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Biological particle type composite adjuvant and preparation method and application thereof

A technology of biological particles and adjuvants, applied in the biological field, can solve the problems of reduced diffusibility of adjuvants, easy diffusion, cytokine storm immune toxicity, etc., and achieve the effect of solving the toxic effect of adjuvants

Pending Publication Date: 2020-10-02
JILIN AGRICULTURAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In general, soluble adjuvants such as small molecule adjuvants and hydrophilic adjuvants are easy to diffuse and enter the circulatory system from the immune site soon after immunization, resulting in the generation of cytokine storm and severe immunotoxicity; and The ease of diffusibility of the adjuvant will be reduced

Method used

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  • Biological particle type composite adjuvant and preparation method and application thereof
  • Biological particle type composite adjuvant and preparation method and application thereof
  • Biological particle type composite adjuvant and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] This embodiment provides a biological particle type composite adjuvant, including bacteria-like particles of lactic acid bacteria and flagellin, the bacteria-like particles of lactic acid bacteria are the peptidoglycan backbone of lactic acid bacteria, and the flagellin passes through the peptidoglycan binding domain PA gene, The anchors are displayed on the surface of bacteria-like particles of lactic acid bacteria.

[0033] In this embodiment, the lactic acid bacteria is Lactococcus lactis, more specifically, Lactococcus lactis MG1363.

[0034] In this embodiment, the flagellin is the flagellin of Bacillus cereus, and the GenBank accession number of the Bacillus cereus is AE016877.1.

[0035] In this embodiment, the nucleotide amino acid sequence of the flagellin is shown in SEQ ID NO:1, and the amino acid sequence of the flagellin is shown in SEQ ID NO:2.

[0036] In this embodiment, the peptidoglycan binding domain PA gene is the C-terminal cell wall binding activi...

Embodiment 2

[0039]This embodiment provides the preparation method of the bioparticle type composite adjuvant as described in Example 1, comprising the following steps:

[0040] S1. Construction of recombinant flagellin comprising the peptidoglycan binding domain PA gene.

[0041] S2. Expressing and purifying the recombinant flagellin obtained in step S1.

[0042] S3, lactic acid bacteria are treated with hot acid to obtain bacteria-like particles containing only peptidoglycan skeleton;

[0043] S4. The recombinant flagellin obtained in step S2 is co-incubated with the bacteria-like particles obtained in step S3, and the PA gene of the peptidoglycan binding domain is bound to the surface of the bacteria-like particles to obtain a biological particle-type composite adjuvant.

[0044] In this embodiment, the specific process of constructing the recombinant flagellin containing the peptidoglycan binding domain PA gene in step S1 is as follows:

[0045] Synthesize the flagellin (BcFlg) gene ...

Embodiment 3

[0084] This example provides application examples and comparative examples of the above-mentioned bioparticle-type compound adjuvant vaccine.

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Abstract

The invention discloses a biological particle type composite adjuvant and a preparation method and application thereof. A novel biological particle type composite adjuvant produced by combining BLPs and BcFlg can simultaneously activate TLR2 and TLR5 signal pathways and activate an innate immune system more efficiently. However, different from the direct mixing of different TLR ligands, a biological particle material constructed based on a BLP surface display technology can perfectly solve the problem of the adjuvant toxicity and long-term retention at an antigen site. The use of biological particle type delivery tools can simultaneously deliver different adjuvant molecules to antigen-presenting cells to control the adjuvant density and the ratio of the adjuvant to immune cells, and simulate the characteristics of real pathogen particles in terms of size, shape, composition and the like. Therefore, the biological particle type composite adjuvant is an effective form of a novel high-efficiency adjuvant.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a biological particle type composite adjuvant and its preparation method and application. Background technique [0002] Vaccine adjuvant is a kind of substance that can non-specifically change or enhance the body’s specific immune response to antigens and play an auxiliary role. It can enhance the body’s immune response to antigens or change the immune response after being injected into the body before the antigen or mixed with the antigen. type. There is no uniform standard for the classification of adjuvants in the world. According to the different chemical components, they can be divided into aluminum salt adjuvant, protein adjuvant, nucleic acid adjuvant, lipid-containing adjuvant and mixed adjuvant. According to their physical properties, they can be divided into granular adjuvants and non-granular adjuvants. According to research, adjuvants mainly include several modes of act...

Claims

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Application Information

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IPC IPC(8): A61K39/39A61P37/04A61K39/145A61P31/16
CPCA61K39/39A61P37/04A61K39/12A61P31/16A61K2039/55594A61K2039/55516A61K2039/543A61K2039/552C12N2760/16134
Inventor 王建忠王春凤杨锐杨桂连杨文涛姜延龙康元环黄海斌石春卫
Owner JILIN AGRICULTURAL UNIV