High-throughput, nanoliter-scale macromolecular crystallization at UCSD

加州大学圣地亚哥分校的高通量、纳升级大分子结晶

• 批准号: 8052145
• 负责人: PARTHO GHOSH
• 金额: $ 14.3万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-04-01 至 2012-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8052145
• 关键词: Automation; Biochemistry; Biological; California; Chemistry; Core Facility; Crystallization; Crystallography; Data Collection; Drops; Engineering; Equipment; Fees; Financial Support; Growth; Housing; Human; Human Resources; Image; Manuals; Outcome; Pharmacologic Substance; Pharmacy Schools; Process; Proteins; Research; Research Personnel; Robot; Robotics; Roentgen Rays; Sampling; Science; Solutions; Structure; System; Therapeutic; United States National Institutes of Health; Universities; X-Ray Crystallography; base; design; experience; insight; instrumentation; macromolecule; nanolitre scale; operation; programs; structural biology; success

DESCRIPTION (provided by applicant): The structures of biological macromolecules determined by X-ray crystallography provide unparalleled functional insights and make possible the design of therapeutics aimed at treating human illnesses. Crystallization of proteins and other biological macromolecules, such as ordered RNAs, is limited in many cases by the quantity of purified sample available for crystallization trials. Crystallization remains an empirical process, meaning that success generally depends on the number of different conditions tried. Thus, a large number of precipitants must be screened with a limiting quantity of biological sample. The solution to this problem is robotic automation of the crystallization process, which requires ~10-fold less sample than manual means due to the reliable ability of robots to set up nanoliter-scale drops. Robotic automation also enables a vast number of conditions to be tried in high-throughput fashion. This proposal seeks to acquire integrated crystallization robotics for NIH-supported investigators at the University of California, San Diego (UCSD). Specifically, we propose to acquire an automated solution mixing machine for formulating crystallization solutions, an automated dispenser of these solutions into crystallization trays, robotics to set up the crystallization drop, and an automated imaging system for examining and recording the outcome of crystallization trials. The equipment will be incorporated into an existing core facility, the Macromolecular Crystallography Facility, which currently houses an X-ray data collection setup and has an experienced staff engineer who manages daily operations. The equipment will be administered by the existing Macromolecular Crystallography Facility Committee and daily operations will be managed by the existing staff engineer. UCSD has a long tradition of X-ray crystallography, and this commitment continues today. This commitment is evidenced by financial support for personnel to manage the crystallization robotics from the Department of Chemistry & Biochemistry and the Skaggs School of Pharmacy and Pharmaceutical Sciences. The long-term support of the instrumentation will come on a recharge basis through user fees as part of the Macromolecular Crystallography Facility. UCSD has a substantial number of X-ray crystallographers whose research efforts would be markedly enhanced by regular access to crystallization robotics. Significantly, the instrumentation, as part of a core facility, will also enable investigators at UCSD whose expertise is outside of X-ray crystallography to move their research programs towards structure determination. The detailed level of information gained from X-ray crystallography is likely to permit growth of these research topics towards therapeutic solutions. In short, the proposed instrumentation is crucial to the campus-wide success of structural biology at UCSD. PUBLIC HEALTH RELEVANCE: The structures of biological macromolecules determined by X-ray crystallography provide unparalleled functional insights and make possible the design of therapeutics aimed at treating human illnesses. This proposal seeks to obtain an integrated robotic system for the crystallization of biological macromolecules at the University of California, San Diego. This equipment will enable a large group of biomedical scientists at this campus to pursue fundamental questions related to human diseases, and to apply the basic understanding of proteins and ordered RNAs gained from structure determination to therapeutics.

描述(由申请人提供)： 由X射线结晶学确定的生物大分子的结构提供了无与伦比的功能洞察，并使旨在治疗人类疾病的疗法的设计成为可能。蛋白质和其他生物大分子(如有序RNA)的结晶在许多情况下受到可用于结晶试验的纯化样品的数量的限制。结晶仍然是一个经验过程，这意味着成功通常取决于所尝试的不同条件的数量。因此，必须用有限数量的生物样品来筛选大量的沉淀物。这个问题的解决方案是结晶过程的机器人自动化，由于机器人能够可靠地建立纳升尺度的液滴，因此需要的样品比人工方法少约10倍。机器人自动化还能够以高通量的方式尝试大量的条件。这项提议寻求为加州大学圣地亚哥分校(UCSD)NIH支持的研究人员获得集成结晶机器人技术。具体地说，我们建议购买一台用于配制结晶溶液的自动溶液混合机，一台将这些溶液自动分配到结晶托盘中的机器，用于设置结晶液滴的机器人，以及用于检查和记录结晶试验结果的自动成像系统。这些设备将被并入现有的核心设施--高分子结晶设施，该设施目前拥有一套X射线数据收集装置，并拥有一名经验丰富的员工工程师来管理日常操作。这些设备将由现有的大分子结晶设施委员会管理，日常操作将由现有的员工工程师管理。加州大学圣迭戈分校在X射线结晶学方面有着悠久的传统，这一承诺今天仍在继续。化学和生物化学系以及斯卡格斯制药和制药科学学院为管理结晶机器人的人员提供的财政支持证明了这一承诺。该仪器的长期支持将在充值的基础上通过作为大分子结晶设施的一部分的使用费来提供。加州大学圣迭戈分校有相当数量的X射线结晶学家，他们的研究工作将通过定期接触结晶机器人而得到显着加强。值得注意的是，作为核心设施的一部分，该仪器还将使UCSD的研究人员能够将他们的研究计划转向结构确定，这些研究人员的专业知识不是X射线结晶学。从X射线结晶学获得的详细信息水平可能允许这些研究主题向治疗解决方案发展。简而言之，拟议的仪器对于加州大学圣地亚哥分校结构生物学的全校园成功至关重要。 与公共健康相关：由X射线结晶学确定的生物大分子结构提供了无与伦比的功能洞察力，并使旨在治疗人类疾病的疗法的设计成为可能。这项提议寻求在加州大学圣地亚哥分校获得一个用于生物大分子结晶的集成机器人系统。这一设备将使这一校园中的一大批生物医学科学家能够探索与人类疾病相关的基本问题，并将从结构确定中获得的对蛋白质和有序RNA的基本理解应用于治疗学。