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Swine fever virus chimeric type virus-like particle, preparation method, application thereof and vaccine

A swine fever virus, chimeric technology, applied in veterinary vaccines, botanical equipment and methods, biochemical equipment and methods, etc. Poor immunity, large dose of vaccination, etc., to achieve the effect of significant immune effect, wide application range and obvious effect

Inactive Publication Date: 2019-03-29
NOVARTIS BIOTECH WUHAN +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

For example, nucleic acid vaccines are slow in inducing antibodies, and the inoculation dose is large, which has the potential risk of integration and transformation of chromosomes; for humans, live virus vector vaccines cannot control this type of virus because they do not fully understand the natural occurrence and mutation mechanism of the virus. The future trend of the future; the subunit vaccine immunogenicity is usually worse than the virus particle antigen, and cannot provide effective protection; the gene deletion vaccine is usually a live attenuated vaccine, and its long-term use may change the original ecological environment and virus community of CSFV. Make the swine fever epidemic strain evolve away from the vaccine virus, and it is difficult to meet the requirements of modern prevention and control and eradication of swine fever

Method used

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  • Swine fever virus chimeric type virus-like particle, preparation method, application thereof and vaccine
  • Swine fever virus chimeric type virus-like particle, preparation method, application thereof and vaccine
  • Swine fever virus chimeric type virus-like particle, preparation method, application thereof and vaccine

Examples

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Effect test

Embodiment 1

[0041] Embodiment 1: Construction and detection of expression vector CSF-ALVGag, CSF-HIVGag recombinant plasmid

[0042] A chimeric virus-like particle of classical swine fever virus according to the present invention, the chimeric virus-like particle comprises Gag precursor protein of retrovirus, surface envelope protein E0 of classical swine fever virus and surface envelope protein E2 of classical swine fever virus . Wherein the retrovirus Gag precursor protein is chicken leukemia virus precursor protein ALV-Gag or human immunodeficiency virus precursor protein HIV-Gag. Envelope proteins E2 and E0 of classical swine fever virus are derived from envelope proteins E2 and E0 of all genotype strains of classical swine fever virus, preferably, envelope proteins E2 and E0 contained in chimeric virus-like particles of classical swine fever virus are derived from Strain of classical swine fever virus genotype 2.1. The amino acid sequence of the surface envelope protein E0 of class...

Embodiment 2

[0058] Embodiment 2: the preparation method of the chimeric virus-like particle of classical swine fever virus

[0059] This example provides a method for expressing and preparing chimeric virus-like particles of CSFV and detecting the expressed target protein.

[0060] 1. In this embodiment, the expression and preparation of CSF-ALVGag chimeric virus-like particles and the detection of the expressed target protein are taken as an example to illustrate. The specific methods are as follows:

[0061] Insect cells sf-9 were plated in a 6-well cell culture plate, and the recombinant baculovirus plasmid CSF-ALVGag was transfected by lipid method, and the cell supernatant was collected six days later to obtain the first generation of recombinant baculovirus species, 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 inf...

Embodiment 3

[0068] Example 3: Purification of CSFV chimeric virus-like particles

[0069] This example provides the purification method and electron microscope observation results of chimeric virus-like particles of CSFV.

[0070] This example illustrates the purification and electron microscope results of the prepared CSF-ALVGag chimeric virus-like particles as an example. The specific methods are as follows:

[0071]CSF-ALVGag was 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 5 ml of the above-mentioned overnight PBS suspension to the top l...

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Abstract

The present invention provides a swine fever virus chimeric type virus-like particle, a preparation method thereof, an application thereof and a vaccine, and belongs to the technical fields of bioengineering and virus vaccines. The swine fever virus chimeric type virus-like particle selects a Gag precursor protein of retroviruses as a matrix protein and swine fever virus envelope proteins E0 and E2 as surface membrane proteins to self-assemble into the ideal and functional virus-like particle, weaknesses of a subunit protein vaccine are overcome, and immunogenicity, safety and stability of thevaccine are effectively improved. The swine fever virus chimeric type virus-like particle can be prepared in large quantities by constructing expression vectors to transfect cells. The swine fever virus chimeric type virus-like particle can be applied to prepare the vaccine for preventing swine fever epidemic diseases, a swine fever virus antibody detection preparation or a swine fever virus epidemic disease monitoring preparation, and has a relatively good practical application value.

Description

technical field [0001] The invention relates to the technical fields of bioengineering and virus vaccines, in particular to a chimeric virus-like particle of classical swine fever virus, a preparation method and its application, and a vaccine. Background technique [0002] Swine fever is an acute, febrile, and highly contagious infectious disease of pigs caused by swine fever virus. It can be divided into acute, subacute, chronic, and atypical swine fever. Acute swine fever is caused by virulent strains and generally results in high morbidity and mortality in pigs. In addition to causing sepsis, classical swine fever virus can also cause a series of clinical manifestations, such as abortion in pregnant sows, fetal malformation, chronic nutrient consumption, lymphocyte and thrombocytopenia and other immune system lesions, and are prone to secondary and concurrent bacterial or other viral infections . Swine fever has caused huge economic losses to the swine industry in the w...

Claims

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Application Information

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IPC IPC(8): C07K14/18C07K14/15C07K14/155C12N15/866A61K39/187A61P31/14G01N33/68G01N33/569
CPCA61K39/12G01N33/56983G01N33/6854C12N15/86C07K14/005G01N2469/20C12N2740/16023C12N2710/14043C12N2740/11023C12N2770/24323C12N2770/24334C12N2800/105A61K2039/552A61K2039/5258G01N2333/183
Inventor 许雁诺曼﹒吉利卡李改夏燕
Owner NOVARTIS BIOTECH WUHAN
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