The relationship between the positioning of ligands on the surface of nanoparticles and the structural features of nanoconjugates has been underestimated for a long time, albeit of primary importance to promote specific biological recognition at the nanoscale. In particular, it has been formerly observed that a proper molecular orientation can play a crucial role, first optimizing ligand immobilization onto the nanoparticles and, second, improving the targeting efficiency of the nanoconjugates. In this work, we present a novel strategy to afford peptide-oriented ligation using genetically modified cutinase fusion proteins, which combines the presence of a site-directed "capture" module based on an enzymatic unit and a "targeting" moiety consisting of the ligand terminal end of a genetically encoded polypeptide chain. As an example, the oriented presentation of U11 peptide, a sequence specific for the recognition of urokinase plasminogen activator receptor (uPAR), was achieved by enzyme-mediated conjugation with an irreversible inhibitor of cutinase, an alkylphosphonate p-nitrophenol ester linker, covalently bound to the surface of iron oxide nanoparticles. The targeting efficiency of the resulting protein-nanoparticle conjugates was assessed using uPAR-positive breast cancer cells exploiting confocal laser scanning microscopy and quantitative fluorescence analysis of confocal images. Ultrastructural analysis of transmission electron micrographs provided evidence of a receptor-mediated pathway of endocytosis. Our results showed that, despite the small average number of targeting peptides presented on the nanoparticles, our ligand-oriented nanoconjugates proved to be very effective in selectively binding to uPAR and in promoting the uptake in uPAR-positive cancer cells.
Galbiati, E., Cassani, M., Verderio, P., Martegani, E., Colombo, M., Tortora, P., et al. (2015). Peptide-nanoparticle ligation mediated by cutinase fusion for the development of cancer cell-targeted nanoconjugates. BIOCONJUGATE CHEMISTRY, 26(4), 680-689 [10.1021/acs.bioconjchem.5b00005].
|Citazione:||Galbiati, E., Cassani, M., Verderio, P., Martegani, E., Colombo, M., Tortora, P., et al. (2015). Peptide-nanoparticle ligation mediated by cutinase fusion for the development of cancer cell-targeted nanoconjugates. BIOCONJUGATE CHEMISTRY, 26(4), 680-689 [10.1021/acs.bioconjchem.5b00005].|
|Tipo:||Articolo in rivista - Articolo scientifico|
|Carattere della pubblicazione:||Scientifica|
|Presenza di un coautore afferente ad Istituzioni straniere:||No|
|Titolo:||Peptide-nanoparticle ligation mediated by cutinase fusion for the development of cancer cell-targeted nanoconjugates|
|Autori:||Galbiati, E; Cassani, M; Verderio, P; Martegani, E; Colombo, M; Tortora, P; Mazzucchelli, S; Prosperi, D|
GALBIATI, ELISABETTA (Primo)
MAZZUCCHELLI, SERENA (Corresponding)
PROSPERI, DAVIDE (Ultimo)
|Data di pubblicazione:||2015|
|Digital Object Identifier (DOI):||http://dx.doi.org/10.1021/acs.bioconjchem.5b00005|
|Appare nelle tipologie:||01 - Articolo su rivista|