Affinity chromatography of guanyloribonuclease from streptomyces aureofaciens

Abstract

2',3'-O-2-Carboxyethylideneinosine 5'-phosphate (Ia), 5'-phosphorothioate (IIa), 5'-O-carboxymethylinosine 2'(3')-phosphate (IIIa) and 2'(3')-phosphorothioate (IVa) were bound to 6-aminohexyl-Sepharose 4B by the method of mixed anhydrides with ethyl chloroformate. 5'-O-(2-Aminoethylamidocarbonylmethyl)inosine 2'(3')-phosphorothioate (Va), O-(4-aminophenyl)inosine 5'-phosphorothioate (VIa) and O-(4-aminophenyl)-O-phenyl phosphorothioate (VIIa) were bound to Sepharose 4B activated with cyanogen bromide. The resulting modified supports IIa-VIb bind the ribonuclease of Streptomyces aureofaciens within the pH range 6.5-7.5. The specific affinity bond manifests itself in derivatives with ligands of type IVb and Vb, on which a 50-70fold purification degree can be achieved, with respect to the crude preparation.