Journal of Membrane Biology

, Volume 221, Issue 3, pp 141–152 | Cite as

Transferrin-Associated Lipoplexes as Gene Delivery Systems: Relevance of Mode of Preparation and Biophysical Properties

  • Nuno Penacho
  • Ana Filipe
  • Sérgio Simões
  • Maria C. Pedroso de Lima
Article

Abstract

The successful application of gene therapy depends highly on understanding the properties of gene carriers and their correlation with the ability to mediate transfection. An important parameter that has been described to improve transfection mediated by cationic liposomes involves association of ligands to cationic liposome–DNA complexes (lipoplexes). In this study, ternary complexes composed of 1,2-dioleoyl-3-(trimethylammonium) propane:cholesterol, plasmid DNA and transferrin (Tf, selected as a paradigm of a ligand) were prepared under various conditions, namely, in medium with different ionic strengths (HEPES-buffered saline [HBS] or dextrose), at different lipid/DNA (+/–) charge ratios and using different modes for component addition. We investigated the effect of these formulation parameters on transfection (in the absence and presence of serum), size of the complexes, degree of DNA protection and extent of their association with cells (in terms of both lipid and DNA). Our results show that all the tested parameters influenced to some extent the size of the complexes and their capacity to protect the carried genetic material, as well as the levels of cell association and transfection. The best transfection profile was observed for ternary complexes (Tf-complexes) prepared in high ionic strength solution (HBS), at charge ratios close to neutrality and according to the following order of component addition: cationic liposomes–Tf–DNA. Interestingly, in contrast to what was found for dextrose–Tf-complexes, transfection mediated by HBS-Tf-complexes in the presence of serum was highly enhanced.

Keywords

Cationic liposome Transfection Gene delivery Biophysical property Ternary complex Serum 

Notes

Acknowledgements

N. P. is the recipient of a fellowship from the Portuguese Foundation for Science and Technology (SFRH/BD/6135/2001). This work was partially financed by a grant from the Portuguese Foundation for Science and Technology (PTDC/BIO/65627/2006).

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Copyright information

© Springer Science+Business Media, LLC 2008

Authors and Affiliations

  • Nuno Penacho
    • 1
    • 3
  • Ana Filipe
    • 1
    • 2
  • Sérgio Simões
    • 1
    • 2
  • Maria C. Pedroso de Lima
    • 1
    • 3
  1. 1.Center for Neuroscience and Cell BiologyUniversity of CoimbraCoimbraPortugal
  2. 2.Laboratory of Pharmaceutical Technology, Faculty of PharmacyUniversity of CoimbraCoimbraPortugal
  3. 3.Department of Biochemistry, Faculty of Science and TechnologyUniversity of CoimbraCoimbraPortugal

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