Acquisition of triple quadrupole mass spectrometer for quantitative proteomics

获取用于定量蛋白质组学的三重四极杆质谱仪

• 批准号: 8447862
• 负责人: Chang-Gyu Hahn
• 金额: $ 24.37万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-06-20 至 2014-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8447862
• 关键词: Animal Model; Antibodies; Biochemical; Biological Markers; Biological Models; Biological Process; Cell model; Disease; Doctor of Philosophy; Enzyme-Linked Immunosorbent Assay; Equipment; Funding; Genomics; Goals; Human; Ions; Laboratories; Leadership; Mass Spectrum Analysis; Medical; Methodology; Molecular; Pathway interactions; Pennsylvania; Peptides; Post-Translational Protein Processing; Proteins; Proteome; Proteomics; Quantitative Evaluations; Research Personnel; Sampling; Sensitivity and Specificity; Stable Isotope Labeling; Staging; Translational Research; United States National Institutes of Health; Universities; Validation; Western Blotting; base; instrument; interest; mass spectrometer; multiple reaction monitoring; nano; novel therapeutics; programs

DESCRIPTION (provided by applicant): The purpose of this application is to acquire a mass spectrometer, specifically the TSQ Vantage Triple Stage Quadrupole instrument from Thermo Scientific. The TSQ Vantage has unique capabilities to support ongoing studies that investigate pathophysiological pathways of diverse medical illnesses as well as programs that are developing novel therapeutics. This instrument will be used exclusively for quantitative proteomic applications by 10 NIH funded investigators located primarily at the Translational Research Laboratories (TRL) in the University of Pennsylvania. Ongoing projects initiated by these investigators have employed genomic discovery, biochemical, and molecular approaches to discover proteins of interest, posttranslational modifications, and protein networks that execute biological functions. The majority of these projects have advanced past the stages of discovery and specific and practical protein and peptide targets have been identified. A critical next step for these projects is to explore these discoveries in depth by quantifying changes in proteins and peptides that occur in various diseases or treatments. This goal cannot be accomplished with typical antibody based western blot, ELISA or even with currently available linear ion trap mass spectrometers due to lack of sensitivity, specificity, and/or high throughput of these approaches. Targeted peptide quantification is a highly selective and sensitive methodological approach for quantitative evaluation of dynamic changes of proteins and protein networks as well as the validation and utilization of biological markers. Towards this goal the PI and the users of the proposed instrument, have developed and established methodologies that employ stable isotope labeled peptides, proteomes and multiple reaction monitoring (MRM)/mass spectrometry (MS) workflows for the quantification of proteins in human samples and animal and cellular model systems. A TSQ Vantage unit (Thermo-Scientific) combined with an Eksigent Nano 2DLC, which has been operational in the TRL since September 2010, will remain until March 2013 on a lease supported by the University of Pennsylvania. We propose to acquire this remarkably productive unit. Chang-Gyu Hahn, M.D, Ph.D. as the P.I will assume overall responsibility for the equipment. Leadership will be equally divided between Chang-Gyu Hahn, M.D, Ph.D. and Ian Blair, Ph.D. Functioning as Co-P.I.'s. Dr. Hahn will focus on the operational aspects and Dr. Blair will provide scientific oversight.

描述（由申请人提供）：本申请的目的是获得一台质谱仪，特别是 Thermo Scientific 的 TSQ Vantage 三级四极杆仪器。 TSQ Vantage 具有独特的能力来支持正在进行的研究，这些研究调查各种医学疾病的病理生理学途径以及正在开发新疗法的项目。该仪器将由主要位于宾夕法尼亚大学转化研究实验室 (TRL) 的 10 名 NIH 资助的研究人员专门用于定量蛋白质组学应用。这些研究人员发起的正在进行的项目采用了基因组发现、生化和分子方法来发现感兴趣的蛋白质、翻译后修饰和执行生物功能的蛋白质网络。这些项目中的大多数已经过了发现阶段，并且已经确定了特定且实用的蛋白质和肽目标。这些项目的关键下一步是通过量化各种疾病或治疗中发生的蛋白质和肽的变化来深入探索这些发现。由于这些方法缺乏灵敏度、特异性和/或高通量，使用典型的基于抗体的蛋白质印迹、ELISA 甚至当前可用的线性离子阱质谱仪都无法实现这一目标。靶向肽定量是一种高度选择性和灵敏的方法学方法，用于定量评估蛋白质和蛋白质网络的动态变化以及生物标志物的验证和利用。为了实现这一目标，PI 和拟议仪器的用户开发并建立了方法，采用稳定同位素标记的肽、蛋白质组和多反应监测 (MRM)/质谱 (MS) 工作流程来定量人类样本以及动物和细胞模型系统中的蛋白质。 TSQ Vantage 装置 (Thermo-Scientific) 与 Eksigent Nano 2DLC 相结合，自 2010 年 9 月以来一直在 TRL 运行，将在宾夕法尼亚大学支持的租赁下保留到 2013 年 3 月。我们建议收购这个生产力极高的单位。 Chang-Gyu Hahn，医学博士、哲学博士作为 P.I 将对设备承担总体责任。领导权将由医学博士、哲学博士 Chang-Gyu Hahn 平分。和伊恩·布莱尔博士担任 Co-P.I.。哈恩博士将专注于运营方面，布莱尔博士将提供科学监督。