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Co-assembled epitope vaccine and its application

An epitope vaccine and co-assembly technology, applied in the field of biomedical engineering, can solve the problems of ineffective stimulation of Th1 immune response, inability to meet tumor immunotherapy, weak immunogenicity, etc., to improve humoral immunity and enhance cellular immune response , strong specific effect

Active Publication Date: 2022-03-01
INST OF BIOMEDICAL ENG CHINESE ACAD OF MEDICAL SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, this type of vaccine has a short half-life in vivo, is not easily taken up by antigen-presenting cells (APCs), and is difficult to enter the type I major histocompatibility complex (Major histocompatibility complex, MHCI) antigen presentation pathway in APC cells. Therefore, it cannot effectively stimulate Th1 and CTL protective immune responses, showing weak immunogenicity, and cannot meet the requirements of tumor immunotherapy

Method used

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  • Co-assembled epitope vaccine and its application
  • Co-assembled epitope vaccine and its application
  • Co-assembled epitope vaccine and its application

Examples

Experimental program
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Effect test

Embodiment 1

[0035] The preparation method of self-assembly epitope vaccine of the present invention is as follows:

[0036] Step 1. Preparation of epitope polypeptide bond

[0037] The epitope peptide and small molecule polypeptide carrier with a molar ratio of 1 to 3:1 are used as the epitope antigen and carrier, and the polypeptide carrier and epitope polypeptide are synthesized by solid-phase synthesis technology, and the amide bond, amino acid sequence or disulfide bond is selected as the connection Reactive groups such as amino, carboxyl, and sulfhydryl groups on epitope polypeptides and small molecule polypeptide carriers are prepared through amidation, sulfhydryl, and disulfide bond exchange reactions to prepare epitope polypeptide bonds, which are polypeptides containing antigenic epitopes sequence. If you choose the amide bond, it is the solid phase synthesis technique; if you choose the disulfide bond or the polypeptide carrier, it is the epitope peptide modified cysteine, the ...

Embodiment 2

[0042] Example 2 Preparation of co-assembled multivalent epitope vaccine

[0043] The small molecule polypeptides bound to different epitope peptides in Example 1 were dissolved in the aqueous solution at a molar ratio of 1:1, the concentration was adjusted, and nanostructures were formed through the co-assembly of the polypeptides to obtain a polymorphism in the form of hydrogel as a macroscopic expression. Epitope vaccines. For example: We bonded the KWKAKAKAKWK polypeptide with the SIINFEKL epitope, and then bonded the EWEAEAEAEAE polypeptide with the QAVHAAHAEINE epitope to construct two polypeptide sequences. The two polypeptides KWKAKAKAKWKGGGSIINFEK and EWEAEAEAEWEGGGQAVHAAHAEINE can be Spontaneous co-assembly occurs. The results show that as the concentration increases, the co-assembled structure changes from nanofibers to a mixture of three-dimensional porous structures and nanoparticles, and the macroscopic state changes from solution to semi-solid hydrogel.

Embodiment 3

[0044] Example 3: Study on in vitro activity of co-assembled epitope vaccines.

[0045] C57 / BL6 mouse bone marrow cells were isolated, and GM-CSF and IL-4 were added to the culture medium for 6 days, and immature dendritic cells (Dendritic cells, DCs) were harvested, and the solution prepared in Example 2 was The epitope vaccine was added to the medium to stimulate the differentiation of immature DC cells into mature DC cells, and after 24 hours of culture, flow cytometry (FACS) was used to detect the expression of the main surface markers MHCII, CD80, CD86 and CD40 of mature DC cells , using ELISA kits to detect the secretion of cytokines (IL-6, IL-12p40, IFN-γ and TNF-α). Image 6 As shown, the preliminary experimental results show that the epitope vaccine solution bonded to the polypeptide carrier can still maintain the activity of the epitope and effectively stimulate the maturation of DC cells.

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Abstract

The invention discloses a self-assembled and co-assembled epitope vaccine and its application. The epitope peptide and small molecule polypeptide carrier including T cell epitope and B cell epitope are bonded by covalent bonds, self-assembled in aqueous solution, Adjust the concentration, form nanostructures through self-assembly of polypeptides, and then form nanostructures by the co-assembly of polypeptide sequences containing antigenic epitopes bonded to different epitope peptides in aqueous solution to form nanostructures and its role in the preparation / prevention of tumors application, so as to protect the polypeptide, change the existing form of the epitope polypeptide, enhance the recognition and uptake of the epitope polypeptide by antigen-presenting cells, improve the immunogenicity, and finally improve the cellular immune response and tumor immunotherapy effect of the epitope vaccine.

Description

technical field [0001] The invention belongs to the field of biomedical engineering, and in particular relates to self-assembled epitope vaccines and multi-epitope co-assembled multivalent vaccines and applications thereof. Background technique [0002] Compared with traditional vaccines, epitope vaccines have more advantages, and their biggest advantage is that they can overcome the safety problems caused by traditional vaccines caused by virulence recovery or dispersal. In addition, the epitope vaccine itself is non-toxic and stable, which is in line with the direction of future vaccine development. [0003] Among various molecules, stimulating the body with epitope peptides of cytotoxic T lymphocytes (CTL) of tumor antigens expressed on the surface of tumor cells can generate antigen-specific CTL cells to participate in the tumor immune response. The mechanism is that after stimulating the body with CTL epitope peptides, it can cause the clonal proliferation of antigen-s...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K39/385A61K39/00A61K39/395A61P35/00A61P37/02C07K19/00
CPCA61K39/385A61K39/0011A61K39/39558A61P35/00A61P37/02C07K19/00C07K14/705C07K2319/00A61K2039/6031A61K2039/70A61K2300/00
Inventor 宋会娟王伟伟张闯年黄平升孔德领
Owner INST OF BIOMEDICAL ENG CHINESE ACAD OF MEDICAL SCI
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