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Ferritin-based multi-antigen universal influenza vaccine and preparation method and application thereof

A technology of influenza vaccine and ferritin, applied in the field of ferritin-based multi-antigen universal influenza vaccine and its preparation and application, to achieve broad-spectrum immune protection effect

Active Publication Date: 2019-07-05
INST OF PROCESS ENG CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, whether the structural characteristics of Ferritin can be loaded with antigens, how large and how many antigens can be loaded, and under what conditions can be loaded with antigens are rarely studied.

Method used

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  • Ferritin-based multi-antigen universal influenza vaccine and preparation method and application thereof
  • Ferritin-based multi-antigen universal influenza vaccine and preparation method and application thereof
  • Ferritin-based multi-antigen universal influenza vaccine and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

experiment example 1

[0108] A kind of FRT nanoparticle derived from horse spleen (purchased from Sigma-Aldrich Co. LLC., the FRT nanoparticle carrier described in the following experimental examples 2-14 is the same as experimental example 1) as a carrier, the surface of which is covalently bound with M2e antigen The preparation method of antigen influenza vaccine M2e.FRT is as follows:

[0109] 1) Artificial synthesis of M2e polypeptide antigen derived from A / Puerto Rico / 8 / 1934 H1N1 influenza virus (SEQ ID NO. 1).

[0110] 2) Prepare a FRT carrier solution with a final concentration of 1 mg / mL in 0.1M PBS (pH 7.4), and add 10 times the molar amount of NHS-PEG to it n -Mal crosslinking agent, react for 1h in a shaker at 4°C in the dark to modify the amino end of the FRT with maleimide groups, and then take out and remove the unreacted residual crosslinking agent. The 0.5 mg / mL PEG-FRT solution modified by PEG cross-linking agent was mixed with 5 times the molar amount of the FRT subunit M2e polypeptide...

experiment example 2

[0113] A single antigen influenza vaccine NP.FRT with NP antigen distributed on the surface of FRT nanoparticles.

[0114] Compared with Experimental Example 1, in addition to the artificially synthesized NP polypeptide antigen derived from A / Puerto Rico / 8 / 1934H1N1 influenza virus (SEQ ID NO. 2) instead of M2e antigen for covalent cross-linking with FRT, its preparation method Same as experimental example 1. Finally, the NP.FRT single antigen influenza vaccine with NP antigen covalently bound on the surface is prepared, and the covalent binding rate of the surface NP antigen is about 15 NP antigen polypeptide molecules per NP.FRT molecule. (This example serves as a comparative example)

[0115] SEQ ID NO.2: QIASNENMETMESSTL-C

experiment example 3

[0117] A single antigen influenza vaccine HA.FRT with HA full-length protein antigen covalently bound to the surface of FRT as a carrier, the preparation method is as follows:

[0118] 1) Recombinant expression of A / Puerto Rico / 8 / 1934H1N1 influenza virus HA full-length protein (SEQ ID NO. 3), and purification, the final purity is more than 90%.

[0119] 2) According to the method described in Experimental Example 1, prepare a FRT carrier solution with a final concentration of 1 mg / mL with 0.1M PBS (pH 7.4), and add 10 times the molar amount of NHS-PEG to it n -Mal crosslinking agent, react for 1h in a shaker at 4°C in the dark to modify the amino end of the FRT with maleimide groups, and then take out and remove the unreacted residual crosslinking agent. PEG-FRT solution modified by PEG cross-linking agent 0.5mg / mL, the same as 2 times the molar amount of FRT tris(2-carboxyethyl)phosphine-hydrochloride (TCEP·HCl) reduction treatment of recombinant HA antigen Mix and react on a shak...

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Abstract

The invention provides a ferritin-based multi-antigen universal influenza vaccine and a preparation method and application thereof. The ferritin-based multi-antigen universal influenza vaccine comprises ferritin nanoparticle carrier proteins, surface antigens and inner cavity antigens, the surface antigens are distributed on surfaces of ferritin nanoparticles, and the inner cavity antigens are distributed in inner cavities of the ferritin nanoparticles. The ferritin-based multi-antigen universal influenza vaccine structurally simulates natural spatial conformation of influenza viruses, presents various influenza virus antigens and is capable of simultaneously giving play to immunogenicity of the various influenza virus antigens to stimulate comprehensive immunological effects, and broad-spectrum anti-influenza virus immune protection effects can be provided. In addition, a ferritin carrier extensively exists in living organisms including human, mammals and the like, is theoretically high in safety when serving as the vaccine carrier and has probability of applicability to elderly people and children.

Description

Technical field [0001] The invention belongs to the technical field of biomedicine, and relates to an influenza vaccine and a preparation method and application thereof, in particular to a multi-antigen bionic influenza vaccine and a preparation method and application thereof, and mainly relates to a multi-antigen universal influenza vaccine based on ferritin and The preparation method and application thereof specifically relate to an influenza vaccine that loads antigen proteins derived from influenza virus on the outer surface of ferritin nanoparticles and on the inside of the hollow chamber of ferritin nanoparticles to simulate the natural structure of influenza virus and the influenza vaccine and Preparation method and application. Background technique [0002] As one of the most serious infectious diseases, the prevention of influenza has always been a major problem that plagues mankind. At present, the most effective measure to prevent and treat influenza is still inoculat...

Claims

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Application Information

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IPC IPC(8): A61K39/295A61K39/385A61K39/145A61P31/16
CPCA61K39/12A61K39/385A61K2039/6031A61K2039/70A61P31/16
Inventor 张松平苏志国魏江雪李正军杨延丽
Owner INST OF PROCESS ENG CHINESE ACAD OF SCI
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