Monitoring of real-time streptavidin-biotin binding kinetics using droplet microfluidics.
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).
Srisa-Art
Monpichar
M
Department of Chemistry, Imperial College London, South Kensington, London, SW7 2AZ, United Kingdom.
Dyson
Emily C
EC
deMello
Andrew J
AJ
Edel
Joshua B
JB
D001710:Biotin / Q000378:metabolism*; D031541:Fluorescence Resonance Energy Transfer; D007700:Kinetics; D046210:Microfluidic Analytical Techniques / Q000379:methods*; D011485:Protein Binding; D019809:Streptavidin / Q000378:metabolism*; D013997:Time Factors
D001710:Biotin; D019809:Streptavidin