Academic Bibliography

 Search 200 years of publications by Ghent University researchers.
Advanced search
1 file | 21.71 MB
Add to list
  1. Search results
  2. GlycoVHH : optimal sites for introduc...

GlycoVHH : optimal sites for introducing N-glycans on the camelid VHH antibody scaffold and use for macrophage delivery

Loes van Schie (UGent) , Wander Van Breedam (UGent) , Charlotte Roels (UGent) , Bert Schepens (UGent) , Martin Frank, Ahmadreza Mehdipour (UGent) , Bram Laukens, Wim Nerinckx (UGent) , Francis Santens, Simon Devos (UGent) , et al.
(2023) MABS. 15(1).
Author
Organization
Project
Abstract
As small and stable high-affinity antigen binders, VHHs boast attractive characteristics both for therapeutic use in various disease indications, and as versatile reagents in research and diagnostics. To further increase the versatility of VHHs, we explored the VHH scaffold in a structure-guided approach to select regions where the introduction of an N-glycosylation N-X-T sequon and its associated glycan should not interfere with protein folding or epitope recognition. We expressed variants of such glycoengineered VHHs in the Pichia pastoris GlycoSwitchM5 strain, allowing us to pinpoint preferred sites at which Man(5)GlcNAc(2)-glycans can be introduced at high site occupancy without affecting antigen binding. A VHH carrying predominantly a Man(5)GlcNAc(2) N-glycan at one of these preferred sites showed highly efficient, glycan-dependent uptake by Mf4/4 macrophages in vitro and by alveolar lung macrophages in vivo, illustrating one potential application of glyco-engineered VHHs: a glycan-based targeting approach for lung macrophage endolysosomal system delivery. The set of optimal artificial VHH N-glycosylation sites identified in this study can serve as a blueprint for targeted glyco-engineering of other VHHs, enabling site-specific functionalization through the rapidly expanding toolbox of synthetic glycobiology.
Keywords
Immunology, Immunology and Allergy, VHHs, Single-domain antibody scaffold, Pichia pastoris GlycoSwitchM5, N-glycosylation, Macrophage targeting

Downloads

  • Download
    van Schie et al GlycoVHH optimal sites for introducing N-glycans on the camelid VHH antibody scaffold and use for macrophage delivery mAbs 2023.pdf
    • full text (Published version)
    • |
    • open access
    • |
    • PDF
    • |
    • 21.71 MB

Citation

Please use this url to cite or link to this publication:

MLA
van Schie, Loes, et al. “GlycoVHH : Optimal Sites for Introducing N-Glycans on the Camelid VHH Antibody Scaffold and Use for Macrophage Delivery.” MABS, vol. 15, no. 1, Informa UK Limited, 2023, doi:10.1080/19420862.2023.2210709.
APA
van Schie, L., Van Breedam, W., Roels, C., Schepens, B., Frank, M., Mehdipour, A., … Callewaert, N. (2023). GlycoVHH : optimal sites for introducing N-glycans on the camelid VHH antibody scaffold and use for macrophage delivery. MABS, 15(1). https://doi.org/10.1080/19420862.2023.2210709
Chicago author-date
Schie, Loes van, Wander Van Breedam, Charlotte Roels, Bert Schepens, Martin Frank, Ahmadreza Mehdipour, Bram Laukens, et al. 2023. “GlycoVHH : Optimal Sites for Introducing N-Glycans on the Camelid VHH Antibody Scaffold and Use for Macrophage Delivery.” MABS 15 (1). https://doi.org/10.1080/19420862.2023.2210709.
Chicago author-date (all authors)
van Schie, Loes, Wander Van Breedam, Charlotte Roels, Bert Schepens, Martin Frank, Ahmadreza Mehdipour, Bram Laukens, Wim Nerinckx, Francis Santens, Simon Devos, Iebe Rossey, Karel Thooft, Sandrine Vanmarcke, Annelies Van Hecke, Xavier Saelens, and Nico Callewaert. 2023. “GlycoVHH : Optimal Sites for Introducing N-Glycans on the Camelid VHH Antibody Scaffold and Use for Macrophage Delivery.” MABS 15 (1). doi:10.1080/19420862.2023.2210709.
Vancouver
1.
van Schie L, Van Breedam W, Roels C, Schepens B, Frank M, Mehdipour A, et al. GlycoVHH : optimal sites for introducing N-glycans on the camelid VHH antibody scaffold and use for macrophage delivery. MABS. 2023;15(1).
IEEE
[1]
L. van Schie et al., “GlycoVHH : optimal sites for introducing N-glycans on the camelid VHH antibody scaffold and use for macrophage delivery,” MABS, vol. 15, no. 1, 2023.
@article{01H3CK1JFK4G3SKSXB0XK8ES68,
  abstract     = {{As small and stable high-affinity antigen binders, VHHs boast attractive characteristics both for therapeutic use in various disease indications, and as versatile reagents in research and diagnostics. To further increase the versatility of VHHs, we explored the VHH scaffold in a structure-guided approach to select regions where the introduction of an N-glycosylation N-X-T sequon and its associated glycan should not interfere with protein folding or epitope recognition. We expressed variants of such glycoengineered VHHs in the Pichia pastoris GlycoSwitchM5 strain, allowing us to pinpoint preferred sites at which Man(5)GlcNAc(2)-glycans can be introduced at high site occupancy without affecting antigen binding. A VHH carrying predominantly a Man(5)GlcNAc(2) N-glycan at one of these preferred sites showed highly efficient, glycan-dependent uptake by Mf4/4 macrophages in vitro and by alveolar lung macrophages in vivo, illustrating one potential application of glyco-engineered VHHs: a glycan-based targeting approach for lung macrophage endolysosomal system delivery. The set of optimal artificial VHH N-glycosylation sites identified in this study can serve as a blueprint for targeted glyco-engineering of other VHHs, enabling site-specific functionalization through the rapidly expanding toolbox of synthetic glycobiology.}},
  articleno    = {{2210709}},
  author       = {{van Schie, Loes and Van Breedam, Wander and Roels, Charlotte and Schepens, Bert and Frank, Martin and Mehdipour, Ahmadreza and Laukens, Bram and Nerinckx, Wim and Santens, Francis and Devos, Simon and Rossey, Iebe and Thooft, Karel and Vanmarcke, Sandrine and Van Hecke, Annelies and Saelens, Xavier and Callewaert, Nico}},
  issn         = {{1942-0862}},
  journal      = {{MABS}},
  keywords     = {{Immunology,Immunology and Allergy,VHHs,Single-domain antibody scaffold,Pichia pastoris GlycoSwitchM5,N-glycosylation,Macrophage targeting}},
  language     = {{eng}},
  number       = {{1}},
  pages        = {{16}},
  publisher    = {{Informa UK Limited}},
  title        = {{GlycoVHH : optimal sites for introducing N-glycans on the camelid VHH antibody scaffold and use for macrophage delivery}},
  url          = {{http://doi.org/10.1080/19420862.2023.2210709}},
  volume       = {{15}},
  year         = {{2023}},
}
Altmetric
View in Altmetric
Web of Science
Times cited: