Srisa-Art Monpichar, Dyson Emily C, deMello Andrew J, Edel Joshua B
Department of Chemistry, Imperial College London, South Kensington, London, SW7 2AZ, United Kingdom.
Anal Chem. 2008 Sep 15;80(18):7063-7. doi: 10.1021/ac801199k. Epub 2008 Aug 20.
Rapid kinetic measurements are important in understanding chemical interactions especially for biological molecules. Herein, we present a droplet-based microfluidic platform to study streptavidin-biotin binding kinetics with millisecond time resolution. With integration of a confocal fluorescence detection system, individual droplets can be monitored and characterized online to extract kinetic information. Using this approach, binding kinetics between streptavidin and biotin were observed via fluorescence resonance energy transfer. The binding rate constant of streptavidin and biotin was found to be in a range of 3.0 x 10 (6)-4.5 x 10 (7) M (-1) s (-1).
快速动力学测量对于理解化学相互作用非常重要,尤其是对于生物分子。在此,我们展示了一个基于微滴的微流控平台,用于以毫秒级时间分辨率研究链霉亲和素-生物素结合动力学。通过集成共聚焦荧光检测系统,可以在线监测和表征单个微滴,以提取动力学信息。使用这种方法,通过荧光共振能量转移观察到链霉亲和素与生物素之间的结合动力学。发现链霉亲和素与生物素的结合速率常数在3.0×10⁶ - 4.5×10⁷ M⁻¹ s⁻¹范围内。