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记物的发现中，最重要的目标之一是识别和表征存在于相关细胞系中的蛋白质和蛋白质复合体。串联质谱仪(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，就实际可解释的MS/MS数据的比例而言，质谱仪的性能仍然有限。例如，高达70%的由四极飞行时间质谱仪产生的多肽MS/MS光谱，以及高达85%的由离子捕获仪器产生的多肽MS/MS光谱，无法使用任何可用的技术进行解释。此外，大量时间被浪费在获取和尝试解释这些质量不佳的光谱上。根据我们的经验、文献回顾和与业界的对话，很明显，提高蛋白质质谱学性能的主要障碍是缺乏能够实时检测和/或选择用于裂解和MS/MS分析的目标肽离子的快速、高效的算法。这项拟议研究的目标是克服这一挑战，从而提高蛋白质质谱学的吞吐量和效率。这将通过开发(A)用于评估多肽MS/MS图谱质量的有效算法，以及(B)用于使用经过验证的多肽MS/MS图谱在线鉴定蛋白质的快速而准确的算法来实现。拟议的研究将使基于本提案中开发的算法的实时控制方法的开发成为可能，从而显著提高质谱仪的性能。通过为高通量鉴定蛋白质和蛋白质生物标志物提供这种先进的方法，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.