Evaluation of the Flow-Dialysis Technique for Analysis of Protein-Ligand Interactions: An Experimental and a Monte Carlo Study

ABSTRACT Flow dialysis has found widespread use in determining the dissociation constant (K^sub D^) of a protein-ligand interaction or the amount of available binding sites (E^sub 0^). This method has the potency to measure both these parameters in a single experiment and in this article a method to measure simultaneously the K^sub D^ and E^sub 0^ is presented, together with an extensive error analysis of the method. The flow-dialysis technique is experimentally simple to perform. However, a number of practical aspects of this method can have a large impact on the outcome of K^sub D^ and E^sub 0^. We have investigated all sources of significant systematic and random errors, using the …