Sugar matters: improving in vivo clearance rate of highly glycosylated recombinant plasma proteins for therapeutic use

Zeerleder, S.; Engel, R.; Zhang, T.; Roem, D.; Mierlo, G. van; Wagenaar-Bos, I.; ... ; Jongerius, I. (2021)

Article / Letter to editor

Correct glycosylation of proteins is essential for production of therapeutic proteins as glycosylation is important for protein solubility, stability, half-life and immunogenicity. The heavily glycosylated plasma protein C1-inhibitor (C1-INH) is used in treatment of hereditary angioedema attacks. In this study, we used C1-INH as a model protein to propose an approach to develop recombinant glycoproteins with the desired glycosylation. We produced fully functional recombinant C1-INH in Chinese hamster ovary (CHO) cells. In vivo we observed a biphasic clearance, indicating different glycosylation forms. N-glycan analysis with mass spectrometry indeed demonstrated heterogeneous glycosylation for recombinant C1-INH containing terminal galactose and terminal sialic acid. Using a Ricinus Communis Agglutinin I (RCA(120)) column, we could reduce the relative abundance of terminal galactose and increase the relative abundance of terminal sialic acid. This resulted in a fully active protein with a similar in vivo clearance rate to plasmaderived C1-INH. In summary, we describe the development of a recombinant human glycoprotein using simple screening tools to obtain a product that is similar in function and in vivo clearance rate to its plasma-derived counterpart. The approach used here is of potential use in the development of other therapeutic recombinant human glycoproteins.

• C1-inhibitor
• glycosylation
• recombinant protein

• All authors: Zeerleder, S.; Engel, R.; Zhang, T.; Roem, D.; Mierlo, G. van; Wagenaar-Bos, I.; Ham, S.M. van; Wuhrer, M.; Wouters, D.; Jongerius, I.
• Date: 2021-01-01
• Journal: Pharmaceuticals
• Volume: 14
• Issue: 1
• DOI: doi:10.3390/ph14010054
• Link: https://www.mdpi.com/1424-8247/14/1/54/pdf