Quadrupole Time-of-Flight LC-MS

四极杆飞行时间 LC-MS

• 批准号: 10176802
• 负责人: Belinda Belle Willard
• 金额: $ 53万
• 依托单位: CLEVELAND CLINIC LERNER COM-CWRU
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-04-15 至 2022-04-14
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10176802
• 关键词: 3 year old; 8 year old; Acute; Aging; Alcohols; Aortic Aneurysm; Asthma; Biological Markers; Cardiovascular system; Cell Mobility; Cells; Clinic; Data; Degenerative polyarthritis; Dimensions; Event; Extracellular Matrix; Fatty Liver; Glioblastoma; Goals; Laboratories; Liver diseases; Plasma; Proteins; Proteome; Proteomics; Research Institute; Resources; Risk Factors; Running; Sampling; Scanning; System; Time; Wait Time; Workload; base; experimental study; gut microbes; instrument; ion mobility; mouse model; non-alcoholic; trimethyloxamine

Abstract This proposal is for the acquisition of a Bruker timsTOF Pro instrument to be placed in the Proteomics Shared laboratory Resource (SLR) at the Lerner Research Institute at the Cleveland Clinic. The Proteomics SLR is equipped with two LC-MS/MS systems including an eight year old ThermoScientific Orbitrap Elite and a three year old ThermoScientific Fusion Lumos system. The Proteomics SLR is currently running at capacity, with over 45% of the current experiments involving quantitative proteomic projects. The current workload of the Proteomics SLR has resulted in long wait times for proteomic experiments that involve the analysis of more than 20 samples. The large workload and subsequent wait times are exasperated by the low throughput workflow of the Elite and Lumos instruments, each instrument analyzing approximately 8 samples per day. The goal of this proposal is twofold and includes increasing the capacity of the Proteomics SLR and, more importantly, to expand the capabilities of the SLR to include high throughput proteomic experiments. The timsTOF Pro instrument is a fast scanning quadrupole time of flight instrument and was selected for this proposal based on data that indicates that this instrument can analyze between 40-100 samples per day without sacrificing proteome depth, sensitivity, accuracy and robustness. A comparison of a DIA based proteomic analysis of a tryptic digest generated from a cell lysate on the Lumos housed in the Proteomics SLR and the timsTOF Pro instrument showed that the timsTOF Pro quantified more proteins in a 30 minute gradient (over 4800) compared to a 90 minute gradient on the Lumos instrument (3000). This increased proteome depth is due to the higher scan rates, 100 Hz, of the timsTOF Pro, along with an additional dimension of separation (ion mobility), and the capability of this instrument to perform Parallel Accumulation Serial Fragmentation (PASEF) which allows synchronization of the quadrupole mass filter with the tims ion mobility cell. There are several studies that would benefit from access to the timsTOF Pro. These include studies of mouse models of alcohol-associated liver disease and non-alcoholic associated fatty liver disease (Nagy), the analysis of alterations to the sulhydrome that occur in ageing (Hine), a study of the signatures that are acutely altered by the gut microbe-derived metabolites TMA and TMAO (Brown), studies to better understand the mechanisms of glioblastoma (Lathia), identification of risk factors for major cardiovascular events (Hazen), identification of plasma biomarkers of severe asthma (Li), and the study of ECM remodeling that occurs in aortic aneurysms (AAA) and osteoarthritis (Apte).

摘要 该提案旨在收购一台布鲁克timsTOF Pro仪器，用于蛋白质组学研究。 克利夫兰诊所勒纳研究所的共享实验室资源（SLR）。的 Proteomics SLR配备了两个LC-MS/MS系统，包括一个已有八年历史的ThermoScientific Orbitrap Elite和三年前的ThermoScientific Fusion Lumos系统。Proteomics SLR 目前正在满负荷运行，目前超过45%的实验涉及定量蛋白质组学 项目蛋白质组学SLR目前的工作量导致蛋白质组学的等待时间很长， 实验涉及20多个样品的分析。巨大的工作量和随后的 Elite和Lumos仪器的低通量工作流程增加了等待时间， 仪器每天分析大约8个样品。这项建议的目的有两个， 包括增加蛋白质组学SLR的容量，更重要的是， SLR的能力包括高通量蛋白质组学实验。timsTOF Pro仪器 是一种快速扫描四极杆飞行时间仪器， 数据表明，该仪器每天可以分析40-100个样品， 蛋白质组深度、灵敏度、准确度和鲁棒性。基于DIA的蛋白质组学方法的比较 分析Proteomics SLR中Lumos上细胞裂解物产生的胰蛋白酶消化物 timsTOF Pro仪器显示，在30分钟内， 梯度（超过4800）与Lumos仪器（3000）上的90分钟梯度相比。这 增加的蛋白质组深度是由于更高的扫描速率，100 Hz，timsTOF Pro，沿着 分离的额外维度（离子迁移率），以及该仪器执行平行 累积串联裂解（PASEF），允许四极质量过滤器同步 离子迁移率电池。有几项研究将受益于timsTOF的访问 Pro.这些研究包括酒精相关性肝病和非酒精性肝病的小鼠模型的研究。 相关脂肪肝（Nagy），分析衰老过程中发生的巯基变化 （Hine），一项关于肠道微生物衍生代谢物TMA和 TMAO（布朗），研究，以更好地了解胶质母细胞瘤（Lathia）的机制，识别 主要心血管事件的风险因素（哈岑），严重心血管事件的血浆生物标志物的鉴定 哮喘（Li），以及发生在主动脉瘤（AAA）和骨关节炎中的ECM重塑的研究 （Apte）.

项目成果

A novel strategy to characterize the pattern of β-lactam antibiotic-induced drug resistance in Acinetobacter baumannii.

一种表征鲍曼不动杆菌β-内酰胺抗生素诱导耐药性模式的新策略。

• DOI: 10.21203/rs.3.rs-2359505/v1
• 发表时间: 2023
• 期刊: Research square
• 作者: Hillyer,Trae;Benin,BogdanM;Sun,Chuanqi;Aguirre,Noah;Willard,Belinda;Sham,YukYin;Shin,WooShik
• 通讯作者: Shin,WooShik