Process for controlling intracellular glycosylation of proteins
AssigneeThe Regents of the University of Calif.
InventorsJames Paulson, Eryn Ujita-Lee, Jasminder Weinstein
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Abstract
A process for controlling the glycosylation of protein in a cell wherein the cell is genetically engineered to produce one or more enzymes which provide internal control of the cell's glycosylation mechanism. A Chinese hamster ovary (CHO) cell line is genetically engineered to produce a sialyltransferase. This supplemental sialyltransferase modifies the CHO glycosylation machinery to produce glycoproteins having carbohydrate structures which more closely resemble naturally occurring human glycoproteins.
Claims16
A process for altering the glycosylation of a protein produced by a eukaryotic cell, said process comprising the steps of:
- introducing into said eukaryotic cell at least one gene which is capable of expressing at least one enzyme which is selected from the group consisting of glycosyltransferase, fucosyltransferases, galactosyltransferases, .beta.-acetylgalactosaminyltransferases, N-acetylglycosaminyltransferases and sulfotransferases;
- and
- expressing a sufficient amount of at least one of said enzymes in said eukaryotic cell to thereby alter the glycosylation of said protein in said eukaryotic cell.
A process for altering the glycosylation of a protein produced by a cell wherein attachment of the carbohydrate moiety to said protein during glycosylation is dependent upon at least one naturally occurring enzyme present in said cell, said process comprising the steps of:
- introducing into said cell at least one gene which is capable of expressing at least one supplemental enzyme selected from the group consisting of sialyltransferase, fucosyltransferase, galactosyltransferase, N-acetylgalactosaminyltransferase, N-acetylglucosaminyltransferase and sulfotransferase;
- and
- expressing a sufficient amount of at least one of said supplemental enzymes in said cell to produce a cell having both naturally occurring and supplemental enzymes wherein the presence of said supplemental enzyme alters the glycosylation of said protein.
A vector for use in expressing a sialyltransferase in a host cell, said vector comprising cDNA having the nucleotide sequence set forth in FIGS. 3a and 3b of this specification.
Dependencies are inferred from the claim text.