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US 7,517,670 B2Granted
Glycosylation engineering of antibodies for improving antibody-dependent cellular cytotoxicity
AssigneeGlycArt Biotechnology AG
InventorsPablo Umaña, Joël Jean-Mairet
Life of this patent
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Abstract
The present invention relates to the field glycosylation engineering of proteins. More particular, the present invention is directed to the glycosylation engineering of proteins to provide proteins with improved therapeutic properties, e.g., antibodies, antibody fragments, or a fusion protein that includes a region equivalent to the Fc region of an immunoglobulin, with enhanced Fc-mediated cellular cytotoxicity.
Claims67
A method for producing a recombinant antibody having increased Fc mediated cellular cytotoxicity or increased Fc receptor binding affinity, comprising:
- (a) providing a mammalian host cell that expresses a recombinant antibody comprising an IgG Fc region containing N-linked oligosaccharides;
- (b) glycoengineering said host cell, wherein said glycoengineering comprises genetically manipulating said host cell so that said host cell has a decreased level of activity of at least one glycoprotein-modifying glycosyltransferase;
- (c) culturing said glycoengineered host cell under conditions which permit the production of said recombinant antibody;
- and
- (d) isolating said recombinant antibody;
- wherein said recombinant antibody has increased Fc-mediated cellular cytotoxicity or increased Fc receptor binding affinity compared to the corresponding antibody produced by the same host cell that has not been glycoengineered.
3 more elements
A method for producing a recombinant antibody having increased Fc mediated cellular cytotoxicity or increased Fc receptor binding affinity, comprising:
- (a) providing a mammalian host cell that expresses a recombinant antibody comprising an IgG Fc region containing N-linked oligosaccharides;
- (b) glycoengineering said host cell, wherein said glycoengineering comprises genetically manipulating said host cell so that said host cell has an altered level of activity of at least one glycoprotein-modifying glycosyltransferase selected from the group consisting of:
- β(1,4)-N-acetylglucosaminyltransferase III and α-mannosidase II;
- (c) culturing said glycoengineered host cell under conditions which permit the production of said recombinant antibody;
- and
- (d) isolating said recombinant antibody;
- wherein said recombinant antibody has increased Fc-mediated cellular cytotoxicity or increased Fc receptor binding affinity compared to the corresponding antibody produced by the same host cell that has not been glycoengineered.
4 more elements
A method for producing a recombinant antibody having increased Fc mediated cellular cytotoxicity or increased Fc receptor binding affinity, comprising:
- (a) providing a mammalian host cell that expresses a recombinant antibody comprising an IgG Fc region containing N-linked oligosaccharides;
- (b) glycoengineering said host cell so that said host cell has an altered level of activity of at least one glycoprotein-modifying glycosyltransferase;
- (c) culturing said glycoengineered host cell under conditions which permit the production of said recombinant antibody;
- and
- (d) isolating said recombinant antibody;
- wherein said recombinant antibody has increased Fc-mediated cellular cytotoxicity or increased Fc receptor binding affinity compared to the corresponding antibody produced by the same host cell that has not been glycoengineered, and wherein the predominant N-linked oligosaccharide in the Fc region of said antibody produced by said glycoengineered host cell is not a high-mannose structure.
3 more elements
A method for producing a recombinant antibody having increased Fc mediated cellular cytotoxicity or increased Fc receptor binding affinity, comprising:
- (a) providing a mammalian host cell that expresses a recombinant antibody comprising an IgG Fc region containing N-linked oligosaccharides;
- (b) glycoengineering said host cell so that said host cell has an altered level of activity of at least one glycoprotein-modifying glycosyltransferase;
- (c) culturing said glycoengineered host cell under conditions which permit the production of said recombinant antibody;
- and
- (d) isolating said recombinant antibody;
- wherein said recombinant antibody has increased Fc-mediated cellular cytotoxicity or increased Fc receptor binding affinity compared to the corresponding antibody produced by the same host cell that has not been glycoengineered, and wherein the Fc region containing N-linked oligosaccharides in said antibody further comprises an increased proportion of GlcNAc residues compared to the corresponding antibody produced by the same host cell that has not been glycoengineered.
3 more elements
A method for producing a recombinant antibody having increased Fc mediated cellular cytotoxicity or increased Fc receptor binding affinity, comprising:
- (a) providing a mammalian host cell that expresses a recombinant antibody comprising an JgG Fc region containing N-linked oligosaccharides;
- (b) glycoengineering said host cell so that said host cell has an altered level of activity of at least one glycoprotein-modifying glycosyltransferase;
- (c) culturing said glycoengineered host cell under conditions which permit the production of said recombinant antibody;
- and
- (d) isolating said recombinant antibody;
- wherein said recombinant antibody has increased Fc-mediated cellular cytotoxicity or increased Fc receptor binding affinity compared to the corresponding antibody produced by the same host cell that has not been glycoengineered, and wherein said antibody produced by said glycoengineered host cell has an increased proportion of GlcNAc residues in the Fc region relative to the proportion of fucose residues compared to the corresponding antibody produced by the same host cell that has not been glycoengineered, and wherein said antibody has increased Fc-mediated cellular cytotoxicity or increased Fc receptor binding affinity as a result of said glycoengineering.
