Exploring protein-protein ligation approaches for the cytosolic delivery of antigens using AIP56.

ABSTRACT: The intracellular delivery of biologics, particularly large cargoes like proteins, remains a challenge in biotechnology and biomedicine. The modular structure of well-characterized AB toxins allows different cargoes to be grafted, creating a target-specific biotechnological tool capable of cytosolic delivery. In this study, we employed protein-protein fusion strategies-SpyCatcher003, SnoopCatcher, and SnoopLigase-to generate chimeras between the delivery region of AIP56 (AIP56L258-N497) and β-lactamase and performed functional delivery assays. The chimeras were successfully obtained using these strategies and were all able to deliver β-lactamase into the cytosol of J774.A1 macrophages. Cellular fractionation showed that, although most of the β-lactamase remains associated with the endosoma

SUBMITTER: Pinheiro B 

PROVIDER: S-EPMC12364934 | biostudies-literature | 2025

REPOSITORIES: biostudies-literature