Zeerleder, Sacha SergioSacha SergioZeerlederEngel, RuchiraRuchiraEngelZhang, TaoTaoZhangRoem, DorinaDorinaRoemvan Mierlo, GerardGerardvan MierloWagenaar-Bos, InekeInekeWagenaar-Bosvan Ham, Sija MariekeSija Mariekevan HamWuhrer, ManfredManfredWuhrerWouters, DianaDianaWoutersJongerius, IlseIlseJongerius2024-09-212024-09-212021-01-11https://boris-portal.unibe.ch/handle/20.500.12422/45628Correct 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.enC1-inhibitor glycosylation recombinant protein600 - Technology::610 - Medicine & healtharticle10.48350/1568573344084510.3390/ph14010054