USE OF CAPILLARY ZONE ELECTROPHORESIS FOR THE ANALYSIS OF DNA-BINDINGTO A PEPTIDE DERIVED FROM AMYLOID P COMPONENT

Capillary zone electrophoresis was used to characterize the binding in teractions between oligonucleotides and synthetic peptides derived fro m human serum amyloid P component. From a solution containing free and peptide-bound oligonucleotides, free nucleotide was separated from th e complex by means of electrophoresis. In this way, both qualitative a nd quantitative aspects of the binding could be assessed rapidly using minute amounts of unlabelled samples. Minimal structures and sequence specificity of binding of nucleotides and peptides could be character ized and, based on the quantitative output of the electrophoretic anal ysis, binding constants were estimated. In theory, the approach is app licable for any molecular interaction where the charge/mass ratio of c omplexes differ from the free molecules and where at least one of the interacting components is quantitatively recoverable and detectable in the capillary electrophoresis system. As such, it is the only method available for simple and fast estimates of binding parameters for unla belled low molecular weight compounds in solution.