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: 8170857
• 负责人: Catherine E. Costello
• 金额: $ 1.39万
• 依托单位: BOSTON UNIVERSITY MEDICAL CAMPUS
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-06-01 至 2011-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8170857
• 关键词: Biological; Blood capillaries; Caliber; Carbohydrates; Complement; Complex; Computer Retrieval of Information on Scientific Projects Database; Data Analyses; Evaluation; Funding; Grant; Institution; Laboratories; Lipids; Phosphorylation; Process; Proteins; Proteomics; Pulsar; Research; Research Personnel; Research Project Grants; Resources; Robot; Sampling; Source; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; System; United States National Institutes of Health; Water; Work; base; capillary; 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. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. 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. Addition of the Advion TriVersa robot to the Q-Star system has necessitated much work to find optimum conditions, and close interaction with both Advion and Applied Biosystems.

该子项目是利用该技术的众多研究子项目之一 资源由 NIH/NCRR 资助的中心拨款提供。子项目和 研究者 (PI) 可能已从 NIH 的另一个来源获得主要资金， 因此可以在其他 CRISP 条目中表示。列出的机构是 中心，不一定是研究者的机构。 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 系统优化之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。将 Advion TriVersa 机器人添加到 Q-Star 系统需要做大量工作来找到最佳条件，并与 Advion 和 Applied Biosystems 密切互动。