Method for preparing novel influenza vaccine

A technology for influenza and vaccines, which is applied in the field of new inactivated influenza vaccines, and can solve problems such as the source of influenza vaccine safety culture medium

Inactive Publication Date: 2008-04-30
长春百克生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0007] The invention provides a preparation method of a new type of influenza vaccine to solve the existing problems such as the sa

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  • Method for preparing novel influenza vaccine

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Embodiment 1

[0011] 1) Influenza virus subtypes A1, A3 and B were respectively diluted with virus culture medium at a ratio of 1:10 -4 Separately inoculate Vero cells that have grown to a single layer, and culture them at 35°C until the cytopathic pathology (CPE) reaches +++~++++++, then harvest the virus liquid containing influenza virus; the virus culture liquid The ingredients are:

[0012] MEM medium;

[0013] 1% bovine serum albumin; 5μg / ml trypsin;

[0014] Use HEPES to adjust the pH value of the solution to 7.0;

[0015] 200 μg / ml neomycin.

[0016] 2) The influenza virus in the virus liquid harvested in 1) was inactivated with 1:1000 formaldehyde solution, and the inactivated virus antigens were purified separately by chromatography, and purified by DEAE Streamlin chromatography, and the chromatography conditions were: : DEAE Streamline basic buffer: pH6.8, 0.01Mol / L PBS buffer, 0.01Mol / L sodium chloride for elution; obtain virus stock solution;

[0017] 3) Detect the antigen ...

Embodiment 2

[0020] 1) Influenza virus subtypes A1, A3 and B were respectively diluted with virus culture medium at a ratio of 1:10 -7 Separately inoculate Vero cells that have grown to a monolayer, and culture them at 33°C until the cytopathic pathology (CPE) reaches +++~++++++, and harvest the virus liquid containing influenza virus respectively; the virus culture liquid The ingredients are:

[0021] MEM medium;

[0022] 0.01% bovine serum albumin; 20μg / ml trypsin;

[0023] Use 5% sodium bicarbonate solution to adjust the pH value of the solution to 7.5;

[0024] 20 μg / ml neomycin.

[0025] 2) Inactivate the influenza virus in the virus liquid harvested in 1) with a 1:1000 β-propiolactone solution, and purify the inactivated virus antigens separately by chromatography, using DEAE Sepharose FF chromatography Purification, chromatographic conditions: Basic buffer of DEAE SepharoseFF: pH8.0, 0.2Mol / L Tris buffer, elution with sodium chloride concentration of 1Mol / L; obtain virus stock s...

Embodiment 3

[0029] 1) Influenza virus subtypes A1, A3 and B were respectively diluted with virus culture medium at a ratio of 1:10 -1 Separately inoculate Vero cells that have grown to a monolayer, and culture them at 34°C until the cytopathic pathology (CPE) reaches +++~++++++, and harvest the virus liquid containing influenza virus respectively; the virus culture liquid The ingredients are:

[0030] MEM medium;

[0031] 0.1% bovine serum albumin; 2μg / ml trypsin;

[0032] Use 6% sodium bicarbonate solution to adjust the pH value of the solution to 8.0;

[0033] 500 μg / ml neomycin.

[0034] 2) inactivate the influenza virus in the virus liquid harvested in 1) with a 1:10000 formaldehyde solution, and purify the inactivated virus antigens separately by chromatography, and purify by Sepharose 4FF chromatography, and then chromatographically Conditions: Sepharose 4FF elution buffer is pH7.5, 0.1Mol / L PBS buffer, containing 0.1Mol / L sodium chloride; obtain virus stock solution;

[0035] 3)...

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Abstract

The invention relates to a process for preparing a novel influenza vaccine, belonging to the technical field of preparation technology of vaccine. The invention comprises that influenza viruses of A1 and A3 of hypotype and B-type are diluted via virus nutrient solution and are individually inoculated mammal cells which grows to single layer according to 1:10 <-1> -1:10 <-7>, yielding virus liquid which contains the influenza viruses. After being deactivated and purified, each virus primary liquid is obtained. The antigen content of each virus primary liquid is detected. The antigen contents of the virus primary liquids of influenza viruses of A1 and A3 of hypotype and B-type are respectively regulated to prepare the vaccine. The invention has the advantage that by employing the mammal cells to produce the influenza vaccines, the animal immunogenicity experiments of the product show that 15 micrograms of hemagglutinin is capable of enabling serums of all experimental animals to be in seroconversion, and the HI antibody titers are all over 1:40.

Description

technical field [0001] The invention relates to a method for propagating influenza virus in animal cells and a novel inactivated influenza vaccine prepared by the method. In particular, it discloses a method for propagating influenza virus in animal cells, inoculating Vero cells with influenza virus, cultivating and harvesting the virus liquid, and preparing a monovalent stock solution through inactivation and purification. The invention belongs to the technical field of vaccine preparation technology. Background technique [0002] Influenza, referred to as influenza, is an acute respiratory infectious disease caused by influenza virus, referred to as influenza virus. The clinical manifestations are fever, headache, myalgia, fatigue, rhinitis, sore throat and cough, and may have gastrointestinal discomfort. Influenza can aggravate underlying diseases, such as cardiopulmonary disease, or cause secondary bacterial pneumonia or primary influenza viral pneumonia. The elderly an...

Claims

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Application Information

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IPC IPC(8): A61K39/145A61P31/16
Inventor 朱昌林张凤羽徐艳君葛鹏马达沈艳杰
Owner 长春百克生物科技有限公司
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