DEVELOPMENT OF BIOCALORIMETERS FOR SOLUTION AND CELL BIOCHEMICAL STUDIES

黑色金刚石涂层聚丙烯流动池已用于生产一种 停流间歇式微量热计。这台仪器未完成 用于样品插入、数据收集和分析的计算机控制。 目前正在探索几种生化反应，使用 乐器。这些包括粘连蛋白组装、tRNA-转移酶反应。 以及ATCase与各种底物的反应。蛋白质浓度 每次反应只需50微升，即可使用10-6M。四个实验 每小时都可以在这种敏感度下运行。 该项目的意义在于有可能发现新的 由热反应和能力揭示的反应路径 对无氧生物反应进行基本热力学测量 其他检测系统也可用。此外，直接测量 是一种比反应热更精确的方法 用Van‘t Hoff方法测定，即测定反应 在不同温度下的平衡。当使用Van‘t Hoff方法时， 4摄氏度到40摄氏度的温度范围太小，不适合 准确地测定了热焓。因此，为了预测 例如，药物代谢中的反应平衡，直接 量热测量是更好的选择。

Black diamond coated polypropylene flow cells have been used to produce a stopped flow batch microcalorimeter. The instrument is under complete computer control for sample insertion, data collection, and analysis. Several biochemical reactions are presently being explored using the instrument. These include clatherin assembly, tRNA-transferase reactions and ATCase reactions with various substrates. Protein concentrations of 10-6M can be used with only 50 Mul needed per reaction. Four experiments per hour can be run at this level of sensitivity. The significance of the project lies in the possibility of detecting new reaction pathways as revealed by the thermal reactions and in the ability to make basic thermodynamic measurements on biological reactions where no other detection system is available. In addition, the direct measurement of the reaction enthalpy is a much more precise method than the determination using the Van't Hoff method, i.e. measuring of the reaction equilibrium at different temperatures. When the Van't Hoff method is used, the range of temperature, 4 degrees C to 40 degrees C, is too small for an accurate determination of the enthalpy. Thus to predict the change in reaction equilibrium in, for example, drug metabolism, a direct calorimetric measurement is much to be preferred.