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.

所有触觉停止流量微钙化仪，以前报道 （生物物理学杂志49（1986）第87A页； 51（1987）p.443a）现在已经 广泛用于研究DNA-Drug的结合焓 互动。 测得的热合器的分解产生 重建热量表的时间常数大约 三秒钟。 两种方案已用于重建 测得的热合器。 大卫的有限元方法和 Berger（J. Biochem＆Biophys方法。6（1982）P.205-217） 了解量热计的物理结构。 这 Schuette和Mudd改编的迭代方法（J. Biochem＆Biophys 方法，14（1987）p。 167-175）仅要求冲动 以与测量的速度相同的速度知道响应和数字化 热图。 采用两种反卷积方法的结果将是 提出。 微氧化电表能够测量结合 标准偏差为3的30个微孔的焓 Microjoules。 一个反应需要每种试剂的80微升 并在200秒内完成，从而允许典型 每天120-150跑的吞吐量。 高分辨率 仪器可以准确测量反应热量 极度稀释的试剂浓度因此排除了需求 纠正药物和/或DNA聚集的结合焓。