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African swine fever virus chimeric protein, vaccine composition, preparation method and application thereof

A technology of African swine fever virus and vaccine composition, applied in biochemical equipment and methods, viruses, vaccines, etc., can solve the problems of unsatisfactory vaccine immune effect, achieve significant humoral and cellular immunity, solve poor immunogenicity, and facilitate The effect of mass production

Active Publication Date: 2022-08-09
PU LIKE BIO ENG
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] In order to solve the deficiencies in the prior art, the present invention provides a chimeric protein of African swine fever virus. When the chimeric protein of African swine fever virus is used to prepare vaccines, the immune effect is good, which effectively solves the problem of various types of vaccine immunity in the prior art. Problems with unsatisfactory results

Method used

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  • African swine fever virus chimeric protein, vaccine composition, preparation method and application thereof
  • African swine fever virus chimeric protein, vaccine composition, preparation method and application thereof
  • African swine fever virus chimeric protein, vaccine composition, preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0057] Example 1 The expression of African swine fever virus P72 full -length and short protein

[0058] The corresponding encoding sequence list of the corresponding coding sequence of Suzhou Jinwei Zhizhi Biotechnology Co., Ltd. The nucleotide sequence of the P72 protein amino acid sequence is shown in SEQ ID No.1, and the synthetic nucleotide is cloned to the PET28A carrier. Essence

[0059] Similarly, the sequence list SEQ ID No.2 shown in the African swine fever virus P72 domain Ⅰ, serial list SEQ ID No.3, the African swine fever virus P72 domain Ⅱ, serial list SEQ ID No.4 shown in Africa pigs shown in Africa The P72 Domain III is connected through a flexible connection peptide. Among them, the African swine fever virus P72 domain Ⅰ is located on the N -end, the domain III is located on the C end, and the domain II is between the Field Ⅰ and the domain III. Weizhi Biotechnology Co., Ltd. synthesized the corresponding coded nucleotide sequence, and the synthetic nucleotide was...

Embodiment 2

[0063] Example 2 African swine fever virus P72 short -term protein's effect on immunogenicity

[0064] Add the P72 full -length protein and P72 brief protein prepared by the Example 1 to the agent. P72 vaccine composition with short protein. Sales suitable for the present invention can be known to the technical personnel in the art. In the present invention, the agent is used as a bipolar agent (water packaging oil), for example, it can be an adult ISA206 (French Sibik). The specific ratio of each component in the prepared vaccine is shown in Table 1.

[0065] Table 1 African swine fever virus P72 full -length and short -cut protein vaccine composition ingredient ratio

[0066] Component Vaccine 1 Vaccine 2 P72 protein (μg / ml) 200 0 Δp72 protein (μg / ml) 0 200 Two -phase agent (V / V %) 50％ 50％

[0067] The African swine fever virus antigen and antibodies are 15 healthy piglets with a negative weight of about 20kg, which are randomly divided into 3 gr...

Embodiment 3

[0080] Example 3 African swine fever virus chimeric protein skeleton and antigenic protein selection

[0081] The inventor analyzed the protein structure of the African swine fever virus P72 and cut off the creativity, and still maintained the relatively stable protein space structure, and its immune originality was not affected. To this end, the P72 intercept protein was embedded as the invention embedded Skeleton of protein.

[0082] At the same time, after a large number of experimental research, the inventor combined with African swine fever virus virus immunogenic protein antigen surface analysis, the following sequence was screened. The chimeric protein containing African swine fever virus antigenic protein was P54, P30, CD2V and P12 protein. At least one fragment can produce higher immunogenic reactions:

[0083] P54 protein fragments are selected from any one or more shown by SEQ ID No.5, 6, 7, 8, 9, 10, 11, 12;

[0084] P30 protein fragment is selected from any one or mor...

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Abstract

The present invention provides an African swine fever virus chimeric protein, the chimeric protein comprises: (1) African swine fever virus p72 domain I; (2) African swine fever virus p72 domain II; (3) African swine fever virus viral p72 domain III; and (4) African swine fever virus antigenic protein. The chimeric protein provided by the present invention uses the African swine fever virus p72 protein as the backbone for the first time, and better displays the antigenic epitopes of the African swine fever virus antigenic proteins p54, p30, CD2v and p12, has good immune effect, and can produce significant Humoral and cellular immunity.

Description

Technical field [0001] The present invention is a technology field of beastly biological products, which involves a type of African swine fever virus chimiric protein, and a vaccine composition, preparation method and application containing the chimiric protein. Background technique [0002] African Swine Fever (ASF) is an acute, intense, high -contact infectious disease caused by the African swine fever virus (ASFV). For a type of animal disease. The disease was first confirmed in Kenya in Africa in 1921. Since 2007, African swine fever has occurred, spread, and popular in many countries around the world. It was introduced to my country in 2018, bringing huge direct and indirect economic losses. The disease has been discovered for nearly a century, but there are currently no vaccines and special medicines in the world. [0003] The difficulty of developing the African swine fever vaccine is that the virus genome is large, more protein, complex structure, and difficult to cultiva...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K19/00C12N15/62A61K39/295A61K39/12A61P31/20
CPCC07K14/005A61K39/12A61P31/20C07K2319/55C12N2710/12022C12N2710/12034A61K2039/70A61K39/00C12N15/62C07K19/00C12N15/63Y02A50/30A61K2039/55566A61K2039/5252C07K14/01A61K2039/552A61K2039/57C07K2319/00C12N2710/12052C12N2710/12071
Inventor 田克恭逄文强张许科
Owner PU LIKE BIO ENG
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