Proteomics and Metabolomics

蛋白质组学和代谢组学共享资源--Stephen Byers，PhD/Albert Fornace，Jr，MD蛋白质组学和代谢组学共享资源(PMSR)的使命是向研究人员提供最先进的工具、方法和基于质谱学的技术。这种集中的共享资源可供研究人员用于常规和专门的蛋白质组和代谢组学应用。PMSR的目的是提供内部资源，丰富的技术专业知识，促进实验的自由流动 Lombardi综合癌症中心(Lombardi)的知识和合作。PMSR提供一系列基于蛋白质组学和代谢组学的服务。它配备了二维(2D)凝胶电泳仪，包括DGE成像仪和斑点拾取器(GE Healthcare)、4800MALDI TOF-TOF仪(ABI)和QSTAR Elite混合LC MS/MS系统(ABI)，以支持蛋白质组学工作流程。每个人 仪器拥有最新的集成软件和数据库搜索引擎(例如.Protein Pilot、GPS探测器)用于蛋白质识别和数据处理。此外，PMSR还拥有一名QTOF Premier(Waters Corp.)。在线使用UPLC系统进行代谢组分析和药物代谢研究。代谢物生物标记物研究的数据处理由“SIMCA-P”和“随机森林软件”提供支持。PMSR也 管理4000个QTrap，支持小分子代谢物的定量和验证，并允许进行有针对性的蛋白质组分析。PMSR的工作人员包括一名专门的生物信息学家，他与生物统计和生物信息学共享资源(BBSR)以及乔治敦大学的蛋白质信息资源(PIR)密切合作。最常见的实验包括鉴定所含蛋白质。 从电泳胶中分离的条带/斑点，以及使用鸟枪法和稳定同位素标记策略的比较蛋白质组学。我们的质谱学实验具有快速、灵敏和准确的特点，可以实现准确的定性和定量蛋白质组学。代谢组学计划由阿尔伯特·福内斯博士推动，涉及基于UPLC-TOFMS的辐射暴露生物标记物的发现。这项技术现在可以为研究人员提供，用于他们特定的研究兴趣。书面和电子报告 为所有样品提供，平均周转时间为两周。PMSR能够进行检测和表征蛋白质修饰的实验，如磷酸化和乙酰化。这些实验具有挑战性，需要更密集的努力。然而，PMSR提供的补充质谱学方法、工作人员的专业知识和经验以及与调查人员密切合作的能力相结合，使一些困难的项目得以成功完成。PMSR成立于2006年1月，在资源和使用方面都有了显著增长。 仅在PMSR的设备上，Lombardi就投资了100多万美元；因此，该设施不断地向Lombardi内5个项目的许多调查人员提供高质量的数据。PMSR由斯蒂芬·拜尔斯博士和小阿尔伯特·福纳斯博士领导。

Proteomics and Metabolomics Shared Resource-Stephen Byers, PhD /Albert Fornace, Jr, MD The mission of the Proteomics and Metabolomics Shared Resource (PMSR) is to provide state-of-the-art tools, methods, and mass spectrometry-based technology to investigators. This centralized shared resource is available to researchers for routine and specialized proteomic and metabolomic applications. The aim of PMSR is to provide an 'in house' resource, rich in technical expertise that fosters the free flow of experimental knowledge and collaborations across the Lombardi Comprehensive Cancer Center (Lombardi). The PMSR offers an array of proteomics- and metabolomics-based services. It is equipped with two-dimensional (2D) gel electrophoresis apparatus including DIGE imager and spot picker (GE Healthcare), 4800 MALDI TOF-TOF instrument (ABI), and a QSTAR Elite Hybrid LC MS/MS system (ABI) to support the proteomic workflow. Each instrument has the latest integrated software and database search engines (e.g.. Protein Pilot, GPS explorer) for protein identification and data processing. In addition, PMSR houses a QTOF Premier (Waters Corp.) online with an UPLC system for metabolomic profiling and drug metabolism studies. The data processing for metabolite biomarker studies is supported by the 'SIMCA-P' and the 'Random Forest software'. PMSR also manages a 4000 QTrap, which supports quantitation and validation for small-molecule metabolites, and allows targeted proteomic analysis. PMSR staff includes a dedicated bioinformatician who works closely with the Biostatistics and Bioinformatics Shared Resource (BBSR), as well as with the Georgetown University-based Protein Information Resource (PIR). The most common experiments include identification of proteins contained in bands/spots isolated from electrophoretic gels, as well as comparative proteomics using shotgun and stable isotope labeling strategies. The high speed, sensitivity, and accuracy of our mass spectrometry experiments allow accurate qualitative and quantitative proteomics. The metabolomics initiative has been driven by Dr. Albert Fornace, Jr, and involves UPLC-TOFMS based biomarker discovery for radiation exposure. This technology is now available to investigators for their specific research interests. Written and electronic reports are provided for all samples, and the average turn-around time is two weeks. The PMSR is able to carry out experiments that detect and characterize protein modifications such as phosphorylation and acetylafion. These experiments are challenging and require a more intensive effort. However, the combination of the complementary mass spectrometric methods available in PMSR, staff expertise and experience, and the ability to work closely with investigators, have allowed the successful completion of a number of these difficult projects. The PMSR was established in January 2006 and has grown dramatically, both in resources and use. Lombardi has invested over $1 million in equipment alone for the PMSR; as a result, the facility consistently provides high quality data to many investigators from 5 programs within Lombardi. The PMSR is run under the directorship of Drs. Stephen Byers and Albert Fornace Jr.