Enhanced α2-3–linked sialylation determines the extended half-life of CHO-rVWF
Abstract
Abstract The half-life of recombinant human von Willebrand factor (rVWF) expressed in CHO cells (CHO-rVWF; Vonicog alfa; and Vonvendi/Veyvondi) is significantly longer than that of plasma-derived VWF (pdVWF). This finding is intriguing because CHO cells do not generate α2-6 sialylation, which constitutes the majority of human pdVWF sialylation. We hypothesized that glycan differences might regulate the longer half-life of CHO-rVWF. In lectin plate-binding assays and liquid chromatography–mass spectrometry analysis, we confirmed that CHO-rVWF lacked α2-6–linked sialylation. Conversely, however, α2-3–linked sialylation was significantly increased on CHO-rVWF, which also had reduced exposed β-galactose (β-Gal) compared to pdVWF. Consistent with human data, CHO-rVWF clearance was significantly (P < .001) reduced in VWF–/– mice compared to pdVWF. However, clearance of asialo-pdVWF and asialo–CHO-rVWF were identical. In keeping with the in vivo half-life prolongation, CHO-rVWF binding to murine macrophages (P = .012) and HepG2 cells (P = .001) was significantly decreased compared to pdVWF. Furthermore, CHO-rVWF binding to purified macrophage-galactose-type lectin (MGL) receptor and asialoglycoprotein receptor (ASGPR) was also significantly reduced. In contrast to pdVWF, in vivo studies in MGL1–/– mice and Asgr1–/– mice demonstrated that neither MGL nor ASGPR plays significant roles in regulating CHO-rVWF clearance. Together, our findings demonstrate that enhanced α2-3–linked sialylation on CHO-rVWF is responsible for its extended half-life.
Article Details
Authors (19)
Ciara Byrne
1Irish Centre for Vascular Biology, School of Pharmacy and Biomolecular Sciences, Royal College of Surgeons in Ireland, Dublin, Ireland
Soracha Ward
1Irish Centre for Vascular Biology, School of Pharmacy and Biomolecular Sciences, Royal College of Surgeons in Ireland, Dublin, Ireland
Jamie O’Sullivan
1Irish Centre for Vascular Biology, School of Pharmacy and Biomolecular Sciences, Royal College of Surgeons in Ireland, Dublin, Ireland
Alain Chion
1Irish Centre for Vascular Biology, School of Pharmacy and Biomolecular Sciences, Royal College of Surgeons in Ireland, Dublin, Ireland
Patricia Lopes
Bogdan Baci
1Irish Centre for Vascular Biology, School of Pharmacy and Biomolecular Sciences, Royal College of Surgeons in Ireland, Dublin, Ireland
Caoimhe Dowd
1Irish Centre for Vascular Biology, School of Pharmacy and Biomolecular Sciences, Royal College of Surgeons in Ireland, Dublin, Ireland
Darragh Jordan
1Irish Centre for Vascular Biology, School of Pharmacy and Biomolecular Sciences, Royal College of Surgeons in Ireland, Dublin, Ireland
Ross Baker
1Perth Blood Institute, Perth, Australia
Roger J. S. Preston
Marco Monopoli
4Department of Chemistry, Royal College of Surgeons in Ireland, Dublin, Ireland
Peter L. Turecek
6Baxalta Innovations GmbH, a Member of the Takeda Group of Companies, Vienna, Austria
Maximilianos Kotsias
6Ludger Ltd, Abingdon, United Kingdom
Jack Cheeseman
6Ludger Ltd, Abingdon, United Kingdom
Alan B. Moran
Richard A. Gardner
6Ludger Ltd, Abingdon, United Kingdom
Daniel I. R. Spencer
6Ludger Ltd, Abingdon, United Kingdom
Ferdows Atiq
2Irish Centre for Vascular Biology, School of Pharmacy and Biomolecular Sciences, Royal College of Surgeons in Ireland, Dublin, Ireland
James S. O’Donnell