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Bovine A-type foot-and-mouth disease broad-spectrum multi-epitope vaccine, and preparation method and application thereof

A foot-and-mouth disease and multi-epitope technology, which is applied in chemical instruments and methods, pharmaceutical formulations, recombinant DNA technology, etc., can solve the problems of biological security acceleration and achieve the effect of protection against attacks and good application prospects

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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  • Bovine A-type foot-and-mouth disease broad-spectrum multi-epitope vaccine, and preparation method and application thereof
  • Bovine A-type foot-and-mouth disease broad-spectrum multi-epitope vaccine, and preparation method and application thereof
  • Bovine A-type foot-and-mouth disease broad-spectrum multi-epitope vaccine, and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

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

[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 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, the domestic popular strains A / WH / 09, A / XJKT / 58 and GSLX / 62 were compared with the representative strains of bovine type A foot-and-mouth disease virus (NCBI accession number: FJ755067) recently isolated from some countries bordering my count...

Embodiment 2

[0037] Preparation of Example 2 Bovine Type A Foot-and-Mouth Disease Broad-spectrum Multi-epitope Vaccine and Analysis of Immunity Efficacy

[0038] 1. Vaccine preparation:

[0039] The purified protein (shown in SEQ ID NO.6, prepared according to the method in Example 1) was quantified by the Bio-Rad quantitative kit and diluted to an appropriate concentration, alone or with the purified 3D protein fragment (amino acid sequence as shown in SEQ ID NO.8 As shown, the nucleotide sequence encoding it (as shown in SEQ ID NO.7) and the recombinant antigen are configured at a ratio of 1:2 (V / V), and an equal volume of oil adjuvant ISA206 (France) is added 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 ...

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Abstract

The invention discloses a bovine A-type foot-and-mouth disease broad-spectrum multi-epitope recombinant vaccine, and a preparation method and an application thereof, and belongs to the field of veterinary vaccine research. The preparation method comprises the following steps: carrying out reasonable serial connection on the major antigen epitopes of pandemic in China and bovine A-type foot-and-mouth disease virus representative strains recently pandemic in countries bordering China by adopting a brand new reverse vaccinology idea and strategy, coupling with a bovine IgG heavy chain constant region, cloning into a prokaryotic expression vector to construct a recombinant expression vector, transforming Escherichia coli cells to express a recombinant antigen, purifying by adopting Ni-NAT column chromatography, quantifying by a Bio-Rad protein quantification kit, and preparing the vaccine individually or combining with a recombinant foot-and-mouth disease virus 3D protein. A result of animal immune experiments shows that the multi-epitope antigen or 3D protein combined vaccine can stimulate the body to produce a protective antibody with high titer and also can protect immune animals against a virus attack, so the bovine A-type foot-and-mouth disease broad-spectrum multi-epitope recombinant vaccine has a 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 bovine type A foot-and-mouth disease broad-spectrum multi-epitope recombinant vaccine 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 outbreaks. However, the production of such vaccines requires the construction of high-level biosafety production workshops to prevent pathogens from escaping, and to distingu...

Claims

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Application Information

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IPC IPC(8): C07K19/00C12N15/62C12N15/63A61K39/135A61K48/00A61P31/14
Inventor 常惠芸邵军军林彤丛国正独军政高闪电
Owner LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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