title: Gene transfer by histidylated lipopolyplexes: A dehydration method allowing preservation of their physicochemical parameters and transfection efficiency.
abstract: Lipid-Polycation-DNA complexes (LPD) is a promising non-viral system for nucleic acids delivery. Usually, LPD are prepared just before their use. In the present work, we have examined whether dehydration of a new type of LPD (named LPD100) might be a storage option. LPD100 comprises PEGylated histidylated polylysine/pDNA polyplexes and a liposomal formulation made with lipophosphoramidates containing N-methylimidazolium and histamine polar heads. LPD100 were dehydrated by evaporation, and the physicochemical parameters and transfection efficiency (TE) of reconstituted LPD100 were compared to that of fresh LPD100. LPD100 previously dehydrated in the presence of 20% saccharose, displayed comparable size and surface charge as freshly prepared LPD100 but gave a better TE. CryoTEM experiments showed that the reconstituted LPD100 exhibited a shape similar to fresh ones. Moreover, when LPD100 were prepared with dehydrated pDNA/polymer complexes and fresh liposomes, TE was as efficient as with fresh LPD100 while a small increase of their size were observed. These results demonstrate that evaporation of LPD100 in the presence of saccharose is a powerful method to store them for a long period of time.
authors.affiliation: Centre de Biophysique Moléculaire CNRS UPR 4301, University of Orléans and Inserm, rue Charles Sadron, 45071 Orléans Cedex 2, France.
authors.forename: Federico
authors.initials: F
authors.lastname: Perche
authors.forename: Olivier
authors.initials: O
authors.lastname: Lambert
authors.forename: Mathieu
authors.initials: M
authors.lastname: Berchel
authors.forename: Paul-Alain
authors.initials: PA
authors.lastname: Jaffrès
authors.forename: Chantal
authors.initials: C
authors.lastname: Pichon
authors.forename: Patrick
authors.initials: P
authors.lastname: Midoux
mesh_terms: D020285:Cryoelectron Microscopy; D016264:Dextran Sulfate / Q000737:chemistry; D004355:Drug Stability; D004356:Drug Storage / Q000379:methods*; D018014:Gene Transfer Techniques; D057809:HEK293 Cells; D006639:Histidine / Q000031:analogs & derivatives / Q000737:chemistry*; D006801:Humans; D007093:Imidazoles / Q000737:chemistry; D008055:Lipids / Q000737:chemistry*; D008081:Liposomes / Q000138:chemical synthesis / Q000737:chemistry; D049409:Luciferases, Firefly / Q000235:genetics / Q000378:metabolism; D053758:Nanoparticles / Q000737:chemistry* / Q000648:ultrastructure; D010316:Particle Size; D010957:Plasmids / Q000235:genetics; D011092:Polyethylene Glycols / Q000737:chemistry; D011107:Polylysine / Q000737:chemistry; D055672:Static Electricity; D013395:Sucrose / Q000737:chemistry; D013499:Surface Properties; D014162:Transfection
chemical_list: D007093:Imidazoles; D008055:Lipids; D008081:Liposomes; D011107:Polylysine; D011092:Polyethylene Glycols; D006639:Histidine; D013395:Sucrose; D016264:Dextran Sulfate; D049409:Luciferases, Firefly