O-LINKED GLYCOSYLATION OF PEPTIDES
The present invention provides polypeptides that include an O-linked glycosylation site that is not present in the wild-type peptide. The polypeptides of the invention include glycoconjugates in which a species such as a water-soluble polymer, a therapeutic agent of a biomolecule is covalently linked through an intact O-linked glycosyl residue to the polypeptide. Also provided are methods of making the peptides of the invention and methods, pharmaceutical compositions containing the peptides and methods of treating, ameliorating or preventing diseased in mammals by administering an amount of a peptide of the invention sufficient to achieve the desired response.
1 . A method of separating a peptide conjugate from a reaction mixture, said method comprising:
(a) contacting said reaction mixture with a hydrophobic interaction chromatography medium; and
(b) collecting said peptide conjugate from said hydrophobic interaction chromatography medium, thereby separating said peptide conjugate from said reaction mixture.
2 . The method according to claim 1 , wherein said peptide conjugate includes a modifying group which is poly(ethylene glycol).
3 . The method according to claim 2 , wherein said modifying group is attached to said peptide of said peptide conjugate through an intact glycosyl linking group.
4 . A conjugate between a peptide and a modifying group, wherein said conjugate is obtained by a method comprising:
(a) contacting a reaction mixture comprising said conjugate with a hydrophobic interaction chromatography medium; and
(b) collecting said conjugate, thereby separating said peptide conjugate from said reaction mixture.
5 . The conjugate according to claim 4 , wherein said modifying group is poly(ethylene glycol).
6 . The method according to claim 5 , wherein said modifying group is attached to said peptide through an intact glycosyl linking group.