H9 subtype avian influenza virus inactivated vaccine including chicken a-interferon protein

An avian influenza virus, interferon technology, applied in the direction of antiviral agents, peptide/protein components, medical preparations containing active ingredients, etc., can solve problems such as no reports of joint preparation of vaccines, etc. Good level and safety

Active Publication Date: 2015-09-30
YEBIO BIOENG OF QINGDAO
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] According to the existing research data, there is no report on the joint preparation of chicken α-interferon protein and H9 subtype avian influenza virus antigen
In 2004, Jia Lijun and others tested the immunoenhancing effect of expressed chicken gamma interferon on H5 subtype avian influenza vaccine. As a result, the titer of chicken gamma interferon combined immunization group was lower than that of inactivated vaccine alone

Method used

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  • H9 subtype avian influenza virus inactivated vaccine including chicken a-interferon protein
  • H9 subtype avian influenza virus inactivated vaccine including chicken a-interferon protein

Examples

Experimental program
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Effect test

Embodiment 1

[0013] Example 1. Screening of antigenic strains for vaccine production (H9 subtype avian influenza virus QDY strain)

[0014] In 2013, the heart, spleen and other organs, throat and cloacal swabs of the diseased chickens were aseptically collected from the diseased chickens in Chengyang, Shandong Province, and ground in sterile petri dishes under aseptic conditions, and added in a ratio of 1:5. Physiological saline containing 4000 units / ml double antibody was repeatedly freeze-thawed at -20°C for 3 times. The cotton swab was directly put into the normal saline containing 10,000 units of double antibody, and it was treated at 2-8 °C for 4 hours. Both the organs and the cotton swab were centrifuged at 3500 rpm for 10 minutes. Pieces, 0.2ml / piece. The chicken embryos were incubated at 37°C, and checked once a day in the morning and evening. The dead embryos were removed in time and stored in a 4°C refrigerator. The dead embryos within 24 hours were discarded, and all the chicke...

Embodiment 2

[0018] Example 2: Preparation of antigens for seedling production

[0019] 1. Preparation of virus liquid for seedling production Inoculate 10-11-day-old SPF chicken embryos with the virus seed (QDY strain) for production through the allantoic cavity, 0.2 ml per embryo, incubate at 36 ℃ ~ 37 ℃, and illuminate the embryos twice a day, Take the chicken embryos that died after 24 hours, and harvest them at 96 hours regardless of whether they are dead or not, and cool them at 4-8°C for 12-24 hours.

[0020] 2. Preparation of chicken α-interferon protein for seedling production. The fermenter is aerated and cultured. The medium is prepared according to 70% of the volume of the fermenter, and the pH value is adjusted to 7.0-7.2 with 1 mol / L NaOH. Foaming agent. After sterilization, inoculate production strains according to 1% to 2% of the inoculum, add 10% kanamycin sulfate injection according to the proportion of 0.1% of the total volume, incubate at 36 °C, and after 2.5 hours, ta...

Embodiment 3

[0028] 1. Preparation of inactivated vaccine: The semi-finished antigen after passing the inspection is used for vaccine preparation (each liquid component in the following preparation is calculated by volume ratio).

[0029] (1) Preparation of oil phase Take 95 parts of veterinary white oil and 1 part of aluminum stearate, put them in an oil phase preparation tank and heat to 80°C, add 805 parts of Siben-805, until the temperature reaches 115°C, maintain 30min, set aside after cooling.

[0030] (2) Preparation of water phase: Dilute chicken α-interferon protein with normal saline to 2×10 4.0 ~2×10 6.0 Unit / 0.1ml; Dilute the inactivated H9 subtype avian influenza virus solution with physiological saline to not less than 2×10 6.0 EID 50 / 0.1ml. The chicken α-interferon protein liquid and the inactivated H9 subtype avian influenza virus liquid were respectively taken and mixed to prepare an antigen liquid for vaccine preparation. Take 5 parts of sterilized Tween-80, add it ...

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Abstract

The invention provides an H9 subtype avian influenza virus inactivated vaccine including chicken a-interferon protein. The H9 subtype avian influenza virus inactivated vaccine comprises an antigen and a vaccine adjuvant, and the adopted antigen is an inactivated H9 subtype avian influenza virus QDY1 strain including the chicken a-interferon protein. A screened H9 subtype avian influenza virus has the advantages of being small in virulence and high in immunogenicity. The H9 subtype avian influenza virus (QDY strain) is inoculated into a chick embryo, then collects virus liquid, and is mixed with the extracted and purified chicken a-interferon protein after inactivation of formaldehyde solutions, and an oil adjuvant is added for emulsification to manufacture the vaccine. The prepared vaccine can stimulate an organism to generate an antibody fast, the level of the antibody is improved, the persistent period of the antibody is prolonged, and diseases caused by the H9 subtype avian influenza virus is prevented. The vaccine has the advantages of being efficient and good in safety.

Description

technical field [0001] The invention belongs to the technical field of biological veterinary drugs, in particular to a novel H9 subtype avian influenza vaccine comprising chicken alpha-interferon protein. Background technique [0002] H9 subtype avian influenza is a low pathogenic disease, but mixed infection with other pathogens, especially Escherichia coli, can increase its pathogenicity, cause a serious drop in egg production and a significant increase in mortality, and bring serious harm to the poultry industry. The existing avian influenza (H9 subtype) vaccine has been widely used clinically, but there are problems in the aspect of low antibody level and antibody uniformity, which affects the preventive effect of the vaccine. [0003] Interferon is a low molecular weight soluble multifunctional glycoprotein produced by T lymphocytes and NK cells, which can enhance the function of cytotoxic T cells, promote the differentiation and activation of B cells, and induce animal...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/145A61K39/39A61P31/16A61K38/21
Inventor 刘新文杜元钊郭伟伟胡潇王秀丽宫晓
Owner YEBIO BIOENG OF QINGDAO
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