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US 5,514,572 AGranted
Method to maintain the activity in polyethylene glycol-modified proteolytic enzymes
AssigneeConsiglio Nazionale Delle Ricerche
InventorsFrancesco Veronese, Paolo Caliceti, Oddone Schiavon, Sartore Luciana
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Abstract
A method for preparing protease-polyethylene glycol adducts is presented wherein the immobilized reversible inhibitor, benzamidine, prevents reaction of activated polyethylene glycol with the active site of the protease. Improved activity against macromolecular substrates is obtained compared to when the benzamidine is in solution during the conjugation reaction.
Claims5
A method for the preparation of proteolytic enzyme-polyethylene glycol adducts, comprising:
- (A) mixing a proteolytic enzyme selected from the group consisting of trypsin, urokinase, tissue plasminogen activator, plasmin, chymotrypsin, elastase and kallikrein with benzamidine which has been immobilized on a hydrated insoluble polysaccharide;
- (B) reacting the mixture of (A) with activated polyethylene glycol;
- (C) separating the enzyme-polyethylene glycol adduct of (B) from the insoluble polysaccharide;
- and
- (D) recovering the enzyme-polyethylene glycol adduct.
2 more elements
Dependencies are inferred from the claim text.