INSTRUMENTATION FOR PROTEOMICS

在本提案中，我们请求支持购买一个仪器系统，用于在弗吉尼亚大学生物分子研究设施为大学研究人员进行的蛋白质组学项目的半自动化样品处理。具体来说，我们需要一个凝胶成像系统和相关软件，凝胶点选择器，凝胶点消化器和消化点。这些仪器生成的样品将在我们现有的PE Voyager DE Pro MALDI-TOF仪器上进行自动质谱图分析，或在必要时使用该机构的Finnigan LCQ离子阱LC/MS/MS质谱仪进行从头测序。该计划的总体目标是为蛋白质组学相关项目的研究人员提供高通量，低成本的蛋白质鉴定。此外，这项技术将与该设施中的微阵列/基因芯片计划相结合。这些仪器将被安置在弗吉尼亚大学生物分子研究设施，并将支持七个NIH资助的研究人员的研究蛋白质组学项目。对于这些项目，我们预计这些工具将在大约80%的可用时间内使用。剩余时间将用于新的蛋白质组学相关项目的开发。有没有其他质谱蛋白质鉴定服务，高通量或以其他方式，可在大学。生物分子研究机构主任Jay Fox博士将负责仪器的全面管理。Nicholas谢尔曼博士将负责蛋白质组学项目的质谱问题，John Shannon博士将负责蛋白质组学项目的蛋白质化学方面。两人都是生物分子研究机构的工作人员。这三位科学家将在蛋白质组学项目的规划、执行和数据分析方面与研究人员进行磋商。在过去的七年里，该设施提供了质谱蛋白质鉴定和从头序列分析，预计在样品分析的下游端不会出现重大困难。将收取服务费，以帮助支付工作人员薪金和用品费用。生物分子研究设施的监督由医学院院长任命的常务委员会提供。监督委员会直接向Fox博士提供设施管理方面的指导和专业知识，并补充设施工作人员的科学专业知识。该提案的主要研究者之一凯文哈岑博士是该委员会的成员。弗吉尼亚大学生物分子研究设施在过去的20年里一直在运作，并得到弗吉尼亚大学健康科学中心的持续财政支持。健康科学中心的管理部门致力于该设施的长期运营。这种支持，沿着监督委员会和设施工作人员的支持，将有助于确保足够的，适当的仪器使用，以确保仪器的合理和科学上的成功操作，这是为了弗吉尼亚大学NIH资助的研究人员的利益。

In this proposal, we are requesting support for the purchase of an instrument system to semi-automate sample processing for proteomic projects performed in the University of Virginia Biomolecular Research Facility for University investigators. Specifically, we are requesting a gel imaging system and associated software, gel spot picker, gel spot digester and digest spotter. Samples generated by these instruments will be subjected to automated mass mapping on our existing PE Voyager DE Pro MALDI-TOF instrument or if necessary, de novo sequencing with the facility's Finnigan LCQ ion-trap LC/MS/MS mass spectrometer. The overall goal of this program is to provide high throughput, low cost protein identification for investigators with proteomic related projects. Furthermore, this technology will be interfaced with the Microarray/GeneChip Program in the facility. The instruments will be housed in the University of Virginia Biomolecular Research Facility and will support the research proteomic projects of seven NIH-funded investigators. For these projects, we anticipate that the instruments will be in use approximately 80% of the available time. The remaining time will be available for new proteomic-related project development. There is no other mass spectrometric protein identification service, high throughput or otherwise, available at the University. Dr. Jay Fox, Director of the Biomolecular Research Facility, will be responsible for the overall management of the instrumentation. Dr. Nicholas Sherman will be responsible for mass spectrometry issues of proteomics projects and Dr. John Shannon will be responsible for the protein chemistry aspects of the proteomic projects. Both are staff members in the Biomolecular Research Facility. All three scientists will consult with investigators in the planning, execution and data analysis of proteomics projects. For the past seven years, the facility has offered mass spectrometric protein identification and de novo sequence analysis and no major difficulties are expected on the downstream end of sample analysis. A fee-for-service charge will be instituted to help defray the costs of staff salaries and supplies. Oversight of the Biomolecular Research Facility is provided by a standing committee appointed by the Dean of the Medical School. The Oversight Committee provides guidance and expertise directly to Dr. Fox for the management of the facility as well as complementing the scientific expertise of facility staff. One of the principal investigators in this proposal, Dr. Kevin Hazen, is a member of the committee. The University of Virginia Biomolecular Research Facility has been in operation for the past 20 years with continual fiscal support from the University of Virginia Health Sciences Center. The administration of the Health Sciences Center is committed to the long-term operation of the facility. This support, along with that of the Oversight Committee and facility staff, will serve to ensure adequate, appropriate instrument use for the fiscally sound and scientifically successful operation of instrumentation requested for the benefit of NIH-funded investigators at the University of Virginia..