Protein self-assembled novel nanovaccine and preparation method thereof

A nano-vaccine and self-assembly technology, applied in biochemical equipment and methods, drug combinations, pharmaceutical formulations, etc., can solve problems such as high synthesis cost, low antigen density, and complex synthesis process

Active Publication Date: 2017-09-15
TONGJI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0008] The purpose of the present invention is to provide a novel self-nanometer nano-vaccine and its preparation method based on self-nanometer cross-linking of disulfide bonds inside the antigen and its preparation method in order to overcome the above-mentioned defects in the prior art. Existing vaccine technology has high synthesis cost, complex synthesis process, and biological safety issues, and effectively solves

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  • Protein self-assembled novel nanovaccine and preparation method thereof
  • Protein self-assembled novel nanovaccine and preparation method thereof
  • Protein self-assembled novel nanovaccine and preparation method thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0074] Example 1: Nano-vaccine preparation of tumor model antigen

[0075] Dissolve 6mg of OVA powder in 2mL of 0.05M MES buffer at pH 3.5, filter or centrifuge to remove flocculent insoluble matter. Add 50 microliters of 6% sodium dodecyl sulfate aqueous solution to the antigenic protein system, stir for 2 minutes, and mix well. Pour the reaction solution into a screw-top glass bottle and tighten it, place it in an oil bath at 90°C for heating, and stir it on a magnetic stirrer at a speed of 750 rpm, and the reaction time is 4-30 minutes. With different reaction times, nanoparticles with different degrees of opalescence and uniform particle size can be obtained (such as figure 1 As shown, with the difference of the reaction time, the nano-vaccine with good quality, strong controllability and different particle sizes was obtained). After the reaction was completed, the reaction solution was placed in an ice-water bath for 1-2 min to terminate the reaction quickly. Finally, ...

Embodiment 2

[0077] Embodiment 2: Nano vaccine preparation of tumor model antigen

[0078] Dissolve 6mg of OVA powder in 2mL buffer solution with pH of 3.0, 4.0, 5.0, 6.0, filter or centrifuge to remove flocculent insoluble matter. Add 50 microliters of 6% sodium dodecyl sulfate aqueous solution to the antigenic protein system, stir for 2 minutes, and mix well. The reaction solution was poured into a screw-top glass bottle and screwed tightly, heated in an oil bath at 90°C, stirred on a magnetic stirrer at a speed of 750 rpm, and the reaction time was 25 minutes. With different reaction times, nanoparticles with different degrees of opalescence and uniform particle size can be obtained (such as figure 1 shown). After the reaction was completed, the reaction solution was placed in an ice-water bath for 1-2 min to terminate the reaction quickly. Finally, use a 3000 molecular weight ultrafiltration tube to centrifuge, or use a 3000 molecular weight dialysis bag to dialyze for 48 hours to r...

Embodiment 3

[0082] Nano vaccine preparation of embodiment 3 tumor model antigen

[0083] Dissolve 6mg of OVA powder in 2mL of 0.05M MES buffer at pH 3.5, filter or centrifuge to remove flocculent insoluble matter. Add the denaturant to the antigenic protein system, stir for 2 minutes, and mix well. The reaction solution was poured into a screw-top glass bottle and tightened, heated in an oil bath under a temperature gradient of 50°C-120°C, stirred at a speed of 750 rpm on a magnetic stirrer, and the reaction time was 10 minutes. With different reaction times, nanoparticles with different degrees of opalescence and uniform particle size can be obtained (such as figure 1 shown). After the reaction was completed, the reaction solution was placed in an ice-water bath for 1-2 min to terminate the reaction quickly. Finally, use a 3000 molecular weight ultrafiltration tube to centrifuge, or use a 3000 molecular weight dialysis bag to dialyze for 48 hours to remove impurities and unreacted com...

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Abstract

The invention relates to a protein self-assembled novel nanovaccine and a preparation method thereof. The protein self-assembled novel nanovaccine is prepared on the basis of antigen protein self-assembly; in the process of vaccine preparation, a molecular adjuvant is selectively introduced, the antigen content is higher than or equal to 85%, a high-efficiency immune effect can be triggered without needing assistance of an aluminum adjuvant, a Freund's adjuvant and the like, mercapto groups between protein molecules are exposed by virtue of physical regulation and control, and stable protein nanoparticles mainly based on disulfide bond crosslinking are formed through a mercapto/disulfide bond exchange reaction. The defects that the conventional nanovaccine needs to be introduced with an exogenous carrier or a cross-linking agent and the like are overcome, and the immune effect and the biosafety of the vaccine can be improved at the same time; the obtained vaccine granules are tidy in morphology, high in stability, flexible in regulation and control mode and good in repeatability, and can effectively stimulate dendritic cell maturation; the protein self-assembled novel nanovaccine has relatively strong generality and universality, is verified in a series of antigen proteins, and has a potential significant application value in the fields of novel vaccinating methods and biological pharmacy.

Description

technical field [0001] The invention belongs to the field of macromolecule nano-biomedical materials, and in particular relates to a novel protein self-assembled nano-vaccine and a preparation method thereof. Antigen peptides or immune adjuvants are loaded into proteins to form high-density nano-vaccine. Background technique [0002] Vaccines protect humans from many deadly diseases and are the most successful medical measures in human history, such as eradicating smallpox worldwide and effectively reducing infections of other severe infectious diseases, such as: polio, Diphtheria, tetanus, whooping cough, measles, mumps, and rubella virus infection have not only improved the quality of human life, but also greatly extended the average life expectancy of human beings. According to 2010 statistics from the National Institutes of Health, vaccines have prevented at least 2.5 million deaths and countless other cases each year. [0003] The traditional vaccine design uses patho...

Claims

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Application Information

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IPC IPC(8): A61K9/14A61K39/00A61K39/39A61K39/12A61K39/29A61K39/08A61K39/13A61K39/205A61K39/112A61K39/25A61K39/165A61K39/21A61K39/04A61P31/20A61P1/16A61P21/02A61P31/04A61P31/14A61P25/00A61P31/22A61P31/18A61P31/06
CPCA61K9/14A61K39/0011A61K39/0275A61K39/04A61K39/08A61K39/12A61K39/39A61K2039/55511A61K2039/55516A61K2039/55522A61K2039/55561C12N2710/16734C12N2710/20034C12N2710/24034C12N2730/10134C12N2740/16234C12N2760/18434C12N2760/20134C12N2770/32334C12N2770/32434Y02A50/30
Inventor 李永勇王坤任天斌温姝曼董海青
Owner TONGJI UNIV
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