Study of porous cellulose beads as a dye-ligand matrix. Effect of protein admixtures and concentration of immobilized dye in the quantitative analysis of lactate dehydrogenase: Cibacron blue interaction

Abstract

The effect of protein admixtures and concentration of immobilized dye on the interaction of lactate dehydrogenase (LDH) with Cibacron Blue-bead cellulose has been studied by means of zonal chromatography with Cibacron Blue-dextran T 10 as the mobile ligand. The influence of both variables on the values of dissociation constants of complexes immobilized dye-LDH (KI-L) and mobile dye-LDH (KM-L) can be summarized as follows: The values of KI-L remained practically constant and were found to be lower than those of KM-L. The differences found in KI-L and KM-L values should help to estimate the contribution of nonspecific interaction of the matrix to the total interaction between dye-ligand and the enzyme.