A Cost-Effective High-Speed Clinical Diagnostics Instrument for Large Population Screening Based on Novel Liquid AP-MALDI MS Technology

基于先前资助的EPSRC研究（EP/L006227/1）的“开发新型MALDI质谱仪和高灵敏度多电荷离子生成技术”以及随后对该新技术的初步开发，拟议项目将开发一种新仪器，专门满足现代医学所要求的临床诊断关键要求，特别是在新的流行病如COVID-19的时代。临床样品中微生物的准确和快速表征对于启动最佳治疗和限制流行病的爆发至关重要。准确性和时间是患者获得最佳治疗结果的关键，最大限度地缩短恢复时间，更重要的是最大限度地降低发病率和死亡率。特别是，正确和快速地鉴定新发现的微生物病原体或抗微生物剂耐药菌株对于患者的康复非常重要。结合大规模测试的能力，它还将使全球能够更好地应对生物体（生物分型）的基质辅助激光解吸/电离（MALDI）质谱（MS）分析最近已被确立为用于鉴定临床相关微生物的、具有显著提高的分类准确性和分析速度的、优于经典临床微生物测定的上级方法。这已经导致了两个FDA批准的系统，用于通过MALDI MS生物分型进行微生物检测和鉴定。在拟议的项目中，这种方法将通过利用多电荷离子及其与脂质和其他生物分子的共分析在台式MS/MS仪器上得到实质性的改进，该仪器专门针对大规模，廉价的临床分析进行了优化，从而导致下一代的上级MALDI MS生物分型的临床使用和大规模测试。这种新仪器的独特功能和相关技术将是高速度、成本效益和高特异性。结合质谱法无与伦比的分辨率，这项新技术将是诊断测试的一个飞跃，允许在同一测试运行中测试多种疾病，并高度适应新的疾病，而不需要开发疾病/微生物特异性的测试试剂，难以获得，因此昂贵，特别是对于新发现的疾病（参见第10页）。COVID 19）。通过与生物制药行业和分析仪器制造商的合作以及BBSRC资助的研究，我们发现我们的新型AP-MALDI MS离子源为快速分析提供了一个灵敏的平台，并有可能用于微生物疾病的早期检测。拟议的项目将建立在这一初步数据的基础上，开发一种新的量身定制的仪器，供未来的临床使用，并探讨其优点（与目前的MALDI MS生物型相比）在（a）速度，（B）生物基质背景的消除，（c）上级MS/MS分析，（d）更高的离子信号稳定性，（e）多路复用能力和（f）这种新技术可以提供更简单（和具有成本效益）但更灵活的样品制备。

Based on previously funded EPSRC research (EP/L006227/1) for the 'Development of a novel MALDI mass spectrometer and technology for the generation of multiply charged ions at high sensitivity' and subsequent initial exploitation of this new technology, the proposed project will develop a new instrument that specifically fulfils key requirements in clinical diagnostics as demanded by modern medicine, in particular in the age of new pandemics such as COVID-19.Accurate and fast characterisation of microorganisms in clinical samples are crucial for initiating optimal treatment and limiting the outbreaks of pandemics. Both accuracy and time are key to the best treatment outcome for the patient, minimising the time to recovery and more importantly minimising morbidity and mortality. In particular, the correct and rapid identification of newly discovered microbial pathogens or antimicrobial-resistant strains is important for the patient's recovery. Combined with the capability of large-scale testing, it will also allow for a better global response to (microbial) infectious diseases.Matrix-assisted laser desorption/ionisation (MALDI) mass spectrometry (MS) profiling of organisms (biotyping) has recently been established as a superior method to classical clinical microbiology assays for the identification of clinically relevant microbes with substantially increased classification accuracy and speed of analysis. This has already led to two FDA approved systems for microbial detection and identification by MALDI MS biotyping. In the proposed project, this methodology will be substantially advanced by exploiting multiply charged ions and their co-analysis with lipids and other biomolecules on a bench-top MS/MS instrument specifically optimised for large-scale, inexpensive clinical analyses, thus leading to the next generation of superior MALDI MS biotyping for clinical use and mass testing.The unique features of this new instrument and the associated technology will be high speed, cost-effectiveness, and high specificity by MS/MS sequencing. Combined with the unrivalled resolution of mass spectrometry this new technology will be a step-change in diagnostic testing by allowing the testing of multiple diseases within the same test run as well as being highly adaptable to new diseases without the need to develop test reagents that are disease/microbe-specific, difficult to source and therefore expensive, in particular for newly discovered diseases (cf. COVID 19). The aim is to reach a throughput level of 100,000 samples per day at high detection accuracy and low cost per sample.From collaborating with the biopharmaceutical industry and analytical instrument manufacturers, and from research of a BBSRC-funded grant, we found that our novel AP-MALDI MS ion source provides a sensitive platform for rapid assay analysis with the potential for use in the early detection of microbial diseases. The proposed project will build on this preliminary data, develop a new tailor-made instrument for future clinical use and explore the advantages (compared to current MALDI MS biotypers) in (a) speed, (b) the elimination of biological matrix background, (c) superior MS/MS analysis, (d) greater ion signal stability, (e) multiplexing capability and (f) the simpler (and cost-effective) but more flexible sample preparation that this new technology can offer.