PEG-urate oxidase conjugates and use thereof
AssigneesMountain View Pharmaceuticals, Inc., Duke University
InventorsL. David Williams, Michael S. Hershfield, Susan J. Kelly, Mark G. P. Saifer, Merry R. Sherman
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Abstract
A naturally occurring or recombinant urate oxidase (uricase) covalently coupled to poly(ethylene glycol) or poly(ethylene oxide) (both referred to as PEG), wherein an average of 2 to 10 strands of PEG are conjugated to each uricase subunit and the PEG has an average molecular weight between about 5 kDa and 100 kDa. The resulting PEG-uricase conjugates are substantially non-immunogenic and retain at least 75% of the uricolytic activity of the unmodified enzyme.
Claims6
A method for isolating a tetrameric form of uricase from a solution of purified uricase, said purified uricase comprising tetrameric uricase and uricase aggregates, said method comprising:
- separating the solution into fractions on at least one separation column at a pH of 9 to 10.5, wherein said column is selected from the group consisting of an ion-exchange column and a size-exclusion column;
- and
- recovering from said column one or more fractions that contain isolated tetrameric uricase, wherein said one or more fractions contain less than 10% of uricase aggregates larger than tetramers.
Dependencies are inferred from the claim text.