A simple electrochemical method for the determination of association constants between carbohydrates and carbohydrate-binding proteins using cyclic voltammetry (CV) is described. The binding of concanavalin A (Con A) and cholera toxin (CT) to their specific α-mannose and β-galactose derivatives self-assembled on gold electrodes is electrochemically monitored with a redox probe of K3Fe(CN)6/K4Fe(CN)6. Upon binding of the proteins to the carbohydrate-modified electrodes, the redox current in CV decreases. The binding-induced change in electrochemical signal is thus used to construct Langmuir adsorption isotherm for the carbohydrate-protein interactions and to obtain the association constants. The association constants of carbohydrate-protein interactions determined by CV ((5.8 ± 1.2) × 107 M- 1 for mannose-Con A, (2.6 ± 0.5) × 108 M- 1 for galactose-CT) were in good agreement with those measured with electrochemical impedance spectroscopy and quartz crystal microbalance.
All Science Journal Classification (ASJC) codes