Quadrupole-Time-of-Flight Mass Spectrometry System

这项提案的目的是获得资金，用于购买毛细管柱高效液相色谱-电喷雾电离-四极杆飞行时间(Q-TOF)串联质谱仪系统，该系统将用于通过凝胶电泳法分离的蛋白质的自动化、高通量测序和鉴定。该仪器将安装在克利夫兰临床基金会勒纳研究所的一个已建立的蛋白质测序实验室中，用于支持克利夫兰诊所和凯斯西储大学由NIH资助的一组主要用户的研究计划。主要用户的研究计划范围广泛，包括癌症生物学、血管生物学和动脉粥样硬化、心脏病学、神经科学、翻译控制、细胞间通信和细胞内信号传递等领域的项目。主要用户的所有项目都需要质谱仪来在痕量水平上对相对大量的蛋白质进行排序和识别。所要求的仪器所在的蛋白质测序实验室已经包括支持这些分析的其他重要设备，包括基质辅助激光解吸/电离飞行时间质谱仪系统和电喷雾电离-离子陷阱串联质谱仪系统。然而，实验室受到离子陷阱仪器上可用时间的严重限制，因为离子陷阱仪器没有提供自动化。所要求的Q-TOF仪器系统将提供新的样品容量，这对于容纳主要用户并以及时和严格的方式进行他们所需的测序实验至关重要。所要求的仪器还补充和扩展了一个经验丰富和成功的实验室的既定能力，该实验室正在为大量由国家卫生研究院资助的额外用户提供服务。因此，所要求的工具也将对这些计划产生次要的好处。克利夫兰临床基金会已经并将继续为克利夫兰地区的质谱学提供大量资源。这些资源包括翻新的实验室空间、设备、经验丰富的人员、财务支持和使该设施能够长期成功的财务计划。申请的仪器将大大增加这些资源，促进和推进整个克利夫兰地区的研究项目，包括克利夫兰临床基金会、凯斯西储大学、大学医院、MetroHealth医院和克利夫兰州立大学的项目。

The purpose of this proposal is to obtain funding for the purchase of a capillary column HPLC-electrospray ionization-quadrupole-time-of- flight (Q-TOF) tandem mass spectrometry system that will be used for automated, high-throughput sequencing and identification of proteins separated by gel electrophoresis. The instrument will be located in an established protein sequencing laboratory in the Lerner Research Institute at the Cleveland Clinic Foundation and used to support the research programs of a group of NIH-funded Major Users at both the Cleveland Clinic and Case Western Reserve University. The research programs of the Major Users are broad, encompassing projects in the areas of cancer biology, vascular biology and atherosclerosis, cardiology, neuroscience, translational control, cell-to-cell communication, and intracellular signaling. All of the Major User's projects need mass spectrometry to sequence and identify relatively large numbers of proteins at trace levels. The protein sequencing laboratory in which the requested instrument will be located already contains other important equipment to support these analyses, including a matrix-assisted laser desorption/ionization-time-of-flight mass spectrometry system and an electrospray ionization-ion trap tandem mass spectrometry system. However, the laboratory is severely limited by the time available on the ion trap instrument that has no provision for automation. The requested Q-TOF instrument system will provide new sample capacity that is essential to accommodate the Major Users and carry out the sequencing experiments they require in a timely and rigorous manner. The requested instrument also complements and expands the established capabilities of an experienced and successful laboratory that is serving a large number of additional NIH-funded users. Therefore, the requested instrument will have a secondary benefit to these programs as well. The Cleveland Clinic Foundation has provided and continues to provide substantial resources for the availability of mass spectrometry in the Cleveland area. These resources include renovated laboratory space, equipment, experienced personnel, financial support, and a financial plan that will enable the long-term success of the facility. The requested instrument will add significantly to these resources, facilitating and advancing research programs throughout the Cleveland area including programs at the Cleveland Clinic Foundation, Case Western Reserve University, University Hospitals, MetroHealth Hospital, and Cleveland State University.