Multi-User, Isothermal Titration Microcalorimeter

多用户等温滴定微热量计

• 批准号: 8447682
• 负责人: Ivan Julian Dmochowski
• 金额: $ 12.91万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-06-01 至 2015-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8447682
• 关键词: Anesthesiology; Antibodies; Binding; Biochemistry; Biomedical Engineering; Biomedical Research; Calorimetry; Charge; Chemistry; Competitive Binding; Data; Ensure; Environmental Medicine; Faculty; Fluorescence; Funding; Hour; Injection of therapeutic agent; Inorganic Chemistry; Institutes; Ions; Label; Lipids; Measures; Medicine; Metal Ion Binding; Metals; Methods; Mission; Modeling; Oligonucleotides; Organic Chemistry; Performance; Physical Chemistry; Preparation; Proteins; Recovery; Research; Research Personnel; Running; Sampling; Scheme; Schools; Services; Solutions; Speed; Syringes; System; Temperature; Testing; Thermodynamics; Time; Titrations; Training; United States National Institutes of Health; Universities; analytical tool; aqueous; base; cost; density; instrument; member; nanoparticle; operation; polypeptide; small molecule; stoichiometry

DESCRIPTION: Isothermal titration calorimetry (ITC) is one of the most straightforward methods for characterizing the thermodynamics and stoichiometry of molecular interactions involving small molecules, proteins and polypeptides, antibodies, oligonucleotides, lipids and other biomolecules in aqueous solution. ITC is also used routinely for measuring metal ion binding to proteins, nanoparticle-biomolecule interactions, and metal ion-nanoparticle exchange. This is a 'label-free' approach, which simplifies sample preparation and allows the study of molecules in their native state (e.g., without need for fluorescence or radioisotopic labeling). Th GE/MicroCal iTC200 system is versatile, accurate and very sensitive, capable of measuring directly sub-millimolar to nanomolar binding constants~ stronger interactions can be investigated via competitive binding. Technical advances over the previous model enable this instrument to measure samples of roughly 7-fold lower volume (in a 200 mL cuvette), as well as at lower concentration (e.g., only 10 micrograms protein required). The rapid temperature equilibration time allows the iTC200 to analyze up to two samples per hour, while maintaining very high accuracy. This sample throughput is roughly twice that achieved with older ITC systems on UPenn's campus. In our field testing, MicroCal's iTC200 system produced high-quality, reproducible data on a variety of different samples. This ITC instrument has automated injection, syringe washing, and cuvette washing, which is useful for a multi-user instrument. The iTC200 will be a versatile analytical tool for this large users group, particularly enhanced by the greater speed, sensitivity and lower sample requirements of the iTC200 compared to older model ITC systems. We expect this instrument to be in high demand based on the large number of users who wanted to participate in this proposal as well as the high density of NIH-mission-driven biomedical research at University of Penn and the surrounding region. The new Director of the Biological Chemistry Service Center, Dr. Chris Lanci, will oversee day-to-day ITC operations, and will be in charge of keeping the instrument running optimally, training new users, and ensuring that the instrument receives frequent use by a large number of NIH-funded researchers. For the first two years of operation, the iTC200 will be free to all users, and we anticipate that ~ 30 user groups will become trained and collect ITC data during this time. An efficient management and cost-recovery scheme will keep the instrument delivering a high level of performance at reasonable cost to users for the next decade. At the time of installation, the proposed instrument will immediately benefit 16 user groups, ten of whom reside in the University Of Penn Department Of Chemistry and represent all four divisions (Organic, Inorganic, Biological, and Physical Chemistry). Also well represented is the University Of Penn School Of Medicine, with two Anesthesiology faculties and a member of the Institute of Environmental Medicine. Two University of Penn Bioengineering faculties and a member of the nearby Wistar Institute also have research needs for this versatile calorimeter.

描述：等温滴定量热法 (ITC) 是表征水溶液中涉及小分子、蛋白质和多肽、抗体、寡核苷酸、脂质和其他生物分子的分子相互作用的热力学和化学计量的最直接方法之一。 ITC 还经常用于测量金属离子与蛋白质的结合、纳米颗粒-生物分子相互作用以及金属离子-纳米颗粒交换。这是一种“无标记”方法，可简化样品制备并允许研究天然状态的分子（例如，无需荧光或放射性同位素标记）。 GE/MicroCal iTC200 系统用途广泛、准确且非常灵敏，能够直接测量亚毫摩尔至纳摩尔的结合常数~可以通过竞争性结合研究更强的相互作用。与之前型号相比，技术进步使得该仪器能够测量大约 7 倍的体积（在 200 mL 比色皿中）和较低浓度的样品（例如，仅需要 10 微克蛋白质）。快速的温度平衡时间允许 iTC200 每小时分析多达两个样品，同时保持非常高的精度。该样本吞吐量大约是宾夕法尼亚大学校园旧 ITC 系统的两倍。在我们的现场测试中，MicroCal 的 iTC200 系统针对各种不同的样品生成了高质量、可重复的数据。该 ITC 仪器具有自动进样、注射器清洗和比色皿清洗功能，这对于多用户仪器非常有用。 iTC200 将成为这一庞大用户群的多功能分析工具，特别是通过 与旧型号 ITC 系统相比，iTC200 具有更高的速度、灵敏度和更低的样品要求。我们预计该仪器的需求量很大，因为有大量用户希望参与该提案，并且宾夕法尼亚大学及周边地区的 NIH 任务驱动的生物医学研究密度很高。生物化学服务中心新任主任 Chris Lanci 博士将监督 ITC 的日常运营，并负责保持仪器最佳运行、培训新用户并确保仪器得到经常使用大量 NIH 资助的研究人员。在运行的前两年，iTC200 将免费向所有用户开放，我们预计大约 30 个用户组将在此期间接受培训并收集 ITC 数据。高效的管理和成本回收方案将使该仪器在未来十年内以合理的成本为用户提供高水平的性能。安装时，拟议的仪器将立即使 16 个用户组受益，其中 10 个用户组位于宾夕法尼亚大学化学系，代表所有四个部门（有机、无机、生物和物理化学）。宾夕法尼亚大学医学院也很有代表性，该学院拥有两个麻醉学系和一个环境医学研究所的成员。宾夕法尼亚大学生物工程学院的两个院系和附近威斯塔研究所的一个成员也对这种多功能热量计有研究需求。