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A kind of synthetic peptide vaccine and preparation method thereof

A technology for synthesizing peptide vaccines and foot-and-mouth disease, which is applied in the field of vaccines and their preparation, polypeptides of O-type foot-and-mouth disease synthetic peptide vaccines or their polypeptide polymers, and can solve the problems of ineffective protection of animals, affecting the use of new vaccines, and poor effects. , to achieve good application prospects, easy large-scale synthesis, and the effect of enhancing immune effects

Active Publication Date: 2016-07-13
CHINA ANIMAL HUSBANDRY IND
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In terms of research on new vaccines for foot-and-mouth disease, there have been reports on genetically engineered subunit vaccines, foot-and-mouth disease virus vector vaccines, and genetically engineered vaccines for foot-and-mouth disease. use of vaccines
In addition, these vaccines are often less effective against currently circulating strains that have mutated and cannot effectively protect animals

Method used

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  • A kind of synthetic peptide vaccine and preparation method thereof
  • A kind of synthetic peptide vaccine and preparation method thereof
  • A kind of synthetic peptide vaccine and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1、 Embodiment 1

[0036] Example 1, Example 1 Solid-phase synthesis of foot-and-mouth disease synthetic peptide antigen

[0037] The present invention studies the variation of the main antigenic sites of foot-and-mouth disease by determining the sequence of the newly popular strains of foot-and-mouth disease in China and combining with the sequences of the foot-and-mouth disease vaccine strains MYA / 98 and OZK / 93. At the same time, combined with computer-assisted analysis and prediction of antigenic sites of foot-and-mouth disease, chemical synthesis of peptides of possible antigenic sites is carried out. That is, according to the statistical mutation frequency of variable sites, different amino acids are used at these sites to obtain coverage of the current Multiple candidate polypeptide antigens at all possible variant sites. Furthermore, these candidate polypeptide antigens are screened through a large number of animal experiments to obtain polypeptide antigens that can cause an animal's immune ...

Embodiment 2

[0068] Example 2. Preparation of synthetic peptide vaccine

[0069] 1. Preparation of antigen aqueous phase

[0070] First, weigh the three polypeptide antigens synthesized according to the above-mentioned Example 1; then, dilute the synthetic peptide antigen concentration to 50μg / ml with sterile water for injection; filter the antigen solution through a filter with a pore size of 0.2μm to sterilize .

[0071] 2. Preparation of oil phase adjuvant

[0072] The oil phase adjuvant ISA50V was sterilized at 121°C for 30 minutes, and then used.

[0073] 3. Emulsification of synthetic peptide vaccine

[0074] Clean the IKA emulsification equipment (purchased from IKA company, catalog number 200603) 3 times with 2000ml of sterilized distilled water, and then at 20-28 ℃ according to the volume ratio of the oil phase adjuvant and the antigen water phase of 1:1, the oil Add the phase into the emulsification tank, start the motor and stir at 90~150r / m slowly for 2 minutes, then slowly add the wate...

Embodiment 3

[0075] Example 3. Efficacy test of synthetic peptide vaccine

[0076] 1. Materials and methods

[0077] 1. Synthetic peptide vaccine

[0078] According to the above-mentioned Example 1, the polypeptide antigens with SEQIDNO:1, SEQIDNO:2, SEQIDNO:3 sequence were synthesized. The sulfhydryl groups of the two cysteine ​​groups of these polypeptides were oxidized to form a disulfide bond. The reaction forms a covalent connection. These were then formulated according to the method of Example 2 and the corresponding batch numbers were: ZM433A01, ZM433A02, ZM433A03, foot-and-mouth disease O-type synthetic peptide vaccine.

[0079] In addition, all valines in the amino acid sequence of SEQ ID NO: 1 were replaced with norvaline; all leucines were replaced with norleucine, and the antigen synthesis was carried out according to the method provided in the above examples. The same two halves in the antigen The sulfhydryl groups of cystine are oxidized to form a disulfide bond, and the carboxyl g...

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Abstract

The invention provides a synthetic peptide vaccine for a foot-and-mouth disease, and particularly relates to a polypeptide for an O-type synthetic peptide vaccine for a foot-and-mouth disease and a polypeptide polymer thereof, a vaccine containing the polypeptide or the polypeptide polymer thereof, and preparation methods thereof. The polypeptide comprises an amino acid sequence shown in SEQ ID NO.1, SEQ ID NO.2 or SEQ ID NO.3. The candidate polypeptide antigens are screened by a lot of animal tests, the antigen site of a foot and mouth disease virus is optimized according to the screening result, T cell epitope and B cell epitope are effectively combined, and the immune effect of the polypeptide antigen is enhanced. The antigenic variation of the foot-and-mouth virus can be effectively coped with by the O-type synthetic peptide vaccine for the foot-and-mouth disease, and the synthetic peptide vaccine is good in biosecurity, and easy to synthetize at a large scale, and has good application prospect.

Description

Technical field [0001] The present invention relates to a polypeptide or its polypeptide polymer used in a foot-and-mouth disease synthetic peptide vaccine, a vaccine containing the polypeptide or its polypeptide polymer, and a preparation method thereof, in particular to a polypeptide used in a type O foot-and-mouth disease synthetic peptide vaccine Or its polypeptide polymer, as well as vaccines containing the polypeptide or its polypeptide polymer, and their preparation methods. Background technique [0002] Foot and mouth disease (foot and mouth disease, referred to as FMD) is an acute, highly contagious, febrile infectious disease that occurs in cloven-hoofed animals and is widely distributed worldwide. Due to its high infectivity and rapid spread, infection of pigs, cattle, sheep and other livestock causes the death of young animals, and the production capacity of adult animals declines sharply, which seriously endangers the development of animal husbandry and the productio...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/09A61K39/135A61P31/14C07K1/06C07K1/04
CPCA61K39/00A61K39/12A61K2039/552A61K2039/55566C07K14/005C12N2770/32122C12N2770/32134
Inventor 巴利民肖进齐鹏宋芳王楠栗利芳赵洪涛张蕾周强郑应华
Owner CHINA ANIMAL HUSBANDRY IND
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