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Swine fever-porcine circovirus combined subunit vaccine, as well as preparation method and application thereof

A pig ring and dual subunit technology, which is applied in the field of swine fever-porcine ring dual subunit vaccine and its preparation, can solve the problems of difficulty in rational arrangement of immunization procedures for various diseases, hidden dangers of biological safety, immune interference, etc. Prevention and protection of infection, strong immunogenicity, cost-saving effect

Active Publication Date: 2017-10-10
NOVO BIOTECH CORP
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] The technical problems to be solved by the present invention are as follows: one is to develop a vaccine capable of large-scale industrial production, good quality, no immune interference and capable of simultaneously preventing the infection of swine fever virus and porcine circovirus type 2; The problems of multi-injection, multi-dose, and multi-disease immunization procedures in the field are difficult to arrange reasonably; the third is to overcome the possibility of using the live swine fever vaccine to develop combined vaccines that may lead to the remaining nucleic acid in the genome of swine fever virus and other viruses or other vaccines. Recombination causes the mutation of the CSF virus, so there are great biological safety hazards and possible immune interference problems; the fourth is to avoid the need to separate the CSF virus and other disease antigens when using live CSF vaccines to prepare dual vaccines and the trouble of mixing when using

Method used

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  • Swine fever-porcine circovirus combined subunit vaccine, as well as preparation method and application thereof
  • Swine fever-porcine circovirus combined subunit vaccine, as well as preparation method and application thereof
  • Swine fever-porcine circovirus combined subunit vaccine, as well as preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Embodiment 1: Preparation of classical swine fever virus E2 protein and porcine circovirus type 2 Cap protein

[0030] 1.1 Preparation of swine fever virus E2 protein: refer to the preparation of swine fever protein in the invention patent method or other preparation method of swine fever virus E2 in the application number 200810178235.4 or 200310103408.3 or 200810150304.0 or 201310300549.8 or 200810178235.4 or 201610625392.X or 201510187995.1 For E2 protein, full-length CSF E2 protein or truncated CSF E2 protein can be prepared according to actual needs.

[0031] 1.2 Preparation of porcine circovirus type 2 Cap protein: refer to the preparation method of porcine circovirus type 2 Cap protein in the invention patent with application number 201310050003.1 or 201210270504.6 or 201110100331.9 or 201110053536.6 or 201010618223.6 or the preparation method in other patents or literatures to prepare pigs The circovirus type 2 Cap protein can prepare full-length porcine circovi...

Embodiment 2

[0032] Embodiment 2: the preparation of swine fever-porcine circular double subunit vaccine (illustrate with preparation 1ml / head part, totally 220g as example)

[0033] 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.

[0034] 1. Preparation of Montanide GEL 01PR (adjuvant): According to the mass ratio of antigen solution and adjuvant of 10:1, weigh 20g (about 20ml) of adjuvant and place it in a pre-prepared reagent bottle, seal it, and wait for use.

[0035] 2. According to the mass ratio of antigen solution and adjuvant of 10:1, the total mass of antigen solution is 200g (about 200ml). According to the concentration of classical swine fever virus E2 protein and porcine circovirus type 2 Cap protein concentration and the content of each prot...

Embodiment 3

[0040] Embodiment 3: Immunization experiment of swine fever-porcine circular double subunit vaccine

[0041] 3.1 Vaccine preparation: 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:

[0042] Table 2

[0043]

[0044] 3.2 Immunization experiment: 35 piglets aged 28-35 days (negative for CSFV and PCV2 antigen antibodies) were randomly divided into 7 groups with 5 pigs in each group. The blank control group was injected intramuscularly with 1ml of normal saline each time, and the other 6 groups of immunized groups were injected intramuscularly with 1ml of the corresponding vaccine each time. A booster immunization was given three weeks after the initial immunization. Serum was collected before immunization, before the second immunization and 21 days after the second immunization, and tested. Antibody titer.

[0045] Experimental results such as Figure 1a As shown, the test resu...

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Abstract

The invention discloses a swine fever-porcine circovirus combined subunit vaccine, as well as a preparation method and an application thereof, and belongs to the technical fields of animal vaccines and veterinary biological products. The vaccine is prepared from swine fever virus E2 protein, porcine circovirus type 2 cap protein and a pharmaceutically acceptable adjuvant. The preparation method comprises the following steps: (1) preparing swine fever virus E2 protein and porcine circovirus type 2 cap protein; (2) preparing an antigen solution from the swine fever virus E2 protein and porcine circovirus type 2 cap protein prepared in the step (1); and (3) mixing and stirring the antigen solution and Montanide GEL 01PR adjuvant in a mass ratio of 10:1. The vaccine has the advantages of strong immunogenicity, good safety, no immune interference and the like, can be used for preventing potential biological safety hazard for virus variation and fundamentally purifying swine fever virus and porcine circovirus type 2, and can achieve the effect of dual prevention with one injection by immunizing the vaccine to achieve the aims of saving time, labor and cost.

Description

technical field [0001] The invention relates to a swine fever-porcine circular double subunit vaccine and a preparation method and application thereof, belonging to the technical field of animal vaccines and veterinary biological products. Background technique [0002] Classical swine fever is an acute, febrile and fatal disease caused by classical swine fever virus (CSFV). Classical swine fever is highly contagious, acute onset, persistent high fever, and degeneration of small blood vessel walls causing extensive hemorrhage, infarction, and necrosis. Domestic and wild pigs are its only natural hosts. The World Organization for Animal Health (OIE) defines it as a class A infectious disease, and my country's "Animal Epidemic Prevention Law" lists it as a class I infectious disease. Swine fever is one of the main diseases that endanger the development of my country's pig industry. [0003] CSFV is a single-stranded linear RNA virus belonging to the family Flaviviridae and t...

Claims

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Application Information

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IPC IPC(8): A61K39/295A61K39/187A61K39/12A61P31/20A61P31/14
CPCA61K39/12A61K39/39A61K2039/70A61K2039/552C12N2750/10034C12N2770/24334
Inventor 钱泓吴有强查银河
Owner NOVO BIOTECH CORP
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