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Oil-in-water-in-oil type inactivated vaccine for avian influenza virus and preparation method thereof

A bird flu virus, water-in-oil-in-water technology, applied in antiviral agents, pharmaceutical formulas, medical preparations containing active ingredients, etc., can solve problems such as high production costs, biosafety, and slow antibody production

Active Publication Date: 2014-01-08
TIANJIN RINGPU BIO TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the main way to control the incidence of avian influenza virus is to inoculate inactivated whole virus vaccine, which is produced by chicken embryo method, which has high production cost and has biological safety hazards; most inactivated vaccines are water-in-oil type, and the resistance is large when injected , not easy to absorb, slow to produce antibodies

Method used

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  • Oil-in-water-in-oil type inactivated vaccine for avian influenza virus and preparation method thereof
  • Oil-in-water-in-oil type inactivated vaccine for avian influenza virus and preparation method thereof
  • Oil-in-water-in-oil type inactivated vaccine for avian influenza virus and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0014] 1.1 Virus cultivation and harvest

[0015] Take the well-grown MDCK cells in the cell culture flask, digest them with EDTA-trypsin cell dispersion solution to prepare cell suspension, count the cells as 8×10 5 Cell density per mL was inoculated in DMEM medium containing 8% fetal bovine serum, and cultured overnight in shake flasks at 37°C; when the cells covered the monolayer of the medium, the inoculation amount of the seed virus was 0.1 MOI and the virus content was ≥ 10 6.5 TCID 50 / mL HP strain H9 subtype avian influenza virus, control the temperature at 37°C and continue to shake the flask for 48-96 hours, harvest the cell culture, freeze and thaw repeatedly 3 times, and obtain the cell venom containing the supernatant;

[0016] 1.2 Inactivation of virus

[0017] Put the cell venom containing the supernatant obtained above in a sterilized container, add formaldehyde solution according to the volume of the virus liquid and adjust the final concentration of formal...

Embodiment 2

[0050] 1.1 Virus cultivation and harvest

[0051] Take the well-grown MDCK cells in the cell culture flask, digest them with EDTA-trypsin cell dispersion solution to prepare cell suspension, count the cells as 3×10 6 Cell density per mL was inoculated in DMEM medium containing 8% fetal bovine serum, and cultured overnight in shake flasks at 37°C; when the cells covered the monolayer of the medium, the inoculation amount of the seed virus was 0.3 MOI and the virus content was ≥ 10 6.5 TCID 50 / mL HP strain H9 subtype avian influenza virus, control the temperature and continue to shake the flask for 96 hours at 37 ° C, harvest the cell culture, freeze and thaw repeatedly 3 times, and obtain the cell venom containing the supernatant.

[0052] 1.2 Inactivation of virus

[0053] Put the cell venom containing the supernatant obtained above in a sterilized container, add formaldehyde solution according to the volume of the virus liquid and adjust the final concentration of formald...

Embodiment 3

[0057] 1.1 Virus cultivation and harvest

[0058] Take the well-grown MDCK cells in the cell culture flask, digest them with EDTA-trypsin cell dispersion solution to prepare a cell suspension, and count the cells according to 10 6 Cell density per mL was inoculated in DMEM medium containing 8% fetal bovine serum, and cultured overnight in shake flasks at 37°C; when the cells covered the monolayer of the medium, the inoculation amount of the seed virus was 0.05MOI and the virus content was ≥ 10 6.5 TCID 50 / mL HP strain H9 subtype avian influenza virus, control the temperature and continue to shake the flask for 80 hours at 37 ° C, harvest the cell culture, freeze and thaw repeatedly 3 times, and obtain the cell venom containing the supernatant.

[0059] 1.2 Inactivation of virus

[0060] Put the cell venom containing the supernatant obtained above in a sterilized container, add formaldehyde solution according to the volume of the virus liquid and adjust the final concentrat...

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Abstract

The invention provides an oil-in-water-in-oil type inactivated vaccine for an avian influenza virus and a preparation method thereof. The preparation method comprises the following steps: cultivation of the avian influenza virus, inactivation of the virus, virus concentration, vaccine emulsification and the like. Compared with the prior art, the preparation method provided by the invention has the advantages that a local separated avian influenza virus is adopted and bred by using an MDCK (Madin-Darby canine kidney) cell instead of chicken embryo tissue; the production cost of the vaccine is reduced; the quality is easy to balance and stabilize; the yield and the quality of the vaccine are significantly improved. The vaccine is emulsified by using a homogenizer; the prepared vaccine is of an oil-in-water-in-oil type. Compared with the traditional water-in-oil dosage form, the vaccine is low in viscosity, small in particle size and easy to assimilate. The adjuvants used in the oil-in-water-in-oil type inactivated vaccine do not include any surfactant ingredient in the preparation process of the vaccine; the prepared vaccine is good in safety; the side effect due to the fact that the traditional oil adjuvant is difficult to assimilate is avoided.

Description

technical field [0001] The invention relates to the technical field of veterinary vaccines, in particular to an avian influenza virus inactivated water-in-oil-in-water vaccine and a preparation method thereof. Background technique [0002] Avian Influenza (Avian Influenza) is a severe infectious disease of poultry caused by influenza A virus of the family Orthomyxoviridae, and is classified as Class I infectious disease by OIE. According to the antigenicity of hemagglutinin HA and neuraminidase NA, avian influenza viruses are divided into different subtypes, including 17 specific HA subtypes and 9 specific NA subtypes, and different combinations of the two form different subtypes bird flu virus. According to the pathogenicity of strains, avian influenza can be divided into two categories: highly pathogenic avian influenza and low pathogenic avian influenza. H9 subtype avian influenza is a low-pathogenic avian influenza. After infection of egg-laying poultry, it can cause a...

Claims

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Application Information

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IPC IPC(8): A61K39/145A61K9/113A61P31/16
Inventor 张立霞何召庆吴全忠杨保收梁武李守军
Owner TIANJIN RINGPU BIO TECH
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