Procedure optimization for preparing human fecal samples for mass spectrometry analysis******

生物标记物被定义为指示与健康和疾病有关的个体生理状况的改变的分子(例如DNA、RNA、蛋白质)。粪便样本提供了一种非侵入性和生物相关的材料，用于寻找肠道疾病的生物标记物，如结直肠癌(CRC)和炎症性肠病(IBD)。尽管人类粪便蛋白生物标志物具有巨大的医学应用潜力，但到目前为止，对人类粪便蛋白生物标志物的研究仍然相对狭窄。**LC-MS/MS为研究粪便蛋白质组和鉴定生物标志物提供了一种有效的方法。然而，粪便的蛋白质组成非常复杂，利用这项技术开发一种有价值的分析人类粪便蛋白质的方法可能会很复杂。本申请中提出的项目的目标是建立一种协议，利用我们的合作伙伴PhenoSwitch Bioscience Inc.的特定仪器，优化LC-MS/MS检测人类蛋白质的有效性和一致性。将该协议转移到该公司将有助于增加他们的MS服务和未来的商业机会，例如分析来自健康患者、结直肠癌患者和IBD患者的样本，以确定生物标志物候选对象，并验证我们的LC-MS/MS策略。**在健康科学和医学正在迅速朝着更全面地使用生物标记物的背景下，该项目将为开发基于MS的创新测试奠定基础，以改进肠道疾病的筛查和诊断。这种创新对加拿大未来的潜在好处是多方面的，包括减少不必要的结肠镜检查数量和提高CRC患者的存活率，这两项都是加拿大患者和卫生系统所希望的。******

Biomarkers are defined as molecules (e.g. DNA, RNA, proteins) that indicate an alteration of the physiological condition of an individual in relation to health and disease. Stool samples offer a non-invasive and biologically relevant material for the search of biomarkers of intestinal diseases such as colorectal cancer (CRC) and the inflammatory bowel diseases (IBD). Despite a great potential for medical application, the research on human fecal protein biomarkers remains relatively narrow to this date. ** Liquid chromatography-tandem mass spectrometry (LC-MS/MS) offers a potent approach for investigating the fecal proteome and characterizing biomarkers. However, the protein composition of feces is very complex and the development of a worthy protocol for the analysis of human stool proteins with this technology can be complicated. The objective of the project presented in this application is to establish a protocol that optimizes the efficacy and consistency of human protein detection by LC-MS/MS with the specific instrumentation of our partner - PhenoSwitch Bioscience Inc. The transfer of the protocol to this company will serve to increase their MS services and future business opportunities such as analyzing samples from healthy, CRC and IBD patients in order to identify biomarker candidates and to validate our LC-MS/MS strategy. ** In the context where the health sciences and medicine are moving rapidly toward a more comprehensive use of biomarkers, this project will lay the foundation for the development of innovative MS-based tests that improve the screening and diagnosis of intestinal diseases. The future potential benefits of such innovation for Canada are multiple and encompass reducing the number of unnecessary colonoscopies and enhancing the survival rate of patients with CRC, both of which are desirable for Canadian patients and the health system. ******