PURPOSE: To study the stability of polymer-plasmid complexes (polyplexes) both as an aqueous dispersion and in their lyophilized form. METHODS: The characteristics of the polyplexes (size, charge and transfection potential) were monitored at different temperatures. Moreover, we studied possible changes in the secondary and tertiary structure of the plasmid by agarose gel electrophoresis and by CD spectroscopy to gain insight into the mechanism of polyplex degradation. RESULTS: The polyplexes preserved almost their full transfection potential after aging in an aqueous solution of 20 mM Hepes (pH 7.4) containing 10% sucrose at 4 and 20 degrees C for 10 months. On the other hand, the polyplexes aged at 40 degrees C were rather unstable and lost their transfection capability with a half-life of around 2 months. During storage, conformational changes in the secondary and tertiary structure of DNA were observed. When naked plasmid DNA was aged at 40 degrees C as an aqueous solution and complexed with polymer just before the transfection experiment, a slower drop in its transfection capability was observed. The freeze-dried polyplexes using sucrose as lyoprotectant almost fully retained their transfection efficiency, even when aged at 40 degrees C for 10 months. CONCLUSIONS: This study provides information about polyplex stability in aqueous dispersions on storage and demonstrates that freeze-drying is an excellent method to ensure long term stability.
PURPOSE: To study the stability of polymer-plasmid complexes (polyplexes) both as an aqueous dispersion and in their lyophilized form. METHODS: The characteristics of the polyplexes (size, charge and transfection potential) were monitored at different temperatures. Moreover, we studied possible changes in the secondary and tertiary structure of the plasmid by agarose gel electrophoresis and by CD spectroscopy to gain insight into the mechanism of polyplex degradation. RESULTS: The polyplexes preserved almost their full transfection potential after aging in an aqueous solution of 20 mM Hepes (pH 7.4) containing 10% sucrose at 4 and 20 degrees C for 10 months. On the other hand, the polyplexes aged at 40 degrees C were rather unstable and lost their transfection capability with a half-life of around 2 months. During storage, conformational changes in the secondary and tertiary structure of DNA were observed. When naked plasmid DNA was aged at 40 degrees C as an aqueous solution and complexed with polymer just before the transfection experiment, a slower drop in its transfection capability was observed. The freeze-dried polyplexes using sucrose as lyoprotectant almost fully retained their transfection efficiency, even when aged at 40 degrees C for 10 months. CONCLUSIONS: This study provides information about polyplex stability in aqueous dispersions on storage and demonstrates that freeze-drying is an excellent method to ensure long term stability.
Authors: Jacques Bonnet; Marthe Colotte; Delphine Coudy; Vincent Couallier; Joseph Portier; Bénédicte Morin; Sophie Tuffet Journal: Nucleic Acids Res Date: 2009-12-07 Impact factor: 16.971
Authors: Michael D Buschmann; Abderrazzak Merzouki; Marc Lavertu; Marc Thibault; Myriam Jean; Vincent Darras Journal: Adv Drug Deliv Rev Date: 2013-07-18 Impact factor: 15.470