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The Application of Citronellic Acid as a Synergist in Vaccine Production

A technology of citronellic acid and synergist, which is applied in the field of application of citronellic acid as a synergist in the production of viruses or virus vaccines, to achieve the effects of high-efficiency production, increased replication level, and promotion of SVA replication

Active Publication Date: 2022-02-08
LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The above studies mainly focus on the interaction regulation between SVA and key molecules in the innate immune pathway, but there are no reports on the promotion of SVA replication by chemical substances

Method used

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  • The Application of Citronellic Acid as a Synergist in Vaccine Production
  • The Application of Citronellic Acid as a Synergist in Vaccine Production
  • The Application of Citronellic Acid as a Synergist in Vaccine Production

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Embodiment 1 Citronellic acid promotes the effect evaluation experiment of SVA replication

[0029] 1. Preparation of Medium Containing Citronellic Acid

[0030] In DMEM medium, different doses of citronellic acid were added to make the concentrations 0mg / L, 10mg / L and 20mg / L respectively.

[0031] 2. Sample Preparation of Citronellic Acid-Incubated Cells

[0032] The IB cells were incubated with the prepared medium containing citronellic acid, then inoculated with SVA / FJ-201 (1 MOI), and the supernatant was collected 8 hours after inoculation.

[0033] 3. Determination of SVA virus titer

[0034] Determine the TCID of the supernatant obtained in the previous step 50 , for virus titer analysis.

[0035] TCID 50 The assay steps are as follows: inoculate the IB cells into a 96-well plate 16 hours in advance, wash the IB cells with PBS 3 times after the cells form a monolayer, and inoculate the supernatant collected in step 2 (10 -1 -10 -10 ) were used as infection ...

Embodiment 2

[0037] Embodiment 2 Citronellic acid promotes the effect evaluation experiment of SVA replication

[0038] 1. Preparation of Medium Containing Citronellic Acid

[0039] In DMEM medium, different doses of citronellic acid were added to make the concentrations 0mg / L, 10mg / L and 20mg / L respectively.

[0040] 2. Sample Preparation of Citronellic Acid-Incubated Cells

[0041] The PK-15 cells were incubated with the prepared medium containing citronellic acid, then inoculated with SVA / FJ-201 (1 MOI), and the supernatant was collected 12 hours after inoculation.

[0042] 3. Determination of SVA virus titer

[0043] After infection with SVA, the cell supernatant was collected and measured by TCID50 for virus titer analysis. Determination of virus titer: IB cells were inoculated into a 96-well plate 16 hours in advance. After the cells formed a monolayer, the IB cells were washed 3 times with PBS, and the supernatant collected in step 2 (10 -1 -10 -10 ), and set up two columns of ...

Embodiment 3

[0045] The citronellic acid of embodiment 3 different concentrations promotes the duplication of SVA in PK-15 or IB cell

[0046] 1. Preparation of Medium Containing Citronellic Acid

[0047] In the DMEM medium, different doses of citronellic acid were added to make the concentrations respectively 0mg / L, 2.5mg / L, 5mg / L, 10mg / L, 20mg / L, 40mg / L and 50mg / L.

[0048] 2. Sample Preparation of Citronellic Acid-Incubated Cells

[0049] Incubate PK-15 cells with the prepared medium containing the above-mentioned different concentrations of citronellic acid, then inoculate SVA (1MOI), and collect the supernatant 12 hours after inoculation;

[0050] At the same time, the IB cells were incubated with the prepared medium containing the above-mentioned different concentrations of citronellic acid, then inoculated with SVA (1 MOI), and the supernatant was collected 8 hours after inoculation.

[0051] 3. Determination of SVA virus titer

[0052]Cell supernatants were collected after infec...

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Abstract

The invention belongs to the field of biotechnology, and in particular relates to the application of citronellic acid as a synergist for the production of viruses or virus vaccines. The present invention unexpectedly found that adding citronellic acid to the medium for cultivating SVA can significantly increase the replication level of SVA, indicating that citronellic acid has the effect of promoting SVA replication and can be used as a synergist for SVA virus expression or vaccine production. It is used for efficient production of SVA virus or virus vaccine.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to the application of citronellic acid as a synergist for the production of viruses or virus vaccines. Background technique [0002] Senecavirus (Senecavirus A, SVA) is the only member of the newly added Senecavirus genus in the family Picornaviridae. Its clinical symptoms are indistinguishable from those of foot-and-mouth disease. ulcer. As SVA is an emerging infectious disease, there is no commercial vaccine yet, and its pathogenic and immune mechanisms are still not fully understood. [0003] Previous studies have shown that RIG-I is a key molecule for SVA to activate innate immunity, but 2C and 3C of SVA pro The protein can reduce the protein expression of RIG-I, and has the opposite antagonistic function to RIG-I; 2C and 3C of SVA pro It can also activate Caspase-3 and induce apoptosis; 3C of SVA pro Use its own enzyme activity to cleave key natural immune molecules...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N7/00A61K39/125A61P31/14
CPCC12N7/00A61K39/12A61P31/14C12N2770/32051C12N2770/32034A61K2039/552
Inventor 郑海学刘会胜薛巧朱紫祥宋影影薛钊宁
Owner LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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