Development of real-time control methodologies for tandem mass spectrometry

在基因组相关疾病的早期检测和相关生物标志物的发现中，最重要的目标之一是鉴定和表征相关细胞系中存在的蛋白质和蛋白质复合物。串联质谱（Tandem mass spectrometry，MS/MS）是鉴定和比较复杂生物系统中蛋白质的有力工具。然而，质谱仪的性能仍然受限于实际上可以解释的MS/MS数据的比例。例如，由四极杆飞行时间质谱仪产生的高达70%的肽MS/MS谱，以及由离子捕获仪器产生的高达85%的肽MS/MS谱，不能使用任何可用的技术来解释。此外，大量的时间被浪费在获取和试图解释这些质量差的光谱。根据我们的经验、文献综述和与业界的对话，很明显，提高蛋白质质谱性能的主要障碍是缺乏能够实时检测和/或选择靶肽离子进行碎片化和MS/MS分析的快速、有效的算法。该研究的目标是克服这一挑战，从而提高蛋白质质谱的通量和效率。这将通过开发（a）用于评估肽MS/MS谱的质量的有效算法和（B）用于使用经验证的肽MS/MS谱进行在线蛋白质鉴定的快速且准确的算法来实现。拟议的研究将显着提高质谱仪的性能，使开发的实时控制方法的基础上开发的算法在此建议。在提供这种先进的高通量蛋白质和蛋白质生物标志物鉴定方法的同时，该项目也将为加拿大的健康研究做出重大贡献，同时促进加拿大的学术，工业经济活动。

One of the most important goals in the early detection of genomic-related disease, and the discovery ofassociated biomarkers, is to identify and characterize the proteins and protein complexes present in related celllines. Tandem mass spectrometry (MS/MS) is a powerful tool for identifying and comparing the proteinsexpressed in complex biological systems. However, the performance of mass spectrometers remains limited interms of the proportion of MS/MS data that can actually be interpreted. For example, up to 70% of the peptideMS/MS spectra generated by quadrupole-time of flight mass spectrometers, and up to 85% of those producedby ion trapping instruments, cannot be interpreted using any available technique. Furthermore, a great deal oftime is wasted in acquiring and attempting to interpret these poor-quality spectra. Based on our experience,literature review, and conversations with industry, it is apparent that the main obstacle to improvingperformance in protein mass spectrometry is the lack of fast, efficient algorithms capable of real-time detectionand/or selection of target peptide ions for fragmentation and MS/MS analysis. The goal of the proposedresearch is to overcome this challenge, and hence, increase throughput and efficiency for protein massspectrometry. This will be achieved by developing (a) an efficient algorithm for assessing the quality of peptideMS/MS spectra, and (b) a fast and accurate algorithm for on-line protein identification using verified peptideMS/MS spectra. The proposed research will significantly improve the performance of mass spectrometers byenabling the development of real-time control methodologies based upon the algorithms developed in thisproposal. In providing such advanced methodologies for high-throughput identification of proteins and proteinbiomarkers, this project will also make a major contribution to Canadian health research, while promotingacademic, industrial economic activity in Canada.