Quadrupole-Time-of-Flight Mass Spectrometry System

本提案的目的是为购买毛细管柱HPLC-电喷雾电离-四极杆-飞行时间（Q-TOF）串联质谱系统获得资金，该系统将用于对凝胶电泳分离的蛋白质进行自动化、高通量测序和鉴定。该仪器将位于克利夫兰诊所基金会Lerner研究所的一个已建立的蛋白质测序实验室中，并用于支持克利夫兰诊所和凯斯西储大学的一组NIH资助的主要用户的研究计划。主要用户的研究项目范围广泛，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，细胞间通讯和细胞内信号传导等领域的项目。所有主要用户的项目都需要质谱法来测序和识别相对大量的痕量蛋白质。所要求的仪器所在的蛋白质测序实验室已经拥有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-飞行时间质谱系统和电喷雾电离-离子阱串联质谱系统。然而，实验室受到没有自动化设备的离子阱仪器上可用时间的严重限制。所要求的Q-TOF仪器系统将提供新的样本容量，这对于满足主要用户的需求并及时严格地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额外的NIH资助的用户提供服务。因此，所要求的仪器对这些程序也有次要的好处。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可用性提供大量资源。这些资源包括翻新的实验室空间，设备，经验丰富的人员，财务支持和财务计划，这将使该设施的长期成功。所要求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计划，包括克利夫兰诊所基金会，凯斯西储大学，大学医院，MetroHealth医院和克利夫兰州立大学的计划。

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.