Production of glycoproteins using manganese
Culture media comprising manganese and methods of culturing cells to improve sialylation and glycosylation of glycoproteins are provided.
1. A culture medium comprising mammalian host cells and a non-toxic amount of manganese effective to increase the sialylation of a glycoprotein composition, wherein said host cells have been transfected with DNA encoding erythropoiesis-stimulating molecules comprising erythropoietin (SEQ ID NO: 3) or darbepoetin (SEQ ID NO: 2), or analogs thereof with 75% homology to SEQ ID NO: 3 or SEQ ID NO: 2, respectively, and still retaining erythropoietic activity, wherein the concentration of manganese is about 40 μM, and wherein less than about 5% of the secreted erythropoiesis-stimulating molecules are lower sialylated.
2. The culture medium of claim 1 , wherein the medium is essentially serum free.
3. The culture medium of claim 1 , wherein the host cell is a CHO cell.
4. The culture medium of claim 1 , wherein the culture medium further comprises one or more supplementary amino acids selected from the group consisting of asparagine, aspartic acid, cysteine, cystine, isoleucine, leucine, tryptophan, or valine.