Abstract
Poly(N-vinyl pyrrolidone)-based-nanogels (NGs), produced by e-beam irradiation, are conjugated with monoclonal antibodies (mAb) for active targeting purposes. The uptake of immuno-functionalized nanogels is tested in an endothelial cell line, ECV304, using confocal and epifluorescence microscopy. Intracellular localization studies reveal a faster uptake of the immuno-nanogel conjugate with respect to the ‘bare’ nanogel. The specific internalization pathway of these immuno-nanogels is clarified by selective endocytosis inhibition experiments, flow cytometry and confocal microscopy. Active targeting ability is also verified by conjugating a monoclonal antibody which recognizes the αvβ3 integrin on activated endothelial cells. Epifluorescence images of the ‘wound healing assay’ on ECV304 cells provide evidence of nanogels localization only in the target cells. Therefore, the immuno-nanogels produced have the potential to recognize specific cell types in heterogeneous systems, which makes them promising candidates for targeted drug delivery applications.
Acknowledgments
This research was partially supported by the International Atomic Energy Agency (IAEA) Coordinated Research Project (CRP F22064) ‘Nanosized delivery systems for radiopharmaceuticals’ Research Agreement N° 18349/R0. TEM experimental data were provided by Centro Grandi Apparecchiaturee UniNetLab e Università di Palermo funded by P.O.R. Sicilia 2000e2006, Misura 3.15 Quota Regionale. The authors are grateful to Prof G. Giammona for the experimental support on surface charge density measurements.
Funding: Italian Ministry of University and Scientific Research (Grant/Award number: ‘FFR 2012/2013’, ‘PRIN 2010–2011 NANOMED 2010FTPBSH’).
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