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The effect of microfluidization on liposomal surface protein: characterization and immunological studies

Skalko, Natasa; (1995) The effect of microfluidization on liposomal surface protein: characterization and immunological studies. Doctoral thesis (Ph.D), UCL (University College London). Green open access

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Abstract

Liposomes have been shown to act as an immunological adjuvant, both for entrapped and vesicle-surface-linked antigens. In this respect, dehydration-rehydration vesicles (DRV liposomes) have the advantage of simple manufacture under conditions which are not damaging to antigens. Furthermore, coupling procedures can be employed for the attachment of antigens or ligands to the surface of DRVs under conditions where contact of potentially damaging coupling reagents with labile solutes incorporated in liposomes is avoided: antigens or ligands are firstly linked to small unilamellar vesicles (SUV) which are then used to generate DRVs in the presence of solutes destined for entrapment. Much of the protein originally linked to SUVs ends up on the surface of DRV. Microfluidization was used as a method to obtained vesicles of smaller sizes (about 100 nm) from DRVs. During microfluidization for up to 10 cycles only up to 20 % of originally bound protein was released. The presence of proteins (tetanus toxoid or bovine IgG) in SUVs and DRVs was observed by freeze-fracture electron microscopy, which showed protein as particles (10-15 nm in diameter) on the liposomal surface. Cryo-electron microscopy and the small angle X-ray scattering (SAXS) method were applied to characterise liposomes. The effect of vesicle size and phospholipid composition of DRVs (entrapped or surface linked antigens) on immune responses (IgG1, IgG2a and IgG2b) was studied in mice. Moreover the adjuvant activity of DRVs was compared to that of vesicles-in-water-in-oil emulsion and other adjuvants namely niosomes, Freund's complete adjuvant and TiterMax®. No side effects were observed in mice immunised with liposomes, niosomes or vesicles-in-water-in-oil emulsion.

Type: Thesis (Doctoral)
Qualification: Ph.D
Title: The effect of microfluidization on liposomal surface protein: characterization and immunological studies
Open access status: An open access version is available from UCL Discovery
Language: English
Additional information: Thesis digitised by ProQuest.
Keywords: Health and environmental sciences; Immunological; Liposomal; Microfluidization; Protein
URI: https://discovery.ucl.ac.uk/id/eprint/10104859
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