REACTIVE DYES HUMAN SERUM-ALBUMIN (HSA) BINDING BY FRONTAL ANALYSIS AFFINITY-CHROMATOGRAPHY

The binding with human serum albumin (HSA) of Cibacron Blue F3GA, Leva fix Brilliant Blue EB, the structure of which has been carefully inves tigated, Reactive Orange 046, and Reactive Red 022 dyes, immobilised o n sepharose Ct 6B, has been quantitatively investigated by frontal aff inity chromatography. This technique allowed the calculation of the as sociation constants of The ligand-HSA complexes, on the basis of the d ependence of the HSA elution volume on its initial concentration. The values obtained were high and comparable for Cibacron and Levafix dyes (5.4 and 5.7x10(6) M-1, respectively) and significantly smaller for R eactive Orange 046 and Reactive Red 022 (0.23 and 0.4x10(6) M-1). A po ssible interpretation of these data, based on the different structures of the bonded dyes, is proposed.