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Bovini Asia 1/O type foot-and-mouth disease bivalent multi-epitope vaccine and preparation method and application thereof

A multi-epitope, foot-and-mouth disease virus technology, applied in chemical instruments and methods, DNA/RNA fragments, pharmaceutical formulations, etc., can solve problems such as inability to provide cross-immune protection, and achieve reduced immunization costs, maintaining immunogenicity, and comprehensive protection Effect

Active Publication Date: 2014-07-02
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

These are vaccines against a single serotype and do not provide cross-immune protection

Method used

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  • Bovini Asia 1/O type foot-and-mouth disease bivalent multi-epitope vaccine and preparation method and application thereof
  • Bovini Asia 1/O type foot-and-mouth disease bivalent multi-epitope vaccine and preparation method and application thereof
  • Bovini Asia 1/O type foot-and-mouth disease bivalent multi-epitope vaccine and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Example 1 Preparation of bovine Asia1 / O type foot-and-mouth disease virus composite multi-epitope recombinant antigen (Asia1 / O / BCIgG)

[0038] 1. Bioinformatics analysis of O-type foot-and-mouth disease virus VP1 gene:

[0039] 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. In this study, DNAStar biological software was used to analyze the sequence of the representative strain O / China / 99 (GenBank accession number: AF506822) of bovine O-type foot-and-mouth disease virus isolated in my country, and the dominant antigenic epitope was determined to be 140-160 amino acids and 200-213 amino acids For another B antigen epitope. At the same time, 21-40 amino acids of VP1 protein region and 20-34 amino acids of VP4 were selected as T cell e...

Embodiment 2

[0053] Example 2 Preparation of bovine Asia1 / O type foot-and-mouth disease bivalent multi-epitope vaccine and immune efficacy experiment

[0054] 1. Vaccine preparation:

[0055] The purified protein Asia1 / O / BCIgG obtained in Example 1 was quantified by the Bio-Rad quantitative kit and then diluted to 1 mg / ml, either alone or with the purified full-length 3D protein (shown in SEQ ID NO: 10) and recombinant antigen by 1 : 2 (concentration ratio) after preparation, add an equal volume of oil adjuvant ISA206 (France) to emulsify into a vaccine preparation.

[0056] 2. Guinea pig immune efficacy test:

[0057] With the prepared vaccine, 0.5ml of each guinea pig was inoculated by intramuscular route (50μg antigen or 50μg antigen + 25μg 3D protein) to immunize 5 guinea pigs respectively. The results of the immunization experiment showed that after 28 days of the first immunization, whether it was Asia1 / O / BCIgG recombinant Guinea pigs immunized with antigen alone or combined with 3...

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Abstract

The invention discloses a bovini Asia 1 / O type foot-and-mouth disease bivalent multi-epitope vaccine and a preparation method and an application thereof and belongs to the field of veterinary biological vaccines. The bovini Asia 1 / O type foot and mouth disease virus compound multi-epitope recombinant antigen is obtained by coupling dominant epitope of epidemic bovini O and Asia 1 type foot-and-mouth disease virus representative strains connected in series in China with a bovini IgG heavy chain constant region. Immune efficacy experiments verify that the bovini Asia 1 / O type foot-and-mouth disease bivalent multi-epitope vaccine prepared by the invention has good immunogenicity, and high level protective antibodies can be generated by the body induced by immune guinea pig or immune bovine. The inoculated vaccine is safe and harmless to immune animals. The vaccine is a novel vaccine which is abroad in prospect, supports matter storage and technical support to prevention and control of bovini Asia 1 / O type foot-and-mouth disease in China, and is of great importance.

Description

technical field [0001] The invention relates to a bovine foot-and-mouth disease virus recombinant antigen and its preparation method and application, in particular to a bovine Asia1 / O type foot-and-mouth disease bivalent multi-epitope vaccine and its preparation method and application. The invention belongs to the field of veterinary biological vaccines. Background technique [0002] Foot-and-mouth disease, as a major animal disease that infects pigs, cattle and sheep as the main economic livestock, 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, it will suffer heavy losses and have a bad political impact. Inactivated vaccines play a very important role in the prevention and control of FMD outbreaks. However, the production of such vaccines requires the construction of high-level biosafety production workshops to prevent pathogens from escaping...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K19/00C12N15/62C12N15/70A61K39/135A61K48/00A61P31/14
Inventor 邵军军常惠芸
Owner LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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