COATED SILICA AND ITS BEHAVIOR IN DYE-AFFINITY CHROMATOGRAPHY

Cellulose-coated silica as a support for the high-performance affinity liquid chromatography of proteins was prepared. Cibacron Blue 3G-A as an affinity ligand was grafted onto silica beads coated with cellulose in one or two steps. The prepared supports were used for the purification of lactate dehydrogenase from a crude bovine extract using solutions of potassium chloride, NADH and Cibacron Blue-dextran T 10 as eluents of the bound enzyme. The sorption capacities of the dyed cellulose-coated silica for lactate dehydrogenase from bovine muscle were found to be much lower than those of Cibacron Blue-bead cellulose. The comparison of un-coated and coated silica supports showed that the non-specific interactions between the enzyme and the silica coated with cellulose were considerably reduced.