Preparative size exclusion chromatography combined with detergent removal as a versatile tool to prepare unilamellar and spherical liposomes of highly uniform size distribution.
Detergent removal from mixed micelles was combined with preparative size exclusion chromatography (SEC) on Sephacryl S 500 HR to prepare unilamellar and spherical liposomes of defined sizes between 50 and 100 nm with a very narrow size distribution (RSD of vesicle diameter between 13% and 25%). For neutral phosphatidylcholine and negatively charged phosphatidylcholine/phosphatidylglycerol liposome preparations, efficient sizing at the preparative scale was demonstrated by analyzing isolated SEC peak fractions with cryo-transmission electron microscopy and dynamic light scattering. The number-weighted average vesicle diameters obtained using both methods are in very good agreement for fractions of low polydispersity.
Holzer
Martin
M
Department of Pharmaceutical Technology and Biopharmacy, University of Freiburg, Hermann-Herder-Strasse 9, D-79104 Freiburg, Germany. martin.holzer@pharmazie.uni-freiburg.de
Barnert
Sabine
S
Momm
Joachim
J
Schubert
Rolf
R
D002850:Chromatography, Gel / Q000379:methods*; D003902:Detergents / Q000737:chemistry*; D003956:Dialysis; D005960:Glucosides / Q000737:chemistry; D008027:Light; D008081:Liposomes / Q000138:chemical synthesis* / Q000737:chemistry; D046529:Microscopy, Electron, Transmission; D010316:Particle Size; D012542:Scattering, Radiation; D020358:Sodium Cholate / Q000737:chemistry; D053835:Unilamellar Liposomes / Q000138:chemical synthesis* / Q000737:chemistry
D003902:Detergents; D005960:Glucosides; D008081:Liposomes; D053835:Unilamellar Liposomes; C018619:octyl-beta-D-glucoside; D020358:Sodium Cholate