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African swine fever B and T cell tandem epitope fusion vaccine

An African swine fever and fusion vaccine technology, applied in the field of vaccines, can solve the problems of increased antibody levels, lack, and inability to produce antibodies quickly, achieving good safety, good immune effect, and avoiding the risk of accelerating virus infection.

Inactive Publication Date: 2020-04-17
河南省生物工程技术研究中心 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The African swine fever protein engineering vaccine lacks the specific epitope of the p30 protein that has been recognized as effective, so there is a problem of insufficient protective sites in theory. The level does not increase significantly, and the corresponding antibody cannot be produced quickly

Method used

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  • African swine fever B and T cell tandem epitope fusion vaccine
  • African swine fever B and T cell tandem epitope fusion vaccine
  • African swine fever B and T cell tandem epitope fusion vaccine

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Example 1. Obtaining neutralizing epitope peptides of p72 protein, p54 protein, p30 protein, CD2v protein, C-type lectin protein, PP62 protein, p17 protein or p12 protein:

[0033] Using http: / / tools.iedb.org / bcell / , http: / / crdd.osdd.net / raghava / abcpred / ABC_submission.html, SYFPEITHI and BIMAS, etc., combined with existing literature analysis and design, the obtained African swine fever virus The potential epitopes of neutralizing antibodies are as follows:

[0034] 1.1 p72 protein neutralizing epitope peptide:

[0035] SEQ ID NO. 1: MASGGAFCLIANDGKADKI;

[0036] SEQ ID NO. 2: NVNKSYGKPDPEPTLSQIEETHLVHFNAHFKPYVPVGFEYNKVRPHTGTPTLGNKLTFGIPQYGDFFHD;

[0037] SEQ ID NO. 3: HSSWQDAPIQGTSQMGAHGQLQTFPRNGYDWDNQTPLEGAVYTLVDPFGRPIVPGTKNAYRNLVYYCEYPGERL;

[0038] SEQ ID NO. 4: VSVEGTSGPLLCNIHDLHKPHQSKPILTDENDTQRTCSHTNPKFLSQHFPENSHNIQTAGKQDITPITDA;

[0039] SEQ ID NO.5: RRNIRFKPWFIPGVINEISLTNNELYINNLFVTPEIHNLFVKRVRFSLIRVHKTQ1.2p30 protein neutralizing epitope peptide:

[0040]...

Embodiment 2

[0070]Embodiment 2. The specific amino acid sequence of the African swine fever tandem epitope fusion protein of the present invention (only enumerating part as an example,

[0071] Not all possible amino acid sequences of the invention):

[0072] 2.1 When explaining the African swine fever tandem epitope fusion protein, it is characterized by the sequence number representing the amino acid sequence of the neutralizing epitope peptide, and the connecting arm is indicated in bold, NP 147-155 Amino acid sequence for (in italics) and the immunoactive peptide tuftsin TKPR (underlined) indicates that different constituents are connected by connecting arms and "-".

[0073] 2.2 The African swine fever tandem epitope fusion protein of the present invention includes p30, p54, p72 neutralizing epitope peptide and NP 147-155 , its amino acid sequence can be expressed as:

[0074] (1) Including 1 neutralizing antigenic epitope of p30 protein, 1 neutralizing antigenic epitope of p54...

Embodiment 3

[0081] Example 3 According to the amino acid sequence of the African swine fever tandem epitope fusion protein of the present invention, the fusion protein is expressed and purified by genetic engineering

[0082] 3.1 Expression and purification of fusion protein by prokaryotic expression system

[0083] 3.1.1 According to the amino acid sequence of the designed fusion protein (including other necessary fragments without antigenic activity such as purification tag His if necessary), the synthesis of the nucleic acid molecule encoding the fusion protein is carried out by Sangon Bioengineering (Shanghai) Co., Ltd. And the construction of carrier (carry out codon optimization according to the amino acid sequence of fusion protein and the expression system that the nucleic acid molecule of coding this fusion protein uses, synthesize this nucleic acid molecule), prokaryotic expression vector selects pET28a for use, inserts the synthetic nucleic acid between NocI and XhoI Molecules,...

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Abstract

The invention, which belongs to the technical field of vaccines, particularly relates to an African swine fever B and T cell tandem epitope fusion vaccine. The main component of the African swine fever B and T cell tandem epitope fusion vaccine is African swine fever tandem epitope fusion protein. The African swine fever tandem epitope fusion protein comprises a B cell neutralizing epitope peptidefragment and a T cell epitope; and the B cell neutralizing epitope peptide comprises the following fragments: at least one neutralizing epitope peptide of each of p72, p54, p30 proteins. When the African swine fever tandem epitope fusion protein is used as a vaccine, the immune effect is good; and the antibody level significantly higher than that of a control group can be detected after one immunization. Since the non-neutralizing epitope is reduced as much as possible in the fusion protein, the risk of accelerating virus infection (ADE effect and antibody dependence enhancement effect) by anon-neutralizing antibody after immunization can be avoided.

Description

technical field [0001] The invention belongs to the technical field of vaccines, in particular to an African swine fever B and T cell epitope tandem fusion vaccine. Background technique [0002] African swine fever (African swine fever, ASF) is a severe infectious disease of pigs caused by African swine fever virus (ASFV), and is the "number one killer" of the global pig industry. The fatality rate of acute infection is as high as 100%. ASF was introduced into my country for the first time in August 2018. As of June 6, 2019, a total of 137 outbreaks had occurred in 32 provinces, posing a huge threat to my country's society and economy. [0003] ASFV is an enveloped double-stranded DNA virus, which is the only member of the African swine fever virus genus (Asfivirus) in the African swine fever-related virus family (Asfarviridae), and the only DNA arbovirus discovered so far. Extracellular virions are approximately 200 nm in diameter. The full length of the ASFV genome is 1...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K19/00A61K39/12A61P31/20
CPCC07K14/005A61K39/12A61P31/20C12N2710/12022C12N2710/12034C07K2319/40A61K2039/552
Inventor 王云龙毕胜利李玉林伊瑶范雪亭王继创程蕾张怡青宋长绪王国强
Owner 河南省生物工程技术研究中心
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