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Ordinary/chimeric virus-like particle of H7 subtype influenza virus H7N9, preparation method and application of ordinary/chimeric virus-like particle, and vaccine

A technology of influenza A virus and influenza virus, applied in the direction of virus/bacteriophage, biochemical equipment and methods, virus, etc., to achieve high titer, good practical application value, and obvious effect

Active Publication Date: 2018-07-27
NOVARTIS BIOTECH WUHAN +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

It can be seen that the current H7N9 inactivated vaccine has certain safety and environmental risks, and a safer vaccine needs to be developed

Method used

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  • Ordinary/chimeric virus-like particle of H7 subtype influenza virus H7N9, preparation method and application of ordinary/chimeric virus-like particle, and vaccine
  • Ordinary/chimeric virus-like particle of H7 subtype influenza virus H7N9, preparation method and application of ordinary/chimeric virus-like particle, and vaccine
  • Ordinary/chimeric virus-like particle of H7 subtype influenza virus H7N9, preparation method and application of ordinary/chimeric virus-like particle, and vaccine

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0058] This example provides the construction and detection of the expression vectors of common virus-like particles of H7 subtype influenza virus H7N9 and chimeric virus-like particles of H7 subtype influenza virus H7N9; common virus-like particles contain influenza A virus matrix protein M1, H7 subtype influenza virus H7N9 hemagglutinin protein HA, H7 subtype influenza virus H7N9 neuraminidase protein NA.

[0059] Chimeric virus-like particles contain Gag precursor protein, H7N9 influenza virus hemagglutinin protein HA and H7N9 influenza virus neuraminidase protein NA; Gag precursor protein of retrovirus is chicken leukemia virus precursor protein ALV-Gag or Human immunodeficiency virus precursor protein HIV-Gag.

[0060] The gene sequences of hemagglutinin protein HA and neuraminidase protein NA were obtained according to A / Guangdong / 17SF003 / 2017. The gene sequence of matrix protein M1 comes from H5N1 subtype strain A / Indonesia / 5 / 2005 (H5N1)

[0061] The amino acid sequen...

Embodiment 2

[0069] This example provides a method for expressing and preparing common virus-like particles and chimeric virus-like particles.

[0070] Insect cells sf-9 were plated in a 6-well cell culture plate, and the recombinant bacmids HA-NA-M1, HA-NA-ALV-Gag and HA-NA-HIV-Gag were transfected by lipid method After six days, the cell supernatant was collected to obtain the first-generation recombinant baculovirus seed, which is called the P1 generation. Infect insect cells with the P1 virus seed to amplify the virus seed to obtain the second-generation recombinant baculovirus seed, which is called the P2 generation. Insect cells were then infected with the P2 virus seed to amplify the virus seed to obtain the P3 generation. The P3 generation was used as the production virus seed to prepare H7N9 conventional virus-like particles. In short, Sf-9 cells in good growth state were inoculated into shake flasks for suspension culture, and when the cells grew to the logarithmic growth phase...

Embodiment 3

[0079] This embodiment compares the common type virus-like particles (HA-NA-M1 virus-like particles) of H7 subtype influenza virus H7N9 and the chimeric virus-like particles (HA-NA-HIVGag virus-like particles and HA-NA-ALVGag virus-like particles) yield under the same conditions.

[0080] There are two membrane proteins on the surface of avian influenza virus, namely hemagglutinin HA protein and neuraminidase NA protein. The hemagglutinin HA protein can bind to the receptors on the surface of red blood cells to form the phenomenon of red blood cell dispersion without precipitation. This characteristic can be used as a judgment index for the detection of hemagglutination titer. Therefore, chicken erythrocyte agglutination can be used to detect the content of H7N9 virus-like particles in the cell supernatant. In order to compare the yield difference of H7N9 common type VLPs and H7N9 chimeric VLPs, under the condition that the production conditions of the three kinds of VLPs wer...

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Abstract

The invention provides an ordinary / chimeric virus-like particle of an H7 subtype influenza virus H7N9, a preparation method and application of the ordinary / chimeric virus-like particle, and a vaccine,and belongs to the technical field of bioengineering and viral vaccines. The ordinary / chimeric virus-like particle of the H7 subtype influenza virus is used for preparing the vaccine, and accordingly, the safety problems of existing vaccines are well solved; the obtained vaccine has high antibody valence and obvious effect; the ordinary / chimeric virus-like particle has wide application range andhigh actual application value.

Description

technical field [0001] The invention relates to the technical fields of bioengineering and virus vaccines, in particular to common / chimeric virus-like particles of H7 subtype influenza virus H7N9, a preparation method, application and vaccine. Background technique [0002] In nature, influenza A viruses possess enormous diversity due to the antigenicity of their two major viral surface proteins. The virus is usually asymptomatic in waterbirds, its natural host. Poultry infected with the virus can cause a series of clinical diseases, ranging from mild infection known as low pathogenic avian influenza (LPAI) to severe multi-system fatal infection known as highly pathogenic avian influenza (HPAI). H7N9 (Influenza A virus subtype H7N9) is a strain in the H7 subtype of influenza A virus. H7N9 was originally a low-pathogenic influenza virus that was only found in poultry. In 2006, the National Center for Biotechnology Information of the U.S. Department of Health released the fi...

Claims

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Application Information

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IPC IPC(8): C07K14/11C07K14/16C07K14/15A61K39/145A61K39/21A61P31/16G01N33/569
CPCA61K39/12A61P31/16G01N33/56983C07K14/005C12N2760/16123C12N2760/16134C12N2740/11034C12N2740/11023C12N2740/16023C12N2740/16234A61K2039/5258
Inventor 许雁诺曼·吉利卡李改
Owner NOVARTIS BIOTECH WUHAN
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