DEVELOPMENT OF BIOCALORIMETERS FOR SOLUTION AND CELL BIOCHEMICAL STUDIES

开发用于溶液和细胞生化研究的生物量热计

• 批准号: 3920039
• 负责人: R L BERGER
• 金额: --
• 依托单位: NATIONAL HEART, LUNG, AND BLOOD INSTITUTE
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/3920039
• 关键词: DNA; clinical biomedical equipment; drug metabolism; microcalorimetry; optics; reagent /indicator; stereochemistry; stop flow technique; tantalum; thermodynamics; tissue /cell culture

An all tantalum stopped-flow microcalorimeter, previously reported (Biophysical Journal 49(1986) p.87a; 51(1987) p.443a), has now been used extensively to study the binding enthalpies of DNA-drug interactions. Decomposition of the measured thermograms yields reconstructed thermograms with a time constant of approximately three seconds. Two schemes have been used to reconstruct the measured thermograms. The finite element method of Davids and Berger (J. Biochem & Biophys Methods. 6(1982) p.205-217) requires a knowledge of the physical construction of the calorimeter. The iterative method adapted by Schuette and Mudd (J. Biochem & Biophys Methods, 14 (1987) p. 167-175) requires only that the impulse response be known and digitized at the same rate as the measured thermogram. Results employing both deconvolution methods will be presented. The microcalorimeter is capable of measuring binding enthalpies of 30 microjoules with a standard deviation of 3 microjoules. A reaction requires 80 microliters of each reagent and is completed within 200 seconds thus allowing a typical throughput of 120-150 runs per day. The high resolution of this instrument has permitted accurate measurement of reaction heats at extremely dilute reagent concentrations thereby precluding the need to correct the binding enthalpies for drug and/or DNA aggregation.

一个全钽停流微量热计，以前报道 （生物物理学杂志49（1986）p.87a; 51（1987）p.443a），现在已经 广泛用于研究DNA-药物的结合能力 交互. 对测得的热谱图进行分解， 重建的热谱图，时间常数约为 三秒 两种方案已被用于重建 测量的热谱图。 Davids的有限元方法和 Berger（J. Biochem & Biophys Methods. 6（1982）p.205-217）要求 量热计的物理结构知识。 的 Schuette和Mudd采用的迭代方法（生物化学和生物物理学杂志 Methods，14（1987）p.167 -175）仅要求脉冲 响应是已知的，并以与测量的相同的速率数字化。 温度记录图 采用这两种去卷积方法的结果将是 提出了 微量热计能够测量结合 30微焦耳的能量，标准差为3 微焦耳。 一个反应需要80微升的每种试剂 并在200秒内完成，从而允许典型的 生产能力为每天120-150次运行。 这张照片的高分辨率 仪器允许精确测量反应热， 从而排除了需要 以校正药物和/或DNA聚集的结合位点。