African swine fever neutralizing epitope subunit vaccine

A subunit vaccine, African swine fever technology, applied in the field of vaccines, can solve the problems of insignificant increase in antibody levels and inability to quickly produce antibodies, and achieve the effects of good immunity, avoiding accelerated virus infection and improving safety.

Pending 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

In addition, when the fusion protein is used to immunize pigs, the antibody level of the immunized group is not significantly

Method used

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  • African swine fever neutralizing epitope subunit vaccine
  • African swine fever neutralizing epitope subunit vaccine

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] 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:

[0039] 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:

[0040] 1.1 p72 protein neutralizing epitope peptide:

[0041] SEQ ID NO. 1: MASGGAFCLIANDGKADKI;

[0042] SEQ ID NO. 2: NVNKSYGKPDPEPTLSQIEETHLVHFNAHFKPYVPVGFEYNKVRPHTGTPTLGNKLTFGIPQYGDFFHD;

[0043]SEQ ID NO. 3: HSSWQDAPIQGTSQMGAHGQLQTFPRNGYDWDNQTPLEGAVYTLVDPFGRPIVPGTKNAYRNLVYYCEYPGERL;

[0044] SEQ ID NO. 4: VSVEGTSGPLLCNIHDLHKPHQSKPILTDENDTQRTCSHTNPKFLSQHFPENSHNIQTAGKQDITPITDA;

[0045] SEQ ID NO.5: RRNIRFKPWFIPGVINEISLTNNELYINNLFVTPEIHNLFVKRVRFSLIRVHKTQ;

[0046] 1.2 p30 protein neutralizing epitope peptid...

Embodiment 2

[0078] Example 2. The specific amino acid sequence of the African swine fever neutralizing epitope fusion protein of the present invention is exemplified:

[0079] 2.1 When explaining the African swine fever neutralizing epitope fusion protein, the sequence number representing the amino acid sequence of the neutralizing epitope peptide is used for characterization, and the connecting arm is indicated in bold, NP 147-155 The amino acid sequence of TYQRTRAV (in italics) and the immunoactive peptide tuftsin are used TKPR (underlined) indicates that different constituents are connected by connecting arms and "-".

[0080] For example: (1) The amino acid sequence of the African swine fever neutralizing epitope fusion protein represented by SEQ ID NO.1-KK-SEQ ID NO.5-KK-SEQ ID NO.10-GSSSSGSSSG is: MASGGAFCLIANDGKADKI-KK-MEVIFKTDLRSSSQVVFHAG- KK-MDSEFFQPVYPRHYGECLSPVTTPSFFSTHMY-GGGSGGG;

[0081] (2) TKPR The amino acid sequence represented by -KK-SEQ ID NO.6-KK-SEQ ID NO.3-KK-SE...

Embodiment 3

[0115] Example 3 According to the amino acid sequence of the African swine fever neutralizing epitope fusion protein, the method of genetic engineering is used to express and purify the fusion protein

[0116] 3.1 Expression and purification of fusion protein by prokaryotic expression system

[0117] 3.1.1 According to the amino acid sequence of the designed fusion protein (including the purification tag His if necessary), Shanghai Sangong will carry out the synthesis of the nucleic acid molecule encoding the fusion protein and the construction of the vector (according to the amino acid sequence of the fusion protein and the fusion protein encoding the fusion protein). The expression system of nucleic acid molecule application of protein carries out codon optimization, and synthesizes this nucleic acid molecule), prokaryotic expression vector selects pET28a, inserts the nucleic acid molecule of synthesis between NocI and XhoI, and host bacterium selects Bl21(DE)3.

[0118] 3.1...

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Abstract

The invention, which belongs to the technical field of vaccines, particularly relates to an African swine fever neutralizing epitope subunit vaccine that is mainly composed of African swine fever neutralizing epitope fusion protein. The African swine fever neutralizing epitope fusion protein mainly comprises a B cell neutralizing epitope peptide fragment including at least one neutralizing epitopepeptide of p72 protein, at least one neutralizing epitope peptide of p54 protein and at least one neutralizing epitope peptide of p30 protein. With the African swine fever neutralizing epitope fusionprotein, the risk of accelerating virus infection possibly caused by non-neutralizing antibodies can be effectively eliminated and the immune safety of the fusion protein can be effectively improved.Besides, when the African swine fever neutralizing epitope fusion protein is used for immunizing pigs, the antibody level obviously higher than that of a control group can be generated after one immunization.

Description

technical field [0001] The invention belongs to the technical field of vaccines, in particular to an African swine fever neutralizing epitope subunit vaccine. Background technique [0002] African swine fever is an acute, febrile infectious disease of pigs caused by the African swine fever virus ASFV. The International Veterinary Bureau lists it as a class A animal disease, and our country has stipulated this disease as a class I animal infectious disease. ASFV is a complex icosahedral virus with features similar to members of the iridoviridae and poxviridae families. The double-stranded linear DNA genome of ASFV varies in size from 170 to 190 kb depending on the virus strain. ASFV is a very complex virus with a genome of more than 150 open reading frames and a total of more than 100 polypeptides. The results obtained in different laboratories are somewhat different. ASFV virus can encode more than 34 kinds of structural proteins. Although there are many structural protei...

Claims

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Application Information

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IPC IPC(8): C07K19/00A61K39/12A61P31/20
CPCC07K14/005A61K39/12A61P31/20C12N2710/12022C12N2710/12034
Inventor 李玉林毕胜利王云龙伊瑶范雪亭张怡青程蕾王继创宋长绪王国强
Owner 河南省生物工程技术研究中心
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