IMMOBILIZED CIBACRON BLUE F3G-A ON CROSSLINKED POLY(VINYL ALCOHOL) FOR AFFINITY CHROMATOGRAPHY

Immobilized triazine dye affinity chromatography has been widely used for protein purification. In this paper, Cibacron Blue F3G-A was immobilized, through a spacer arm, onto a rigid hydrophilic porous polymer by reacting an epoxy-group-containing poly(vinyl alcohol) with 6-aminohexyl-N(-Cibacron Blue F3G-A, which was obtained by reacting Cibacron Blue F3G-A with excess of 1,6-diaminohexane, in a pH 8.6 buffer. The epoxy-group-containing poly(vinyl alcohol) was prepared by treating macroporous poly(vinyl alcohol) with excess epichlorohydrin in the presence of NaOH in dimethyl sulfoxide. The macroporous poly(vinyl alcohol) was prepared by hydrolysis of macroporous crosslinked poly(vinyl acetate), which was synthesized by suspension copolymerization of vinyl acetate and triallyl isocyanurate in the presence of butyl acetate and n-heptane as diluents. The Cibacron Blue F3G-A-immobilized poly(vinyl alcohol) was packed in a stainless steel column (250×5 mm I. D.) and the chromatographic behaviors of several proteins (cytochrome c, lysozyme, bovine serum albumin, insulin, and lactate dehydrogenase) were determined.