Zeerleder, Sacha; Engel, Ruchira; Zhang, Tao; Roem, Dorina; van Mierlo, Gerard; Wagenaar-Bos, Ineke; van Ham, Sija Marieke; Wuhrer, Manfred; Wouters, Diana; Jongerius, Ilse (2021). Sugar Matters: Improving In Vivo Clearance Rate of Highly Glycosylated Recombinant Plasma Proteins for Therapeutic Use. Pharmaceuticals, 14(1) MDPI 10.3390/ph14010054
|
Text
pharmaceuticals-14-00054-v2.pdf - Published Version Available under License Creative Commons: Attribution (CC-BY). Download (2MB) | Preview |
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.
Item Type: |
Journal Article (Original Article) |
|---|---|
Division/Institute: |
04 Faculty of Medicine > Department of Haematology, Oncology, Infectious Diseases, Laboratory Medicine and Hospital Pharmacy (DOLS) > Clinic of Haematology and Central Haematological Laboratory 04 Faculty of Medicine > Pre-clinic Human Medicine > BioMedical Research (DBMR) > Unit Childrens Hospital > Forschungsgruppe Hämatologie (Erwachsene) |
UniBE Contributor: |
Zeerleder, Sacha Sergio |
Subjects: |
600 Technology > 610 Medicine & health |
ISSN: |
1424-8247 |
Publisher: |
MDPI |
Language: |
English |
Submitter: |
Pierrette Durand Lüthi |
Date Deposited: |
28 Jun 2021 15:51 |
Last Modified: |
05 Dec 2022 15:51 |
Publisher DOI: |
10.3390/ph14010054 |
PubMed ID: |
33440845 |
Uncontrolled Keywords: |
C1-inhibitor glycosylation recombinant protein |
BORIS DOI: |
10.48350/156857 |
URI: |
https://boris.unibe.ch/id/eprint/156857 |
Actions (login required)
 |
Edit item |