There is a growing demand for better delivery systems to improve the stability and efficacy of DNA vaccines. Here we report the synthesis of a non-viral DNA vaccine delivery system using a novel adjuvanted solid lipid nanoparticle (SLN-A) platform as a carrier for a DNA vaccine candidate encoding the Urease alpha (UreA) antigen from Helicobacter pylori. Cationic SLN-A particles containing monophosphoryl lipid A (adjuvant) were synthesised by a modified solvent-emulsification method and were investigated for their morphology, zeta potential and in vitro transfection capacity. Particles were found to bind plasmid DNA to form lipoplexes, which were characterised by electron microscopy, dynamic light scattering and fluorescence microscopy. Cellular uptake studies confirmed particle uptake within 3 h, and intracellular localisation within endosomal compartments. In vitro studies further confirmed the ability of SLN-A particles to stimulate expression of pro-inflammatory cytokine tumor necrosis factor alpha (TNF-α) in human macrophage-like Tohoku Hospital Pediatrics-1 (THP-1) cells. Lipoplexes were found to be biocompatible and could be efficiently transfected in murine immune cells for expression of recombinant H. pylori antigen Urease A, demonstrating their potential as a DNA vaccine delivery system.
Increasing application of nucleic acid vaccines is driving demand for new delivery systems to improve stability and efficacy of DNA vaccines. Solid lipid nanoparticles (SLN) are a particulate carrier system composed of a solid lipid core and a cationic lipid surface suitable for binding negatively charged DNA. SLN delivery systems can be used to bind DNA resulting in an SLN/DNA complex (termed "lipoplex") which can be used as a potential vaccine.In this chapter, the methodologies associated with the use of SLNs as a DNA vaccine nanocarrier are discussed. First, requirements for an effective experimental lipoplex vaccine are discussed along with current and historical examples. Then, flowcharts for design and synthesis of lipoplex vaccines are outlined, followed by detailed materials and methods for synthesis and characterization of lipoplex vaccines.
Increasing emergence of infectious diseases is driving demand for new vaccine technologies capable of improving antigen delivery and protective efficacy. Nanoparticle technology is a modern approach to antigen delivery, capable of stabilizing and increasing the amount of antigen delivered to immune cells. Protein-based nanoparticles are a biodegradable alternative to existing nanomaterials, offering a versatile and biocompatible approach to nanoparticle vaccine delivery. In this chapter, the methods for the synthesis and characterization of protein-based nanocapsule vaccines are discussed. Initially, the requirements for a suitable nanoparticle vaccine are outlined, and finally, methods for the design and synthesis of protein-based nanocapsule vaccines are explained.
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