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A kind of vaccine composition and its application

A vaccine composition and a technology for egg reduction syndrome, which are applied in the directions of vaccines, multivalent vaccines, veterinary vaccines, etc., can solve the problems of low immune efficacy, undeveloped products, and difficulty in cultivating avian adenoviruses, and achieve good immunogenicity. , good biosafety effect

Active Publication Date: 2021-01-15
PULIKE BIOLOGICAL ENG INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Vaccine immunization can effectively prevent the disease. However, in the prior art, because avian adenovirus is not easy to cultivate, it is difficult to obtain high-titer virus, especially the difference between different isolates, which makes it difficult for the prepared vaccine to provide the ideal immune effect. , and the whole virus vaccine also has biosafety risks caused by incomplete inactivation
The antigenic component of the vaccine in the prior art is the hexon (240 / 252) that constitutes the main component of the viral capsid. However, the immune effect has been low all the time, and it has not been developed into a product

Method used

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  • A kind of vaccine composition and its application
  • A kind of vaccine composition and its application
  • A kind of vaccine composition and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment approach

[0021] As an embodiment of the present invention, the avian adenovirus penton protein has a protein sequence encoded by SEQ.ID NO.1.

[0022] As an embodiment of the present invention, the content of the avian adenovirus penton protein is AGP titer ≥ 1:2.

[0023] As a preferred embodiment of the present invention, the content of the avian adenovirus penton protein is AGP titer 1:2-1:16.

[0024] As an embodiment of the present invention, the pharmaceutically acceptable carrier includes an adjuvant, and the adjuvant includes: (1) aluminum gel adjuvant, saponin, avridine, DDA; (2) water-in-oil emulsion , oil-in-water emulsions, water-in-oil-in-water emulsions; or (3) polymers of acrylic acid or methacrylic acid, copolymers of maleic anhydride and alkenyl derivatives; and RIBI adjuvant systems, Block co- One or more of polymer, SAF-M, monophosphoryl lipid A, Avridine lipid-amine adjuvant, Escherichia coli heat-labile enterotoxin, cholera toxin, IMS 1314, muramyl dipeptide, Gel ...

Embodiment 1

[0053] Example 1 Construction of pET28a_FADV_penton expression vector

[0054] 1 Extraction of FADV virus DNA

[0055] The plasmid extraction kit was purchased from Tiangen Biotech; T4 DNA Ligase was purchased from BioLab; the pET28a plasmid was purchased from Novagen; the agarose gel recovery kit was purchased from Tianze Biotech, and other reagents were of analytical grade.

[0056] According to the instructions of the virus DNA extraction kit, take 0.2ml of infected avian adenovirus FAV-HN strain (fowl adenovirus, FAV-HN strain (Fowl aviadenovirus, strain FAV-HN), the preservation number is: CCTCC NO.V201609, and the depository unit is China Typical Culture Collection Center, the preservation address is Wuhan University, Wuhan, China, and the preservation time is February 29, 2016) Chicken liver suspension is placed in a sterile 1.5ml centrifuge tube, 0.4ml VB is added to the sample solution, and vortexed Mix well and let stand at room temperature for 10 minutes. Add 0.45...

Embodiment 2

[0064] The preparation of embodiment 2 penton protein

[0065] Inoculate the pET28a_FADV_penton / E.Coli BL21(DE3) strain containing 50-100 μg / ml kanamycin into the LB medium containing 1% (V / V), shaking at 37°C to cultivate. When OD600=0.4-0.6, place it at 28°C for 30 minutes. Isopropyl-β-D-thiogalactopyranoside (IPTG) was added to make the final concentration 0.1-1.0 mM, and cultured with shaking at 28° C. for 24 hours. After the cultivation, the cells were collected and resuspended with PBS (sodium chloride, 8g, potassium chloride, 0.2g, disodium hydrogen phosphate, 1.44g, potassium dihydrogen phosphate, 0.24g, adjusted to pH 7.4, constant volume 1L) Bacteria were sonicated and centrifuged to obtain the supernatant. The content of soluble target protein in the expression product is high, the AGP titer of penton protein reaches 1:32, and the endotoxin content is 0.56×10 5 EU / ml.

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Abstract

The invention relates to a vaccine composition. The vaccine composition comprises an immune amount of avian adenovirus penton protein or an immune amount of a live carrier containing a recombinant avian adenovirus penton protein gene, and a pharmaceutically acceptable carrier. The vaccine composition can provide effective immune protection against chicken flocks challenged by different serotypes of adenovirus, and can provide 100% protection rate when the content of immunogenic components is low, showing good immune efficacy.

Description

technical field [0001] The invention relates to a vaccine composition and its application, belonging to the field of biomedicine. Background technique [0002] Fowl adenovirus infection (Fowl adenovirus, FADV) is an infectious disease caused by group I poultry adenovirus (FADV). FADV belongs to the family Adenoviridae, and its genome (gemone) is composed of linear double-stranded (double-stranded) DNA without segmentation. [0003] The avian adenovirus has a non-enveloped spherical structure, and its virions are often arranged in a lattice in the infected cell nucleus. Each virion contains a 36kb linear double-stranded DNA, and the linear double-stranded DNA and core protein form a diameter of The pith core of 60-65nm is wrapped in the capsid. The capsid is icosahedrally symmetrical and consists of 252 capsomers with a diameter of 8-10 nm. The capsomers are arranged on a triangular surface, with 6 on each side, of which 240 are hexons (non-vertex capsomers), and the other ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K39/235A61K39/295A61P31/14A61P31/16
CPCA61K39/12A61K2039/5252A61K2039/552A61K2039/70C12N2710/10234C12N2720/10034C12N2760/16134C12N2760/18134C12N2770/20034
Inventor 田克恭逄文强孙进忠张许科
Owner PULIKE BIOLOGICAL ENG INC
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