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US 8,785,212 B2Granted
Oligosaccharide modification and labeling of proteins
AssigneeLife Technologies Corporation
InventorsBrian Agnew, Kyle Gee, Schuyler Corry
Life of this patent
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Abstract
The present invention generally relates to methods of functionalizing proteins, particularly antibodies, at oligosaccharide linkages, methods of humanizing antibodies by modifying glycosylation, as well as to novel antibodies linked to modified oligosaccharides. The invention further relates to kits that may be used to produce the antibodies of the invention.
Claims15
A method of labeling a protein by labeling an oligosaccharide attached to said protein, comprising:
- incubating a protein-producing cell in the presence of an unnatural sugar, wherein the unnatural sugar comprises an azide group, to provide a modified protein, wherein the modified protein produced by the protein-producing cell is attached to an oligosaccharide that comprises the unnatural sugar, wherein the unnatural sugar comprises GalNAz, GlcNAz, Ac4GalNAz, or Ac4GlcNAz;
- and
- attaching the modified protein to a label that comprises an alkyne group or an activated alkyne group by contacting the modified protein with the label under a condition such that a covalent bond is formed between the alkyne or the activated alkyne group of the label and the azide group of the unnatural sugar, thereby labeling the protein.
A method of metabolically modifying a protein by modifying an oligosaccharide group attached to the protein, comprising:
- incubating a protein-producing cell in the presence of an unnatural substance, wherein the unnatural substance comprises an azide group, to provide a modified protein, wherein the modified protein produced by the protein-producing cell is attached to an oligosaccharide that comprises the unnatural substance, wherein the unnatural substance comprises an unnatural sugar selected from GalNAz, GlcNAz, Ac4GalNAz, and Ac4GlcNAz;
- and
- attaching the modified protein to a label that comprises an alkyne group or an activated alkyne group by contacting the modified protein with the label under a condition such that a covalent bond is formed between the alkyne or the activated alkyne group of the label and the azide group of the unnatural sugar, thereby modifying the protein.
Dependencies are inferred from the claim text.