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.
Claims37
A method for modifying the glycosylation of a polypeptide produced by a host cell, comprising introducing into said host cell a nucleic acid comprising a sequence encoding a fusion polypeptide, wherein said fusion polypeptide has β(1,4)-galactosyltransferase activity and comprises a Golgi localization domain selected from the group consisting of:
- the localization domain of mannosidase II, the localization domain of β(1,2)-N-acetylglucosaminyltransferase I, the localization domain of mannosidase I, the localization domain of β(1,2)-N-acetylglucosaminyltransferase II, and the localization domain of α1-6 core fucosyltransferase, 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 host cell, comprising:
- a. culturing a host cell engineered to express at least one nucleic acid encoding a fusion polypeptide having β(1,4)-galactosyltransferase 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 Fc-receptor binding or effector function of said polypeptide produced by said host cell, and wherein said fusion polypeptide having β(1,4)-galactosyltransferase activity comprises a Golgi localization domain selected from the group consisting of:
- the localization domain of mannosidase II, the localization domain of β(1,2)-N-acetylglucosaminyltransferase I, the localization domain of mannosidase I, the localization domain of β(1,2)-N-acetylglucosaminyltransferase II, and the localization domain of α1-6 core fucosyltransferase;
- and
- b. isolating said polypeptide.
A method for producing a polypeptide in a host cell, comprising:
- a. culturing a host cell engineered to express at least one nucleic acid encoding a fusion polypeptide having GalT activity and at least one a 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 GalT activity comprises a Golgi localization domain selected from the group consisting of:
- the localization domain of mannosidase II, the localization domain of β(1,2)-N-acetylglucosaminyltransferase I, the localization domain of mannosidase I, the localization domain of β(1,2)-N-acetylglucosaminyltransferase II, and the localization domain of α1-6 core fucosyltransferase;
- and
- b. isolating said polypeptide.
Dependencies are inferred from the claim text.