Preparation And Preliminary Studies On Control Release Of Chitosan Nanoparticals As DNA Vaccine Vehicle

Chitosan is the only natural alkaline cationic polysaccharide, because of its biodegradability, bio-adhesion, good biocompatibility, and low toxicity is widely application and chitosan/DNA nanoparticles can be prepared by using a complex coacervation process, improve the utilization of nucleic acids, nucleic acids can be protected, so as excellent nucleic acid vector of widespread concern. This chitosan-DNA_ch90 nanoparticles were used complex coacervation to mix chitosan and Candida albicans recombinant Hsp90 and their release in the role of animal experiments conducted in the preliminary study.1,N/P, pH value, Na₂SO₄ concentration, the water bath temperature and DNA_ch90 concentration on the nanoparticle size are positively related; N/P, and DNA_ch90 concentration on the surface potential are positively related; pH value, Na₂SO₄ concentration, and the water bath temperature and particle surface potential is a negative correlation. Optimized conditions to prepare the nanoparticles.2,Preliminary inquiry about nanoparticles on physical and chemical properties and related properties: Scan the nanoparticles to determine the size and distribution of nanoparticle size, surface potential. Results showed size of the uniform diameter of about 166nm, a good dispersion , surface potential is positive; Measure the encapsulation efficiency of nanoparticles and drug loading. Results showed that higher than 90% of the encapsulated particles and the encapsulation rate proportional to the concentration of chitosan;Verify the positive and negative surface potential, enzyme protection experiments. the chitosan concentration,Analysis of gel retardation experiments show that the particle surface with a positive charge, DNA_ch90 wrapped in center; Enzyme protection experiments demonstrate chitosan-DNA_ch90 nanoparticles have a good effect against the degradation.3,Using chitosan-DNA_ch90 nanoparticles as experimental material, two injections of subcutaneous immune ICR mice, Western Blot shows nanoparticles can stimulate body occurs immune response, to produce R-Hsp90 antibodys. ELISA results showed that the nanoparticles with high encapsulation efficiency of delay due to a combination of tightly wrapped in the center of the plasmid. Follow experiments will further interpret chitosan-DNA_ch90 nanoparticles on animals have immune-enhancing effects of control-release effect.