AFFINITY-CHROMATOGRAPHY OF SERINE PROTEASES ON THE TRIAZINE DYE-LIGAND CIBACRON BLUE F3G-A

The interaction between complement component factor B and the triazine dye ligand Cibacron Blue F3G-A coupled to a cross-linked agarose matr ix (Blue Sepharose) was found to involve the Bb part of the molecule, and to be inhibited by benzamidine. Human, chicken and rainbow trout f actor B which had bound to Blue Sepharose could subsequently be eluted with benzamidine. Other serine proteases (C2, factor II, factor IX, t rypsin, chymotrypsin, proteinase 3) also bound to Blue Sepharose but o nly those belonging to the trypsin family could be eluted with benzami dine. Trypsin treated with the active-site inhibitor phenylmethylsulfo nyl fluoride did not bind to Blue Sepharose and pretreatment of Blue S epharose with benzamidine did not influence binding of proteases. We c onclude that trypsin-like serine proteases can be purified on Blue Sep harose and that the interaction of these serine proteases with Blue Se pharose involves the active site of the enzyme. (C) 1998 Elsevier Scie nce B.V. All rights reserved.