Viscometric analysis of DNA-lipid complexes.
DNA-cationic lipid complexes, "lipoplexes", are used as gene carriers for molecular biology and gene therapy applications. Colloidal properties of lipoplexes can be determined by viscometric analysis. (1) The shape parameter of lipoplexes can be one of the factors affecting transfection efficiency; (2) the volume fraction of free liposomes remaining after lipoplex formation can be used as an index of purity of the lipoplex product; (3) the shear dependence of the viscosity of a diluted lipoplex suspension can be used as a macroscopic shape factor: (4) the attraction force parameter between particles can be a colloidal stability factor. These properties should be characterized and specified for process control of lipoplex production and quality control of lipoplex products.We describe an automated mini-capillary viscometer for a sample volume of 0.5 ml, and its application to the characterizations of lipoplexes. We show a procedure for viscosity measurements and provide a calculation using complexes of plant DNA-distearyldimethylammonium chloride (DDAC) at a charged ratio of 1:4 (-/+), in which the amount of DNA is less than 250 microg. The prolate/ellipsoidal axial ratio, a/b, was found to be 70. Determination of the shape parameter with a/b is found to be better than that with other shape parameters, e.g., alpha of the Sakurada equation, because fractionation of the particle size is not necessary. By the proposed method, colloidal parameters of lipoplexes and bioactive polymer complexes are characterized quantitatively.
Hirota
Sadao
S
Department of Material Science, Tokyo Denki University, Japan.
Düzgüneş
Nejat
N
D001703:Biophysics / Q000295:instrumentation / Q000379:methods*; D002412:Cations / Q000737:chemistry; D004247:DNA / Q000008:administration & dosage* / Q000737:chemistry* / Q000302:isolation & purification; D004867:Equipment Design; D008055:Lipids / Q000737:chemistry*; D008081:Liposomes / Q000737:chemistry*; D010944:Plants / Q000737:chemistry; D000644:Quaternary Ammonium Compounds / Q000737:chemistry; D014783:Viscosity
D002412:Cations; D008055:Lipids; D008081:Liposomes; D000644:Quaternary Ammonium Compounds; C514922:distearyldimethylammoniumchloride; D004247:DNA