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）是表征涉及小分子，蛋白质和多肽，抗体，寡核苷酸，寡核苷酸，脂质，脂质和其他生物细菌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还通常用于测量与蛋白质，纳米颗粒 - 双分子相互作用和金属离子 - 纳米颗粒交换的金属离子结合。这是一种“无标签”方法，可简化样品制备，并允许研究其本地状态的分子（例如，无需荧光或放射性分析标记）。 Th ge/Microcal ITC200系统具有多功能，准确且非常敏感，能够通过竞争结合来研究直接测量二摩尔至纳摩尔结合常数〜更强相互作用。对先前模型的技术进步使该仪器能够测量较低体积（在200 mL比色杯中）以及较低浓度（例如，只有10微克蛋白质）的样品。快速的温度平衡时间允许ITC200最多每小时分析两个样品，同时保持非常高的精度。该样本吞吐量大约是Upenn校园中较旧的ITC系统实现的两倍。在我们的现场测试中，Microcal的ITC200系统在各种不同的样本上产生了高质量的可再现数据。该ITC仪器具有自动注入，注射器洗涤和比色杯洗涤，这对于多用户仪器很有用。 ITC200将是该大型用户组的多功能分析工具，尤其是由 与较旧的型号ITC系统相比，ITC200的速度，灵敏度和较低的样本要求。我们希望该工具的需求量很高，因为希望参与该提案的大量用户以及宾夕法尼亚大学及其周边地区的NIH-Mession驱动生物医学研究的高密度。生物化学服务中心的新主任克里斯·兰奇（Chris Lanci）将监督日常的ITC操作，并将负责保持乐器的运行最佳，培训新用户，并确保该仪器通过大量NIH资助的研究人员经常使用。在操作的前两年，ITC200将对所有用户免费，我们预计在此期间，约有30个用户组将受到培训并收集ITC数据。有效的管理和成本回收计划将使工具在未来十年内以合理的成本以合理的成本提供高度的性能。在安装时，拟议的工具将立即使16个用户群体受益，其中10个居住在宾夕法尼亚大学化学系，代表所有四个部门（有机，无机，生物学和物理化学）。宾夕法尼亚大学医学院，有两个麻醉学和环境医学研究所的成员也有很好的代表。宾夕法尼亚大学生物工程学院和附近的Wistar Institute成员也有对这种多功能热量计的研究需求。