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 该提议是为了获得将放置在蛋白质组学中的bruker timstof pro仪器 克利夫兰诊所Lerner研究所共享实验室资源（SLR）。这 蛋白质组学SLR配备了两个LC-MS/MS系统，包括八年的热科学 Orbitrap Elite和三年历史的热科学融合Lumos系统。蛋白质组学SLR是 目前以容量运行，当前超过45％的实验涉及定量蛋白质组学 项目。蛋白质组学SLR的当前工作量导致蛋白质组学的等待时间很长 涉及分析20多个样本的实验。大型工作量和随后的 等待时间因精英和Lumos仪器的低吞吐量工作流程而感到愤怒 每天分析大约8个样品的仪器。该提议的目标是双重的， 包括增加蛋白质组学SLR的能力，更重要的是，扩展 SLR的功能包括高吞吐量蛋白质组学实验。 Pro乐器的Timstof 是快速扫描飞行器的四极时间，并根据此提案选择 表明该仪器每天可以分析40-100个样品而无需牺牲的数据 蛋白质组深度，灵敏度，准确性和鲁棒性。基于DIA的蛋白质组学的比较 分析来自蛋白质组学SLR中腔的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分钟内量化了更多的蛋白质 梯度（超过4800），与Lumos仪器的90分钟梯度相比（3000）。这 蛋白质组深度升高是由于timstof Pro的较高的扫描速率，以及 分离的其他维度（离子迁移率）以及该仪器的能力进行平行的能力 累积序列碎片（PASEF），该碎片允许四极质量滤波器同步 带有tims离子迁移池。有几项研究将受益于进入Timstof Pro。其中包括研究酒精相关肝病和非酒精的小鼠模型 相关的脂肪肝病（NAGY），分析衰老中发生的磺胺的改变 （Hine），一项对肠道微生物代谢物TMA急性改变的特征的研究 tmao（棕色），更好地了解胶质母细胞瘤（lathia）机制的研究，鉴定 重大心血管事件的危险因素（HAZE），严重的血浆生物标志物的鉴定 哮喘（LI）以及在主动脉动脉瘤（AAA）和骨关节炎中发生的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