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 这项提议是为了购买一台Bruker timsTOF Pro仪器，放在蛋白质组学中 克利夫兰诊所勒纳研究所的共享实验室资源(SLR)。这个 蛋白质组学SLR配备了两个LC-MS/MS系统，其中包括已有八年历史的ThermoScience Orbitrap Elite和一个使用了三年的热科学聚变Lumos系统。蛋白质组学SLR是 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。目前蛋白质组学SLR的工作量导致蛋白质组的等待时间很长 涉及分析20多个样本的实验。庞大的工作负载和后续 等待时间因Elite和Lumos乐器的低吞吐量工作流而变得恼火 每天分析大约8个样品的仪器。这项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，扩大 SLR的能力，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，并被选为这项建议的基础上 数据表明，该仪器每天可以分析40-100个样本而不牺牲 蛋白质组的深度、敏感性、准确性和稳健性。一种基于DIA的蛋白质组学方法比较 蛋白质组学SLR中腔上细胞裂解物产生的胰酶消化分析 而timsTOF Pro仪器显示，timsTOF Pro在30分钟内定量了更多的蛋白质 倾斜(超过4800)与Lumos仪器90分钟的倾斜(3000)形成鲜明对比。这 蛋白质组深度的增加是由于timsTOF Pro更高的扫描频率(100赫兹)，以及 额外的分离维度(离子迁移率)，以及该仪器并行执行的能力 累加序列分段(PASEF)，允许同步四极质量滤光器 使用TIMS离子迁移率单元。有几项研究将受益于访问timsTOF 职业选手。这些研究包括对酒精相关性肝病和非酒精相关性肝病小鼠模型的研究。 相关性脂肪性肝病(NAGY)，分析在衰老过程中发生的硫氢酶变化 (Hine)，一项关于肠道微生物衍生代谢物TMA和TMA急剧改变签名的研究 TMAO(Brown)，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(HAZEN)，重症患者血浆生物标志物的识别 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