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).

抽象的 该提案旨在购买布鲁克 timsTOF Pro 仪器，并将其放置在蛋白质组学中 克利夫兰诊所勒纳研究所的共享实验室资源 (SLR)。这 蛋白质组学 SLR 配备了两个 LC-MS/MS 系统，其中包括一个已有 8 年历史的 ThermoScientific Orbitrap Elite 和使用三年的 ThermoScientific Fusion Lumos 系统。蛋白质组学 SLR 是 目前正在满负荷运行，超过 45% 的当前实验涉及定量蛋白质组学 项目。目前蛋白质组SLR的工作量导致蛋白质组的等待时间很长 涉及 20 多个样品分析的实验。工作量大，后续 Elite 和 Lumos 仪器的低通量工作流程加剧了等待时间 仪器每天分析大约 8 个样品。该提案的目标是双重的 包括提高蛋白质组学 SLR 的容量，更重要的是，扩大 SLR 的功能包括高通量蛋白质组实验。 timsTOF Pro 仪器 是一种快速扫描四极杆飞行时间仪器，被选用于本提案的基于 数据表明该仪器每天可以分析 40-100 个样品而不牺牲 蛋白质组深度、灵敏度、准确性和稳健性。基于 DIA 的蛋白质组学比较 在蛋白质组学 SLR 中的 Lumos 上分析细胞裂解物产生的胰蛋白酶消化物 timsTOF Pro 仪器显示 timsTOF Pro 在 30 分钟内定量了更多蛋白质 梯度（超过 4800）与 Lumos 仪器上的 90 分钟梯度（3000）相比。这 蛋白质组深度的增加是由于 timsTOF Pro 的扫描速率更高（100 Hz）以及 分离的附加维度（离子淌度）以及该仪器执行并行操作的能力 累积串行碎片 (PASEF) 允许四极杆质量过滤器同步 与 tims 离子淌度池。有几项研究将受益于 timsTOF 的访问 亲。其中包括对酒精相关性肝病和非酒精性肝病小鼠模型的研究 相关脂肪肝病（Nagy），对衰老过程中发生的硫氢酶改变的分析 (Hine)，一项针对肠道微生物衍生代谢物 TMA 和 TMA 急剧改变的特征的研究 TMAO（布朗），研究以更好地了解胶质母细胞瘤（Lathia）的机制，鉴定 主要心血管事件的危险因素（Hazen），严重心血管事件血浆生物标志物的鉴定 哮喘 (Li)，以及主动脉瘤 (AAA) 和骨关节炎中发生的 ECM 重塑的研究 （阿普特）。

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