Use of cationic biodegradable ceramic polymer microparticles for vaccine delivery
A cationic, microparticle technology, applied in the direction of microorganisms, drug delivery, protozoan antigen components, etc., can solve the problems of lack of CD8T lymphocyte immune response, biological toxicity, etc.
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Embodiment 1
[0028] Preparation of Dengue Protein Vaccine with Surface Modified Calcium Hydrogen Phosphate (MCHP)
[0029]Microparticles of surface-modified calcium hydrogen phosphate were weighed and transferred to 1.5 milliliter (ml) microtubes, rinsed with ice phosphate buffered solution (PBS) and resuspended in iced phosphate buffered solution, each dose was 0.25 - 10 mg (mg) surface-modified calcium hydrogen phosphate microparticles were suspended in 200 microliters of ice phosphate buffer solution, and then 1-10 micrograms (μg) of dengue virus envelope domain III (D2-ED3) recombinant protein was added to the aforementioned surface Modified dibasic calcium phosphate microparticle suspension was mixed overnight at 4°C in a vertical rotation for protein adsorption. After the protein adsorption process, calcium dibasic phosphate-type II dengue virus envelope domain III (D2- ED3) Protein vaccine; In this embodiment, preferably, it is prepared in 200 microliters of phosphate buffer solutio...
Embodiment 2
[0032] In vitro protein adsorption and controlled release in vivo of calcium hydrogen phosphate protein vaccine
[0033] (1) Protein adsorption rate test
[0034] In the experiment, 10 mg of cationic polylactic acid-glycolic acid copolymer microparticles or 10 mg of calcium hydrogen phosphate microparticles were mixed with 100 micrograms of ovalbumin (ovalbumin; OVA) overnight in 300 microliters of phosphate buffer solution at 4 °C. , and then remove the unadsorbed ovalbumin, then wash the microparticles with 300 microliters of phosphate buffer solution, then settle the microparticles, and set them in 100 microliters of eluting buffer (containing 0.1 mol / L NaOH and 1% mass fraction SDS). Elute overnight on a rotator to obtain ovalbumin adsorbed on microparticles, and then, after centrifugation, quantify the concentration of ovalbumin in the supernatant by BCA protein assay.
[0035] figure 2 In order to show the mean value and standard deviation after ovalbumin quantificati...
Embodiment 3
[0040] Optimizing the dosage of surface-modified calcium hydrogen phosphate microparticles to enhance the immunogenicity of vaccines
[0041] In the experiment, C57BL / 6 mice were subcutaneously injected with 4 different samples, respectively (1) 0.5 μg ovalbumin, (2) 10 mg calcium hydrogen phosphate-0.5 μg ovalbumin vaccine, (3) 1 mg hydrogen phosphate Calcium-0.5 μg ovalbumin vaccine and (4) 0.25 mg dibasic calcium phosphate-0.5 μg ovalbumin vaccine, both samples were dissolved in 200 μl phosphate buffered solution and mice were inoculated with the same vaccine two weeks after injection of the samples , Sacrifice the mouse one week after vaccination and obtain spleen cells, then analyze the specific T lymphocyte response to ovalbumin in the spleen cells by ELISPOT, and analyze the specific T lymphocytes respectively The production of interferon-γ and interleukin-4 produced by cells, the number of spot-forming cells (spot-forming cells; SFC) specific to interferon-γ and interl...
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