proteome coverage when combined with bottom-up separation systems; 2) the ability to probe higher energy fragmentation pathways, which will offer complementary sequence coverage to conventional methods; 3) the ability to recapture and mass-analyze low mass product ions, which is highly beneficial for sequencing peptides and performing quantitative proteomics; 4) the ability to increase the size of precursor ions that can be fragmented in QITs, thereby expanding capabilities to top-down proteomics. The new instrument will utilize a method of collisional activation, called dynamic collision-induced dissociation (DCID), whereby precursor ions can be selectively fragmented during the mass acquisition scan. The majority of this work will involve software development in order to run an ion trap in the non-conventional method required to accomplish DCID. Programs will also be written to perform sequence-looped experiments that are capable of examining or optimizing the influence of different experimental parameters in an automated manner. To help streamline the implementation of this development into future instruments, considerable effort will be placed on determining a set or sets of optimal experimental parameters that will be universally applicable to a wide range of precursor ions. Because the application of this novel fragmentation method does not require any hardware modifications, developments made through this work will be readily available for implementation into existing or new quadrupole or linear ion traps, and could therefore find immediate use in diverse biological applications. Mass spectrometry is a central technology for proteomics. Proteomics, the large-scale study of proteins, is vital to understanding organisms. In addition to this scientific benefit, the award will have valuable educational benefits. Undergraduate and graduate students will learn first-hand about instrumentation development and the various methods of mass spectrometry that are used in biological research. The award will support two female graduate students and also undergraduate students during the summer quarters. Students will learn technical and engineering skills related to instrumentation development that are uncommon and highly sought-after in the instrumentation industry.