Separation of active complexes

C - Chemistry – Metallurgy – 12 – N

Patent

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Details

C12N 15/87 (2006.01) A61K 9/127 (2006.01) A61K 47/48 (2006.01) A61K 48/00 (2006.01) C12Q 1/68 (2006.01)

Patent

CA 2223934

The invention separates defined, active complexes by a characteristic from defined, active complexes that share a particular physicochemical characteristic such as density, surface charge or particle size are separated from complexes fromed by the association of a polynucleotide with a transfecting component that increases transfection activity, such as a lipid, cationic lipid, liposome, peptide, cationic peptide, dendrimer or polycation. In a preferred embodiment, polynucleotide-transfecting component complexes are ultracentrifuged to resolve one or more bands corresponding to complexes having a specific polynucleotide-transfecting component interaction. Polynucleotide complexes having a cationic liposome transfecting component resolve into two primary bands corresponding to complexes formed either under excess lipid conditions or under excess polynucleotide conditions. In an alternate embodiment, polynucleotide-transfecting component complexes are resolved using cross-flow electrophoresis to identify complexes having specific interactions and to separate them from excess initial components.

La présente invention concerne la séparation de complexes actifs définis à l'aide d'une caractéristique physicochimique particulière telle que la densité, la charge superficielle ou la grosseur des particules, de complexes formés par l'association d'un polynucléotide avec un constituant transfectant qui accroît l'activité de transfection, comme un lipide, un lipide cationique, un liposome, un peptide, un peptide cationique, un dendrimère ou un polycation. Dans un mode de réalisation préférentiel, les complexes polynucléotide-constituant transfectant subissent une ultracentrifugation pour diviser une ou plusieurs bandes correspondant à des complexes qui ont une interaction spécifique polynucléotide-constituant transfectant. Les complexes de polynucléotides ayant un constituant transfectant de type liposome cationique se divisent en deux bandes primaires qui correspondent à des complexes formés soit dans des conditions d'excédent de lipides, soit dans des conditions d'excédent de polynucléotides. Dans un autre mode de réalisation, les complexes polynucléotide-constituant transfectant sont divisés grâce à l'utilisation de l'électrophorèse transversale pour identifier les complexes ayant des interactions spécifiques et les séparer des constituants de départ en excédent.

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