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Broad-spectrum multi-epitope recombinant vaccine for bovine foot-and-mouth disease virus type A strain epidemic abroad, and preparation method and application thereof

A technology of foot-and-mouth disease vaccine and epidemic strains, which is applied in the field of veterinary vaccine research and can solve the problems of biosafety acceleration

Active Publication Date: 2014-10-29
LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Uncertainty in the biosafety of traditional inactivated vaccines has accelerated the process of researching new vaccines against foot-and-mouth disease

Method used

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  • Broad-spectrum multi-epitope recombinant vaccine for bovine foot-and-mouth disease virus type A strain epidemic abroad, and preparation method and application thereof
  • Broad-spectrum multi-epitope recombinant vaccine for bovine foot-and-mouth disease virus type A strain epidemic abroad, and preparation method and application thereof
  • Broad-spectrum multi-epitope recombinant vaccine for bovine foot-and-mouth disease virus type A strain epidemic abroad, and preparation method and application thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] The preparation of embodiment 1 broad-spectrum multi-epitope recombinant antigen of bovine type A foot-and-mouth disease epidemic strain abroad

[0030] 1. Bioinformatics analysis of VP1 gene of type A foot-and-mouth disease virus:

[0031] The structural protein VP1 of foot-and-mouth disease virus is the dominant antigen of the virus. Whether it is the isolated and purified natural VP1 protein or the recombinant expression product, it can induce the body to produce protective neutralizing antibodies, which is type-specific. The VP1 gene of type A foot-and-mouth disease virus has a full length of 639 nucleotides, encoding a protein consisting of 213 amino acids, and its main antigenic epitopes are concentrated in the 140-160 amino acid and 200-213 amino acid segments. In this study, DNAStar biological software was used to analyze the sequence of representative bovine type A foot-and-mouth disease virus strains (NCBI accession numbers: FJ755067, HQ666886 and AY59371) rec...

Embodiment 2

[0037] Example 2 Preparation of broad-spectrum multi-epitope vaccine of bovine type A foot-and-mouth disease epidemic strains abroad and analysis of immune efficacy

[0038] 1. Vaccine preparation:

[0039] The purified protein was quantified by the Bio-Rad quantitative kit and diluted to an appropriate concentration, alone or with the purified 3D protein full-length fragment (the amino acid sequence is shown in SEQ ID NO.8, and the nucleotide sequence encoding it is shown in SEQ ID NO .7) and the recombinant antigen at a ratio of 1:2 (V / V), then added an equal volume of oil adjuvant ISA206 (France) to emulsify into a vaccine preparation.

[0040] 2. Guinea pig immune efficacy test:

[0041] Using the prepared vaccine, 0.2ml of each guinea pig was inoculated by intramuscular route (20μg antigen or 20μg antigen + 10μg 3D protein) to immunize 5 guinea pigs respectively. The results of the immunization experiment showed that no matter whether the recombinant antigen alone or com...

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Abstract

The invention discloses a broad-spectrum multi-epitope recombinant vaccine for bovine foot-and-mouth disease virus type A strain epidemic abroad and a preparation method and application thereof, and belongs to the field of veterinary vaccine research. Adopting the ''antigenized antibody'' strategy and a novel reverse vaccine operation technique and strategy, the method is as below: conducting reasonable combination of the dominant antigen epitopes of representative strains of bovine foot-and-mouth disease virus type A recently epidemic in countries bordering with China; conducting coupling of the epitopes with bovine IgG heavy chain constant region; cloning into a prokaryotic expression vector to construct a recombinant expression vector; transforming into E.coli competent cell to express the recombinant antigen, and purifying by using an Ni-NAT chromatographic column; quantifying by a Bio-Rad protein quantification kit; preparing the vaccine by using the antigen individually or combining the antigen with a recombinant foot-and-mouth disease virus 3D protein. Animal immune experiment results show that the recombinant protein or vaccine after compatibility can stimulate the body to produce protective antibody with high potency and at the same time protect the immune animals against homologous virus attacks; therefore, the invention has good application prospect.

Description

technical field [0001] The invention relates to a bovine type A foot-and-mouth disease vaccine and its preparation method and application, in particular to a broad-spectrum multi-epitope recombinant vaccine of bovine type A foot-and-mouth disease epidemic strains abroad and its preparation method and application, belonging to the field of veterinary vaccine research. Background technique [0002] Foot-and-mouth disease is a major animal disease of pigs, cattle and sheep as the main economic livestock, which not only seriously threatens the healthy development of animal husbandry, but also involves the safety of animal-derived food and its foreign trade export. Once an epidemic occurs, the losses will be heavy and the political impact will be bad. Inactivated vaccines play a very important role in the prevention and control of FMD, but due to the biological safety hazards in the production of such vaccines, it is necessary to distinguish between vaccine immunity and naturally...

Claims

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Application Information

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IPC IPC(8): C07K19/00C12N15/62C12N15/63C12N1/21A61K39/135A61K48/00A61P31/14
Inventor 邵军军常惠芸陈建文
Owner LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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