Newcastle disease virus chimeric virus-like particle, vaccine and preparation method

A technology for Newcastle disease virus and chimeric virus is applied in the field of Newcastle disease virus chimeric virus-like particles, preparation and prevention of Newcastle disease virulent vaccines, which can solve problems such as mismatch of vaccine strains, achieve elimination of insect cell fusion, Improves synthesis and increases the number of cells growing

Active Publication Date: 2020-02-11
NOVARTIS BIOTECH WUHAN +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0013] Another object of the present invention is to provide a vaccine for preventing virulent Newcastle disease. The vaccine is a chimeric virus-like particle vaccine, which is a novel vaccine prepared by modern biological principles and methods. Virus strain gene type VII is used as a vaccine strain, which solves the problem that the vaccine strain does not match the epidemic strain; secondly, the vaccine has high antigen content and is easy to produce; the post-translational processing of the expressed product is similar to the structural protein of Newcastle disease virus, and the particles Newcastle disease virus membrane protein on the surface retains its native structure, biological activity and immunogenicity

Method used

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  • Newcastle disease virus chimeric virus-like particle, vaccine and preparation method
  • Newcastle disease virus chimeric virus-like particle, vaccine and preparation method
  • Newcastle disease virus chimeric virus-like particle, vaccine and preparation method

Examples

Experimental program
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Effect test

Embodiment 1

[0058] Construction and Detection of Newcastle Disease Virus (NDV) Chimeric Virus-like Particle Protein Vector

[0059] According to the epidemic trend of virulent strains of Newcastle disease virus combined with antigen analysis and codon optimization at the same time, the genes of F protein and HN protein of gene type VII of virulent Newcastle disease virus strains were artificially synthesized, and two retroviruses (chicken leukemia virus ALV, Gag precursor protein gene of human immunodeficiency virus HIV-1).

[0060] Newcastle disease virus fusion protein F and hemagglutinin-neuraminidase HN are used as the membrane glycoprotein of Newcastle disease virus (NDV) chimeric virus-like particles, and have the function of virus surface antigen of virulent strain gene type VII. The Gag precursor protein will be used as the matrix protein of the NDV chimeric virus-like particle, released from the host cell in the form of budding, and self-assembled into the chimeric virus-like par...

Embodiment 2

[0072] Expression and self-assembly of NDV chimeric virus-like particle protein

[0073] Insect cell sf9 was transfected with recombinant bacmid bacmid with F gene, HN gene and Gag gene (from chicken leukemia virus ALV, or human immunodeficiency virus HIV-1), and the cell supernatant was collected six days later to obtain The first generation of recombinant baculovirus strains 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. Then use the P2 generation virus seeds to amplify to obtain the P3 generation. The NDV chimeric virus-like particles were prepared by using the P3 generation as the production virus seed. Briefly, Sf-9 cells in good growth state were inoculated into shake flasks, and when the cells grew to the logarithmic growth phase, the cell concentration was diluted to 3.0×10 6 cells. Inoculate the virus into cell shake f...

Embodiment 3

[0083] Purification and Electron Microscopic Observation of NDV Chimeric Virus-like Particles

[0084] NDV chimeric virus-like particles were purified by discontinuous sucrose ultracentrifugation. The collected 20mL cell supernatant was centrifuged at 4°C and 100000xg for 1 hour in an ultracentrifuge with an angle rotor, discarded the centrifuged supernatant, and added 5ml PBS to suspend and dissolve overnight. The sucrose solutions were prepared, and the mass percentage concentrations were respectively adjusted to 20%, 45% and 60%, and carefully loaded into ultracentrifuge tubes to form a discontinuous sucrose density gradient. Add overnight 5ml PBS suspension to the top layer, and perform ultracentrifugation with a horizontal rotor at 4°C, 100000xg, for 1 hour. There is a band at the junction of 20% and 45% and at the junction of 45% and 60%, and the band at the junction of 20% and 45% is the most prominent (see Figure 5 ). The bands here were collected, which contained ...

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Abstract

The invention discloses virus like particles of newcastle disease virus, a vaccine and a preparing method. The vaccine is a virus like particle vaccine. The virus like particles are chimeric newcastle disease virus like particles. The vaccine is a novel vaccine prepared by the adoption of modern biology principles and methods. The vaccine adopts currently popular virulent newcastle disease virus strain gene type VII as a vaccine strain, and solves the problem that existing vaccine strains are not matched with epidemic strains; besides, the vaccine is high in antigen content and easy to produce; post-translational processing of an expression product is similar to that of structural protein of newcastle disease virus, and newcastle disease virus membrane protein on the surfaces of the particles maintains a natural structure, biological activity and immunogenicity.

Description

technical field [0001] The invention relates to the technical field of virus vaccines, in particular to a chimeric virus-like particle of Newcastle disease virus, a preparation method and a vaccine for preventing virulent Newcastle disease virus. Background technique [0002] Newcastle disease (ND) is an acute and highly contagious infectious disease. The disease spreads rapidly, the morbidity and mortality rate can reach more than 85%, and the infection caused by strong virus strains often reaches 100%. It is one of the most serious poultry infectious diseases in the world today. Since it first appeared in West Asia, Indonesia, and Newtown, England in 1926, in the following 90 years of history, Newcastle disease has had many pandemics in the world. Experiments have shown that more than 250 kinds of birds can be infected by Newcastle disease virus (NDV) Infect. In many areas of our country, the disease occurs every year, and Newcastle disease virus is also isolated from wa...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N7/04A61K39/17A61P31/14G01N33/569
CPCA61K39/12A61K2039/5258A61K2039/552C12N7/00C12N2760/18123C12N2760/18134G01N33/56983
Inventor 许雁王志亮戈胜强诺曼·吉利卡李改
Owner NOVARTIS BIOTECH WUHAN
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