Advancing intact proteome separation for top-down and bottom-up mass spectrometry

想象一下，从一滴血中获得自己的病史是什么感觉。通过分析蛋白质分子（负责执行几乎所有的生化功能），医生可以诊断疾病的早期发作，甚至可能在你生病之前就开处方治疗以预防疾病！这是蛋白质组学的梦想，这是一门新兴的科学，利用先进的分析技术来表征极其复杂的生物混合物（在一个实验中检测到数千种蛋白质）。 不幸的是，尽管在过去十年左右取得了显着的进步，蛋白质组学的梦想还没有完全实现。与科学中的其他问题一样，理解蛋白质组学的关键在于改进技术的发展，以简化和标准化蛋白质分析策略。这是我目前研究计划的重点。 蛋白质组学已经看到了爆炸性的增长，因为质谱法的适用性，以加速表征的样品。现代蛋白质组学实验室将质谱法与多种分析技术相结合，以实现对样品中许多蛋白质组分的正确检测。利用NSERC所要求的资金，我的目标是了解和促进蛋白质样品的制备，这将使质谱检测成为可能。研究重点是蛋白质组分析中最具挑战性的两个问题：（1）低丰度蛋白质的定量分析;（2）在新兴的自上而下蛋白质组学领域的背景下，完整蛋白质的操作。我提出的方法，以促进直接分析完整的蛋白质的质谱，并加强蛋白质的分离，以进一步提高其表征。 我还提出了战略，在完整的水平上标记蛋白质的基础上，使定量蛋白质组分析。预计通过这些创新，蛋白质组分析将成为一种常规程序，从而可以用于临床实践，以改善人类健康。

Imagine what it would be like to obtain your own medical history from a single drop of blood. By analyzing protein molecules (responsible for carrying out practically all biochemical functions), a physician could diagnose the early onset of disease, and perhaps even prescribe treatments to prevent the illness, even before you get sick! This is the dream of PROTEOMICS, an emerging science that utilizes advanced analytical techniques to characterize extremely complex biological mixtures (several thousand proteins detected in a single experiment). Unfortunately, despite the remarkable advances made over the last decade or so, the dream of proteomics has not been fully realized. Like other problems in science, the key to understanding proteomics lies in the development of improved technologies, both to simplify and to standardize the protein analysis strategy. This is the focus of my current research proposal. Proteomics has seen explosive growth since the applicability of mass spectrometry to accelerate characterization of the sample. Modern proteomics laboratories couple mass spectrometry with a multitude of analytical technologies to achieve proper detection of the many protein components in the sample. With the requested funds from NSERC, I aim to understand and facilitate protein sample preparation which will enable mass spectrometry detection. Research emphasis is on two of the most challenging problems in proteome analysis: (1) quantitative analysis of low abundance proteins; (2) manipulation of intact proteins, within the context of the emerging field of top-down proteomics. I propose methods to facilitate the direct analysis of intact proteins by mass spectrometry, and enhance the separation of proteins to further improve their characterization. I also propose strategies, based on labelling proteins at the intact level, to enable quantitative proteome profiling. It is anticipated through these innovations that proteome profiling will become a routine procedure that can thus be brought into clinical practice in order to improve human health.