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.

先前报道的全钽停流微量热计 (Biophysical Journal 49(1986) p.87a; 51(1987) p.443a)，现已 广泛用于研究 DNA 药物的结合焓 互动。 测得的热分析图产量的分解 重建的热分析图的时间常数约为 三秒钟。 已使用两种方案来重建 测量的热分析图。 Davids 的有限元方法和 Berger (J. Biochem & BiophysMethods. 6(1982) p.205-217) 要求 了解热量计的物理结构。 这 Schuette 和 Mudd (J. Biochem & Biophys) 采用的迭代方法 方法，14 (1987) p。 167-175）仅要求脉冲 响应以与测量相同的速率已知并数字化 热分析图。 采用两种反卷积方法的结果将是 提出。 微量热计能够测量结合 30 微焦的焓，标准差为 3 微焦耳。 一次反应需要每种试剂 80 微升 并在 200 秒内完成，从而允许典型的 每天运行 120-150 次的吞吐量。 这个的分辨率很高 仪器可以准确测量反应热 极稀的试剂浓度，从而无需 校正药物和/或 DNA 聚集的结合焓。