abstract |
The present invention relates to a method for producing a recombinant glycoprotein having a high sialic acid content, and more particularly, arginine at amino acid 263 with leucine or arginine at amino acid 263 with leucine and arginine at amino acid 266. The UDP-GlcNAc 2-epimerase / ManNAc kinase (GNE / MNK) enzyme, which was substituted with glutamine or tryptophan to induce point mutations, was used to determine epimerase (regardless of the concentration of cytidine monophosphate (CMP) -sialic acid). epimerase activity is sustained and cells overexpressed increase the content of CMP-sialic acid in the cells, and in particular, GNE / MNK, human alpha-2,3 sialic acid sugar transferase, Intracellular CMP-sialic acid and glycoproteins in glycoprotein (erythropoietin or thrombopoietin) producing host cells simultaneously overexpressing the CMP-sialic acid transport protein gene Since the sialic acid content of the vagina is increased, overexpression in recombinant glycoprotein-producing host cells with sialic acid binding of the three genes can be useful for the preparation of glycoproteins with increased sialic acid content. |