Patent ID: 6569661
Filing Date: 2003-05-27
Classification: A61K,A61P,C12N,C12Y

Abstract:
A method of purifying human recombinant &agr;-L-iduronidase having a purity of equal to or greater than about 99% purity, comprising the steps of:(a) harvesting and filtering fluid obtained from a culture of Chinese hamster ovary cells transformed with nucleic acids encoding said human recombinant &agr;-L-iduronidase; (b) adjusting the pH of the fluid to an acidic pH wherein any potential virus is inactivated and said human recombinant &agr;-L-iduronidase is not harmed, followed by filtration through a 0.2 micron to 0.54 micron filter; (c) passing the fluid from step (b) through a Cibracon blue dye interaction chromatography column to capture said human recombinant &agr;-L-iduronidase; (d) passing the fluid through a copper chelation chromatography column to remove contaminating Chinese hamster ovary proteins; (e) passing the fluid through a phenyl hydrophobic interaction chromatography column to reduce residual leached Cibacron blue dye and copper ions canned over from previous columns; and (f) concentrating and diafiltering the purified human recombinant &agr;-L-iduronidase; wherein said purity of equal to or greater than about 99% purity of human recombinant &agr;-L-iduronidase is measured by &mgr;g of contaminating Chinese hamster ovary protein per mg of total protein.