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Trastuzumab-polyethylenimine-polyethylene glycol conjugates for targeting Her2-expressing tumors.
- Source :
-
Bioconjugate chemistry [Bioconjug Chem] 2006 Sep-Oct; Vol. 17 (5), pp. 1190-9. - Publication Year :
- 2006
-
Abstract
- In this study, we describe the synthesis and characterization of a conjugate consisting of poly(ethylene glycol 2,000 Da)(10)-graft-poly(ethylene imine 25 kDa) (PEG-PEI) covalently coupled to Trastuzumab (Herceptin) via N-succinimidyl-3-(2-pyridyldithio)propionate (SPDP) for specific gene delivery to Her2-expressing cell lines. The efficiency of DNA condensation was studied using an ethidium bromide exclusion assay and demonstrated negligible differences compared to PEG-PEI. Conjugate complex sizes were determined by dynamic light scattering to be in the range 130-180 nm. zeta potentials at different N/P ratios were close to neutral. Flow cytometry and confocal microscopy revealed efficient binding and uptake of Trastuzumab-PEI-PEG complexes using Her2-positive SK-BR-3 cells. In contrast, binding and uptake into Her2-negative OVCAR-3 cells was negligible. In good correlation with these findings, reporter gene expression using targeted complexes in SK-BR-3 cells was up to sevenfold higher than that of unmodified PEG-PEI complexes. With the use OVCAR-3 cells, no significant difference in expression efficiencies could be observed between conjugate and PEG-PEI complexes. Inhibition experiments with free Trastuzumab showed a significant decrease in reporter gene expression using SK-BR-3 cells but no decrease using OVCAR-3 cells, strongly supporting a specific Her2-receptor-mediated uptake mechanism. Our results suggest that Trastuzumab-PEI-PEG might be a promising new bioconjugate for targeted gene transfer to Her2-positive tumor cells in vivo.
- Subjects :
- Antibodies, Monoclonal metabolism
Antibodies, Monoclonal, Humanized
Antineoplastic Agents chemical synthesis
Antineoplastic Agents metabolism
Cell Line, Tumor
Genes, Reporter
Genetic Therapy methods
Humans
Molecular Structure
Neoplasms metabolism
Particle Size
Trastuzumab
Antibodies, Monoclonal chemistry
Antineoplastic Agents chemistry
Gene Transfer Techniques
Polyethylene Glycols chemistry
Polyethyleneimine chemistry
Receptor, ErbB-2 metabolism
Subjects
Details
- Language :
- English
- ISSN :
- 1043-1802
- Volume :
- 17
- Issue :
- 5
- Database :
- MEDLINE
- Journal :
- Bioconjugate chemistry
- Publication Type :
- Academic Journal
- Accession number :
- 16984128
- Full Text :
- https://doi.org/10.1021/bc0601119