Methods for preparing polynucleotide transfection complexes

Details Methods for preparing polynucleotide transfection complexes Methods for preparing polynucleotide transfection complexes

C - Chemistry, Metallurgy

12

N

C12N 15/64 (2006.01) C12N 15/88 (2006.01)

Patent

CA 2348675

Methods are provided for the preparation of transfection complexes of polynucleotides and polycations, especially cationic lipids, suitable for delivering polynucleotides to cells. In particular, methods are provided for preparing transfection complexes using a reduced-volume, dual-feed process. Complexes are formed upon the collision of two feed stream, containing polynucleotides and polycation, respectively, under conditions resulting in turbulent mixing conditions in minimal volume, and removal of transfection complexes under laminar flow conditions. Alternatively, the components are mixed in a static mixer. The process is easily scaleable and highly reproducible.

L'invention concerne des méthodes permettant de préparer des complexes de transfection de polynucléotides et de polycations, notamment des lipides cationiques, aptes à assurer un apport de polynucléotides dans des cellules. En particulier, l'invention concerne des méthodes de préparation de complexes de transfection par un procédé de double-alimentation à volume réduit. Les complexes sont formés par la collision de deux flux d'alimentation, renfermant respectivement des polynucléotides et des polycations, dans des conditions induisant des conditions de mélange par turbulence de volume minimum, et par élimination des complexes de transfection dans des conditions d'écoulement laminaire. Selon un autre mode de réalisation, les composants sont mélangés dans un mélangeur statique. Le procédé présente une grande flexibilité d'échelle et des possibilités étendues de reproduction.

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