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Influenza virus subunit vaccine and preparation method thereof

A subunit vaccine and influenza virus technology, applied in the new influenza virus subunit vaccine and the field of preparation thereof, can solve problems such as intractable induration, itching, etc., and achieve the effects of easy and rapid preparation and reduction of redness and swelling.

Inactive Publication Date: 2015-09-09
ZHONGYI ANKE BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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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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  • Influenza virus subunit vaccine and preparation method thereof

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Effect test

Embodiment 1

[0030] The invention relates to an influenza virus subunit vaccine, wherein each dose of the influenza virus subunit vaccine contains three influenza hemagglutinins of type A1 (H1N1), type A3 (H3N2) and type B (B) with a content of more than 80%.

[0031] Further, the influenza virus subunit vaccine does not contain adjuvants such as CpG ODN adjuvant or aluminum hydroxide, and the vaccine does not contain preservatives such as thimerosal.

[0032] (1) Virus inoculation: Inoculate A1 (H1N1), A3 (H3N2) and B (B) influenza virus working seeds into the allantoic cavity of 9-11 day-old healthy chicken embryos respectively to obtain chicken embryos after inoculation. ;

[0033] (2) Virus proliferation culture: transfer the inoculated chicken embryos to a 33-35° C. incubator for 48-72 hours to propagate influenza virus;

[0034] (3) Harvesting of allantoic fluid: screen live chicken embryos, place cold embryos at 2-8°C for 10-24 hours, and harvest allantoic fluid;

[0035] (4) Clar...

Embodiment 2

[0051] This embodiment is a preferred solution based on Example 1. This embodiment is a method for producing a monovalent stock solution of a subunit vaccine of A1 (H1N1) type influenza virus, which comprises the following steps in turn:

[0052] (1) Virus inoculation: Inoculate the A1 (H1N1) type influenza virus working seeds in the allantoic cavity of 10-day-old healthy chicken embryos;

[0053] (2) Virus propagation and cultivation: transfer the inoculated chicken embryos to a 34° C. incubator for 48 hours to propagate the influenza virus;

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

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

[0056] (5) Inactivation: add formaldehyde with a final concentration not higher than 200 μg / ml to the monovalent virus harvest solution, and inactivate at 2-8°C for 80 hours;

[0057] (...

Embodiment 3

[0061] This embodiment is a preferred solution based on Example 1. This embodiment is a method for producing a monovalent stock solution of a subunit vaccine of A3 (H3N2) type influenza virus, which comprises the following steps in turn:

[0062] (1) Virus inoculation: Inoculate the A3 (H3N2) type influenza virus working seeds in the allantoic cavity of 10-day-old healthy chicken embryos;

[0063] (2) Virus propagation and cultivation: transfer the inoculated chicken embryos to a 34° C. incubator for 48 hours to propagate the influenza virus;

[0064] (3) Harvesting of allantoic fluid: screen live chicken embryos, place cold embryos at 2-8°C for 16 hours, and harvest allantoic fluid;

[0065] (4) Clarification: remove most impurities such as chicken embryo red blood cells by centrifugation.

[0066] (5) Inactivation: add formaldehyde with a final concentration not higher than 200 μg / ml to the monovalent virus harvest solution, and inactivate at 2-8°C for 100 hours;

[0067] ...

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Abstract

The invention discloses an influenza virus subunit vaccine and a preparation method thereof. The influenza virus subunit vaccine is formed by further purifying cracked viral proteins through a cracking agent and a new purification method; each agent of subunit vaccine comprises more than 80% of H1N1, H3N2 and B type influenza hemagglutinin, and does not comprise adjuvant, thiomersal or other corrosion removers. The invention further provides the preparation method of the influenza virus subunit vaccine. The preparation method of the influenza virus subunit vaccine comprises the following steps of virus inoculation, virus multiplication culture, allantonic fluid harvesting, clarification, inactivation, ultrafiltration and concentration, cracking and overspeed centrifugal purification, gel filtration chromatography purification, mixture, filtration sterilization, subpackage, packaging and the like. The influenza virus subunit vaccine can improve safety, eliminate untoward effects caused by adjuvant and eliminate the toxic and side effects caused by thiomersalate.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a novel influenza virus subunit vaccine and a preparation method thereof. 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...

Claims

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Application Information

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IPC IPC(8): A61K39/295A61P31/16
Inventor 孙卫华阮承迈孟东方王志刚关瑜
Owner ZHONGYI ANKE BIOTECH CO LTD
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