3 more elements
A method for producing a recombinant antibody having increased Fc mediated cellular cytotoxicity or increased Fc receptor binding affinity, comprising:
- (a) providing a mammalian host cell that expresses a recombinant antibody comprising an IgG Fc region containing N-linked oligosaccharides;
- (b) glycoengineering said host cell, wherein said glycoengineering comprises genetically manipulating said host cell so that said host cell has an altered level of activity of at least one glycoprotein-modifying glycosyltranferase;
- (c) culturing said glycoengineered host cell under conditions which permit the production of said recombinant antibody;
- and
- (d) isolating said recombinant antibody;
- wherein said recombinant antibody has increased Fc-mediated cellular cytotoxicity or increased Fc receptor binding affinity compared to the corresponding antibody produced by the same host cell that has not been glycoengineered, and wherein said recombinant antibody produced by said glycoengineered host cell exhibits at least an 80% increase in maximal ADCC activity compared to the same antibody produced by the same host cell under identical culture and purification conditions, but which has not been glycoengineered.
3 more elements
A method for producing a recombinant antibody having increased Fc mediated cellular cytotoxicity or increased Fc receptor binding affinity, comprising:
- (a) providing a mammalian host cell that expresses a recombinant antibody comprising an IgG Fc region containing N-linked oligosaccharides;
- (b) glycoengineering said host cell, wherein said glycoengineering comprises genetically manipulating said host cell so that said host cell has an altered level of activity of at least one glycoprotein-modifying glycosyltranferase;
- (c) culturing said glycoengineered host cell under conditions which permit the production of said recombinant antibody;
- and
- (d) isolating said recombinant antibody;
- wherein said recombinant antibody has increased Fc-mediated cellular cytotoxicity or increased Fc receptor binding affinity compared to the corresponding antibody produced by the same host cell that has not been glycoengineered, and wherein said recombinant antibody has an increased proportion of nonfucosylated oligosaccharides in the Fc region as a result of said glycoengineering compared to the corresponding antibody produced by the same host cell that has not been glycoengineered.
3 more elements
A method for producing a recombinant antibody having increased Fc mediated cellular cytotoxicity or increased Fc receptor binding affinity, comprising:
- (a) providing a mammalian host cell that expresses a recombinant antibody comprising an IgG Fc region containing N-linked oligosaccharides;
- (b) glycoengineering said host cell so that said host cell has an altered level of activity of at least one glycoprotein-modifying glycosyltranferase;
- (c) culturing said glycoengineered host cell under conditions which permit the production of said recombinant antibody;
- and
- (d) isolating said recombinant antibody;
- wherein said recombinant antibody has increased Fc-mediated cellular cytotoxicity or increased Fc receptor binding affinity compared to the corresponding antibody produced by the same host cell that has not been glycoengineered, and wherein the predominant N-linked oligosaccharide in the Fc region of the antibody produced by said glycoengineered host cell is nonfucosylated.
3 more elements
A method for producing a recombinant antibody having increased Fc mediated cellular cytotoxicity or increased Fc receptor binding affinity, comprising:
- (a) providing a mammalian host cell that expresses a recombinant antibody comprising an IgG Fc region containing N-linked oligosaccharides;
- (b) glycoengineering said host cell so that said host cell has an altered level of activity of at least one glycoprotein-modifying glycosyltranferase;
- (c) culturing said glycoengineered host cell under conditions which permit the production of said recombinant antibody;
- and
- (d) isolating said recombinant antibody;
- wherein said recombinant antibody has increased Fc-mediated cellular cytotoxicity or increased Fc receptor binding affinity compared to the corresponding antibody produced by the same host cell that has not been glycoengineered, and wherein the majority of the N-linked oligosaccharides in the Fc region of said antibody produced by said glycoengineered host cell are bisected.
3 more elements
A method for producing a recombinant antibody having increased Fc mediated cellular cytotoxicity or increased Fc receptor binding affinity, comprising:
- (a) providing a mammalian host cell that expresses a recombinant antibody comprising an IgG Fc region containing N-linked oligosaccharides;
- (b) glycoengineering said host cell so that said host cell has an altered level of activity of at least one glycoprotein-modifying glycosyltranferase;
- (c) culturing said glycoengineered host cell under conditions which permit the production of said recombinant antibody;
- and
- (d) isolating said recombinant antibody;
- wherein said recombinant antibody has increased Fc-mediated cellular cytotoxicity or increased Fc receptor binding affinity compared to the corresponding antibody produced by the same host cell that has not been glycoengineered, and wherein the majority of the N-linked oligosaccharides in the Fc region of said antibody produced by said glycoengineered host cell are nonfucosylated.
3 more elements
A method for producing a recombinant antibody having increased Fc mediated cellular cytotoxicity or increased Fc receptor binding affinity, comprising:
- (a) providing a mammalian host cell that expresses a recombinant antibody comprising an IgG Fc region containing N-linked oligosaccharides;
- (b) glycoengineering said host cell so that said host cell has an altered level of activity of at least one glycoprotein-modifying glycosyltranferase;
- (c) culturing said glycoengineered host cell under conditions which permit the production of said recombinant antibody;
- and
- (d) isolating said recombinant antibody;
- wherein said recombinant antibody has increased Fc-mediated cellular cytotoxicity or increased Fc receptor binding affinity compared to the corresponding antibody produced by the same host cell that has not been glycoengineered, and wherein at least 45% of the oligosaccharides in the Fc region of said antibody produced by said glycoengineered host cell are complex structures.
3 more elements
Dependencies are inferred from the claim text.