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. Author manuscript; available in PMC: 2026 Oct 3.
Published in final edited form as: Nat Protoc. 2025 Nov 19;21(5):1841–1868. doi: 10.1038/s41596-025-01272-3

Fig. 1 |. Overview of the proposed protocol for directional conjugation of monoclonal antibodies to nanoparticles.

Fig. 1 |

The protocol includes three main stages: (1) mild oxidation of the carbohydrate anybody moiety (Steps 1–5), (2) conjugation of the heterobifunctional aminooxy-PEG-DBCO linker to the antibody’s Fc region (Steps 6–10) and (3) antibody conjugation to azide-functionalized nanoparticles (Steps 11–17). Created in BioRender. DBCO, dibenzocyclooctyne; EGFR, epidermal growth factor receptor; FRα, folate receptor-α; RT, room temperature; TEM, transmission electron microscopy.