A novel sugar analog enhances sialic acid production and biotherapeutic sialylation in CHO cells. Issue 8 (8th May 2017)

• Record Type: Journal Article
• Authors: Yin, Bojiao; Wang, Qiong; Chung, Cheng‐Yu; Bhattacharya, Rahul; Ren, Xiaozhi; Tang, Juechun; Yarema, Kevin J.; Betenbaugh, Michael J.
• Is Part Of: Biotechnology and bioengineering. Volume 114:Issue 8(2017)
• Page Start: 1899
• Page End: 1902
• Publication Date: 2017-05-08
• Subjects: metabolic oligosaccharide engineering -- sialylation -- glycosylation -- N‐acetylmannosamine (ManNAc) analog -- Chinese hamster ovary (CHO) media; Biotechnology -- Periodicals; Bioengineering -- Periodicals; 660.6
• DOI: 10.1002/bit.26291
• Languages: English
• ISSNs: 0006-3592

Abstract

ABSTRACT: A desirable feature of many therapeutic glycoprotein production processes is to maximize the final sialic acid content. In this study, the effect of applying a novel chemical analog of the sialic acid precursor N‐acetylmannosamine (ManNAc) on the sialic acid content of cellular proteins and a model recombinant glycoprotein, erythropoietin (EPO), was investigated in CHO‐K1 cells. By introducing the 1, 3, 4‐ O ‐Bu3 ManNAc analog at 200–300 µM into cell culture media, the intracellular sialic acid content of EPO‐expressing cells increased ∼8‐fold over untreated controls while the level of cellular sialylated glycoconjugates increased significantly as well. For example, addition of 200–300 µM 1, 3, 4‐ O ‐Bu3 ManNAc resulted in >40% increase in final sialic acid content of recombinant EPO, while natural ManNAc at ∼100 times higher concentration of 20 mM produced a less profound change in EPO sialylation. Collectively, these results indicate that butyrate‐derivatization of ManNAc improves the capacity of cells to incorporate exogenous ManNAc into the sialic acid biosynthetic pathway and thereby increase sialylation of recombinant EPO and other glycoproteins. This study establishes 1, 3, 4‐ O ‐Bu3 ManNAc as a novel chemical supplement to improve glycoprotein quality and sialylation levels at concentrations orders of magnitude lower than alternative approaches. Biotechnol. Bioeng. 2017;114: 1899–1902. © 2017 Wiley Periodicals, Inc. Abstract : A novel sugar analog (1, 3, 4‐ O ‐Bu3 ManNAc) of the sialic acid precursor N‐acetylmannosamine was developed as a media supplement to enhance the sialic acid content in CHO cells. The authors evaluated the capacity of this new analog to affect the sialylation of cellular proteins and a model secreted recombinant glycoprotein erythropoietin (EPO). This compound serves as a more effective chemical media supplement to improve sialylation of recombinant proteins at concentrations orders of magnitude lower than alternative approaches.