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

Abstract

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 (RCA120) 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.