差示扫描荧光及其在蛋白质研究中的应用

Differential scanning fluorimetry and its application in the study of protein

差示扫描荧光(differential scanning fluorimetry,DSF)是在荧光定量PCR仪上缓慢加热样品,在加热的过程中检测荧光染料与结构发生改变的蛋白质相结合的量,来评价蛋白质热稳定性的方法。该方法具有数据准确、高通量、蛋白质样品损耗较小和温度变化范围广的优点,目前已被广泛应用于蛋白质稳定性、结构和蛋白质-配体相互作用研究等领域。本文主要介绍了DSF的技术原理、仪器设备与反应体系、数据分析及应用进展,并对其发展趋势和前景进行评价,旨在为相关领域的研究提供一定的参考。

Differential Scanning Fluorimetry（DSF） is a method to evaluate the thermal stability of protein by the determination on the quantity of fluorescence dye bound to the unfold protein during the slow heating of protein sample on a real-time PCR instrument. This method, which has the advantages of accuracy, high throughput, low loss of protein sample and operating over large temperature ranges, is widely used in the research on thermal stability and structure of protein, interaction of protein-ligand and so on. In this paper, in order to offer the reference for the researches in related fields, the technical principle, instruments and reaction system, data analysis and application progress of DSF were introduced, meanwhile the development tendency and prospect of DSF were evaluated.