Building research capacity with high-throughput Ultra-performance liquid chromatography high resolution Q-TOF

生物分子的质谱分析(LC-MS)能够在一次测量中识别和定量数千种代谢物、脂质和蛋白质。在过去的10-15年里，仪器的灵敏度和分辨率发生了一场革命，使人们能够更深入地了解活细胞内的分子过程。在伦敦帝国理工学院，我们拥有世界领先的从临床和研究样本中分析代谢物和脂肪的能力，并开创了对临床组织(如癌症活检)的分子足迹进行成像的工作流程。然而，我们缺乏类似蛋白质组学工作流程的技术工具。我们希望建立一个最先进的多用户平台，通过尖端的蛋白质组学应用和小分子检测来支持健康研究。对新的LC-MS平台的投资将使临床样本具有全面的分子足迹的独特能力，使我们更好地了解赋予我们生命的化学反应，以及治疗疾病的新方法。我们已经确认的仪器(EvoSep-7600 ZenoTOF)具有独特的属性，使我们能够识别不同类别的分子(如胆汁酸种类)，并以高通量方式(每个样本约20分钟)分析数千种蛋白质，以支持神经退行性变、癌症、营养、实验医学和抗微生物等关键领域的研究。神经退行性变：帝国理工学院痴呆症研究所拥有数百个血液和大脑样本，用于研究阿尔茨海默病(AD)等疾病。特别是，了解不同的基因变异如何导致阿尔茨海默病，将在一定程度上通过对&gt；260大脑的蛋白质组分析来确定，这将导致对这种目前无法治愈的疾病的更深入了解。其他优先领域包括亨廷顿氏病和慢性疼痛。癌症：我们的研究小组率先对癌症组织进行了成像质谱分析，揭示了肿瘤内可能导致对化疗不同反应的不同区域。我们可以使用激光来解剖这些不同的区域(有时只有几个细胞)，并通过高度敏感的蛋白质组学和代谢组学分析来进一步表征分子途径。我们希望将其应用于更好地了解癌症及其对药物的反应，特别是乳房和胶质母细胞瘤。营养学和实验医学：检测营养可获得性的变化对于维持健康的生理至关重要。要了解营养感知，需要检测生物液中丰度较低的不同类别的分子(胆汁酸和信号肽)。7600 ZenoTOF的灵敏度将允许量化血液中低丰度的信号肽，而独特的EAD碎片特征将表征胆汁酸，以确定与肠道微生物区系的新关系。肠道微生物区系在胃肠道健康中起着基础性的作用，这一平台将有助于了解肠道微生物区系与我们之间的相互作用以及肠道微生物区系移植对健康有益的机制。抗菌素耐药性：最近的大流行突显了对新的抗病毒药物的迫切需求。使用LC-MS，我们将通过应用“标记”与病毒遗传物质接触的蛋白质的技术来调查病毒与其宿主(人类)之间的关系。通过这种方式，我们检测到病毒在分子水平上与什么相互作用以复制自己。了解这一点可以通过生产破坏这些相互作用的抗病毒药物来发现治疗病毒的新方法。它还将支持开发一种新的非抗生素类抗微生物药物的研究，这种药物可以破坏细菌与我们身体的病理性相互作用。

The analysis of biological molecules by mass spectrometry (LC-MS) enables the identification and quantification of many thousands of metabolites, lipids, and proteins in a single measurement. Over the last 10-15 years, there has been a revolution in instrument sensitivity and resolution allowing deeper insight into the molecular processes inside living cells. At Imperial College London, we have world-leading capabilities for metabolite and lipid characterisation from clinical and research samples and have pioneered workflows to image the molecular footprint of clinical tissue (such as cancer biopsies). However, we lack the technical instrumentation for similar proteomics workflows. We wish to establish a state-of-the-art multi-user platform to support health research with cutting-edge proteomics applications and small-molecule detection. Investment in a new LC-MS platform will enable the unique capacity for comprehensive molecular footprinting of clinical samples, leading to a greater understanding of the chemical reactions that give us life, and new ways of treating disease. The instrument that we have identified (EvoSep - 7600 ZenoTOF) has unique attributes to allow us to identify different classes of molecules (such as bile-acid species) and analyse thousands of proteins in a high-throughput manner (~20 minutes per sample) to support research in key areas such as neurodegeneration, cancer, nutrition, experimental medicine and anti-microbials.Neurodegeneration: The Dementia Research Institute in Imperial College have hundreds of blood and brain samples for studying diseases such as Alzheimer's disease (AD). In particular, understanding how different genetic variants can lead to AD will be determined, in part, through proteomic analysis of >260 brains, leading to greater insight into this currently incurable disease. Other areas of priority areas include Huntington's disease and chronic pain.Cancer: Imaging mass spectrometry of cancer tissue has been pioneered by our research group, revealing distinct regions within tumours that may cause different responses to chemotherapies. We can use a laser to dissect these distinct regions (sometimes only a few cells) and further characterise the molecular pathways by highly sensitive proteomics and metabolomic profiling. We wish to apply this to better understand cancers and their response to drugs, in particular breast and glioblastoma.Nutrition and experimental medicine: Detecting changes in nutrient availability is essential to maintaining healthy physiology. Understanding nutrient-sensing requires detecting distinct classes of molecules (bile acids and signalling peptides) at low abundance in biological fluids. The sensitivity of the 7600 ZenoTOF will permit quantification of the low abundance of signalling peptides within blood while the unique EAD fragmentation feature will characterise bile acids to identify new relationships with gut microbiota. Gut microbiotas play a fundamental role in gastrointestinal health, this platform will contribute to understanding the interaction between gut microbiota and ourselves as well as the mechanisms behind the health benefits of intestinal microbiota transplantation.Antimicrobial resistance: The recent pandemic has highlighted the urgent need for new antivirals. Using LC-MS, we will investigate the relationship between viruses and their host (humans) by applying technologies that "label" proteins coming into contact with the viral genetic material. In this way, we detect what the virus interacts with at a molecular level to replicate itself. Understanding this can lead to the discovery of new ways to treat viruses, by producing antivirals that disrupt these interactions. It will also support research into the development of a new non-antibiotic class of anti-microbials that disrupt how bacteria pathologically interact with our body.