Fusion constructs and use of same to produce antibodies with increased Fc receptor binding affinity and effector function
AssigneeRoche Glycart AG
InventorsPablo Umaña, Peter Bruenker, Claudia Ferrara, Tobias Suter
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Abstract
The present invention relates to the field of glycosylation engineering of proteins. More particularly, the present invention relates to nucleic acid molecules, including fusion constructs, having catalytic activity and the use of same in glycosylation engineering of host cells to generate polypeptides with improved therapeutic properties, including antibodies with increased Fc receptor binding and increased effector function.
Claims33
A method for modifying the glycosylation profile of a polypeptide produced by a mammalian host cell, comprising introducing into said host cell an isolated nucleic acid comprising a sequence encoding a fusion polypeptide, wherein said fusion polypeptide has β(1,4)-N-acetylglucosaminyltransferase III activity and comprises the Golgi localization domain of mannosidase II, and wherein said modified polypeptide has increased Fc-receptor binding or effector function as a result of said modification.
A method for modifying the glycosylation profile of a polypeptide produced by a mammalian host cell, comprising introducing into said host cell an expression vector which comprises an isolated nucleic acid comprising a sequence encoding a fusion polypeptide, wherein said fusion polypeptide has β(1,4)-N-acetylglucosaminyltransferase III activity and comprises the Golgi localization domain of mannosidase II, and wherein said modified polypeptide has increased Fc-receptor binding or effector function as a result of said modification.
A method for producing a polypeptide in a mammalian host cell, comprising:
- a. culturing a mammalian host cell engineered to express at least one nucleic acid encoding a fusion polypeptide having β(1,4)-N-acetylglucosaminyltransferase III activity under conditions which permit the production of a polypeptide selected from the group consisting of a whole antibody molecule, an antibody fragment, and a fusion protein that includes a region equivalent to the Fc region of an immunoglobulin, wherein said fusion polypeptide is expressed in an amount sufficient to modify the oligosaccharides in the Fc region and increase the Fc-receptor binding or effector function of said polypeptide produced by said host cell and wherein said fusion polypeptide having β(1,4)-N-acetylglucosaminyltransferase III activity comprises the Golgi localization domain of mannosidase II;
- and
- b. isolating said polypeptide.
A method for producing a polypeptide in a mammalian host cell, comprising:
- a. culturing a mammalian host cell engineered to express at least one nucleic acid encoding a fusion polypeptide having GnT III activity and at least one nucleic acid encoding a polypeptide having Man II activity under conditions which permit the production of a polypeptide selected from the group consisting of a whole antibody molecule, an antibody fragment, and a fusion protein that includes a region equivalent to the Fc region of an immunoglobulin, wherein said fusion polypeptide is expressed in an amount sufficient to modify the oligosaccharides in the Fc region and increase the Fc-receptor binding or effector function of said polypeptide produced by said host cell and wherein said fusion polypeptide having GnT III activity comprises the Golgi localization domain of mannosidase II;
- and
- b. isolating said polypeptide.
Dependencies are inferred from the claim text.