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Duck flavivirus subunit vaccine, as well as a preparation method and application thereof

A subunit vaccine, duck flavivirus disease technology, applied in the fields of botanical equipment and methods, biochemical equipment and methods, applications, etc., can solve the problems that need to be deepened, there is no vaccine, etc., and reduce production costs and antigenic epitopes. The effect of high and increasing antigen expression

Active Publication Date: 2012-11-28
HARBIN VETERINARY RES INST CHINESE ACADEMY OF AGRI SCI +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Since this pathogen is a newly emerged pathogen, relevant research needs to be in-depth
In particular, there is currently no vaccine available for the prevention and control of the disease

Method used

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  • Duck flavivirus subunit vaccine, as well as a preparation method and application thereof
  • Duck flavivirus subunit vaccine, as well as a preparation method and application thereof
  • Duck flavivirus subunit vaccine, as well as a preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Embodiment 1 Construction and detection of a recombinant cell line stably expressing duck yellow virus prM-E protein

[0038] 1 Materials and methods

[0039] 1.1 Plasmids, strains and cells

[0040] The expression vector plasmid pCAGneo, DH5α competent cells and BHK21 cells were preserved by the State Key Laboratory of Veterinary Biotechnology of Harbin Veterinary Research Institute, the plasmid extraction kit and RNA extraction kit were produced by QIAGEN, and the gel recovery kit was purchased from Shanghai Huashun Biotechnology Co., Ltd., G418 was purchased from Gibco, trypsin was purchased from Hyclone, Reverse Transcriptase M-MLV, PrimeSTAR TM HS DNA Polymerase﹑SalI﹑XhoI﹑BamHI﹑T4 DNA ligase was purchased from TaKaRa Company, anti-PrM protein and anti-E protein monoclonal antibodies were prepared by our research group, goat anti-mouse IgG antibody labeled with infrared fluorescent dye was purchased from KPL Company, FITC-labeled goat anti-mouse IgG was purchased fr...

Embodiment 2

[0069] Preparation of Example 2 Duck Flavivirus Subunit Vaccine and Immunization Test to Ducks

[0070] 1. Vaccine preparation:

[0071] When the recombinant cell line BHK-BYD-ME cells constructed and screened in Example 1 grew to 90% full after normal passage, the serum-free medium was changed to continue culturing for 4-6 days, and the cell culture was harvested and frozen and thawed three times at -20°C. The vaccine antigen solution is obtained by adding 0.02% thimerosal to the stock solution containing cell culture after repeated freezing and thawing three times. The oil adjuvant vaccine preparation method is to mix the vaccine antigen solution with the French SEPPIC company ISA 50V2 oil adjuvant at a volume ratio of 1:1, fully emulsify and store at 4°C for later use.

[0072] 2. Animal grouping and immunization:

[0073]Experimental animals SPF-grade Shaoxing ducks were provided by the Experimental Animal Center of Harbin Veterinary Research Institute, Chinese Academy o...

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Abstract

The invention discloses a duck flavivirus subunit vaccine, as well as a preparation method and an application thereof. The duck flavivirus subunit vaccine contains stable transfected cell line expressed duck flavivirus prM-E protein and an adjuvant. The invention also discloses a method for establishing a stably expressed duck flavivirus prM-E protein cell line and culturing cell line expressed protein, and in particular relates to an expressed duck flavivirus prM-E protein cell line BHK-BYD-ME which is preserved in China General Microbiological Culture Collection Center with the number of CGMCC No.6342. The duck flavivirus subunit vaccine has high safety and a good immunization effect, and virus-neutralizing antibodies can be induced after a duck is immunized. Experiments prove that the vaccine can remarkably improve the resistance of a duck to the duck flavivirus, and can be used for preventing and controlling the duck flavivirus.

Description

technical field [0001] The invention relates to a subunit vaccine for duck flavin virus disease expressed by a recombinant cell line. Specifically, the recombinant cell line involved in the invention is BHK-BYD-ME, and its strain preservation number is: CGMCC No.6342. The invention also discloses a method for preparing the recombinant cell line and the application of the recombinant cell line in preparing a vaccine for preventing duck yellow virus disease. The invention belongs to the technical field of animal vaccines and veterinary biological products. Background technique [0002] Duck yellow virus disease (tentative name), also known as duck hemorrhagic oophoritis, or duck egg drop syndrome, is a new duck disease first discovered in southeastern China in 2010. The disease is mainly pathogenic to laying ducks, but also occurs in breeding geese and meat ducks. The clinical manifestations of the disease of laying ducks are unstable limbs, difficulty walking, decreased fee...

Claims

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Application Information

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IPC IPC(8): A61K39/12A61P31/14C12N5/10C12N15/40C12N15/85G01N33/569C12R1/91
Inventor 华荣虹步志高陈化兰柳金雄
Owner HARBIN VETERINARY RES INST CHINESE ACADEMY OF AGRI SCI
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