Enhanced Sialylation of Albumin-erythropoietin through Increased Synthesis of UDP-N-acetylglucosamine in CHO Cell Cultures
2021
Han, H.J. | Lee, J.H. | Cha, H.M. | Lim, J.H. | Kim, D.I.
Sialylation is a major factor to determine half-life of glycoproteins and sialic acid content of glycoprotein was effectively increased by supplementation of nucleotide sugar precursors during cell cultures. So it is necessary to investigate the effect on glycan may be different when two or more precursors are fed in combination. Albumin-erythropoietin (Alb-EPO) can form a tetra-antennary structure and up to 14 sialic acids can be attached depending on the antennary structure of glycan. It is possible to increase the sialic acid contents of Alb-EPO by promoting the formation of the antennary structure of glycans. In this study, uridine and N-acetylglucosamine (GlcNAc) were added to Chinese hamster ovary (CHO) cell cultures to enhance the synthesis of uridine diphosphate N-acetylglucosamine (UDP-GlcNAc). Also, manganese ion, which is a cofactor of glycosyltransferases or UDP-GlcNAc biosynthesis, was added in the form of MnCl₂. By optimizing the feeding concentrations of uridine and GlcNAc with response surface methodology based on central composite design to reduce the negative impact on cell growth and productivity. Cell growth was observed similar and then productivity and sialic acid content of Alb-EPO were enhanced 1.21-fold and 1.69-fold higher than that of the control, respectively. These results suggest that the combined feeding of uridine, GlcNAc, and MnCl₂ is an effective strategy to enhance the sialic acid content of glycoproteins in CHO cell cultures.
Afficher plus [+] Moins [-]Informations bibliographiques
Cette notice bibliographique a été fournie par Korea Agricultural Science Digital Library
Découvrez la collection de ce fournisseur de données dans AGRIS