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

该子项目是利用由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。子项目和研究者（PI）可能从另一个NIH来源获得主要资金，因此可以在其他CRISP条目中表示。列出的机构是用于资助的，不一定是研究者的机构。说明（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。该仪器将主要配置用于SUPREX数据的采集。SUPREX（Stability of Unpurified Proteins from Rates of H/D Exchange）是一种新的基于H/D交换和MALDI的方法，用于测量溶液中蛋白质的热力学稳定性。SUPREX技术诞生于五年前杜克大学菲茨杰拉德和Oas实验室的合作。在过去的五年中，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-配体复合物的定量热力学分析。共有14个已发表的作品的SUPREX分析选定的模型蛋白质系统已被用来定义的准确性，精密度和通用性的技术。 随着SUPREX实验方案和数据分析方法的确立，SUPREX数据生成和分析的瓶颈已成为MALDI质谱的获取。完整的SUPREX分析蛋白质或蛋白质-配体系统需要大约500个质谱;在组合化学应用中使用SUPREX可能需要获得数百万个MALDI质谱。所要求的MALDI质谱仪系统的高通量能力对于该提案的Co-PI在其NIH资助的项目中利用SUPREX的独特优势（例如，能够快速分析纯化和未纯化形式的pmol数量的蛋白质）是必不可少的，这些项目需要对蛋白质和蛋白质-配体复合物进行热力学测量。杜克校区SUPREX用户数量的不断增加以及SUPREX在组合文库筛选项目中的拟议开发，迫切需要一种新的高通量MALDI-TOF质谱仪，专门用于杜克校区的SUPREX实验。

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.