Ubi-tagged vaccine conjugates T cell activation assays

This data collection contains the raw data collected at the RadboudUMC of the associated open access publication "Site-directed multivalent conjugation of antibodies to ubiquitinated payloads". The DOI is provided in the associated publication field. This dataset contains the raw data for the main figures 4-6. For the raw data of the other figures, please refer to m.verdoes1@lumc.nl to obtain the data generated at the LUMC. A short overview of the publication: this work introduces the novel chemoenzymatic conjugation technique ubi-tagging, which utilizes the ubiquitination machinery in vitro to site-specifically conjugate ubiquitin-tagged molecules. We demonstrated the successful conjugation of antibodies, antibody fragments, nanobodies, peptides, and small molecules using ubi-tagging. Furthermore, we applied this technology to synthesize a DC-targeted antibody fragment-antigen conjugate and characterized its efficacy. A significant increase in ex vivo and in vivo T cell activation induced by the ubi-tagged vaccine is observed in comparison to vaccines lacking the ubiquitin. We partly explain this effect by demonstrating that ubi-tagged vaccines achieve higher uptake in CD11c+ splenocytes in vivo compared to analogues prepared by sortase-mediated conjugation. In addition, we hypothesize the contribution of alternative intracellular routing and processing by the ubiquitin containing vaccines to explain the increased efficacy ex vivo. Lastly, we utilized ubi-tagging to prepare a library of notoriously hydrophobic MHC I epitopes conjugated to a DC-targeting nanobody. We demonstrated ex vivo efficacy for one of the clinically relevant epitopes using human primary cells as proof-of-concept for clinical translation.