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US 10,352,943 B2Granted
A-fucosylation detection in antibodies
AssigneeRoche Glycart AG
InventorsChristiane Jaeger, Hans Koll, Peter Sondermann, Pablo Umana
Life of this patent
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Abstract
This invention describes a new analytical method to determine the quantity and distribution of fucose per Fc within an antibody preparation.
Claims10
A method for detecting the presence or absence of fucose residues within a homogenously deglycosylated immunoglobulin G antibody, said method comprising:
- (a) providing an antibody preparation comprising a glycosylated immunoglobulin G antibody,
- (b) removing heterogeneous saccharide residues from the glycosylated immunoglobulin G antibody with the enzyme endoglycosidase S and the enzyme endoglycosidase H, thereby obtaining said homogenously deglycosylated immunoglobulin G antibody,
- (c) removing other heterogenous residues from said homogenously deglycosylated immunoglobulin G antibody with an enzyme, and
- (d) analyzing said homogenously deglycosylated immunoglobulin G antibody for the presence or absence of fucose residues, wherein the linkage information on the presence or absence of fucose residues within the homogenously deglycosylated immunoglobulin G antibody is maintained.
1 more element
A method for detecting the presence or absence of fucose residues within a homogenously deglycosylated immunoglobulin G antibody, said method comprising:
- (a) providing an antibody preparation comprising a glycosylated immunoglobulin G antibody,
- (b) removing heterogeneous saccharide residues from the glycosylated immunoglobulin G antibody with the enzyme endoglycosidase S and the enzyme endoglycosidase H, thereby obtaining said homogenously deglycosylated immunoglobulin G antibody,
- (c) removing C-terminal lysine residues from said homogenously deglycosylated immunoglobulin G antibody with carboxypeptidase B, and
- (d) analyzing said homogenously deglycosylated immunoglobulin G antibody for the presence or absence of fucose residues by liquid-chromatography-mass spectrometry analysis, capillary electrophoresis-sodium dodecyl sulfate molecular weight analysis, and electrospray ionization mass spectrometry analysis, wherein the linkage information on the presence or absence of fucose residues within the homogenously deglycosylated immunoglobulin G antibody is maintained.
1 more element
Dependencies are inferred from the claim text.