Multi-User, Isothermal Titration Microcalorimeter

描述：等温滴定量热法(ITC)是表征水溶液中小分子、蛋白质和多肽、抗体、寡核苷酸、脂类和其他生物分子相互作用的热力学和化学计量学最直接的方法之一。ITC还常规用于测量金属离子与蛋白质的结合、纳米颗粒与生物分子的相互作用以及金属离子与纳米颗粒的交换。这是一种“无标记”的方法，简化了样品制备，并允许研究处于天然状态的分子(例如，不需要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，能够直接测量亚毫米到纳米分子的结合常数~通过竞争结合可以研究更强的相互作用。与以前型号相比的技术进步使该仪器能够测量体积大约小7倍的样品(在200毫升试管中)，以及更低的浓度(例如，只需要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。这个样本吞吐量大约是宾夕法尼亚大学校园内较旧ITC系统的两倍。在我们的现场测试中，MicroCal的iTC200系统在各种不同的样品上产生了高质量、可重现的数据。这台ITC仪器具有自动注射、注射器清洗和试管清洗功能，这对于多用户仪器非常有用。对于这一庞大的用户群体，iTC200将是一个多功能的分析工具，尤其是通过 与较旧型号的ITC系统相比，iTC200的速度更快、灵敏度更高、样品要求更低。我们预计，基于希望参与这项计划的大量用户，以及宾夕法尼亚大学和周边地区由NIH使命驱动的高密度生物医学研究，这种仪器的需求将会很高。新任生物化学服务中心主任Chris Lanci博士将监督ITC的日常运作，并将负责保持仪器的最佳运行状态，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。在运行的头两年，iTC200将向所有用户免费开放，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未来十年内以合理的成本向用户提供高水平的性能。在安装时，建议的仪器将立即使16个用户组受益，其中10个用户组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、生物和物理化学)。宾夕法尼亚大学医学院也有很好的代表，该学院有两个麻醉学系和一名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维斯塔尔研究所的一名成员也对这种多功能热量计有研究需求。

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.