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 Streptomyces orientalis and carbonic anhydrase B (CAB, EC 4.2.1.1). In this technique the capillary is first partially-filled with ligand followed by a sample of receptor and non-interacting standard and electrophoresed. Analysis 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 those estimated using other binding and ACE techniques. Data demonstrating the quantitative potential of this method is presented.
Heintz
J
J
Department of Chemistry and Biochemistry, California State University, Los Angeles 90032-8202, USA.
Hernandez
M
M
Gomez
F A
FA
D000900:Anti-Bacterial Agents / Q000737:chemistry; D002256:Carbonic Anhydrases / Q000032:analysis; D019075:Electrophoresis, Capillary / Q000379:methods*; D007700:Kinetics; D008024:Ligands; D011955:Receptors, Drug / Q000737:chemistry*; D014640:Vancomycin / Q000737:chemistry
D000900:Anti-Bacterial Agents; D008024:Ligands; D011955:Receptors, Drug; D014640:Vancomycin; D002256:Carbonic Anhydrases