title: Capillary electrophoresis method to determine siRNA complexation with cationic liposomes.
abstract: Small interfering RNA (siRNA) inducing gene silencing has great potential to treat many human diseases. To ensure effective siRNA delivery, it must be complexed with an appropriate vector, generally nanoparticles. The nanoparticulate complex requires an optimal physiochemical characterization and the complexation efficiency has to be precisely determined. The methods usually used to measure complexation in gel electrophoresis and RiboGreen®fluorescence-based assay. However, those approaches are not automated and present some drawbacks such as the low throughput and the use of carcinogenic reagents. The aim of this study is to develop a new simple and fast method to accurately quantify the complexation efficiency. In this study, capillary electrophoresis (CE) was used to determine the siRNA complexation with cationic liposomes. The short-end injection mode applied enabled siRNA detection in less than 5 min. Moreover, the CE technique offers many advantages compared with the other classical methods. It is automated, does not require sample preparation and expensive reagents. Moreover, no mutagenic risk is associated with the CE approach since no carcinogenic product is used. Finally, this methodology can also be extended for the characterization of other types of nanoparticles encapsulating siRNA, such as cationic polymeric nanoparticles.
authors.affiliation: Laboratory of Pharmaceutical Technology and Biopharmacy, Department of Pharmaceutical Sciences - CIRM, University of Liege, Liege, Belgium. tania.furst@ulg.ac.be.
authors.forename: Tania
authors.initials: T
authors.lastname: Furst
authors.affiliation: Laboratory for the Analysis of Medicines, Department of Pharmaceutical Sciences - CIRM, University of Liege, Liege, Belgium.
authors.forename: Virginie
authors.initials: V
authors.lastname: Bettonville
authors.affiliation: Laboratory for the Analysis of Medicines, Department of Pharmaceutical Sciences - CIRM, University of Liege, Liege, Belgium.
authors.forename: Elena
authors.initials: E
authors.lastname: Farcas
authors.affiliation: Laboratory of Pharmaceutical Technology and Biopharmacy, Department of Pharmaceutical Sciences - CIRM, University of Liege, Liege, Belgium.
authors.forename: Antoine
authors.initials: A
authors.lastname: Frere
authors.affiliation: Laboratory of Pharmaceutical Technology and Biopharmacy, Department of Pharmaceutical Sciences - CIRM, University of Liege, Liege, Belgium.
authors.forename: Anna
authors.initials: A
authors.lastname: Lechanteur
authors.affiliation: Laboratory of Pharmaceutical Technology and Biopharmacy, Department of Pharmaceutical Sciences - CIRM, University of Liege, Liege, Belgium.
authors.forename: Brigitte
authors.initials: B
authors.lastname: Evrard
authors.affiliation: Laboratory for the Analysis of Medicines, Department of Pharmaceutical Sciences - CIRM, University of Liege, Liege, Belgium.
authors.forename: Marianne
authors.initials: M
authors.lastname: Fillet
authors.affiliation: Laboratory of Pharmaceutical Technology and Biopharmacy, Department of Pharmaceutical Sciences - CIRM, University of Liege, Liege, Belgium.
authors.forename: Géraldine
authors.initials: G
authors.lastname: Piel
authors.affiliation: Laboratory for the Analysis of Medicines, Department of Pharmaceutical Sciences - CIRM, University of Liege, Liege, Belgium.
authors.forename: Anne-Catherine
authors.initials: AC
authors.lastname: Servais
mesh_terms: D002138:Calibration; D002412:Cations / Q000032:analysis / Q000737:chemistry; D019075:Electrophoresis, Capillary / Q000379:methods*; D006801:Humans; D008081:Liposomes / Q000032:analysis / Q000737:chemistry*; D053758:Nanoparticles / Q000032:analysis / Q000737:chemistry*; D034741:RNA, Small Interfering / Q000032:analysis / Q000737:chemistry*
chemical_list: D002412:Cations; D008081:Liposomes; D034741:RNA, Small Interfering
keywords: Capillary electrophoresis; Complexation efficiency; Liposomes; siRNA