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Trivalent influenza virus subunit vaccine and preparation method thereof

A subunit vaccine and influenza virus technology, applied in the field of preparation of influenza virus subunit vaccines, can solve problems such as stubborn induration, itching and the like, and achieve the effects of easy process, high production efficiency and rapid preparation

Inactive Publication Date: 2018-01-05
ZHONGYI ANKE BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

It is also reported that the use of DPT vaccine adsorbed on aluminum hydroxide produced by the Statens Serum Institute in Denmark resulted in 546 serious adverse reactions of intractable itching at the injection site.

Method used

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  • Trivalent influenza virus subunit vaccine and preparation method thereof

Examples

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Embodiment 1

[0031] A 1 (H1N1) type influenza virus subunit vaccine monovalent stock solution production method, including the following steps in sequence:

[0032] (1). Virus inoculation: inoculate the working seeds of A1 (H1N1) influenza virus in the allantoic cavity of 10-day-old healthy chicken embryos;

[0033] (2). Virus propagation culture: transfer the inoculated chicken embryos to a 34°C incubator for 48 hours to propagate influenza virus;

[0034] (3). Harvesting of allantoic fluid: screen live chicken embryos, place the embryos at 2-8°C for 16 hours, and then harvest the allantoic fluid;

[0035] (4). Clarification: remove most impurities such as chicken embryo red blood cells by centrifugation.

[0036] (5). Concentration by ultrafiltration: The allantoic harvest fluid of influenza virus after fully inactivated is concentrated by ultrafiltration with an ultrafiltration membrane with a molecular weight cut-off of 1000kD;

[0037] (6). Inactivation: add formaldehyde with a fina...

Embodiment 2

[0041] A 3 (H3N2) type influenza virus subunit vaccine monovalent stock solution production method, including the following steps in sequence:

[0042] (1). Virus inoculation: Inoculate the working seeds of A3 (H3N2) influenza virus into the allantoic cavity of 10-day-old healthy chicken embryos;

[0043] (2). Virus propagation culture: transfer the inoculated chicken embryos to a 34°C incubator for 48 hours to propagate influenza virus;

[0044] (3). Harvesting of allantoic fluid: screen live chicken embryos, place the embryos at 2-8°C for 16 hours, and then harvest the allantoic fluid;

[0045] (4). Clarification: remove most impurities such as chicken embryo red blood cells by centrifugation.

[0046] (5). Concentration by ultrafiltration: Concentrate the allantoic harvested fluid of influenza virus fully inactivated by ultrafiltration with an ultrafiltration membrane with a molecular weight cut-off of 300kD;

[0047] (6). Inactivation: add formaldehyde with a final concent...

Embodiment 3

[0051] The method for producing monovalent stock solution of type B (B) influenza virus subunit vaccine comprises the following steps in sequence:

[0052] (1). Virus inoculation: inoculate the working seeds of type B (B) influenza virus in the allantoic cavity of 10-day-old healthy chicken embryos;

[0053] (2). Virus propagation culture: transfer the inoculated chicken embryos to a 34°C incubator for 72 hours to propagate influenza virus;

[0054] (3). Harvesting of allantoic fluid: screen live chicken embryos, place the embryos at 2-8°C for 16 hours, and then harvest the allantoic fluid;

[0055] (4). Clarification: remove most impurities such as chicken embryo red blood cells by centrifugation.

[0056] (5). Concentration by ultrafiltration: The allantoic harvest fluid of influenza virus after fully inactivated is concentrated by ultrafiltration with an ultrafiltration membrane with a molecular weight cut-off of 1000kD;

[0057] (6). Inactivation: Add formaldehyde with a...

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Abstract

The invention discloses a trivalent influenza virus subunit vaccine and a preparation method thereof, wherein virus protein after lysis is further purified by using a lysis agent and a new purification method to prepare a tetravalent influenza virus subunit vaccine, the content of three influenza hemagglutinins such as influenza A virus H1N1, influenza A virus H3N2 and influenza B virus in each dose of the vaccine is more than 80%, and the trivalent influenza virus subunit vaccine does not contain adjuvant and does not contain thimerosal and other preservatives. The invention further providesa preparation method of the trivalent influenza virus subunit vaccine, wherein the preparation method comprises: virus inoculation, virus proliferation culture, allantoic fluid harvesting, clarification, ultra-filtration concentration, inactivation, lysis and ultracentrifugation purification, gel filtration chromatography purification (ultra-filtration), blending, filtration sterilization, sub-packaging, packaging and other steps. According to the present invention, the trivalent influenza virus subunit vaccine can improve the safety of the influenza vaccine, can eliminate the adverse reactioncaused by the adjuvant, and can eliminate the toxic-side effects caused by thimerosal.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a preparation method of a novel influenza virus subunit vaccine. Background technique [0002] Influenza is caused by influenza virus, which can cause large-scale epidemics of acute respiratory diseases transmitted by humans and animals. Influenza viruses are divided into three types: A, B, and C. Among them, type A has the strongest pathogenicity and can infect animals and humans and cause epidemics and even worldwide pandemics; type B is less pathogenic and can cause local epidemics. Influenza viruses often undergo antigenic drift and antigenic shift, evading the defenses of the body's immune system, which is also the cause of the pandemic. Influenza viruses are highly contagious and spread through the air through droplets. After a short incubation period, high fever and chills develop rapidly, and the body temperature can reach as high as 40°C within 1 to 2 days, ac...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/145A61P31/16
Inventor 高辉
Owner ZHONGYI ANKE BIOTECH CO LTD
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