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（MS/MS）是鉴定和比较复杂生物系统中表达的蛋白质的有力工具。然而，质谱仪的性能在实际可以解释的MS/MS数据的比例方面仍然有限。例如，由四极杆飞行时间质谱仪产生的高达70%的肽MS/MS光谱，以及由离子捕获仪器产生的高达85%的肽MS/MS光谱，不能使用任何可用的技术进行解释。此外，在获取和试图解释这些质量差的光谱时浪费了大量时间。根据我们的经验、文献综述和与行业的对话，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。拟议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光谱质量的有效算法和（B）使用经验证的肽MS/MS光谱进行在线蛋白质鉴定的快速准确算法来实现。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。通过提供这种先进的蛋白质和蛋白质生物标志物的高通量鉴定方法，该项目还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和工业经济活动。

One of the most important goals in the early detection of genomic-related disease, and the discovery of associated biomarkers, is to identify and characterize the proteins and protein complexes present in related cell lines. Tandem mass spectrometry (MS/MS) is a powerful tool for identifying and comparing the proteins expressed in complex biological systems. However, the performance of mass spectrometers remains limited in terms of the proportion of MS/MS data that can actually be interpreted. For example, up to 70% of the peptide MS/MS spectra generated by quadrupole-time of flight mass spectrometers, and up to 85% of those produced by ion trapping instruments, cannot be interpreted using any available technique. Furthermore, a great deal of time 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 improving performance in protein mass spectrometry is the lack of fast, efficient algorithms capable of real-time detection and/or selection of target peptide ions for fragmentation and MS/MS analysis. The goal of the proposed research is to overcome this challenge, and hence, increase throughput and efficiency for protein mass spectrometry. This will be achieved by developing (a) an efficient algorithm for assessing the quality of peptide MS/MS spectra, and (b) a fast and accurate algorithm for on-line protein identification using verified peptide MS/MS spectra. The proposed research will significantly improve the performance of mass spectrometers by enabling the development of real-time control methodologies based upon the algorithms developed in this proposal. In providing such advanced methodologies for high-throughput identification of proteins and protein biomarkers, this project will also make a major contribution to Canadian health research, while promoting academic, industrial economic activity in Canada.