TWO DIMENSIONAL MASS SPECTROMETER FOR PROTEOMIC RESEARCH

描述（由申请人提供）：在过去的二十年中，质谱在生命科学中的影响越来越受到关注。随着人们对质谱应用的兴趣越来越广泛，质谱技术对各个研究领域的渗透越来越深，也带来了更多的挑战。蛋白质组学研究的挑战之一是处理更复杂的样品，具有更高的通量和更宽的动态范围。现有的仪器和技术还远远不能满足研究人员的需求。本研究的目标是开发一种新的质谱仪和相关方法，以更高的通量为低检测限的复杂蛋白质样品，特别是低丰度蛋白质样品提供更好的肽覆盖。本项目将重点研究串联二维质谱仪及其应用。新仪器将获得完整的MS/MS信息，包括所有前体的碎片离子及其亲子连锁。离子源中的离子不会被过滤掉或进行歧视性选择，而是将每个离子碎片化以获得MS/MS谱，从而大大提高了离子的利用效率。在第一阶段的研究中，建立了一个用于二维质谱/质谱实验的混合Qit-q-TOF仪器原型。研究了各组分之间的界面，特别是离子阱的离子喷射。计算机模拟和实验结果均表明，所提出的反向扫描共振弹射法，弹射离子的动能足够低，可以与TOF分析仪相连接，进一步证明了Qit-q-TOF概念在二维质谱/质谱中是可行的。在第二期资助下，我们将继续改善性能，最终达到商业工具的要求。通过与前沿研究小组的合作，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.