Vaccine composition as well as preparation method and application thereof

一种疫苗组合物、减蛋综合征的技术，应用在生物化学设备和方法、疫苗、多价疫苗等方向，能够解决免疫效力低下、疫苗难以提供免疫效果、未开发产品等问题，达到生物安全性好、良好预防和控制效果、免疫原性好的效果

Active Publication Date: 2018-06-08
PULIKE BIOLOGICAL ENG INC +1
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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 subunit 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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  • Vaccine composition as well as preparation method and application thereof
  • Vaccine composition as well as preparation method and application thereof
  • Vaccine composition as well as preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment approach

[0023] As an embodiment of the present invention, the avian adenovirus fiber-2 protein is the protein encoded by SEQ.ID NO.1.

[0024] As an embodiment of the present invention, the protein content of the avian adenovirus fiber-2 is AGP titer ≥ 1:2.

[0025] As a preferred embodiment of the present invention, the protein content of the avian adenovirus fiber-2 is AGP titer 1:2-1:16.

[0026]The term "veterinary acceptable carrier" refers to all other components in the vaccine composition of the present invention except the adenovirus antigen, a carrier or diluent that does not stimulate the body and does not hinder the use of the biological activity and properties of the compound, Adjuvants are preferred. The term "adjuvant" may include aluminum gel adjuvants; saponins, such as Quil A, QS-21 (Cambridge Biotech Incorporation, Cambridge MA), GPI-0100 (Galenica Pharmaceuticals Incorporation, Birmingham AL); water-in-oil emulsions; Oil emulsions; water-in-oil-in-water emulsions;...

Embodiment 1

[0060] Example 1 Construction of pET28a_FADV_fiber-2 expression vector

[0061] 1 Extraction of FADV virus DNA

[0062] 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.

[0063] 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.) The chicken liver suspension is placed in a sterile 1.5ml centrifuge tube, and 0.4ml VB is added to the sample solution. Vortex to mix and let stand at room temperature for 10 minutes. Add 0....

Embodiment 2

[0071] Example 2 Preparation of fiber-2 protein

[0072] Inoculate the pET28a_FADV_fiber-2 / E.Coli BL21(DE3) strain containing prepared in Example 1 into the LB medium containing 50-100 μg / ml kanamycin, the inoculum size is 1% (V / V), 37 Cultivate with shaking. 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 relatively high, the AGP titer of fiber-2 protein reaches 1:64, and the endotoxin content is 0.48×10 5 EU / ml.

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Abstract

The invention relates to vaccine composition. The vaccine composition comprises an immunizing dose of FADV (fowl adenovirus) fiber-2 protein or an immunizing dose of a live vector containing an FADV fiber-2 protein gene, as well as a pharmaceutically acceptable vector. The vaccine composition can provide effective immunoprotection against attack of different serotypes of adenoviruses of chicken flocks, 100% protection rate can be realized when content of immunogenic components is low, and good immunization efficiency is shown.

Description

technical field [0001] The invention relates to an anti-avian adenovirus vaccine composition and its preparation method and 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 60-65nm pith core is wrapped in the capsid. The capsid is icosahedral and composed of 252 capsicles with a diameter of 8-10 nm. The capsicles are arranged on a triangular surface, with 6 on each side, of which 240 are hexons (non-vert...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/235A61K39/39A61K39/295A61P31/14A61P31/20A61K39/17A61K39/145A61K39/215A61K39/12
CPCA61K9/0019A61K39/12A61K39/39A61K2039/70A61K2039/5252A61K2039/552C12N2710/10234C12N2720/10034C12N2760/18134C12N2760/16134C12N2770/20034A61K39/295A61P31/14A61K2039/55511C12N2710/10034C12N2710/10071A61P31/20A61K39/235A61K39/17A61K39/215A61K39/145
Inventor 田克恭逄文强孙进忠张许科
Owner PULIKE BIOLOGICAL ENG INC
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