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Subunit vaccine composition against African swine fever as well as preparation method and application thereof

A vaccine composition, African swine fever virus technology, applied in biochemical equipment and methods, vaccines, veterinary vaccines, etc., can solve the problems of virus mutation, immune interference, hidden dangers of biological safety, etc. Immune interference, good safety effect

Active Publication Date: 2022-01-18
NOVO BIOTECH CORP
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] Technical problems to be solved by the present invention: one is to provide a subunit vaccine with strong immunogenicity, good safety, and the ability to fundamentally purify African swine fever Composition; the second is to overcome the virus mutation caused by the nucleic acid recombination of the live vaccine and the natural African swine fever virus genome, so that there are great biological safety hazards and possible immune interference problems; the third is to provide a preparation for African swine fever virus The method of swine fever subunit vaccine and its application in the prevention of African swine fever

Method used

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  • Subunit vaccine composition against African swine fever as well as preparation method and application thereof
  • Subunit vaccine composition against African swine fever as well as preparation method and application thereof
  • Subunit vaccine composition against African swine fever as well as preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029]Example 1: Preparation of African swine fever virus outer envelope CD2V protein and p72 protein

[0030] 1.1 The reference company’s application number is 201910004596.5 or 201910069838.9 The method for preparing the CD2V protein in the outer layer of the African swine fever envelope in the invention patent or the outer layer in other patents or documents (expressed by eukaryotic expression systems, such as CHO, 293T cells, etc.) Method for preparing envelope CD2V protein.

[0031] 1.2 References are Liu Q, Ma B, Qian N, et al. Structure of the African swinefever virus major capsid protein p72. Cell Res. 2019; 29(11): 953-955. Or other protein expression methods (such as prokaryotic expression system expression, insect baculovirus expression, CHO cell expression system, etc.) or the preparation method of African swine fever virus p72 protein in other patents or literatures, its trimer content reaches more than 40%. like figure 1 Shown: From the comparison of the p72 pr...

Embodiment 2

[0033] Embodiment 2: the preparation of African swine fever virus subunit vaccine (to prepare 2ml / head portion, totally 200ml is example illustration)

[0034] The consumables and materials used for the preparation of vaccines need to undergo aseptic treatment in advance, and the preparation process is completed in a biological safety cabinet or other instruments or environments that can ensure that the entire preparation process is sterile.

[0035] 1. Preparation of ISA 201VG: According to the volume ratio of antigen solution and adjuvant of 46:54, measure 108ml of adjuvant into a pre-prepared reagent bottle, seal it, and preheat it in a 33°C water bath for about 30 minutes.

[0036] 2. According to the volume ratio of antigen solution and adjuvant is 46:54, the total volume of aqueous phase is 92ml. According to the African swine fever outer envelope CD2V protein and p72 protein concentration and the total protein content in the vaccine, calculate the volume of African swi...

Embodiment 3

[0042] Embodiment 3: The immune challenge experiment of African swine fever two-component subunit vaccine

[0043] 3.1 Vaccine preparation

[0044] Proteins and vaccines were prepared according to the methods of Examples 1 and 2, and the specific vaccine information is shown in Table 2 below:

[0045] Table 2

[0046]

[0047] 3.2 Immunization experiment

[0048] 25 piglets aged 35-40 days (negative for African swine fever virus) were screened and randomly divided into 5 groups, 5 pigs in each group. One group was used as the blank control group, and the other 5 groups were used as the immunization group, and were immunized with vaccines 1 to 4 respectively. The blank control group was injected intramuscularly with 2ml of normal saline each time, and the other 4 groups of immunized groups were injected with 2ml of the corresponding vaccine intramuscularly each time, and boosted immunization once three weeks after the initial immunization. Three weeks after the second imm...

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Abstract

The invention discloses a subunit vaccine composition against African swine fever as well as a preparation method and an application thereof, and belongs to the technical field of animal vaccines and veterinary biological products. The vaccine comprises an African swine fever outer envelope CD2V protein, an African swine fever outer capsid p72 protein and a pharmaceutically acceptable adjuvant. The preparation method of the vaccine comprises the following steps: 1) preparing African swine fever outer envelope CD2V protein and African swine fever outer capsid p72 protein; 2) mixing the African swine fever outer envelope CD2V protein prepared and the African swine fever outer capsid p72 protein prepared in the step 1) to prepare an antigen solution; and 3) mixing the antigen solution and ISA 201 VG in volume ratio of 46: 54 for emulsification. The vaccine has the advantages of high immunogenicity, good safety and no immune interference. The vaccine can effectively prevent and protect pigs from being infected by African swine fever.

Description

technical field [0001] The invention relates to an African swine fever subunit vaccine composition and a preparation method and application thereof, belonging to the technical field of animal vaccines and veterinary biological products. Background technique [0002] African swine fever (African swine fever, ASF) is an acute, febrile, highly contagious infectious disease of pigs caused by African swine fever virus (ASFV). Pigs infected with African swine fever virus are clinically characterized by skin congestion, internal organ bleeding, and high fever, and the morbidity and mortality are as high as 100%. The disease has been listed as a Class A disease by the World Organization for Animal Health. my country is a large livestock and poultry breeding country, with hog breeding accounting for 56.6% of the world's total hog breeding, and pork consumption accounting for 49.6% of the world's pork consumption. Poultry production is second only to the United States, and it is the ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/12A61P31/20
CPCA61K39/12A61P31/20A61K2039/552A61K2039/5256C12N2710/12034C07K14/005C12N2710/12022A61K2039/55566A61K2039/55522C07K14/01C12N7/00
Inventor 张强钱泓吴有强吴素芳车影
Owner NOVO BIOTECH CORP
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