Synthetic peptide vaccine as well as preparation method and application thereof

一种合成肽疫苗、口蹄疫的技术，应用在疫苗及它们的制备,O型口蹄疫合成肽疫苗的多肽或其多肽聚合物领域，能够解决不能有效地保护动物、效果差、影响新型疫苗使用等问题，达到良好应用前景、增强免疫效果、易于大规模合成的效果

Active Publication Date: 2016-08-31
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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  • Synthetic peptide vaccine as well as preparation method and application thereof
  • Synthetic peptide vaccine as well as preparation method and application thereof
  • Synthetic peptide vaccine as well as preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1、 Embodiment 1

[0040] Embodiment 1, the solid-phase synthesis of embodiment 1 foot-and-mouth disease synthetic peptide antigen

[0041]The present invention studies the variation of the main antigenic sites of the FMD by determining the sequence of the newly popular strains of the domestic FMD and combining the sequences of the MYA / 98 and OZK / 93 vaccine strains of the FMD, and counts the variation frequency of the main mutated amino acid sites , combined with computer-aided analysis and prediction of antigenic sites of FMD, chemically synthesized peptides of possible antigenic sites, that is, according to the statistical variation frequency of variable sites, different amino acids are used in these sites to obtain coverage of the current Multiple candidate peptide antigens for all possible variant sites. Further, these candidate polypeptide antigens are screened through a large number of animal experiments, and the polypeptide antigens that can cause the immune response of animals, and have ...

Embodiment 2

[0072] Embodiment 2, the preparation of synthetic peptide vaccine

[0073] 1. Preparation of antigen aqueous phase

[0074] First, weigh the three peptide antigens synthesized according to the above-mentioned Example 1; then, dilute the concentration of the synthesized peptide antigen to 50 μg / ml with sterile water for injection; filter the antigen solution through a filter with a pore size of 0.2 μm, and sterilize .

[0075] 2. Preparation of oil phase adjuvant

[0076] Sterilize the oil phase adjuvant ISA50V at 121°C for 30 minutes, and set aside.

[0077] 3. Emulsion of synthetic peptide vaccine

[0078] Wash the IKA emulsification equipment (purchased from IKA Company, product number 200603) with 2000ml of sterilized distilled water for 3 times, and then under the condition of 20-28°C, the oil phase adjuvant and the antigen water phase are in a volume ratio of 1:1, and the oil Add the phase into the emulsification tank, start the motor and stir at a slow speed of 90-15...

Embodiment 3

[0079] Embodiment 3, efficacy test of synthetic peptide vaccine

[0080] 1. Materials and methods

[0081] 1. Synthetic peptide vaccine

[0082] Synthesize polypeptide antigens having the sequences of SEQ ID NO: 1, SEQ ID NO: 2, and SEQ ID NO: 3 according to the above-mentioned Example 1. The sulfhydryl groups of the two cysteines of these polypeptides are oxidized and linked together to form a disulfide bond, and The reaction between the carboxyl and hydroxyl groups of the head and tail amino acids forms a covalent linkage. These are then prepared respectively according to the method of embodiment 2 and corresponding batch number is: the foot-and-mouth disease O type synthetic peptide vaccine of ZM433A01, ZM433A02, ZM433A03.

[0083] In addition, all the valines in the amino acid sequence in SEQ ID NO: 1 were replaced with norvaline; all the leucines were replaced with norleucine, and the antigen was synthesized according to the method provided in the above examples, and th...

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

Description

[0001] This application is a divisional application of a patent application with the application number "201410081709.9" and the title of the invention "a synthetic peptide vaccine and its preparation method" technical field [0002] The present invention relates to a polypeptide or its polypeptide polymer used for foot-and-mouth disease synthetic peptide vaccines, vaccines containing the polypeptide or its polypeptide polymer and their preparation methods, in particular to a polypeptide used for O-type foot-and-mouth disease synthetic peptide vaccines 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 (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, it infects pigs, cattle, sheep and other livestock, resulti...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/09A61K39/135A61P31/14
CPCA61K39/00A61K39/12A61K2039/552A61K2039/55566C07K14/005C12N2770/32122C12N2770/32134
Inventor 肖进巴利民宋芳王楠齐鹏栗利芳赵洪涛张蕾郑应华
Owner CHINA ANIMAL HUSBANDRY IND
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