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Asia I synthetic peptide vaccine of foot and mouth disease

A technology for synthesizing peptide vaccines and foot-and-mouth disease, which is applied in the field of vaccines and their preparation, and can solve the problems of limited gene expression products, complicated processing and disappearance of genetically engineered vaccines, etc.

Active Publication Date: 2010-02-10
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, the Asian type 1 FMD virus is one of the main FMD viruses prevalent in my country. The antigenicity of the FMD strain is still changing. As time goes by, the effectiveness of the original vaccine weakens or even disappears. The prevention and control work has brought great difficulties
Moreover, the degree of antigenic variation within the same serotype is so great that an FMD vaccine effective against one subtype may not be protective against another subtype within the same serotype

Method used

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  • Asia I synthetic peptide vaccine of foot and mouth disease
  • Asia I synthetic peptide vaccine of foot and mouth disease
  • Asia I synthetic peptide vaccine of foot and mouth disease

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Example 1: Solid phase synthesis of synthetic peptide vaccine polypeptide antigen

[0035] 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 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. Preparation of synthetic raw materials

[0037] The polypeptide antigen sequence of the synthetic peptide vaccine is shown in SEQ ID NO:1. In addition, the following polypeptides are also included:

[0038] The amino acid sequence is 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.

[0039] The amino acid sequence is a dimer of the polypeptide with the a...

Embodiment 2

[0052] Embodiment 2: the preparation of synthetic peptide vaccine

[0053] Dilute the three polypeptide antigen solutions prepared in Example 1 to 50 μg / ml with water for injection as the water phase; sterilize the 50V white 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 Asian type 1 foot-and-mouth disease virus synthetic peptide vaccine after packaging, that is, the peptide vaccine of SEQ ID NO: 1, and the peptide substituted by SEQ ID NO: 1 Vaccines and Polypeptide Vaccines of SEQ ID NO: 1.

Embodiment 3

[0054] Example 3: Study on the immune effect of synthetic peptide vaccine on guinea pigs

[0055] Healthy guinea pigs weighing 300-500 g were randomly divided into five groups, six guinea pigs in each group, and intramuscularly injected into the hind legs. See Table 1 for the injection reagents and doses of each group. The second immunization was carried out four weeks after the first immunization, and the third week was challenged after the second immunization.

[0056] Table 1: Immunization antigens and immunization doses of each test group

[0057]

[0058] 1. Detection of FMDV specific antibody

[0059] 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 measur...

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Abstract

The invention provides an Asia I synthetic peptide vaccine of a foot and mouth disease, in particular to a polypeptide or a polypeptide polymer thereof for the Asia I synthetic peptide vaccine of thefoot and mouth disease, the vaccine containing the polypeptide or the polypeptide polymer thereof, and a preparation method of both, wherein, the amino acid sequence of the polypeptide is shown as SEQID NO:1. In the invention, variation situation of main antigen sites of the foot and mouth disease is researched by determining a sequence of an epidemic strain of the foot and mouth disease, and meanwhile analysis prediction of the antigen sites of the foot and mouth disease is performed followed by chemical synthesis on possible peptide segments of the antigen sites combined with a computer auxiliary method; and candidate polypeptide antigens are screened through a large quantity of animal experiments to finally obtain the polypeptide antigens with high immune reaction level which can protect well an animal from being attacked by the epidemic strain of the foot and mouth disease. The vaccine prepared from the polypeptide antigens has the advantages of no biological safety problem, easylarge-scale synthesis, good application prospect and good ability of effectively dealing with antigen variation of foot and mouth disease virus.

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): C07K14/09C07K1/04A61K39/135A61P31/14
CPCY02P20/55
Inventor 肖进齐鹏郭丽清李丙首巴利民
Owner CHINA ANIMAL HUSBANDRY IND
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