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Preparation method and application of antigen and adjuvant co-delivery nano vaccine based on protamine as carrier

A nano-vaccine, protamine technology, applied in the field of immunology, can solve the problems of low antigen delivery efficiency, affecting the effect of antigen immunotherapy, poor immunogenicity, etc., achieving effective immunotherapy effect, realizing immunotherapy effect, and convenient preparation. Effect

Active Publication Date: 2021-03-19
WEIFANG MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0012] The immunogenicity of soluble protein and polypeptide antigens in the prior art is poor and the antigen delivery efficiency in vivo is low, which seriously affects the immunotherapy effect of such antigens

Method used

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  • Preparation method and application of antigen and adjuvant co-delivery nano vaccine based on protamine as carrier
  • Preparation method and application of antigen and adjuvant co-delivery nano vaccine based on protamine as carrier
  • Preparation method and application of antigen and adjuvant co-delivery nano vaccine based on protamine as carrier

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0061] Preparation of Nanovaccine

[0062] Dissolving PRT in PBS buffer solution, the concentration of PRT is 0.1-20 mg / mL to form a PRT solution. Ovalbumin (OVA, as a model antigen) was dissolved in a PBS buffer solution with a concentration of 0.1-5 mg / mL to form an OVA solution. CpG is dissolved in PBS buffer solution, and the concentration of CpG is 0.1-5 mg / mL to form a CpG solution. The PRT solution, the OVA solution and the CpG solution were mixed, vortexed for 20 seconds, and then left to stand at room temperature for 25 minutes to obtain the nanovaccine PRT / CpG / OVA.

Embodiment 2

[0064] Nano vaccine particle size distribution

[0065] The nano-vaccine prepared in Example 1 was studied for particle size distribution. The average particle size, Zeta potential and particle size dispersion index (PDI) were determined by nanometer particle size and Zeta potential analyzer. Refer to Table 1 for the results.

[0066] Potential, particle size and PDI test result of table 1 embodiment 1

[0067]

Embodiment 3

[0069] Effect experiment

[0070] (1) Endocytosis of nanovaccine in antigen-presenting cells

[0071] Mouse bone marrow-derived dendritic cells (BMDCs) were selected, bone marrow cells were obtained from mice, stimulated and induced in vitro, and differentiated into BMDCs after seven days of culture. After culturing BMDCs for seven days, use a pipette to absorb the medium and gently blow the cells, collect the cell suspension and place it in a 15mL centrifuge tube, centrifuge at 1000rpm for 3min to collect the cells; 6 The density of cells was seeded in 6-well plates and cultured overnight in an incubator containing 5% carbon dioxide by volume at 37°C. Then add sample materials (the samples are: PBS, CpG / OVA-FITC and PRT / CpG / OVA-FITC respectively, the final concentration of OVA added is 5 μg / mL), continue to cultivate for 4 hours, and then use flow cytometry to detect the antigen and endocytosis of adjuvants.

[0072] The above-mentioned OVA-FITC is OVA labeled with fluores...

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Abstract

The invention provides an antigen and adjuvant co-delivery nano vaccine based on protamine as a carrier. The nano vaccine comprises a carrier, an antigen and an adjuvant, wherein the carrier is protamine sulfate; The invention also provides a preparation method and application of the vaccine. The nano vaccine prepared by the invention can effectively improve uptake efficiency of antigen presentingcells on antigens and adjuvants, efficiently stimulate maturation of the antigen presenting cells, assist the antigens in realizing cross presentation, significantly improve in-vivo transfer efficiency of the antigens and the adjuvants, stimulate a body to produce high-efficiency humoral immunity and cellular immunity, and at the same time produce an immune memory effect. In-vivo treatment is carried out on mouse subcutaneous melanoma, and it is proved that the nano vaccine has a good anti-tumor immunotherapy effect. The nano vaccine is simple in structure, convenient to prepare and excellentin immunotherapy performance, and has good application potential.

Description

technical field [0001] The invention relates to the technical field of immunology, in particular to a preparation method and application of a nano-vaccine based on protamine as a carrier. Background technique [0002] Tumor has become a disease that seriously endangers human health. Traditional tumor treatment methods such as chemotherapy, radiotherapy, and surgical resection are difficult to inhibit tumor metastasis and recurrence in the body, and it is difficult to achieve tumor eradication. Tumor immunotherapy shows great promise in completely eradicating tumors by stimulating and enhancing the body's immune function to kill tumor cells. [0003] In recent years, immunotherapy has received great attention in the field of anti-tumor treatment. Currently, commonly used methods of immunotherapy include: adoptive lymphocyte therapy (T cells, NK cells), cytokine therapy, gene therapy, anti-tumor antibody therapy, immune examination Point (PD-1 / PD-L1, CTLA-4) blocking therapy ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/00A61K39/39A61K47/42A61P35/00
CPCA61K39/0011A61K39/39A61K47/42A61P35/00A61K2039/55561A61K2039/5154A61K2039/575A61K2039/54Y02A50/30
Inventor 关秀文姜明霞张维芬
Owner WEIFANG MEDICAL UNIV
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