Use of a partial-filling technique in affinity capillary electrophoresis for determining binding constants of ligands to receptors

This work evaluates the concept of a partial-filling technique in affinity capillary electrophoresis (ACE) using two model systems: vancomycin from St reptomyces orientalis and carbonic anhydrase B (CAB, EC 4.2.1.1). In this t echnique the capillary is first partially-filled with ligand followed by a sample of receptor and non-interacting standard and electrophoresed. Analys is of the change in the mobility ratio, M, of the receptor, relative to the non-interacting standard, as a function of the concentration of the ligand , yields a value for the binding constant. These values agree well with tho se estimated using other binding and ACE techniques. Data demonstrating the quantitative potential of this method is presented. (C) 1999 Elsevier Scie nce BN. All rights reserved.