Linkup of a Fast Protein Liquid Chromatography System with a Stirred Thermostated Cell for Sterile Preparation of Liposomes by the Proliposome–Liposome Method: Application to Encapsulation of Antibiotics, Synthetic Peptide Immunomodulators, and a Photosensitizer

• Published in: Analytical biochemistry Vol. 249; no. 2; pp. 131 - 139
• Main Authors: Turánek, Jaroslav, Záluská, Dana, Neča, Jiřı́
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 01.07.1997
• Subjects: Adjuvants, Immunologic - administration & dosage; Adjuvants, Immunologic - chemical synthesis; Anti-Bacterial Agents - administration & dosage; Chromatography, High Pressure Liquid - methods; Drug Carriers - chemical synthesis; Drug Contamination; Freezing; Liposomes - chemical synthesis; Peptides - administration & dosage; Peptides - chemical synthesis; Photosensitizing Agents - administration & dosage; Temperature
• ISSN: 0003-2697; 1096-0309
• DOI: 10.1006/abio.1997.2146

The proliposome–liposome method is based on the conversion of the initial proliposome preparation into a liposome dispersion by dilution with the aqueous phase. This technique is characterized by an extremely high entrapment efficiency and is suitable for the encapsulation of a wide range of drugs with different water and alcohol solubility. A description of a home-made stirred thermostated cell and its linkup with an FPLC system for a rapid and automated preparation of multilamellar liposomes under strictly controlled conditions (temperature, dilution rate, and schedule) is presented. The highly reproducible procedure yields multilamellar liposomes with a high encapsulation efficiency for various drugs. Carboxyfluorescein, as a model hydrophilic compound, was entrapped with an efficiency of 81 ± 2%. The antibiotics neomycin and gentamycin were entrapped with efficiencies of 65 and 69%, respectively. Synthetic immunomodulators adamantylamide dipeptide, muramyl dipeptide, and β-d-GlcNAc-norMurNAc-l-Abu-d-isoGln were entrapped with efficiencies of 87, 62, and 85%, respectively. The photosensitizer mesotetra-(parasulfophenyl)-porphin was entrapped with an efficiency of 65%. The cell has been designed for laboratory-scale preparation of liposomes (300–1000 mg of phospholipid per run) in a procedure taking less than 90 min. The method can be readily scaled up and linked with secondary processing methods, such as pressure extrusion through polycarbonate filters.