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 problems such as inability to effectively protect animals, poor effect, and influence on the use of new vaccines. , to achieve good application prospects, enhance immune effect, and facilitate large-scale synthesis.

Active Publication Date: 2016-08-03
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 engin...

Method used

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  • Synthetic peptide vaccine and preparation method thereof
  • Synthetic peptide vaccine and preparation method thereof
  • Synthetic peptide vaccine and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1、 Embodiment 1

[0039] Example 1, Example 1 Solid-phase synthesis of foot-and-mouth disease synthetic peptide antigen

[0040] 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 the sequences of the foot-and-mouth disease vaccine strains MYA / 98 and OZK / 93, and calculates the frequency of the variation of the main variable amino acid sites. At the same time, combined with computer-aided analysis and prediction of the 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 the vulnerable 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 ...

Embodiment 2

[0071] Example 2. Preparation of synthetic peptide vaccine

[0072] 1. Preparation of antigen aqueous phase

[0073] 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 .

[0074] 2. Preparation of oil phase adjuvant

[0075] The oil phase adjuvant ISA50V was sterilized at 121°C for 30 minutes, and then used.

[0076] 3. Emulsification of synthetic peptide vaccine

[0077] Clean the IKA emulsification equipment (purchased from IKA, Catalog No. 200603) 3 times with 2000 ml 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 water phase a...

Embodiment 3

[0078] Example 3. Efficacy test of synthetic peptide vaccine

[0079] 1. Materials and methods

[0080] 1. Synthetic peptide vaccine

[0081] According to the above-mentioned Example 1, the polypeptide antigens with SEQIDNO:1, SEQIDNO:2, SEQIDNO:3 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.

[0082] In addition, all valines in the amino acid sequence in 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-mentioned examples. The sulfhydryl groups of cystine are oxidized to form a disulfide bond, and the carboxyl group of the head and tail amino ac...

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Abstract

The invention provides a synthetic peptide vaccine and particularly relates to polypeptide or a polypeptide polymer thereof used for the synthetic peptide vaccine for O type foot-and-mouth disease, the vaccine containing the polypeptide or the polypeptide polymer thereof and preparation methods of the polypeptide or the polypeptide polymer thereof and the vaccine, wherein the polypeptide has the amino acid sequence as indicated in SEQ ID No .2 .According to the synthetic peptide vaccine and the preparation method thereof, candidate peptide antigen is screened through a great number of animal tests, foot and mouth disease virus antigen sites are 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 synthetic peptide vaccine for O type foot-and-mouth disease can effectively cope with antigen variation of foot-and-mouth disease virus, biosecurity is good, large-scale synthesis can be conducted easily, and good application prospect is achieved.

Description

[0001] This application is a divisional application of the patent application with the application number "201410081709.9" and the invention title "A synthetic peptide vaccine and its preparation method" Technical field [0002] 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 [0003] Foot and mouth disease (foot and mouth disease, FMD for short) is an acute, highly contact, and febrile infectious disease that occurs in cloven-hoofed animals and is widely distributed worldwide. Because of its high infectivity and rapid spread, infection of pigs, cattle...

Claims

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Application Information

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IPC IPC(8): C07K14/09C07K1/34C07K1/06C07K1/04A61K39/135A61P31/14
CPCA61K39/12A61K2039/54A61K2039/552C07K14/005C12N2770/32134
Inventor 肖进巴利民齐鹏宋芳王楠栗利芳赵洪涛张蕾郑应华
Owner CHINA ANIMAL HUSBANDRY IND
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