Asia I-type aftosa synthetic peptide vaccine

A technology for synthesizing peptide vaccines and foot-and-mouth disease, which is applied in the field of vaccines and their preparation, can solve the problems of complex processing of genetically engineered vaccines, limited gene expression products, and high cost, and achieve good application prospects, high immune response levels, and easy large-scale synthesis Effect

Inactive 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

However, due to the limited amount of gene expression products and the inability to obtain the three-dimensional structure of the antigen, it is difficult to improve the immune efficacy
In 2003, Zheng Zhaoxin et al. concatenated the T-cell epitope and B-cell epitope genes on VP1 and VP4 to develop a genetically engineered vaccine against Asia 1 type foot-and-mouth disease, but the test results showed that it could not reach the target level under virulent attack. 100% protection, at the same time as a vaccine, the post-processing of genetically engineered vaccines is too complicated, the cost is high, and the potency is low
At present, a synthetic peptide vaccine against porcine O-type foot-and-mouth disease virus has been successfully developed, but there is no synthetic peptide vaccine against Asian type-1 foot-and-mouth disease virus in the world, and Asian type-1 foot-and-mouth disease virus is one of the most prevalent foot-and-mouth disease viruses in my country. A synthetic peptide vaccine against Asian type 1 foot-and-mouth disease virus with good efficacy, high safety, stable production process and low cost has become an urgent and important task

Method used

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  • Asia I-type aftosa synthetic peptide vaccine
  • Asia I-type aftosa synthetic peptide vaccine
  • Asia I-type aftosa synthetic peptide vaccine

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Example 1: Solid phase synthesis of synthetic peptide vaccine polypeptide antigen

[0034] The polypeptide antigen of the present invention can be prepared by ABI 433A full-automatic polypeptide synthesizer by Merrifield solid-phase synthesis method, wherein Fmoc modified amino acid is used, and the solid-phase carrier is RinkAmide MBHA resin. The production process usually includes solid-phase synthesis of polypeptide antigens, cleavage of polypeptides, purification of antigens and sterilized storage.

[0035] 1. Preparation of synthetic raw materials

[0036] The peptide antigen sequences of the synthetic peptide vaccine are shown in SEQ ID NO: 1, SEQ ID NO: 2, and SEQ ID NO: 3. In addition, the following polypeptides are also included:

[0037] A peptide having an amino acid sequence in which all leucine and valine in the sequence of SEQ ID NO: 1 are substituted with norleucine and norvaline.

[0038] A dimer of a polypeptide having the amino acid sequence shown i...

Embodiment 2

[0051] Embodiment 2: the preparation of synthetic peptide vaccine

[0052] Dilute the five polypeptide antigen solutions prepared in Example 1 to 50 μg / ml with water for injection as the water phase; sterilize the SEPPIC MONTANIDE ISA 50V oil adjuvant at 121°C for 30 minutes, and use it as the oil phase for later use. Under the condition of 22°C, according to the volume ratio of antigen water phase and sterilized 50V as 1:1, first add the oil phase into the emulsification tank, stir at a slow speed of 100 rpm for 2 minutes, slowly add the water phase, after the addition is complete Stir for 30 minutes, then stir at a high speed of 9000 rpm for 20 minutes, then let it stand for 5 minutes, and then obtain the synthetic peptide vaccine against Asian type 1 foot-and-mouth disease virus after packaging, that is, the peptide vaccine of SEQ ID NO: 1, and the peptide of SEQ ID NO: 2 Vaccines, SEQ ID NO: 3 polypeptide vaccine, SEQ ID NO: 1 substituted polypeptide vaccine and SEQ ID NO:...

Embodiment 3

[0053] Example 3: Study on the immune effect of synthetic peptide vaccine on guinea pigs

[0054] 300-500g healthy guinea pigs were randomly divided into seven groups, six guinea pigs in each group, and intramuscularly injected in the hind legs. The injection reagents and doses of each group are shown in Table 1. The second immunization was carried out four weeks after the first immunization, and the third week was challenged after the second immunization.

[0055] Table 1: Injection reagents and doses of experimental animals in each group

[0056]

[0057] 1. Detection of FMDV specific antibody

[0058] Determination of OD of serum antibody by indirect ELISA method 492 On a 96-well microtiter plate, the Asia 1 type FMDV Jiangsu virus was used as the coating antigen (1:10 dilution), the tested guinea pig serum was used as the primary antibody, and the peroxidase-labeled rabbit anti-guinea pig IgG was used as the secondary antibody. The OD value of each well was measured ...

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Abstract

The invention provides Asia I-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 Asia I-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 of aftosa and combining with a computer-aided method to carry out aftosa antigen site analytical prediction; and candidate polypeptide antigens are screened out by carrying out large numbers of animal experiments to obtain polypeptide antigens which have high immunoreaction level and can protect animals from the attack of aftosa epidemic strains. The Asia I-type aftosa synthetic peptide vaccine prepared by the polypeptide antigens can effectively cope with the antigen variation of aftosa virus without causing biosafety problems and has 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 synthetic peptide vaccine for Asian type I foot-and-mouth disease Polypeptide or its polypeptide polymer, 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. Foot-and-mouth disease is caused by foot-and-mouth disease virus, which belongs to the genus of foot-and-mouth disease virus in the family Picornaviridae. It is spherical in shape with a diameter of 30nm and no envelope. Foot-and-mouth disease virus has a simple structure and is composed of single-stranded RNA and protein. It is most abundant in the blister sk...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/00C07K1/34C07K1/06C07K1/04A61K39/00A61K39/39A61P31/14
Inventor 齐鹏肖进郭丽清栗利芳李丙首巴利民
Owner CHINA ANIMAL HUSBANDRY IND
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