Production Of Glycoproteins Using Manganese

Abstract

Culture media comprising manganese and methods of culturing cells to improve sialylation and glycosylation of glycoproteins are provided.

1 - 22 . (canceled) 
     
     
         23 . A method for producing an erythropoiesis-stimulating molecule by growing a manganese-responsive host cell transfected with a nucleic acid encoding the amino acid sequence of SEQ ID NO: 2 in a culture medium containing an amount of manganese effective to increase the O-site occupancy of said erythropoiesis-stimulating molecule, wherein the concentration of manganese in said culture medium ranges from about 0.01 to about 10 μM. 
     
     
         24 . The method of  claim 23  wherein the culture medium is essentially serum-free. 
     
     
         25 . The method of  claim 23  wherein the host cell is a mammalian cell. 
     
     
         26 . The method of  claim 23  wherein the host cell is a CHO cell. 
     
     
         27 . The method of  claim 23  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. 
     
     
         28 . The method of  claim 23  wherein the host cells are grown in roller bottles. 
     
     
         29 . The method of  claims 23  wherein the manganese is added after a rapid cell growth phase.