LC ESI QOTOF METHOD DEVELOPMENT AND NEW INSTRUMENTATION EVALUATION

LC ESI QOTOF 方法开发和新仪器评估

• 批准号: 8365494
• 负责人: Catherine E. Costello
• 金额: $ 1.23万
• 依托单位: BOSTON UNIVERSITY MEDICAL CAMPUS
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-06-01 至 2012-08-09
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8365494
• 关键词: Biological; Biology; Blood capillaries; Caliber; Carbohydrates; Complement; Complex; Data Analyses; Evaluation; Funding; Grant; Laboratories; Lipids; Mass Spectrum Analysis; Medicine; National Center for Research Resources; Phosphorylation; Principal Investigator; Process; Proteins; Proteomics; Pulsar; Research; Research Infrastructure; Research Project Grants; Resources; Sampling; Source; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; System; United States National Institutes of Health; Water; base; capillary; cost; instrumentation; mass spectrometer; member; method development; microbial alkaline proteinase inhibitor

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. Our laboratory uses two Applied Biosystems API QSTAR(tm) Pulsar i QqTOF mass spectrometers with Waters Acuity LCs and an Agilent chip-LC Q-TOF mass spectrometer for the characterization of complex biological samples obtained from collaborative research projects by ESI and MALDI. Currently, our focus is on establishing routine capillary LC-IDA-MS/MS for proteomics, glycomics and lipidomics applications, in addition to characterization of the large variety of biological samples that are submitted to the MS Resource. Use of automated IDA MS/MS functions are continuously being optimized for typical protein digests, lipids and carbohydrates, and batch parameter files have been generated for ease of use by the members within the laboratory. Processing scripts have been optimized for use on workstations for data analysis. The use of the smaller diameter column assemblies offers a significant increase in sensitivity, enabling samples of low concentrations or limited quantity to be analyzed. Optimization of the LC systems is followed by method development studies to complement the current research undertaken within the MS resource. Method development is being pursued for glycomics, lipidomics and proteomics-based studies, including phosphorylation analysis. The use of online ESI-MS for analysis of TLC-separated samples is being explored.

该子项目是利用资源的众多研究子项目之一 由 NIH/NCRR 资助的中心拨款提供。子项目的主要支持 并且子项目的主要研究者可能是由其他来源提供的， 包括其他 NIH 来源。 子项目可能列出的总成本 代表子项目使用的中心基础设施的估计数量， NCRR 赠款不直接向子项目或子项目工作人员提供资金。 我们的实验室使用两台配有 Waters Acuity LC 的 Applied Biosystems API QSTAR(tm) Pulsar i QqTOF 质谱仪和一台安捷伦芯片-LC Q-TOF 质谱仪，对 ESI 和 MALDI 合作研究项目中获得的复杂生物样品进行表征。目前，我们的重点是建立用于蛋白质组学、糖组学和脂质组学应用的常规毛细管 LC-IDA-MS/MS，以及对提交给 MS 资源的各种生物样品进行表征。自动 IDA MS/MS 功能的使用不断针对典型的蛋白质消化、脂质和碳水化合物进行优化，并且已生成批次参数文件，以便于实验室成员使用。处理脚本已经过优化，可在工作站上用于数据分析。使用较小直径的色谱柱组件可显着提高灵敏度，从而能够分析低浓度或有限数量的样品。 LC 系统优化后进行方法开发研究，以补充 MS 资源中当前进行的研究。正在为糖组学、脂质组学和蛋白质组学研究（包括磷酸化分析）开发方法。正在探索使用在线 ESI-MS 来分析 TLC 分离的样品。