title: Incorporation of ovalbumin into ISCOMs and related colloidal particles prepared by the lipid film hydration method.
abstract: The aim of this study was to investigate the incorporation of a model antigen, fluorescently labelled ovalbumin (FITC-OVA), into various colloidal particles including immune stimulating complexes (ISCOMs), liposomes, ring and worm-like micelles, lamellae and lipidic/layered structures that are formed from various combinations of the triterpene saponin Quil A, cholesterol and phosphatidylethanolamine (PE) following hydration of PE/cholesterol lipid films with aqueous solutions of Quil A. Colloidal dispersions of these three components were also prepared by the dialysis method for comparison. FITC-OVA was conjugated with palmitic acid (P) and PE to produce P-FITC-OVA and PE-FITC-OVA, respectively. Both P-FITC-OVA and PE-FITC-OVA could be incorporated in all colloidal structures whereas FITC-OVA was incorporated only into liposomes. The incorporation of PE-FITC-OVA into all colloidal structures was significantly higher than P-FITC-OVA (P < 0.05). The degree of incorporation of protein was in the order: ring and worm-like micelles < liposomes and lipidic/layered structures < ISCOMs and lamellae. The incorporation of protein into the various particles prepared by the lipid film hydration method was similar to those for colloidal particles prepared by the dialysis method (provided both methods lead to the formation of the same colloidal structures). In the case of different colloidal structures arising due to the preparation method, differences in encapsulation efficiency were found (P < 0.05) for formulations with the same polar lipid composition. This study demonstrates that the various colloidal particles formed as a result of hydrating PE/cholesterol lipid films with different amounts of Quil A are capable of incorporating antigen, provided it is amphipathic. Some of these colloidal particles may be used as effective vaccine delivery systems.
authors.affiliation: Drug Delivery Solutions, New Zealand National School of Pharmacy, University of Otago, Dunedin.
authors.forename: Patrick H
authors.initials: PH
authors.lastname: Demana
authors.forename: Nigel M
authors.initials: NM
authors.lastname: Davies
authors.forename: Bianca
authors.initials: B
authors.lastname: Berger
authors.forename: Thomas
authors.initials: T
authors.lastname: Rades
mesh_terms: D002784:Cholesterol / Q000737:chemistry; D003102:Colloids; D016503:Drug Delivery Systems; D016650:Fluorescein-5-isothiocyanate / Q000737:chemistry; D005456:Fluorescent Dyes / Q000737:chemistry; D016882:ISCOMs / Q000737:chemistry*; D010047:Ovalbumin / Q000008:administration & dosage / Q000737:chemistry*; D010714:Phosphatidylethanolamines / Q000737:chemistry; D062605:Quillaja Saponins; D012503:Saponins / Q000737:chemistry; D013678:Technology, Pharmaceutical
chemical_list: D003102:Colloids; D005456:Fluorescent Dyes; D016882:ISCOMs; D010714:Phosphatidylethanolamines; D062605:Quillaja Saponins; D012503:Saponins; C483858:phosphatidylethanolamine; C046386:Quil A; D010047:Ovalbumin; D002784:Cholesterol; D016650:Fluorescein-5-isothiocyanate