PURCHASE OF A HIGH-THROUGHPUT MALDI TOF/TOF: PROTEOMICS

这个子项目是利用由NIH/NCRR资助的共享仪器赠款提供的资源的许多研究子项目之一。子项目和调查员(PI)可能从另一个NIH来源获得了主要资金，因此可能会出现在其他CRISE条目中。列出的机构是用于拨款的，而不一定是用于调查人员的机构。说明(由申请人提供)：申请资金用于购买高通量基质辅助激光解吸/电离(MALDI)质谱仪。该仪器将主要配置并专门用于获取SUPREX数据。SUPREX(稳定性来自H/D交换率)是一种基于H/D交换率和MALDI的新方法，用于测量蛋白质在溶液中的热力学稳定性。Suprex技术诞生于五年前杜克大学菲茨杰拉德实验室和美洲国家组织实验室之间的合作。在过去的五年里，SUPREX技术已经发展成为一种强大的分析方法，用于蛋白质和蛋白质-配体复合体的定量热力学分析。共有14篇发表的关于选定模型蛋白质系统的SUPREX分析的著作已经确定了该技术的精确度、精密度和一般性。随着SUPREX实验协议和数据分析方法的确立，SUPREX数据的产生和分析的瓶颈已经成为MALDI质谱图的获取。完整的SUPREX分析需要大约500个质谱图；在组合化学应用中使用SUPREX可能需要获取数百万个MALDI质谱图。所要求的MALDI质谱计系统的高通量能力对于这项提议的Co-PI在他们的NIH资助的需要对蛋白质和蛋白质配体复合体进行这种热力学测量的项目中利用SUPREX的独特优势(例如，能够快速分析仅pmol数量的纯化和未纯化的蛋白质的能力)是至关重要的。随着杜克大学校园中SUPREX用户数量的不断增加，以及在组合图书馆筛选项目中对SUPREX的拟议开发，迫切需要一台专门用于杜克大学校园中的SUPREX实验的新型高通量MALDI-TOF质谱仪。

This subproject is one of many research subprojects utilizing the resources provided by a Shared Instrumentation 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 grant, which is not necessarily the institution for the investigator. DESCRIPTION (provided by applicant): Funds are requested for the purchase of a high-throughput matrix-assisted laser desorption/ionization (MALDI) mass spectrometer. This instrument will be primarily configured for and dedicated to the acquisition of SUPREX data. SUPREX (Stability of Unpurified Proteins from Rates of H/D Exchange) is a new H/D exchange- and MALDI-based method for measuring the thermodynamic stability of proteins in solution. The SUPREX techique was born out of a collaboration between the Fitzgerald and Oas laboratories at Duke University five years ago. Over the past five years the SUPREX technique has been developed into a robust analytical method for the quantitative thermodynamic analysis of proteins and protein-ligand complexes. A total of 14 published works on the SUPREX analysis of selected model protein systems have served to define the accuracy, precision, and generality of the technique. With the experimental protocols and data analysis methods of SUPREX firmly established, the bottleneck in the generation and analysis of SUPREX data has become the acquisition of MALDI mass spectra. Approximately 500 mass spectra are required for the complete SUPREX analysis a protein or protein-ligand system; and the use of SUPREX in combinatorial chemistry applications can require the acquisition of millions of MALDI mass spectra. The high-throughput capacity of the requested MALDI mass spectrometer system is essential for the Co-PI's on this proposal to exploit the unique advantages of SUPREX (e.g. the ability to rapidly analyze just pmol quantities of protein in both purified and unpurifed form) in their NIH-funded projects that require such thermodynamic measurements on proteins and protein-ligand complexes. The increasing number of SUPREX users on the Duke campus and the proposed exploitation of SUPREX in combinatorial library screening projects create a critical need for a new high-throughput MALDI-TOF mass spectrometer dedicated to SUPREX experiments on the Duke campus.