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.

产品说明：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学的最直接的方法之一。ITC也经常用于测量金属离子与蛋白质的结合、纳米粒子-生物分子相互作用和金属离子-纳米粒子交换。这是一种“无标记”方法，其简化了样品制备并允许研究处于其天然状态的分子（例如，而不需要荧光或放射性同位素标记）。GE/MicroCal iTC 200系统是通用的、准确的和非常灵敏的，能够直接测量亚毫摩尔到纳摩尔的结合常数，可以通过竞争性结合来研究更强的相互作用。相对于先前型号的技术进步使该仪器能够测量体积低约7倍的样品（在200 mL比色皿中）以及较低浓度的样品（例如，仅需10微克蛋白质）。快速的温度平衡时间允许iTC 200每小时分析多达两个样品，同时保持非常高的精度。这个样品吞吐量大约是宾夕法尼亚大学校园内旧ITC系统的两倍。在我们的现场测试中，MicroCal的iTC 200系统对各种不同的样品产生了高质量的可重复数据。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。iTC 200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，iTC 200具有更高的速度、灵敏度和更低的样品要求。我们预计，基于大量希望参与该提案的用户以及宾夕法尼亚大学及其周边地区NIH任务驱动的生物医学研究的高密度，该仪器的需求量很大。生物化学服务中心的新任主任Chris Lanci博士将负责ITC的日常运营，并将负责保持仪器的最佳运行，培训新用户，并确保该仪器得到大量NIH资助的研究人员的频繁使用。在运营的头两年，iTC 200将免费向所有用户开放，我们预计在此期间将有约30个用户组接受培训并收集ITC数据。有效的管理和成本回收计划将使该仪器在今后十年内以合理的成本向用户提供高水平的性能。在安装时，拟议的仪器将立即受益16个用户组，其中10人居住在宾夕法尼亚大学化学系，并代表所有四个部门（有机，无机，生物和物理化学）。宾夕法尼亚大学医学院也有很好的代表性，有两个麻醉学系和一个环境医学研究所的成员。宾夕法尼亚大学的两个生物工程学院和附近的Wistar研究所的一名成员也对这种多功能热量计有研究需求。