Synthetic peptide vaccine and preparation method thereof

A technology for synthesizing peptide vaccines and vaccines, which is applied in the field of vaccines and their preparation, peptides of O-type foot-and-mouth disease synthetic peptide vaccines or their polypeptide polymers, 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.

Inactive Publication Date: 2014-04-02
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 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

[0034] The solid-phase synthesis of embodiment 1 foot-and-mouth disease synthetic peptide antigen

[0035] The polypeptide antigen of the present invention can be prepared by a Merrifield solid-phase synthesis method using an automatic polypeptide synthesizer, wherein 9-fluorenylmethoxycarbonyl (Fmoc)-modified amino acids are used, and the solid-phase carrier is Rink Amide MBHA resin. The production process usually includes solid-phase synthesis of polypeptide antigens, cleavage of polypeptides, purification of antigens and sterilized storage.

[0036] 1.1 Solid phase synthesis of polypeptide antigen

[0037] 1.1.1 Preparation of synthetic raw materials

[0038] The sequences of the synthetic polypeptide antigens are respectively shown in SEQ ID NO: 1, SEQ ID NO: 2, and SEQ ID NO: 3.

[0039] According to the sequence of the antigen and the synthesis scale, 1 mmol of the appropriate Fmoc-modified amino acid was prepared and added to the corresponding amino acid vial. Also w...

Embodiment 2

[0065] Example 2. Preparation of synthetic peptide vaccines

[0066] 2.1 Preparation of antigen aqueous phase

[0067] 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 .

[0068] 2.2 Preparation of oil phase adjuvant

[0069] Sterilize the oil-phase adjuvant at 121°C for 30 minutes, and set aside.

[0070] 2.3 Emulsion of synthetic peptide vaccine

[0071] Clean the IKA emulsification equipment with 2000ml of sterilized distilled water for 3 times, then put the oil phase into the emulsification tank at 20-28°C according to the volume ratio of oil phase adjuvant and antigen water phase of 1:1, and start the motor to After stirring at a slow speed of 90-150r / m for 2 minutes, slowly add the water-phase antigen at the same time, stir f...

Embodiment 3

[0072] Efficacy test of embodiment 3 synthetic peptide vaccine

[0073] 1. Materials and methods

[0074] 1.1 Synthetic peptide vaccine

[0075] Synthesize peptide antigens with SEQ ID NO: 1, SEQ ID NO: 2, and SEQ ID NO: 3 sequences according to the above-mentioned examples, and then prepare the corresponding batch numbers: ZM433A01, ZM433A02, and ZM433A03 FMD O-type synthetic peptide vaccines .

[0076] 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 was synthesized according to the method provided in the above examples. The batch number of the prepared vaccine was: ZM433A04.

[0077] According to the sequence of SEQ ID NO: 1, the dimer antigen of this sequence was synthesized by conventional synthetic peptide technology, and the vaccine was prepared to obtain the synthetic peptide vaccine with batch number: ZM433A05.

[0078] 1.2 Test animals

[0079] Select 27 ...

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Abstract

The invention provides aftosa synthetic peptide vaccine, more specifically to polypeptide or polypeptide polymer used for O type aftosa synthetic peptide vaccine, vaccine containing the polypeptide orpolypeptide polymer and a preparation method thereof. The polypeptide has amino acid sequence represented by SEQ ID NO.1, SEQ ID NO.2 or SEQ ID NO.3. By sequencing of recent aftosa strain in China an d combination of aftosa vaccine strain OS / 99 and OZK / 93 sequence, the invention researches variation case of main antigen site, carries out antigen site analysis and forecast assisted by computer, andcarries out chemical synthesis to possible antigen site peptide segment. The invention screens polypeptide antigen through a large amount of animal tests, optimizes aftosa virus antigen site accordin g to screening result, effectively combines T cell epitope and B cell epitope, and enhances immunization effect of the polypeptide antigen. The O type aftosa synthetic peptide vaccine can effectively reply antigenic variation of aftosa virus, has good biological safety, is easy for large scale production, and has better application prospect.

Description

technical field [0001] 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 [0002] 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, resulting in the death of young animals and a sharp decline in the production capacity of adult animals, seriously endangering the development of animal husbandry and the production and supply of meat and animal produc...

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
Inventor 郭丽清齐鹏肖进巴利民宋芳董春娜郑应华
Owner CHINA ANIMAL HUSBANDRY IND
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