Integrated Instrumentation for Proteomics

蛋白质组学集成仪器

• 批准号: 6440421
• 负责人: HELEN KIM
• 金额: $ 44.65万
• 依托单位: UNIVERSITY OF ALABAMA AT BIRMINGHAM
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-05-01 至 2004-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6440421
• 关键词: biomedical equipment purchase; gel electrophoresis; high throughput technology; mass spectrometry; matrix assisted laser desorption ionization; proteomics

DESCRIPTION (provided by applicant): The objective of this grant is to purchase an integrated instrumentation system that includes high-resolution and high-throughput, two-dimensional electrophoresis and dedicated mass spectrometry to analyze patterns of protein expression (proteomics) relevant to human disease. The requested instrumentation will provide state-of-the-art proteomics analytical capabilities for a large NIH-funded biomedical research community at the University of Alabama at Birmingham. Analysis of single protein changes and modifications has been a major utilization of the mass spectrometry facility, principally through use of the matrix-assisted laser-desorption ionization-time-of-flight mass spectrometer (MALDI-TOF MS), and the Q-TOF. In this postgenome era, once a polypeptide gel "spot" is excised, trypsin-digested and analyzed by MALDI-TOF, the availability of genome sequence data enables the rapid identification of unknown polypeptides through various software. The requested instrumentation will address the major factors that limit proteomic analysis: (1) enhance the detection of a relevant polypeptide(s) in a 2-D gel "display," (2) allow greater resolution of a display for a given sample, and (3) enable high-throughput processing of gel "spots" such that this step is not rate limiting. Accompanying software ensure seamless integration among various instruments, and allow sample tracking from beginning step, isoelectric focusing, to the last step, MALDI-TOF identification of protein based on automated peptide mass fingerprinting. In the last year, the principal investigator directed an initial developmental phase of 2-D electrophoresis in conjunction with the mass spectrometry facility, utilizing mini gels and Coomassie-or silver stained mini gels for the second dimension. The conclusion is that it is essential to go to larger-format gels, with fluorescence detection, robotic excision and in-gel digestions of gel "spots," as well as spotting onto MALDI-TOF plates, ending with automated MALDI-TOF mass spectrometry The requested integrated system (BioRad-Micromass) will provide the instrumentation to take our proteomics analytical capabilities to the level that will sustain the high quality of biomedical research that is a hallmark of UAB.

描述（由申请人提供）：这笔赠款的目的是购买 集成仪器系统，包括高分辨率和 高通量、二维电泳和专用质量 光谱法分析与相关的蛋白质表达模式（蛋白质组学） 人类疾病。所需的仪器将提供最先进的 NIH 资助的大型生物医学研究的蛋白质组学分析能力 阿拉巴马大学伯明翰分校社区。单项分析 蛋白质的变化和修饰一直是质量的主要利用 光谱测定设施，主要通过使用基质辅助 激光解吸电离飞行时间质谱仪（MALDI-TOF MS）， 和Q-TOF。在这个后基因组时代，一旦多肽凝胶“点” 切除、胰蛋白酶消化并通过 MALDI-TOF 分析，基因组的可用性 序列数据能够通过以下方式快速识别未知多肽 各种软件。所需的仪器将解决主要因素 限制蛋白质组分析：（1）增强相关相关蛋白的检测 二维凝胶“展示”中的多肽，(2) 允许更高的分辨率 显示给定样品，并且 (3) 实现凝胶的高通量处理 “点”，使得该步骤不受速率限制。配套软件保证 各种仪器之间的无缝集成，并允许样品跟踪 起始步骤，等电聚焦，到最后一步，MALDI-TOF 基于自动肽质量指纹分析的蛋白质鉴定。在 去年，首席研究员指导了初步发展 二维电泳与质谱相结合 设施，利用迷你凝胶和考马斯或银染迷你凝胶进行 第二维度。结论是，有必要采用更大的格式 凝胶，具有荧光检测、机器人切除和凝胶内消化 凝胶“点”，以及点样到 MALDI-TOF 板上，最后以自动化的方式结束 MALDI-TOF 质谱法 所需的集成系统 (BioRad-Micromass) 将提供仪器来发挥我们的蛋白质组学分析能力 达到维持高质量生物医学研究的水平 UAB 的一个标志。