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O-type aftosa synthetic peptide vaccine

A technology for synthesizing peptide vaccines and foot-and-mouth disease, which is applied in the field of polypeptides or their polypeptide polymers, can solve problems affecting the use of new vaccines, achieve good application prospects, high immune response levels, and enhance immune effects

Active Publication Date: 2010-03-03
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 engineered vaccines for foot-and-mouth disease. use of vaccines

Method used

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  • O-type aftosa synthetic peptide vaccine

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] The solid-phase synthesis of embodiment 1 foot-and-mouth disease synthetic peptide antigen

[0034] The polypeptide antigen of the present invention can be prepared by using ABI 433A automatic polypeptide synthesizer and Merrifield solid-phase synthesis method, wherein 9-fluorenylmethoxycarbonyl (Fmoc) modified amino acid is 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.

[0035] 1.1 Solid phase synthesis of polypeptide antigen

[0036] 1.1.1 Preparation of synthetic raw materials

[0037] The sequences of the synthetic polypeptide antigens are respectively shown in SEQ ID NO: 1, SEQ ID NO: 2, and SEQ ID NO: 3.

[0038] 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. ...

Embodiment 2

[0063] Example 2. Preparation of synthetic peptide vaccines

[0064] 2.1 Preparation of antigen aqueous phase

[0065] 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 .

[0066] 2.2 Preparation of oil phase adjuvant

[0067] Sterilize the oil-phase adjuvant at 121°C for 30 minutes, and set aside.

[0068] 2.3 Emulsion of synthetic peptide vaccine

[0069] 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

[0070] Efficacy test of embodiment 3 swine foot-and-mouth disease O type synthetic peptide vaccine

[0071] 1. Materials and methods

[0072] 1.1 Synthetic peptide vaccine

[0073] Synthesize polypeptide 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 swine foot-and-mouth disease O-type synthetic peptides vaccine.

[0074] 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.

[0075] 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 n...

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Abstract

The invention provides O-type aftosa synthetic peptide vaccine, and in particular relates to polypeptide or polypeptide polymer thereof used in the vaccine as well as the vaccine containing the polypeptide or the polypeptide polymer thereof and a preparation method thereof. The polypeptide has amino acid sequences shown in SEQ ID No.1, SEQ ID No.2 and SEQ ID No.3. The O-type aftosa synthetic peptide vaccine carries out chemical synthesis of potential antigen site peptide segments by carrying out sequencing of domestic aftosa epidemic strains to study the variation of the main antigen sites ofaftosa and combining with computer assistant to carry out antigen site analytical prediction. Candidate polypeptide antigens are screened out by carrying out large numbers of animal experiments and aftosa virus antigen sites are optimized according to the screening result; and T cell epitope and B cell epitope are effectively combined to improve the immune effects of the polypeptide antigens. TheO-type aftosa synthetic peptide vaccine can effectively cope with the antigen variation of aftosa virus and has ideal biosafety and easy large-scale synthesis, thereby having a good 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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IPC IPC(8): C07K14/00C07K1/06C07K1/04A61K39/00A61P31/14
Inventor 齐鹏肖进栗利芳郭丽清宋芳
Owner CHINA ANIMAL HUSBANDRY IND
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