Preparation of liposomes via detergent removal from mixed micelles by dilution. The effect of bilayer composition and process parameters on liposome characteristics

Abstract

Liposomes were prepared from mixed micelles by a dilution method. Mixed micellar solutions, containing constant octyl glucoside and egg phosphatidylcholine concentrations and varying amounts of cholesterol and/or a charged compound, were diluted at defined rates. After dilution, the resulting liposome dispersions were sequentially concentrated, washed or dialysed, and filtered. The effect of lipid composition and experimental conditions on physicochemical characteristics was studied. Fairly homogeneous liposome dispersions with mean diameters ranging from 100 to over 200 nm could be obtained. The particle size was dependent on cholesterol content and surface charge, and could be reproducibly controlled by adjustment of the dilution rate. Liposomes with a mean diameter below 100 nm could also be obtained, but were heterodisperse and unstable. The incorporation of charged compounds was monitored by microelectrophoresis. 31P-NMR measurements indicated that the liposomes were unilamellar. Dialysis appeared to be more convenient than washing to remove octyl glucoside.