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Quadrivalent subunit influenza vaccine and preparation method thereof

An influenza vaccine and subunit technology, applied in the field of tetravalent subunit influenza vaccine, can solve problems such as limited influenza prevention, and achieve the effects of low adverse reactions, low formaldehyde and lysing agent residues, and a wide range of defense protection.

Inactive Publication Date: 2017-05-17
AB&B BIO TECH CO LTD JS
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The protection of trivalent vaccine against influenza is obviously limited

Method used

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  • Quadrivalent subunit influenza vaccine and preparation method thereof
  • Quadrivalent subunit influenza vaccine and preparation method thereof
  • Quadrivalent subunit influenza vaccine and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0059] A quadrivalent subunit influenza vaccine, wherein each dose of the quadrivalent influenza subunit vaccine contains A1 (H1N1) type, A3 (two lineages of (H1N3) type and B (B) type), each valent antigen content It is 30~36μg / mL.

[0060] 1. Virus Inoculation and Culture

[0061] After diluting the batch virus species of the working seeds to a final concentration within the range of 2.0lgEID50 / mL to 5.0lgEID50 / mL (various types of influenza strains should be inoculated according to the same virus titer), inoculate chickens at a dose of 0.2mL / embryo. Embryo allantoic cavity, cultured at 33-35°C for 48-72 hours, unused working seeds to batch virus species, shall not be thawed for further use.

[0062] 2. Virus Harvesting

[0063] Screen live chicken embryos, place them in cold embryos at 2-8°C for 12-24 hours, harvest the allantoic fluid and take samples for testing: microbial limit, hemagglutination titer.

[0064] 3. Centrifuge to clarify and combine

[0065] The allant...

experiment example 1 4

[0077] Experimental Example 1: Research on semi-finished product technology of quadrivalent influenza virus subunit vaccine

[0078] 1. Materials

[0079] Monovalent stock solution of four types of influenza virus

[0080] 2. Method

[0081] According to the hemagglutinin content of each monovalent stock solution, the semi-finished products of various types of influenza viruses were prepared according to the same hemagglutinin content (the amount of hemagglutinin preparation can be within the range of 30-36 μg / mL, and each type of influenza virus strain should be prepared according to the same hemagglutinin content every year). Lectin content for preparation), that is, semi-finished products. According to the volume of the required monovalent stock solution diluent as V and the monovalent stock solution hemagglutinin content C0, calculate the volume V0=V / (C0 / hemagglutinin preparation amount) of the required monovalent stock solution, 0.01mol / L phosphate buffer saline The vo...

Embodiment 2

[0153] On the basis of Example 1, the density gradient centrifugation technique was replaced by the hydrophobic chromatography technique.

[0154] The specific method is: prepare the lysing agent Triton N101 with a concentration not higher than 1.5%, place it at 15-25° C., and lyse the virus for 0.5-2 hours. Afterwards, the different serotype samples were separated and purified using a hydrophobic chromatography column Butyl-S-Sepharose 6ff (C4). The chromatography medium is butylff, Octylff or PhenylF, and the loading buffer is 50mmol / L PBS+1.5mol / L (NH 4 ) 2 SO 4 (pH7.0), containing 100% ammonium sulfate, and set the ammonium sulfate content in 50mmol / L PBS as 0. After loading an appropriate amount of sample, the flow rate to ensure the column pressure will be eluted from the set ammonium sulfate content of 100% to 0 at a constant speed. , monitored at a wavelength of 280nm, and collected protein peaks.

[0155] The samples after hydrophobic chromatography were analyzed,...

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Abstract

The invention relates to the field of biological pharmacy, and particularly discloses a quadrivalent subunit influenza vaccine and a preparation method thereof. Used virus seeds are respectively of an H1N1 type (NYMC X-179A), an H3N2 type (NIB-88), a BY type (B / Phuket), and a BV type (NYMC BX-35). The preparation method comprises the following steps: performing chick embryo cultivation on the virus seeds; preparing a mono-valent virus stock solution by the steps of allantoic fluid collection, clarification, concentration, purification and the like; and proportioning four mono-valent virus stock solutions with different serotypes according to certain dilution points to prepare a quadrivalent subunit influenza vaccine sample. A relatively low content of ovalbumin and relatively low residual quantities of formaldehyde and a cracking agent in the quadrivalent subunit influenza vaccine sample ensure that adverse reactions of a quadrivalent subunit vaccine are lower, and the safety of the vaccine is improved. Meanwhile, compared with an existing trivalent vaccine, the quadrivalent subunit vaccine additionally has a BV serotype, thereby having a broader protection scope for popularity of influenzas with different serotypes.

Description

technical field [0001] The invention relates to the field of biopharmaceuticals, in particular to a quadrivalent subunit influenza vaccine. Background technique [0002] Influenza is caused by influenza virus, which can cause large-scale epidemics of acute respiratory diseases. Influenza can occur suddenly in a short period of time and spread rapidly, causing different degrees of epidemics, including world pandemics, local outbreaks, and dissemination. High-risk groups are elderly and infirm or patients with chronic diseases. After each influenza epidemic, there will always be different degrees of excess deaths in the population. Therefore, the prevention of influenza has been highly valued by countries all over the world. The most effective way to prevent influenza is to get a flu vaccine. [0003] At present, the influenza vaccines used all over the world are trivalent, including two types of A (H1N1, H3N2) and one type B (type B) influenza virus antigen components. Th...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/145A61P31/16
CPCA61K39/12A61K2039/70C12N2760/16134C12N2760/16234
Inventor 安有才付作申张玉辉于海龙陈辉黎刘鹏刘冰冰安蕊徐奇冷文娜陈晓芬
Owner AB&B BIO TECH CO LTD JS
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