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Bioconjugate chemistry v.28 no.1, 2017년, pp.222 - 229   SCI SCIE
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Binding and Uptake into Human Hepatocellular Carcinoma Cells of Peptide-Functionalized Gold Nanoparticles

Jha, Satadru (Dipartimento di Scienze Chimiche, Università ); Ramadori, Federico (di Padova, via Marzolo 1, 35131, Padova, ); Quarta, Santina (Dipartimento di Scienze Chimiche, Università ); Biasiolo, Alessandra (di Padova, via Marzolo 1, 35131, Padova, ); Fabris, Enrica (Dipartimento di Medicina, Univeristà ); Baldan, Paola (di Padova, via Giustiniani, 2, 35128 Padova, ); Guarino, Gaetano (Dipartimento di Medicina, Univeristà ); Ruvoletto, Mariagrazia (di Padova, via Giustiniani, 2, 35128 Padova, ); Villano, Gianmarco (Dipartimento di Scienze Chimiche, Università ); Turato, Cristian (di Padova, via Marzolo 1, 35131, Padova, ); Gatta, Angelo (Dipartimento di Scienze Chimiche, Università ); Mancin, Fabrizio (di Padova, via Marzolo 1, 35131, Padova, ); Pontisso, Patrizia (Dipartimento di Scienze Chimiche, Università ); Scrimin, Paolo (di Padova, via Marzolo 1, 35131, Padova, );
  • 초록  

    One of the most daunting challenges of nanomedicine is the finding of appropriate targeting agents to deliver suitable payloads precisely to cells affected by malignancies. Even more complex is the ability to ensure that the nanosystems enter those cells. Here, we use 2 nm (metal core) gold nanoparticles to target human hepatocellular carcinoma (HepG2) cells stably transfected with the SERPINB3 (SB3) protein. The nanoparticles were coated with a 85:15 mixture of thiols featuring, respectively, a phosphoryl choline (to ensure water solubility and biocompatibility) and a 28-mer peptide corresponding to the amino acid sequence 21–47 of the hepatitis B virus-PreS1 protein (PreS1(21–47)). Conjugation of the peptide was performed via the maleimide–thiol reaction in methanol, allowing the use of a limited amount of the targeting molecule. This is an efficient procedure also in the perspective of selecting libraries of new targeting agents. The rationale behind the selection of the peptide is that SB3, which is undetectable in normal hepatocytes, is overexpressed in hepatocellular carcinoma and in hepatoblastoma and has been proposed as a target of the hepatitis B virus (HBV). For the latter, the key recognition element is the PreS1(21–47) peptide, which is a fragment of one of the proteins composing the viral envelope. The ability of the conjugated nanoparticles to bind the target protein SB3, expressed in liver cancer cells, was investigated by surface plasmon resonance analysis and in vitro via cellular uptake analysis followed by atomic absorption analysis of digested samples. The results showed that the PreS1(21–47) peptide is a suitable targeting agent for cells overexpressing the SB3 protein. Even more important is the evidence that the gold nanoparticles are internalized by the cells. The comparison between the surface plasmon resonance analysis and the cellular uptake studies suggests that the presentation of the protein on the cell surface is critical for efficient recognition. Graphic Abstract ACS Electronic Supporting Info


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