Figure 6
From: A rebinding-assay for measuring extreme kinetics using label-free biosensors
Comparison of the competitive kinetics assay with the rebinding assay in terms of sensitivity and parameter estimation, evaluated using Monte Carlo simulations seeded with pairs of pseudo-random kinetic values. (a) Affinity space plot for competitive kinetics with contour curves connecting regions of equal response (response (RU) is inset on contour curves). (b) As in (a) but replicated for the rebinding assay format. (c) Correlation of true_ka versus ka returned from fitting the competitive kinetic model, associated with the parent Monte Carlo simulation, for pseudo-random ka/kd combinations spanning 10-orders with limits 4 ≤ Log(ka) ≤ 9 and − -6 ≤ Log(kd) ≤ 4. The diagonal, or unit slope, indicates the accuracy of parameter return and the SE associated with the parameter fit is indicated by the ± error bars. (d) Data set given in (c) were ka/kd combinations containing transient kd values were eliminated by restricting the kd limit to − -6 ≤ Log(kd) ≤ 1. (e) As in (d) but given in terms of the fitted kd versus the true-kd. (f) Correlation of fitted ka versus the true_ka for the rebinding assay format. Assay parameters for the Monte Carlo simulations were matched with competitive kinetics in (c) and are shown over the full kinetic range given by limits 4 ≤ Log(ka) ≤ 9 and − 6 ≤ Log(kd) ≤ 4. (g) Correlation of fitted kd versus the true_kd for the rebinding assay in (f), where kd was restricted to transient kd values over the limit 1 ≤ Log(kd) ≤ 4. See methods section for more details.
