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Seneca virus type A genetic engineering composite epitope protein, vaccine and application of Senecavirus type A genetic engineering composite epitope protein and vaccine

A compound epitope and genetic engineering technology, applied in the direction of antiviral immunoglobulin, positive-sense single-stranded RNA virus, virus, etc.

Active Publication Date: 2021-10-22
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

At present, the research on SVA epitope vaccine has not been reported in the literature, and the research and development of safe and efficient SVA epitope vaccine has important scientific and application value

Method used

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  • Seneca virus type A genetic engineering composite epitope protein, vaccine and application of Senecavirus type A genetic engineering composite epitope protein and vaccine
  • Seneca virus type A genetic engineering composite epitope protein, vaccine and application of Senecavirus type A genetic engineering composite epitope protein and vaccine
  • Seneca virus type A genetic engineering composite epitope protein, vaccine and application of Senecavirus type A genetic engineering composite epitope protein and vaccine

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preparation example Construction

[0048] The invention provides a preparation method of the type A Seneca virus genetic engineering composite epitope protein vaccine, comprising the following steps:

[0049] Dissolving the type A Seneca virus genetically engineered complex epitope protein with PBS buffer, mixing with adjuvant and emulsifying to obtain the vaccine.

[0050] In the present invention, after the dissolution, the concentration of the Seneca virus type A genetically engineered compound epitope protein solution is 500-600 μg / mL, more preferably 500 μg / mL. The volume ratio of the type A Seneca virus genetic engineering compound epitope protein solution and the adjuvant is preferably 1:1. The method of the emulsification is not particularly limited in the present invention, and emulsification methods well known in the art can be used.

[0051] The invention also provides a type A Seneca virus neutralizing antibody, which is obtained by immunizing piglets with the type A Seneca virus genetic engineerin...

Embodiment 1

[0058] 1. Design of SVA compound epitope gene

[0059] Epitope is the basis of protein antigenicity, and is the basic structural and functional unit of antigen molecule to induce specific immune response. The use of antigenic epitopes with precise positioning and short amino acid sequences can not only be effectively recognized and presented by the immune system, but also induce the body to produce specific humoral and cellular immune responses. Epitope vaccines are a genetically engineered vaccine. Strong, safe and reliable, convenient for large-scale production, and has broad application prospects.

[0060] According to literature reports, the SVA VP1 protein has a full length of 264 amino acids, including a BC loop, a CD loop, and a GH loop; the SVA VP2 protein has a full length of 284 amino acids, including an EF loop, and a VP2 protein 141 -The key motif LDV at position 143 and the key motif DGK at positions 146-148; the SVA VP3 protein has a full length of 239 amino aci...

Embodiment 2

[0088] Method for expression, purification and identification of SVA composite epitope protein

[0089] Small-scale expression and identification of SVA complex epitope protein: entrusted Nanjing GenScript Biotechnology Co., Ltd. to optimize and synthesize the designed three sets of SVA multi-epitope genes, and added NdeI and XhoI enzymes to both ends of them respectively Cutting site, the synthetic sequence is connected to the pET30(a) vector through the NdeI / XhoI restriction site for the expression of complex epitope proteins, and the recombinant plasmids are named pET30(a)-rP1, pET30(a)-rP2 and pET30 (a)-rP3. Transform the recombinant plasmid into BL21(DE3) competent cells for small-scale expression and identification, namely: select a single colony containing the recombinant plasmid into 5mL LB liquid medium (Kana-resistant), culture at 37°C overnight, and then store at -20°C Then select a single colony containing the recombinant plasmid and put it in 5mL LB liquid medium...

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Abstract

The invention provides a Seneca virus type A genetic engineering composite epitope protein, a vaccine and application of the Seneca virus type A genetic engineering composite epitope protein and the vaccine, and belongs to the technical field of veterinary biological products. The invention provides an amino acid sequence of a Seneca virus A genetic engineering composite epitope protein as shown in SEQ ID NO: 14. After the recombinant composite epitope protein is obtained through in-vitro recombinant expression, the vaccine prepared by mixing the recombinant composite epitope protein with an adjuvant is used for immunizing healthy susceptible piglets, the generation of a specific antibody and a neutralizing antibody can be detected in 28 days, and the composite epitope protein rP2 protected by the application has a poison attack protection rate of 80% or more, and has a higher poison attack protection effect compared with a control composite epitope protein. The prepared novel Seneca virus A gene engineering composite epitope protein vaccine has a wide application prospect in the aspect of preventing and controlling the Seneca virus A.

Description

technical field [0001] The invention belongs to the technical field of veterinary biological products, and in particular relates to a type A Seneca virus genetically engineered complex epitope protein, a vaccine and applications thereof. Background technique [0002] Seneca virus disease is a viral infectious disease mainly infecting pigs caused by type A Seneca virus (Seneca virus A, SVA). The clinical symptoms are very similar to foot-and-mouth disease, swine vesicular disease and vesicular stomatitis. It is characterized by blisters, ulcers, and even shedding of hoof shells on the snout, oral mucosa, and hoof crowns of pigs. Diarrhea symptoms are occasionally seen, and the condition is rapid. Newborn piglets ( Within 4 days of age) the mortality rate after onset is as high as 30% to 70%. The disease has a great potential threat to the production and economic benefits of the pig industry, so attention should be paid to the prevention and control of the disease. On June 2...

Claims

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Application Information

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IPC IPC(8): C07K19/00C12N15/62A61K39/125A61K39/42A61P31/14C07K16/10G01N33/569
CPCC07K14/005A61K39/12A61P31/14C07K16/1009G01N33/56983C12N2770/32022C12N2770/32034C07K2319/40C07K2317/76A61K2039/552G01N2333/085Y02P20/55Y02A50/30
Inventor 张中旺潘丽
Owner LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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