Robotic spot picker & liquid handling device/proteomics

描述（由申请人提供）：Kresge中心蛋白质组学设施（KCPF）由本基金的PI指导，是哈佛公共卫生学院（HSPH）的NIEHS和NSF赞助的资源，为全系统水平的蛋白质分析提供基础设施（=“蛋白质组学”）。KCPF提供基于二维凝胶电泳（2DGE）和液相色谱和串联质谱（LC/MS/MS）的蛋白质鉴定的自动化蛋白质组学过程。在这项拨款申请中，申请资金用于购买一个组合式机器人点选器和液体处理器，该机器人将集成到现有的自动化流程链中，提供高质量的蛋白质分析数据。 KCPF为蛋白质组学研究提供了两个主要的应用，这将大大受益于所要求的设备：1。蛋白质表达谱，和2.从复杂样品中纯化的蛋白质组装体的鉴定。这项提案的资助所产生的研究活动广泛利用这些应用程序，以回答疾病基础生物学中的主要问题。这些项目使用来自进化阶梯上的生物体的样本，包括病毒、细菌、酵母、寄生虫、转基因小鼠、哺乳动物组织培养细胞和人体器官。研究活动的范围很广，包括大规模分析泛素依赖的蛋白水解系统，分析细胞对自由基和氧化应激反应的决定因素，鉴定早产胎盘功能障碍中涉及的蛋白质，确定控制金属和肽生长因子膜转运的因素，分析细胞谱系确定中的信号传导和转录激活途径，确定蛋白质的相互作用，包括建立细胞极性。 这些领域正在进行的项目表明，迫切需要快速处理2D凝胶上鉴定的大量蛋白质点的能力。这包括从凝胶上切除斑点、从凝胶塞中提取蛋白质、酶促消化和纯化肽用于MS分析。最近开发的现场采摘和液体处理设备要求在此授权申请结合了所有这些能力在一个单一的单位。

DESCRIPTION (provided by applicant): The Kresge Center Proteomics Facility (KCPF), which is directed by the PI of this grant, is an NIEHS and NSF sponsored resource at the Harvard School of Public Health (HSPH) that offers an infrastructure for protein analysis at a system-wide level (= "proteomics"). KCPF offers automated proteomics processes based on 2-dimensional gel electrophoresis (2DGE) and protein identification by liquid chromatography and tandem mass spectrometry (LC/MS/MS). In this grant application funds are requested for the acquisition of a combined robotic spot picker and liquid handler, which will be integrated in the existing chain of automated processes that deliver high quality protein analysis data. KCPF offers two major applications for proteomics research that will substantially benefit from the requested equipment: 1. Protein expression profiling, and 2. Identification of protein assemblies purified from complex samples. The research activities resulting from funding of this proposal make extensive use of these applications in order to answer major questions in the basic biology of disease. The projects use samples derived from organisms across the evolutionary ladder, including viruses, bacteria, yeast, parasites, transgenic mice, mammalian tissue culture cells, and human organs. The scope of research activities is wide-ranging, and includes large scale analysis of the ubiquitin-dependent proteolysis system, analyzing determinants of cellular responses to radicals and oxidative stress, identifying proteins involved in placental dysfunction in preterm deliveries, determining factors controlling membrane transport of metals and peptide growth factors, analyzing signaling and transcriptional activation pathways in cell lineage determination, determining protein interactions involved in establishing cell polarity, among many others. Ongoing projects in these areas have revealed an acute need for the capability to rapidly process an extensive number of protein spots identified on 2D gels. This includes spot excision from gels, extraction of the proteins from gel plugs, enzymatic digestions, and purification of the peptides for MS analysis. The recently developed spot picking and liquid handling device requested in this grant application combines all of these capabilities in a single unit.