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)由这笔赠款的PI指导，是哈佛大学公共卫生学院(HSPH)由NIEHS和NSF赞助的资源，为全系统水平的蛋白质分析(=“蛋白质组学”)提供基础设施。KCPF提供基于双向凝胶电泳法(2DGE)的自动化蛋白质组学过程，以及通过液相色谱和串联质谱仪(LC/MS/MS)鉴定蛋白质。在这项赠款申请中，申请资金用于购买组合的机器人斑点拾取器和液体处理机，这将被整合到提供高质量蛋白质分析数据的现有自动化过程链中。 KCPF为蛋白质组学研究提供了两个主要应用，这两个应用将从所要求的设备中获得实质性的好处：1.蛋白质表达图谱，以及2.从复杂样品中纯化的蛋白质组合的鉴定。这项提案的资助所产生的研究活动广泛利用这些应用，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。研究活动范围广泛，包括大规模分析依赖泛素的蛋白质分解系统，分析细胞对自由基和氧化应激的反应决定因素，确定早产时参与胎盘功能障碍的蛋白质，确定控制金属膜运输的因素和肽生长因子，分析细胞谱系确定中的信号和转录激活途径，确定参与确定细胞极性的蛋白质相互作用等等。 这些领域正在进行的项目揭示了对快速处理2D凝胶上确定的大量蛋白质斑点的能力的迫切需求。这包括从凝胶中去除斑点，从凝胶塞中提取蛋白质，酶消化，以及纯化用于MS分析的多肽。最近开发的现场拾取和液体处理设备在本赠款申请中要求将所有这些功能结合在一个单元中。

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.