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J. Biol. Chem., Vol. 275, Issue 38, 29533-29538, September 22, 2000

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Biophysical and Structural Properties of DNA·diC₁₄-amidine Complexes
INFLUENCE OF THE DNA/LIPID RATIO^*

Véronique Pector^§, Jan Backmann^¶, Dominique Maes^¶, Michel Vandenbranden, and Jean-Marie Ruysschaert

From the  Laboratoire de Chimie Physique des Macromolécules aux Interfaces, Université Libre de Bruxelles, Campus Plaine CP 206/2, B-1050 Brussels, Belgium and the ^¶ Département Ultrastructuur, Vrije Universiteit Brussel, Paardenstraat 65, B-1640, Sint-Genesius-Rode, Belgium

Cationic liposomes are used as vectors for gene delivery both in vitro and in vivo. Comprehension of both DNA/liposome interactions on a molecular level and a description of structural modifications involved, are prerequisites to an optimization of the transfection protocol and, thus, successful application in therapy. Formation and stability of a DNA/cationic liposome complex were investigated here at different DNA:lipid molar ratios (). Isothermal titration calorimetry (ITC) of cationic liposomes with plasmid DNA was used to characterize the DNA-lipid interaction. Two processes were shown to be involved in the complex formation. A fast exothermic process was attributed to the electrostatic binding of DNA to the liposome surface. A subsequent slower endothermic reaction is likely to be caused by the fusion of the two components and their rearrangement into a new structure. Fluorescence and differential scanning calorimetry confirmed this interpretation. A kinetic model analyzes the ITC profile in terms of DNA/cationic liposome interactions.

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