Quantitative and Qualitative Differential Expression Proteomics

定量和定性差异表达蛋白质组学

• 批准号: 7794613
• 负责人: Martin Arthur Moseley
• 金额: $ 49.97万
• 依托单位: DUKE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-07-15 至 2011-07-14
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7794613
• 关键词: Address; Antibodies; Area; Biological Assay; Biological Markers; Biomedical Research; Blood; Body Fluids; Cell Line; Cells; Clinical; Clinical Research; Clinical Trials; Commit; Communicable Diseases; Core Facility; Critical Pathways; Data Analyses; Degenerative polyarthritis; Development; Disease; Funding; Genome; Health; Heart Diseases; Human; Institutes; Lead; Liver diseases; Medical; Mission; Patients; Principal Investigator; Proteins; Proteomics; Reporting; Reproducibility; Request for Proposals; Research; Research Personnel; Research Project Grants; Resource Sharing; Resources; Sampling; Science Policy; System; Time; Tissues; United States Food and Drug Administration; Universities; base; design; improved; innovation; instrument; liquid chromatography mass spectrometry; medical schools; multiple reaction monitoring; oncology; programs; protein complex; protein expression; public health relevance; research study; success

DESCRIPTION (provided by applicant): the Food and Drug Administration announced the development of the Critical Path Initiative in order to address the decreasing numbers of innovative medical products being submitted for approval. The first topic addressed by this report was the pressing need for the development of new biomarkers to improve clinical trials and improve medical therapy. In 2007, the School of Medicine and the Institute for Genome Sciences & Policy at Duke University collaborated to create the Duke Proteomics Core Facility, a shared resource facility with a mission to provide protein characterization resources and expertise in support of the basic and clinical research programs of Duke Investigators. The Proteomics Core Facility was originally designed to provide capabilities for liquid-chromatography-mass spectrometry (LC/MS) based proteomics for protein identification and protein quantitation, including biomarker discovery (open LC/MS/MS) and biomarker verification (targeted LC/MS/MS with multiple reaction monitoring) experiments, and has recently (February 2009) added the capability for multiplexed antibody assays for these biomarker verification studies. The Duke Proteomics Facility came on-line in September 2007, and has successfully served the proteomics needs of 44 Principal Investigators across the School of Medicine. The majority of the projects have been differential expression projects, providing both qualitative and quantitative information on a wide variety of sample types, including cell lines, tissue lysates, protein complexes, and biofluids. Of particular relevance is the success in biomarker discovery experiments in human clinical samples, which has lead to a significant increase in the number of projects submitted for biomarker discovery proteomics. Because of the large number of biomarker discovery/verification samples in the queue in 2009 from projects that have committed funds for their analyses (>1,300 LC/MS analyses), our Facility has reached maximum capacity, with new projects of this type being delayed indefinitely or turned away due to lack of LC/MS instrument time. Therefore, this S10 proposal requests an open 'omic LC/MS system effectively identical to the system we have in place in our facility to support the research projects from NIH-funded investigators described in this proposal. Thus, the focus is on adding capacity to our Facility, with the additional capacity coming from an instrument platform where we have a proven track record of success, one which has demonstrated excellent quantitative reproducibility and sufficient system ruggedness for clinical-scale biomarker studies, as is necessary to achieve the research aims of the studies described in this proposal. PUBLIC HEALTH RELEVANCE: The proposed LC/MS system will significantly increase the capacity of the Duke Proteomics Facility to conduct differential protein expression analyses of biologic materials such as cell and tissue lysates, blood, and other body fluids. The research projects enabled by this proposal aim to advance our understanding of disease areas including oncology, heart disease, liver disease, osteoarthritis, and infectious diseases, and ultimately will improve clinical trials and patient therapies through the discovery and characterization of protein 'biomarkers' of health and disease. This LC/MS system will essentially double the instrument capacity available for such studies at Duke and enable us to leverage the extensive expertise, robust data analysis capabilities, and plethora of important biomedical research opportunities at Duke to the fullest extent.

描述（由申请人提供）：美国食品和药物管理局宣布制定关键路径计划，以解决提交审批的创新医疗产品数量不断减少的问题。本报告讨论的第一个主题是迫切需要开发新的生物标志物以改进临床试验和改善药物治疗。 2007年，杜克大学医学院和基因组科学与政策研究所合作创建了杜克蛋白质组学核心设施，这是一个共享资源设施，其使命是提供蛋白质表征资源和专业知识，以支持杜克大学研究人员的基础和临床研究项目。蛋白质组学核心设施最初设计用于提供基于液相色谱-质谱 (LC/MS) 的蛋白质组学功能，用于蛋白质鉴定和蛋白质定量，包括生物标志物发现（开放式 LC/MS/MS）和生物标志物验证（具有多反应监测的靶向 LC/MS/MS）实验，最近（2009 年 2 月）添加了多重抗体测定的功能 这些生物标志物验证研究。杜克大学蛋白质组学设施于 2007 年 9 月上线，并成功满足了医学院 44 名主要研究人员的蛋白质组学需求。大多数项目都是差异表达项目，提供各种样品类型的定性和定量信息，包括细胞系、组织裂解物、蛋白质复合物和生物流体。特别重要的是人类临床样本中生物标志物发现实验的成功，这导致提交生物标志物发现蛋白质组学的项目数量显着增加。由于 2009 年队列中有大量生物标志物发现/验证样品，这些样品来自已承诺分析资金的项目（> 1,300 次 LC/MS 分析），我们的设施已达到最大容量，此类新项目被无限期推迟或由于缺乏 LC/MS 仪器时间而被拒绝。因此，本 S10 提案要求开放的“组学 LC/MS 系统”与我们设施中现有的系统实际上相同，以支持本提案中描述的 NIH 资助的研究人员的研究项目。因此，重点是增加我们设施的容量，额外的容量来自我们拥有良好成功记录的仪器平台，该平台已证明了临床规模生物标志物研究的出色定量再现性和足够的系统耐用性，这是实现本提案中所述研究目标所必需的。 公共健康相关性：拟议的 LC/MS 系统将显着提高杜克大学蛋白质组学设施对细胞和组织裂解物、血液和其他体液等生物材料进行差异蛋白表达分析的能力。该提案启动的研究项目旨在增进我们对肿瘤学、心脏病、肝病、骨关节炎和传染病等疾病领域的了解，并最终通过发现和表征健康和疾病的蛋白质“生物标志物”来改善临床试验和患者治疗。该 LC/MS 系统基本上将使杜克大学可用于此类研究的仪器容量增加一倍，并使我们能够最大程度地利用杜克大学广泛的专业知识、强大的数据分析能力和大量重要的生物医学研究机会。