Glycosylation engineering of antibodies for improving antibody-dependent cellular cytotoxicity
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.
1 - 85 . (Cancelled).
86 . A glycoengineered, recombinant antibody comprising an Fc region containing N-linked oligosaccharides, wherein said antibody has been engineered to have an altered pattern of glycosylation in the Fc region and wherein said antibody has increased Fc-mediated cellular cytotoxicity as a result of said altered glycosylation.
87 . A glycoengineered, recombinant antibody comprising an Fc region containing N-linked oligosaccharides, wherein said antibody has been engineered to have an altered pattern of glycosylation in the Fc region and wherein said antibody has increased Fc receptor binding affinity as a result of said altered glycosylation.
88 . A glycoengineered, recombinant antibody according to claim 86 or claim 87 , wherein said antibody is a chimeric antibody.
89 . A glycoengineered, recombinant antibody according to claim 86 or claim 87 , wherein said antibody is a humanized antibody.
90 . A glycoengineered, recombinant antibody according to claim 86 or claim 87 , wherein said antibody is an antibody fragment that contains the Fc region.
91 . A glycoengineered, recombinant antibody according to claim 86 or claim 87 , wherein said antibody is a fusion protein that includes a Fc region of an immunoglobulin.
92 . A glycoengineered, recombinant antibody according to claim 86 or claim 87 , wherein said altered pattern of glycosylation is an increase in the proportion of GlcNAc residues compared to the nonglycoengineered recombinant antibody.
93 . A glycoengineered, recombinant antibody according to claim 86 or claim 87 , wherein said altered pattern of glycosylation is a decrease in the proportion of fucose residues compared to the nonglycoengineered recombinant antibody.
94 . A glycoengineered, recombinant antibody according to claim 86 or claim 87 , wherein said altered pattern of glycosylation is an increase in the proportion of GlcNAc residues relative to the proportion of fucose residues compared to the nonglycoengineered recombinant antibody.
95 . A glycoengineered recombinant antibody according to claim 92 , wherein said oligosaccharides are bisected.
96 . A glycoengineered, recombinant antibody according to claim 92 , wherein said oligosaccharides are bisected, complex.
97 . A glycoengineered, recombinant antibody according to claim 92 , wherein said oligosaccharides are bisected, hybrid.
98 . A glycoengineered, recombinant antibody according to claim 86 or claim 87 , wherein said antibody is isolated from an engineered host cell selected from the group consisting of an engineered CHO cell, an engineered BHK cell, an engineered NS0 cell, an engineered SP2/0 cell, an engineered yeast cell, and an engineered plant cell.
99 . A glycoengineered, recombinant antibody according to claim 98 , wherein said antibody is isolated from an engineered CHO cell.
100 . A glycoengineered, recombinant antibody according to claim 86 or claim 87 , wherein said antibody is a therapeutic antibody.
101 . A glycoengineered, recombinant antibody according to claim 100 , wherein said antibody selectively binds to a cancer antigen.
102 . A glycoengineered, recombinant antibody according to claim 100 , wherein said antibody is a monoclonal antibody.
103 . A glycoengineered recombinant antibody according to claim 100 , wherein said antibody is a selected from the group consisting of: an anti-CD20 antibody, an anti-human neuroblastoma antibody, an anti-human renal cell carcinoma antibody, an anti-HER2 antibody, an anti-human colon, lung, and breast carcinoma antibody, an anti-human 17-1A antigen antibody, a humanized anti-human colorectal tumor antibody, an anti-human melanoma antibody, and an anti-human squamous-cell carcinoma antibody.
104 . A glycoengineered, recombinant antibody according to claim 86 or claim 87 , wherein said antibody is IgG.
105 . A glycoengineered, recombinant antibody according to claim 86 or claim 87 , wherein the majority of the N-linked oligosaccharides are bisected.
106 . A glycoengineered, recombinant antibody according to claim 86 or claim 87 , wherein the majority of the N-linked oligosaccharides are nonfucosylated.
107 . A glycoengineered, recombinant antibody according to claim 86 or claim 87 , wherein the majority of the N-linked oligosaccharides are bisected, nonfucosylated.
108 . A glycoengineered, recombinant antibody produced by a process comprising:
(a) providing a glycoengineered host cell comprising at least one nucleic acid encoding a recombinant antibody and at least one nucleic acid encoding a glycosyltransferase;
(b) culturing said host cell under conditions which permit expression of said recombinant antibody having an altered pattern of glycosylation in the Fc region of said antibody; and
(c) isolating said recombinant antibody;
wherein said recombinant antibody has increased Fc-mediated cellular cytotoxicity as a result of said altered glycosylation.