O6-alkylguanine-DNA transferase (SNAP) as capture module for site-specific covalent bioconjugation of targeting protein on nanoparticles

Serena Mazzucchelli; Miriam Colombo; Elisabetta Galbiati; Fabio Corsi; Josè M. Montenegro; Wolfgang J. Parak; Davide Prosperi

doi:10.1117/12.2001648

22 February 2013 O₆-alkylguanine-DNA transferase (SNAP) as capture module for site-specific covalent bioconjugation of targeting protein on nanoparticles

Serena Mazzucchelli, Miriam Colombo, Elisabetta Galbiati, Fabio Corsi, Josè M. Montenegro, Wolfgang J. Parak, Davide Prosperi

Author Affiliations +

Proceedings Volume 8595, Colloidal Nanocrystals for Biomedical Applications VIII; 859502 (2013) https://doi.org/10.1117/12.2001648
Event: SPIE BiOS, 2013, San Francisco, California, United States

Abstract

A bimodular genetic fusion comprising a delivery module (scFv) and a capture module (SNAP) is proposed as a novel strategy for the biologically mediated site-specific covalent conjugation of targeting proteins to nanoparticles. ScFv800E6, an scFv mutant selective for HER2 antigen overexpressed in breast cancer cells was chosen as targeting ligand. The fusion protein SNAP-scFv was irreversibly immobilized on magnetofluorescent nanoparticles through the recognition between SNAP module and pegylated O₆-alkylguanine derivative. The targeting efficiency of the resulting nanoparticle against HER2-positive breast cancer cells was assessed by flow cytometry and immunofluorescence.

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Serena Mazzucchelli, Miriam Colombo, Elisabetta Galbiati, Fabio Corsi, Josè M. Montenegro, Wolfgang J. Parak, and Davide Prosperi "O₆-alkylguanine-DNA transferase (SNAP) as capture module for site-specific covalent bioconjugation of targeting protein on nanoparticles", Proc. SPIE 8595, Colloidal Nanocrystals for Biomedical Applications VIII, 859502 (22 February 2013); https://doi.org/10.1117/12.2001648

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