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A kind of protein self-assembly novel nano-vaccine and preparation method thereof

A nano-vaccine and self-assembly technology, applied in biochemical equipment and methods, drug combinations, pharmaceutical formulations, etc., can solve the problems of high synthesis cost, low antigen density, complex synthesis process, etc., and achieve high antigen density and simple components. , the effect of a simple synthesis method

Active Publication Date: 2021-05-11
TONGJI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

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 the traditional problems of exogenous carriers, chemical cross-linking agents, low antigen density, and no surface display of antigens in the field of nano-vaccine design. , and the structure is stable, easy to store, and has the characteristics of highly imitating the structure of viruses, realizing the dual role of the carrier as the antigen, and at the same time, the nanoparticles have the dual role of immune adjuvant

Method used

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  • A kind of protein self-assembly novel nano-vaccine and preparation method thereof
  • A kind of protein self-assembly novel nano-vaccine and preparation method thereof
  • A kind of protein self-assembly novel nano-vaccine and preparation method thereof

Examples

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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 novel protein self-assembly nano-vaccine and a preparation method thereof. It is prepared based on the self-assembly of antigenic protein, and a molecular adjuvant is selectively introduced in the vaccine preparation process. The antigen content is ≥ 85%, and aluminum adjuvant and Freund With the help of adjuvants and other aids, an efficient immune effect can be induced. Physical regulation is used to expose the sulfhydryl groups between protein molecules, and through the thiol / disulfide bond exchange reaction, stable protein nanoparticles mainly composed of disulfide bond crosslinks are formed. The invention overcomes the defects that the traditional nano-vaccine needs to introduce foreign carrier or cross-linking agent, etc., and can improve the immune effect and biological safety of the vaccine at the same time. The obtained vaccine particles have regular shape, strong stability, flexible regulation mode, good repeatability, and can efficiently stimulate the maturation of dendritic cells. It has strong versatility and universality, has been verified on a series of antigenic proteins, and has potential significant application value in the field of new vaccine methods and biopharmaceuticals.

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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Patent Type & Authority Patents(China)
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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