Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Mixed virus-like particles of swine influenza virus and foot and mouth disease virus, preparation method and application thereof

A swine foot-and-mouth disease virus, influenza virus technology, applied in chemical instruments and methods, botanical equipment and methods, microorganism-based methods, etc., can solve the problems of long production cycle, leakage and spread of live viruses, and high production conditions, and achieve strong immunity. , good immune effect, long-lasting effect

Active Publication Date: 2014-06-11
SUN YAT SEN UNIV
View PDF3 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] 1. Long production cycle: It takes 5-8 months from the acquisition of new subtype virus strains to the production and marketing of vaccines, making it difficult to contain new outbreaks
[0006] 2. High production conditions: In order to prevent artificial pollution of the environment and the leakage and spread of the live virus produced, the whole set of production must be carried out under the conditions of a biosafety level 3 laboratory according to regulations, and the inactivation of the virus must be guaranteed to be complete and thorough
[0007] 3. Inability to distinguish vaccinated pigs from virus-infected pigs
The currently used foot-and-mouth disease vaccines are mainly two types of whole virus inactivated vaccines and peptide vaccines, which cannot provide ideal immune protection due to potential safety hazards or poor immune effects

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Mixed virus-like particles of swine influenza virus and foot and mouth disease virus, preparation method and application thereof
  • Mixed virus-like particles of swine influenza virus and foot and mouth disease virus, preparation method and application thereof
  • Mixed virus-like particles of swine influenza virus and foot and mouth disease virus, preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 2

[0058] Example 2: Construction of insect baculovirus expression plasmids expressing M1, NP, HA, NA, and HF genes and synthesis of recombinant insect baculoviruses in insect cells

[0059] (1) Construction of recombinant plasmids expressing M1 and NP genes

[0060] Insect baculovirus plasmid PFastBac-dual (product of Invitrogen Company) was digested with restriction endonuclease Sal I / Hind III at 37°C for 3 hours, and the digested plasmid PFastBac-dual was recovered and purified with a gel extraction kit. Under the action of T4 DNA ligase, the digested plasmid and the digested M1 DNA fragment were ligated overnight at 16°C. The reaction system is as follows: 1ml of 10×T4 ligation buffer, 3ml of DNA fragment recovered by M1 digestion, 1ul of product recovered from enzyme digestion of PFastBac-dual plasmid, 1ul of T4 DNA ligase, and 10ul of ddH2O. Use the heat shock method to transfer the ligation product into Top10 competent cells, add it to a small plastic centrifuge tube, mix...

Embodiment 3

[0076] Example 3: Expression of M1, NP, HA, HF and NA genes in co-transfected suspension cultured insect cells sf-9

[0077] 200ml of sf-9 cell mixture was suspended and cultured in a 1-liter Erlenmeyer shaker flask, the cell culture medium was serum-free sf-900II (or Grace insect medium from Invitrigen), the shaking speed of the shaker was 100rpm, and the temperature was constant at 27°C. When the cell concentration reaches 2×10 6Cells / ml, sf9 cells were co-transfected with Bac-M1, Bac-NP, Bac-HA-NA, Bac-HF-NA insect baculovirus. The MOI ratio of the virus was 3 (Bac-M1 and Bac-NP): 1 (Bac-HA-NA): 1 (Bac-HF-NA). After the transfected cells were shaken and cultured at constant temperature for 3 days, all the samples were collected, centrifuged at 4° C. for 30 minutes at a spin speed of 3000 rpm, and the supernatant was collected. The centrifuged cell pellet was treated with cell lysate, and centrifuged at 4°C for 10 minutes at a centrifugal speed of 10,000 rpm. Keep the su...

Embodiment 4

[0078] Example 4: Purification of virus-like particles, indirect immunofluorescence detection and electron microscope observation.

[0079] Put the cell supernatant collected by the above centrifugation into a 13ml ultracentrifuge tube, weigh, balance, and seal the tube, put it into an ultracentrifuge (product of Bechmem Company), centrifuge at 100,000rpm at 4°C for 1 hour, and then take out the centrifuge tube , pour off the supernatant carefully, and keep the sediment at the bottom of the centrifuge tube. Add 5ml of PBS, put it in a refrigerator at 4°C, and dissolve for 24 hours. The next day, in another 13ml ultracentrifuge tube, first carefully add 1ml of 60% sucrose solution, then sequentially add 1ml of 30% and 3ml of 20% sucrose solution, and finally put 5ml of the dissolved sample solution on it. After accurate weighing and balance, seal the tube and put it into an ultracentrifuge. Centrifuge at 100,000 rpm for 1 hour at 4°C. The centrifuge tube was taken out, and t...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

PropertyMeasurementUnit
diameteraaaaaaaaaa
sedimentation rateaaaaaaaaaa
diameteraaaaaaaaaa
Login to View More

Abstract

The invention provides a mixed virus-like particles, containing stromatin M1 of influenza virus, surface antigen erythrohemagglutinin HA of influenza virus and a nexus protein containing main antigenic epitope contained capsid protein of foot and mouth disease virus and a transmembrane zone and an inner membrane zone of the erythrohemagglutinin HA of influenza virus. The main antigenic epitope contained capsid protein of foot and mouth disease virus substitutes a basically same length outer membrane zone of a 5' terminal of the erythrohemagglutinin HA of influenza virus. Surface antigen erythrohemagglutinin HA of influenza virus and the main antigenic epitope contained capsid protein of foot and mouth disease virus are expressed simultaneously on surfaces of the mixed virus-like particles. The invention also provides a divalent vaccine of swine influenza and foot and mouth disease; and the vaccine contains the mixed virus-like particles and adjuvants. The invention also provides a method for preparing the mixed virus-like particles.

Description

technical field [0001] The invention relates to a mixed virus-like particle of swine influenza virus and porcine foot-and-mouth disease virus, as well as its preparation and application. Background technique [0002] Swine influenza is an acute, febrile and highly contagious respiratory infectious disease caused by swine influenza virus (SIV) in pigs of different ages, sexes and breeds. Coughing, dyspnea, prostration, and death are the hallmarks. Pigs of all ages, sexes, and breeds can be infected, and the incidence rate is high. Infection with swine influenza virus alone can lead to a decline in pig production performance, increased feed-to-meat ratio, and slowed weight gain, causing certain economic losses to pig farms. What's more serious is that pigs are "mixers" for the recombination and replication of poultry, human, and swine influenza viruses. Swine influenza viruses have the ability to infect humans to the maximum without recombination. Moreover, early human infl...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Patents(China)
IPC IPC(8): A61K39/295A61K39/145A61K39/135C07K19/00C12N15/866A61P31/14A61P31/16C12R1/93
Inventor 曹永长刘大才薛春宜
Owner SUN YAT SEN UNIV
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com