Hybrid Triple Quadrupole Mass Spectrometer for Quantitative Mass Spectrometric Ap

用于定量质谱应用的混合三重四极杆质谱仪

• 批准号: 7794609
• 负责人: HARRY ISCHIROPOULOS
• 金额: $ 50万
• 依托单位: CHILDREN'S HOSP OF PHILADELPHIA
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-08-01 至 2012-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7794609
• 关键词: Animals; Antibodies; Biochemical; Biological Markers; Biological Models; Biological Process; Cell model; Childhood; Collaborations; Development; Diagnosis; Disease; Enzyme-Linked Immunosorbent Assay; Funding; Generations; Genomics; Goals; Human; Hybrids; Ions; Knowledge; Liquid Chromatography; Methodology; Molecular; Pathway interactions; Pediatric Hospitals; Peptides; Philadelphia; Physiological; Physiological Processes; Post-Translational Protein Processing; Proteins; Proteolytic Processing; Quantitative Evaluations; Research; Research Institute; Research Personnel; Research Project Grants; Sampling; Sensitivity and Specificity; Stable Isotope Labeling; Staging; Techniques; Translational Research; United States National Institutes of Health; Validation; Western Blotting; base; insight; instrument; instrumentation; interest; mass spectrometer; multiple reaction monitoring; novel; outcome forecast; programs; public health relevance; research study; tandem mass spectrometry

DESCRIPTION (provided by applicant): The purpose of this application is to acquire a hybrid triple quadrupole-ion trap mass spectrometer, specifically the QTRAP 5500 from Applied Biosystems, with unique capabilities to support actively ongoing studies that aim to uncover critical physiological pathways and causes of pediatric diseases. This instrument will be used exclusively for quantitative mass spectrometric applications by 9 NIH funded investigators at the Joseph Stokes, Jr. Research Institute of the Children's Hospital of Philadelphia. Ongoing projects initiated by these investigators have employed genomic discovery platforms, biochemical, molecular, different mass spectrometric and orthogonal experiments to discover proteins of interest, posttranslational modifications, proteolytic processing and networks of proteins that execute biological functions. Therefore all these projects are well advanced past the stages of discovery and specific and practical protein and peptide targets have been identified. The immediate challenge for these projects is to move beyond qualitative descriptions to explore these discoveries in depth by quantifying changes in proteins and peptides. This goal cannot be accomplished with typical antibody based western blot, ELISA or even with our current linear ion trap mass spectrometers since these approaches are often neither feasible nor practical due to lack of sensitivity, specificity, and/or throughput. Targeted peptide quantification is becoming the clear choice as a highly selective and sensitive methodological approach for quantitative evaluation of dynamic changes of proteins, protein networks, posttranslational modifications, validation and utilization of biological markers. Towards this goal the PI and co-Investigator in collaboration with the users of the proposed instrument are actively developing methodologies that employ stable isotope labeled peptides and multiple reaction monitoring (MRM) workflows for the quantification of proteins in human samples as well as in animal and cellular model systems. Multiple reaction monitoring is emerging as the standard technique for quantitative liquid chromatography tandem mass spectrometry (LC/MS/MS) experiments; however generation and application of these MRM workflows with our current ion trap platforms is limited. Therefore the installation and use of the proposed mass spectrometer is vital for enabling the development and implementation of methodologies for targeted peptide quantification to generate sizeable new knowledge and insights. Overall, the proposed projects although biologically diverse are united by the need to develop, validate and implement quantitative hypothesis-driven mass spectrometric approaches. Thus, while each research project stands alone in scientific value and integrity the added value of this combined proposal is substantial. PUBLIC HEALTH RELEVANCE: The instrument will have a major and immediate impact on the research programs of 9 NIH-funded investigators that are engaged in both basic and translational research aimed to uncover fundamental physiological processes and mechanisms of disease at the Children's Hospital of Philadelphia. The requested instrumentation will facilitate the quantification of potentially novel biological markers for the prognosis and diagnosis of pediatric diseases.

描述（由申请人提供）：本申请的目的是获取混合三倍四极杆-ION陷阱质谱仪，特别是来自Applied Biosystems的QTRAP 5500，具有独特的功能，以支持积极持续的研究，旨在揭露儿童疾病的关键关键生理途径和儿童疾病的原因。该仪器将专门用于费城儿童医院研究所约瑟夫·斯托克斯（Joseph Stokes）的9HIH资助研究人员的定量质谱应用。这些研究者发起的正在进行的项目采用了基因组发现平台，生化，分子，不同的质谱和正交实验，以发现感兴趣的蛋白质，翻译后修饰，蛋白水解处理，蛋白质的网络以及蛋白质的网络，这些蛋白质的蛋白质是执行生物学功能的。因此，所有这些项目都超过了发现的阶段，并且已经确定了特定和实用的蛋白质和肽靶标。这些项目的直接挑战是超越定性描述，通过量化蛋白质和肽的变化来深入探索这些发现。通过典型的基于抗体的蛋白质印迹，ELISA或我们当前的线性离子陷阱质谱仪，无法实现此目标，因为由于缺乏灵敏度，特异性和/或吞吐量，这些方法通常既不是可行的也不是可行的。作为高度选择性和灵敏的方法学方法，有针对性的肽定量成为对蛋白质，蛋白质网络，翻译后修饰，生物学标记验证和利用的动态变化进行定量评估的一种明确的选择。为了实现这一目标，PI和共同研究器与所提出的仪器的使用者合作开发了使用稳定的同位素标记的肽和多个反应监测（MRM）工作流程的方法，以量化人类样品中的蛋白质以及动物和细胞模型系统中的蛋白质。多重反应监测已成为定量液相色谱串联质谱法（LC/MS/MS）实验的标准技术。但是，这些MRM工作流对我们当前的离子陷阱平台的生成和应用是有限的。因此，所提出的质谱仪的安装和使用对于实现目标肽量化的方法的开发和实施至关重要，以生成庞大的新知识和见解。总体而言，拟议的项目尽管生物学上的多样化是由于需要开发，验证和实施定量假设驱动的质谱方法而团结的。因此，尽管每个研究项目都以科学价值和完整性为单独，但该组合提案的附加值却很重要。 公共卫生相关性：该工具将对9位由NIH资助的研究人员的研究计划产生重大和直接的影响，这些研究计划旨在揭示费城儿童医院的基本生理过程和疾病的基本生理过程和机制。所需的仪器将促进对潜在的新型生物学标记物进行预后和诊断儿科疾病的量化。