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Preparation method of broad-spectrum vaccine

A vaccine and broad-spectrum technology, applied in the field of preparation of broad-spectrum vaccines, can solve problems such as the inability to present antigens, the denaturation of the original structure of viral antigens, and the instability of protein antigens

Pending Publication Date: 2022-03-04
张公义
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0014] Second, due to the instability of protein antigens in vivo (protein antigens are unstable in vitro or in vivo), follicular dendritic cells may no longer be able to present antigens in their native form when needed (antigens have been denatured in vivo or be digested)
[0015] Finally, the traditional vaccine preparation process, especially where formalin or propiolactone is used, takes several days or more due to the long inactivation process, which can denature the original structure of the viral antigen

Method used

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  • Preparation method of broad-spectrum vaccine

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0092] Example 1 - Generation of stable protein antigens resistant to extreme harsh conditions

[0093] Applicants have analyzed and elucidated many fundamental mechanisms of protein folding and unfolding. The disruption of hydrogen bonds was found to be the driving force for disrupting the three-dimensional structure of proteins (Wang et al., 2014). It was later found that hydrogen bonds are the main driving force for the initial folding of proteins (Lee, 2017). However, the three-dimensional structure of proteins is mostly maintained by weak hydrophobic interactions (van der Waals forces). These findings suggest that the three-dimensional structure of proteins is fragile and therefore susceptible to environmental changes that lose their original, normal structure. Small perturbations in the protein environment can lead to denaturation of the protein's three-dimensional structure. Formalin (formaldehyde), β-propiolactone (BPL), extreme pH values, certain solution component...

Embodiment 2

[0104] Example 2—Preparation of vaccines with stable antigens of influenza A strains.

[0105] Next, Applicants investigated treating influenza A virus with glutaraldehyde to produce a vaccine with stable antigens.

[0106] Specific steps are as follows:

[0107] (1), obtain H1N1 influenza virus, this H1N1 influenza virus comes from Charles River (product number: 10100374, Influenza A / PR / 8 / 34);

[0108] (2), the H1N1 influenza virus obtained in step (1) is diluted with a low-temperature buffer, and enters step (3) after its concentration is below 1.0 mg / ml;

[0109] (3), the cross-linking agent is diluted with a low-temperature buffer, and the mass concentration of the cross-linking agent is 0.005% before entering step (4);

[0110] (4), the H1N1 influenza virus dilution obtained in step (2) is diluted twice with the cross-linking agent dilution obtained in step (3), and enters step (5) after the H1N1 influenza virus concentration is 0.01mg / ml;

[0111] (5), place the patho...

Embodiment 3

[0143] Example 3—Preparation of Antigen-stabilized Type B (B) Influenza Strain Vaccines

[0144] A method similar to Example 2 was used to prepare type B (B) strain influenza vaccine. In addition, vaccines were produced using traditional methods (ie, using formaldehyde to kill viruses and prepare vaccines) as a control. Type B (B) strain virus was also obtained from Charles River (USA) company (product number: 10100379, B / Lee / 40).

[0145] Two batches of virus were prepared as follows:

[0146] Preparation of the first batch of influenza B virus vaccines - adopt the preparation method disclosed by the invention to prepare the influenza B (B) influenza strain vaccine of stable antigen, comprising the following steps:

[0147] (1), obtain the live type B (B) strain virus of purification (purchase from Charles River Company);

[0148] (2), the live virus obtained in step (1) is diluted with a low-temperature buffer, and enters step (3) after its concentration is below 1.0 mg / m...

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Abstract

The invention relates to a preparation method of a broad-spectrum vaccine. The preparation method comprises the following steps: combining a cross-linking agent with a pathogen; or contacting a cross-linking agent with a recombinant protein from a pathogen and allowing them to form a new protein internal covalent bond; if the pathogen is adopted, inactivation needs to be carried out, then the cross-linking agent which is not combined with the protein is removed, and if the recombinant protein from the pathogen is adopted, the cross-linking agent which is not combined with the protein is directly removed; separating the cross-linking agent modified pathogen or recombinant protein; and combining the cross-linking agent modified pathogen or recombinant protein with an adjuvant to obtain the broad-spectrum vaccine. In the maturing and hypermutation periods of B cells in a hair growth center, follicular dendritic cells present antigens with stable original ecological structures to the B cells in the hair growth center. Therefore, the B cells of the hair growth center have sufficient time to repeatedly circulate and mutate in the hair growth center, and finally a broad-spectrum immunoglobulin G antibody with high affinity with the original antigen is generated.

Description

technical field [0001] The invention belongs to the technical field of biological medicines, and in particular relates to a preparation method of a broad-spectrum vaccine. Background technique [0002] Judging from the records of the last two centuries, influenza epidemics have brought disasters to mankind. The 1918 epidemic infected about 500 million people and killed about 50 to 100 million people worldwide. [0003] In the United States, current flu vaccines cost about $2.5 to $3 billion per year and are expected to reach about $5 billion by 2025. Although corresponding influenza vaccines have also been developed against influenza, however, due to the limitations of these vaccines, they are only about 50% effective. In this regard, annual direct medical costs in the United States for treating people with influenza infection can reach approximately $10.4 billion, and annual indirect costs are approximately $87 billion. Worldwide costs can be as much as 10 times these fi...

Claims

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Application Information

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IPC IPC(8): A61K39/215A61K39/145A61K39/21A61K39/00A61K39/02A61K39/12A61K39/39A61K41/17A61K47/18A61P31/04A61P31/10A61P31/12A61P31/14A61P31/16
CPCA61K39/12A61K39/02A61K39/0002A61K39/39A61K41/17A61K47/183A61P31/12A61P31/14A61P31/16A61P31/04A61P31/10A61K2039/55505C12N2770/20034C12N2760/16134C12N2760/16234
Inventor 张公义
Owner 张公义
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