Figure 3
From: A rebinding-assay for measuring extreme kinetics using label-free biosensors
Estimation of ka from individual inhibition curves at two values of ka′. (a) Fitted inhibition curves for moderate- (left panel Da = 8.7) and high- (right panel, Da = 87) mass transport limitation. Inhibition curves (black) were fit (red) to Eq. (4), where ka was fit locally, giving independent estimates of ka for each curve, while all other parameters were held constant. (b) Relative error in ka.A as a function of true ka.A. All values where the absolute relative error was < 0.2 are shown. Relative error = 1-(true_ka.A/ka.A). (c) 95% confidence intervals expressed as a fraction of estimated ka.A and plotted versus true ka.A and were < 0.05. Details of the numerical simulations in (a) are as follows. At t = 0 s, B is pre-bound to the hydrogel to a concentration equivalent to R0 = 300 RU = 0.1Rmax. At t = 10 s, 0.5 mM A is injected inhibiting rebinding of B thereby accelerating escape of B. The inhibition curves were simulated over serial 1.5-fold increasing ka and included a blank inhibition curve, where A = 0. Other simulation parameters were as follows, kd = 0.001 s−1, kd′ = 0.05 s−1, h = 10 μm and νc = 0.15 m/s. Inhibition curves were simulated using the virtual instrument and finite element analysis engine described in the Method section.
