Monitoring of real-time streptavidin-biotin binding kinetics using droplet microfluidics
Monitoring of real-time streptavidin-biotin binding kinetics using droplet microfluidics
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DOI：
10.1021/ac801199k
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发表时间：
2008-09-15
影响因子：
7.4
通讯作者：
Edel, Joshua B.
中科院分区：
文献类型：
--
作者：
Srisa-Art, Monpichar;Dyson, Emily C.;Edel, Joshua B.
快速动力学测量对于理解化学相互作用（尤其是生物分子）非常重要。在此，我们提出了一种基于液滴的微流体平台，以毫秒时间分辨率研究链霉亲和素-生物素结合动力学。通过集成共焦荧光检测系统，可以在线监测和表征单个液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链霉亲和素和生物素之间的结合动力学。发现链霉亲和素和生物素的结合速率常数在3.0 x 10(6)-4.5 x 10(7) M-1 S-1范围内。
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.