National Cancer Institute and DOE: Protein and Peptide Production.

美国国家癌症研究所和能源部：蛋白质和肽生产。

• 批准号: 7713842
• 金额: $ 65万
• 依托单位: NATIONAL CANCER INSTITUTE
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/7713842
• 关键词: National; Cancer; Institute; DOE; Protein

Article 3 Program will be modified to include the additional project milestones: ¿ Construct expression clones for a majority of the 1261 candidate biomarkers specified in Polanski and Anderson (2006) manuscript and screen them for expression and solubility. ¿ Carry out scale-up production of milligram quantities of N 15 labeled and non-labeled proteins constituting a subset of these proteins as specified by Nd. ¿ Carry out additional expression and solubility screening on the 1261 candidate biomarkers using alternate vectors to increase the success rate for production of soluble protein products. ¿ Carry out preliminary experiments to determine the feasibility of producing phosphorylated proteins by treating proteins expressed in E. coli with kinases. The Director of the Clinical Proteomic Technologies for Cancer programs at the NCI will be responsible for oversight of this project from the NCI. Other approved NCI staff will be responsible for the management of the project. The Principal Investigator of this DOE project will be responsible for oversight of the project from ANL. ANL will provide proteins and peptides as a resource to support clinical proteomic technology development and assessment for cancer research. The objective of this agreement is to support the development of human proteins and peptides for use by the research community that will enable development of assays, and standard reagents. NCI will make these resources available to the research community to foster discovery research. The proteins and peptides to be produced from this project will be those that have been identified. as important factors in cancer biological processes. The NCI has major initiatives to support the development and assessment of novel protein detection technologies. These technologies are designed to standardize proteomic technologies, reference reagents and standards, measurement protocols, instrumentation, and sample acquisition, preparation and analysis platforms. A critical component needed to achieve the goals of these initiatives will be the development and validation of proteomic reagents including peptides/proteins and antibodies. DOE has established unique capabilities and expertise in the production, qualification, and characterization of peptides and proteins. This experience currently supports the production of proteins that are used to develop crystal structures for the resolution of their 3-dimensional structures. Such high quality, well characterized, and readily available proteins and peptides will be a key resource in advancing proteomic technologies and capabilities and will be essential for developing highly specific antibodies to proteins associated with cancer. The goal of the NCI protein production program is the production, characterization, and distribution of peptides and proteins for use in the development and validation of clinical cancer proteomic technology platforms. Protein targets will be selected based on scientific rationale that considers the biological relevance of the discovery of the targets in the cancer proteome. A candidate list of peptides and proteins for the proposed project will be selected by NCI in consultation with ANL scientists. DOE will support the expression of the proteins through a variety of strategies using E. coli expression systems that employ a target analysis system to select protein targets based on informatics-based selection or target categorization (e.g. size, localization signals).

第3条计划将进行修改，以包括额外的项目里程碑： 构建Polanski和安德森（2006）手稿中指定的1261种候选生物标志物中大多数的表达克隆，并筛选它们的表达和溶解度。 按照Nd的规定，进行毫克量的N15标记和非标记蛋白质的放大生产，这些蛋白质构成这些蛋白质的子集。 使用替代载体对1261个候选生物标志物进行额外的表达和溶解度筛选，以提高可溶性蛋白产品生产的成功率。 进行初步实验，以确定通过处理在E.大肠杆菌中的激酶。 NCI癌症临床蛋白质组学技术项目主任将负责监督NCI的该项目。其他经批准的NCI工作人员将负责项目的管理。本DOE项目的主要研究者将负责监督ANL的项目。 ANL将提供蛋白质和肽作为支持临床蛋白质组学技术开发和癌症研究评估的资源。该协议的目的是支持研究界使用的人类蛋白质和肽的开发，从而能够开发检测试剂和标准试剂。NCI将向研究界提供这些资源，以促进发现研究。从该项目中生产的蛋白质和肽将是那些已经鉴定的蛋白质和肽。作为癌症生物学过程中的重要因素。 NCI有重大举措来支持新蛋白质检测技术的开发和评估。这些技术旨在标准化蛋白质组学技术、参考试剂和标准品、测量方案、仪器以及样品采集、制备和分析平台。实现这些倡议目标所需的一个关键组成部分是开发和验证蛋白质组试剂，包括肽/蛋白质和抗体。DOE在肽和蛋白质的生产、鉴定和表征方面建立了独特的能力和专业知识。这一经验目前支持用于开发晶体结构以解析其三维结构的蛋白质的生产。这种高质量、良好表征和容易获得的蛋白质和肽将是推进蛋白质组学技术和能力的关键资源，并且对于开发针对与癌症相关的蛋白质的高度特异性抗体至关重要。 NCI蛋白质生产计划的目标是生产，表征和分配用于临床癌症蛋白质组学技术平台开发和验证的肽和蛋白质。蛋白质靶点的选择将基于科学原理，考虑癌症蛋白质组中靶点发现的生物学相关性。NCI将与ANL科学家协商，为拟议项目选择肽和蛋白质的候选名单。 DOE将通过使用E.大肠杆菌表达系统，其采用靶分析系统来基于基于信息学的选择或靶分类（例如大小、定位信号）选择蛋白质靶。