Advanced tandem quadrupole mass spectrometer with an automated sample extractor for a multiuser interdisciplinary translational research facility

已经有数十亿英镑用于研究新的疾病测试（生物标志物），但只有少数人将其用于临床。原因是研究生物标志物的研究实验室通常缺乏直接的临床问题，并且通常缺乏可靠的方法开发和验证，因此可以在临床上使用。货车Geest Multi-OMIC设施在生物标志物的发现和翻译方面处于世界领先地位。我们与莱斯特大学医院合作开发了国家依从性测试中心（NCAT），用于评估患者是否依从处方药。这项工作不仅对患者结局和国际指南产生了影响，而且对临床试验的设计产生了影响。货车Geest Multi-OMIC设施参加了DHSC登月项目。该国家方案为快速开发、验证和实施用于检测拭子中SARS-CoV-2病毒的三重四极杆质谱仪方法建立了框架。该方案花了大约5个月的时间开发出一种有效的临床使用方法。LC-MS仪器是最先进的，由制造商沃茨提供一年的最低费用，并在我们的实验室现场。LC-MS具有精致的灵敏度和令人难以置信的仅鉴定目标蛋白质的能力（特异性）。这方面的一个例子是使用LC-MS鉴定优秀运动员的兴奋剂。在将样品加载到LC-MS之前，需要对其进行制备。这些手动的净化和分离步骤很慢，可能会导致错误。因此，在本提案中，我们还要求使用Agilent Bravo液体处理装置，以便我们能够以快速、可重现和稳健的方式进行各种样品制备方法。我们请求购买上述LC-MS，以开发四个主要的临床项目，这些项目可以利用从Moonshot计划和NCAT中吸取的经验教训转化为临床相关的检测。该设备物超所值，因为沃茨提供53.5%的折扣。第一个项目是开发一种方法，可以快速、可扩展地检测冬季出现的多种病毒，如流感和新冠肺炎。我们还可以使用MS来识别病毒的变体，并看看这是否可以用于确定长期COVID中的病毒持续性。第二个项目将为NCAT提供大大改进的方法。它将分析时间从80分钟缩短到45秒至3.5分钟。目前，由于所需的时间，该方法主要适用于高血压诊所。新的方法将使使用扩大到其他条件，包括心力衰竭和类风湿性关节炎，在住院患者和那些看到他们的家庭医生。我们还将开发研究大规模研究人群的能力，并根据个人的药物血液水平调整药物。第三个项目将率先开发10种方法来研究血压调节的关键途径--肾素-血管紧张素醛固酮系统（RAAS）。这些方法将有助于了解高血压的亚组和对药物的反应。第四个项目将允许远程病人监测长期疾病，如糖尿病，甲状腺疾病和心脏病的病人。患者将通过手指穿刺自行采集血液，我们将开发用于测量常规检查（如肝脏和肾脏功能）的方法。成功完成这些项目将有助于远程患者管理的稳健性-特别是在未来的流行病期间，并在患者时间和NHS中释放资源。这些项目是项目类型的典范，使我们能够快速验证并获得鉴定，然后直接进入NHS实验室。

There have been billions of pounds spend in research on new tests for diseases (biomarkers) but only a handful have made it to clinical use. The reasons are that there is often a lack of direct clinical question asked by research labs studying biomarkers and often a lack of robust method development and validation such that it can be used clinically. The van Geest Multi-OMIC facility is a world leader in the discovery and translation of biomarkers. We, in joint partnership with the University Hospitals of Leicester have developed the National Centre of Adherence Testing (NCAT), for assessing whether patients are adhering to their prescribed medications. This work has had impact not just on patient outcomes and international guidelines but to the design of clinical trials. The van Geest Multi-OMIC facility participated in the DHSC Moonshot Project. This national programme established a framework for rapid development, validation and implementation of a triple quadrupole mass spectrometer (LC-MS) method for the SARS-CoV-2 virus in swabs. The programme took about 5 months to develop a valid method for use clinically. The LC-MS instrument is state of the art and was provided with minimal fee for a year by the manufacturer Waters and is on site in our laboratory.LC-MS have exquisite levels of sensitivity and incredible levels of ability to identify only the protein of interest (specificity). An example of this is the use of LC-MS in identification of doping by elite athletes. Before samples can be loaded on the LC-MS they need to be prepared. These manual steps of purifying and isolating are slow and can lead to errors. Therefore, in this proposal we are requesting also an Agilent Bravo liquid handling unit to enable us to do a wide range of sample preparation approaches in a swift, reproducible and robust manner. We request the purchase of the above LC-MS to develop four main clinical projects that can be translated into clinically relevant assays using lessons learnt from the Moonshot programme and NCAT. The equipment offers very good value for money as it is being offered at a 53.5% discount by Waters. The first project is to develop a method that can detect multiple viruses seen in the winter such as influenza and Covid-19- in a rapid and scalable manner. We can also use MS to identify variants of viruses and look to see whether this can be used in determining viral persistence in Long COVID.The second project will deliver vastly improved methods for NCAT. It will reduce analysis time from 80 minutes to between 45 seconds to 3.5 minutes. Currently, due to the time taken, the method is available mainly for hypertension clinics. The new methods will enable the use to be expanded to other conditions including heart failure and rheumatoid arthritis, in hospitalised patients and those seeing their GP. We will also develop capabilities to study large research populations and the tailoring of medications according to an individual's medication blood level.The third project will lead a pioneering effort to develop ten methods to study a key pathway in blood pressure regulation- the renin-angiotensin aldosterone system (RAAS). These methods will help understand subgroups of hypertension and response to medications.The fourth project will allow remote patient monitoring of patients with long term conditions such as diabetes, thyroid disease and heart disease. Patients will self-collect their blood by finger prick, and we will develop methods for measurements of routine tests such their liver and kidney function. Successful completion will help provide robustness in patient management remotely- especially during future pandemics and free resources both in patient time and of the NHS.These projects are exemplars of the type of project that will enable us to swiftly validate and gain accreditation for assays that can then be taken straight into NHS laboratories.