TWO DIMENSIONAL MASS SPECTROMETER FOR PROTEOMIC RESEARCH

描述(申请人提供)：在过去的二十年里，质谱学在生命科学中的影响越来越引起人们的关注。随着人们对质量规范应用的兴趣越来越广泛，技术越来越深入到每个研究领域，也产生了更多的挑战。蛋白质组学研究的挑战之一是处理更复杂的样本，具有更高的吞吐量和更宽的动态范围。现有的仪器和技术仍然远远不能满足研究人员的需求。这项工作的目标是开发一种新型的质谱计及其相关方法，能够以更高的吞吐量提供更好的多肽覆盖率，以更低的检测限检测复杂的蛋白质样品，特别是对低丰度的蛋白质。本项目主要研究串联式二维质谱计及其应用。新仪器将获得完整的MS/MS信息，包括所有具有父子链接的前体离子的裂解离子。来自离子源的离子不会被过滤掉或在歧视性选择下，而是将每个离子进行碎片处理以获得MS/MS光谱，从而产生更高的离子利用效率。在第一阶段的研究中，建立了一台用于二维MS/MS实验的混合QIT-Q-TOF仪器样机。研究了各组元之间的界面，特别是离子陷阱的离子喷射。计算机模拟和实验结果都表明，利用所提出的反向扫描共振喷射，发射离子的动能足够低，可以与TOF分析仪对接，这进一步证实了QIT-Q-TOF概念在技术上是可行的。在二期资助下，我们将继续改进性能，最终达到商用仪器的要求。通过与尖端研究小组的合作，将开发关键应用程序和信息学工具。

DESCRIPTION (provided by applicant): The impact of mass spectrometry in life science has been increasingly draw attentions in the last two decades. As interests to mass spec applications getting broader and the techniques penetrate into each research area getting deeper, more challenges have been created. One of the challenges from proteomic researches is dealing with more complex samples with higher throughputs and wider dynamic range. Existing instruments and technologies are still far from meeting researchers demand. The goal of the proposed work is to develop a novel mass spectrometer and the associate methods which could provide much better peptide coverage at higher throughput for complex protein samples with lower detection limits, especially for low abundant proteins. This project will focus on tandem two-dimensional mass spectrometer and its applications. The new instrument will obtain complete MS/MS information, include fragmentation ions from all precursors with their parent-daughter linkage. The ions from the ion source will not be filtered out or under discriminatory selection, instead each of them will be fragmented to get MS/MS spectra, resulting much higher ion usage efficiency. Under phase I investigation, a prototype hybrid Qit-q-TOF instrument was built for 2D MS/MS experiments. Interfaces between each components have been studied, especially for ion trap ion ejection. Results from both computer simulation and experiments demonstrated the kinetic energy of ejected ions are low enough for interfacing TOF analyzer by using proposed reverse scan resonance ejection, which further approved Qit-q-TOF concept is technically feasible for 2D MS/MS. Under phase II funding, we will continue to improve the performance, eventually achieve the requirements of a commercial instrument. With the collaboration to cutting edge research groups, key applications and informatics tools will be developed.