Development of New Technique to Separate DNA Related Molecules on Affinity Capillary Electrophoresis

Abstract

The binding properties of several DNA ligands with calf thymus DNA have been studied by using capillary zone electrophoresis (CZE). The peaks of the ligands retarded with increasing concentration of DNA in carrier solution. The binding constants of the ligands with DNA were determined easily with high reproducibility from the slope of the (t-t₁)^-1 vs. [DNA-p]^-1, where t₁ and t are the migration time of the ligands in the absence and in the presence of DNA, respectively. In this procedure, only small quantities of the ligands and DNA samples were required. This binding assay was very convenient compared with conventional methods such as spectroscopic methods and equilibrium dialysis.