Subunit vaccine against feline infectious peritonitis virus and application

A subunit vaccine, peritonitis virus technology, applied in vaccines, viruses, viral peptides, etc., can solve the problems of few drugs showing enough good therapeutic effect, and achieve the effects of easy expression and purification, high safety, and simple preparation method

Inactive Publication Date: 2021-04-23
INST OF ANIMAL SCI OF CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Over the past 40 years, several studies have investigated potential treatments for FIP. Antivirals, immunostimulatory drugs, and immunosuppressive drugs have been used experimentally in the treatment of FIP, but few drugs have shown to be good enough The therapeutic effect of

Method used

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  • Subunit vaccine against feline infectious peritonitis virus and application
  • Subunit vaccine against feline infectious peritonitis virus and application
  • Subunit vaccine against feline infectious peritonitis virus and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Construction of FIPV antigen (RBD-FTH1)

[0037] The schematic diagram of the fusion protein RBD-FTH1 structure is as follows: figure 1 shown. The specific construction method is as follows: add RBD-FTH1 to the start codon and stop codon, and insert the prokaryotic expression plasmid pBV220 ( Bam H I) Between restriction sites, plasmid mass spectrum such as figure 2 shown. DH5αcompetent cells were transformed with the recombinant plasmid, cultured overnight at 37°C, and positive clones were identified by screening and PCR. After being verified by sequencing, it is used for the expression of nanoantigen protein. The plasmid was transformed into expression strain BL21, and the expression of nanoantigen protein was induced at 42°C.

[0038] 2. Purification of RBD-HFn nanoantigens

[0039] OD 280 =0.8 strains were sonicated and expressed in the supernatant after identification, and filtered through a 0.22 μm filter membrane to remove cell debris. After ultrafiltr...

Embodiment 2

[0041] Mouse Immunization Experiment

[0042] The RBD-FTH1 obtained in Example 1 was diluted to 100 μg / ml with physiological saline, and injected directly into mice. Then the 6-8 week-old Bal / C mice were immunized in groups. By intraperitoneal injection, each mouse was immunized three times on day 0, week 3 (day 21), and week 14 (day 108), with an inoculation volume of 100ul (10μg) each time. On the 10th, 31st, and 108th days, the mice were subjected to orbital blood collection. The mouse serum was left at room temperature for 2 hours, centrifuged at 3000 rpm for 15 minutes at 4°C, and immediately used in the FIPV neutralization assay.

[0043]

[0044] Neutralization experiment

[0045] Dilute the purified antibody 2-fold and mix with TCID 50 The final concentration of virus was mixed and incubated at 37°C for 1 hour. The mixture was added to a 96-well plate of Vero cells grown to about 80%. After 48 hours, the culture medium was discarded, and the cell lysate was wa...

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Abstract

The invention discloses a subunit vaccine against feline infectious peritonitis virus and an application thereof. The vaccine is a fusion protein RBD-FTH formed by connecting a surface protein epitope sequence containing FIPV particles and ferritin through a flexible Linker. According to the fusion protein RBD-FTH1, RBD-FTH1 monomers can be assembled into spherical 24-mer nanoparticles through the self-assembly effect of ferritin, and antigens in a RBD region are displayed on the surfaces of the nanoparticles, thereby causing stronger immune response of a receptor. According to the invention, the RBD of the FIPV is used as antigen fragments to achieve multimerization of the antigens based on the ferritin, thereby overcoming insufficient immunogenicity of RBD monomers, effectively causing stronger immune response and significantly improving antibody neutralization level of a host against FIPV.

Description

technical field [0001] The invention relates to the biological field, in particular to a feline infectious peritonitis virus subunit vaccine and its application. Background technique [0002] Feline infectious peritonitis (FIP), first reported four decades ago, is one of the deadliest infectious diseases of kittens. The disease is caused by feline coronavirus (FCoV) infection, has no effective treatment, and has a case fatality rate of approximately 100%. FCoV can be serologically divided into 2 types: serotypes I and II. Serotype I FCoV is the most prevalent virus epidemiologically, accounting for more than 70% of circulating virus isolates worldwide. FIP is difficult to isolate, which limits the research progress of FIP, and the molecular transmission mechanism is still unclear. Cats are one of the most popular pet species for humans, and they are also the main host of coronaviruses. Several large-scale coronavirus epidemics since the new century remind us that the risk...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/215A61K39/385A61P31/14C07K14/005C12N15/70
CPCA61K39/385A61K39/12A61P31/14C12N15/70C07K14/005C12N2770/20022C12N2770/20034C07K2319/00A61K2039/6031A61K2039/552
Inventor 梁瑞英成建伟崔尚金汤新明梁琳
Owner INST OF ANIMAL SCI OF CHINESE ACAD OF AGRI SCI
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