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

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 Man₅GlcNAc₂-glycans can be introduced at high site occupancy without affecting antigen binding. A VHH carrying predominantly a Man₅GlcNAc

SUBMITTER: van Schie L 

PROVIDER: S-EPMC10208171 | biostudies-literature | 2023 Jan-Dec

REPOSITORIES: biostudies-literature