Nanobodies Outperform Antibodies – Rapid Functionalization with Equal In Vivo Targeting Properties

Abstract Highly specific targeting of dendritic cells in vivo is crucial for the development of effective tumor nanovaccines. This group recently presented an antibody‐functionalized nanocarrier system able to maintain its targeting properties when transferred from in vitro to in vivo studies. However, producing this system requires long synthesis times and involves high expenses due to the involved site‐specific enzymatic multi‐step modification procedure of the antibody. Consequently, improving the previously proposed system is necessary in order to accelerate the development. Here, a novel system utilizing nanobodies for the targeting of dendritic cells is presented. A C‐terminal cysteine tag facilitates an easy attachment of the nanobody to the nanocarrier via a thiol‐maleimide conjugation technique. This reduces the functionalization time from several days to mere hours. Using in vitro and in vivo assays, it is shown that the optimized system possesses equal targeting properties as the antibody‐based system. As a result, nanobodies and the coupling chemistry are found to be a superior strategy for the in vivo targeting of dendritic cells when compared to antibodies, due to their rapid attachment to nanocarriers and equal targeting specificity. This would replace antibodies as the current “gold standard” of targeting moieties.