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Recombinant pseudorabies virus vaccine heat-resistant freeze-drying protectant and preparation method thereof

A technology for recombinant pseudorabies and virus vaccines, applied in antiviral agents, biochemical equipment and methods, vaccines, etc., can solve the problem of unsatisfactory protection performance of heat-resistant freeze-drying protective agents, long-term storage and long-distance transportation restrictions of vaccines, and storage period There are only 3-6 months to achieve the effect of protecting live vaccines, good physical properties, and mass production

Active Publication Date: 2018-03-20
ZHAOQING DAHUANONG BIOLOGIC PHARMA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the domestic conventional protective agents are mainly composed of skim milk, sucrose, gelatin and other components. The composition is simple, but the thermal stability and protection function are poor, and the storage time is short. Under the condition of 2°C-8°C, the storage period is only 3-3. 6 months, and most of them need to be stored below -15°C, which greatly limits the long-term storage and long-distance transportation of vaccines
Chinese patent CN102727904A discloses a heat-resistant protective agent for highly pathogenic porcine reproductive and respiratory syndrome-pseudorabies dual live vaccine. ~6%, glycine 0.5~1.5%, sucrose 3~5%, trehalose 2~3%, polyvinylpyrrolidone 1~2%, mannitol 0.5~1%, vitamin E0.1~0.2% and water for injection High-temperature sterilization, microporous membrane filtration sterilization, and preparation by mixing steps have good protective functions, but its ingredients contain gelatin, which is the main source of vaccine endotoxins, and can easily cause fever reactions in vaccinated animals
Existing heat-resistant freeze-dried protective agents for vaccines have unsatisfactory multi-protection performance, or contain animal-derived components such as gelatin, serum albumin, and tryptone, which may affect the safety of the vaccine

Method used

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  • Recombinant pseudorabies virus vaccine heat-resistant freeze-drying protectant and preparation method thereof
  • Recombinant pseudorabies virus vaccine heat-resistant freeze-drying protectant and preparation method thereof
  • Recombinant pseudorabies virus vaccine heat-resistant freeze-drying protectant and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Example 1 Recombinant pseudorabies virus vaccine heat-resistant lyoprotectant of the present invention

[0023] Recombinant pseudorabies virus vaccine heat-resistant lyoprotectant, which is made of the following components by weight percentage: trehalose 7%, sucrose 1%, sorbitol 0.6%, polyvinylpyrrolidone K-30 1%, soybean isolate Protein 6%, wheat protein hydrolyzate 0.7%, and the balance is water for injection.

[0024] Preparation:

[0025] S1. Take wheat protein powder, add purified water 15 times its weight, stir evenly, add 10mol / L HCL to the suspension to adjust the pH value of the solution to 3.0, add 1800U / g acid protease, enzymolyze at 40°C for 8h, use 2mol / L HCL or NaOH to keep the pH value of the solution unchanged. After the enzymolysis is completed, the enzyme is extinguished at 95°C for 20 minutes. After cooling, the supernatant is collected by centrifugation, concentrated under reduced pressure, and dried under reduced pressure to obtain the enzymatic hy...

Embodiment 2

[0029] Example 2 Recombinant pseudorabies virus vaccine heat-resistant lyoprotectant of the present invention

[0030] Recombinant pseudorabies virus vaccine heat-resistant lyoprotectant, which is made of the following components by weight percentage: trehalose 5%, sucrose 1%, sorbitol 0.6%, polyvinylpyrrolidone K-30 1.5%, soybean isolate Protein 5%, wheat protein hydrolyzate 0.8%, and the balance is water for injection.

[0031] Preparation:

[0032] S1. Take wheat protein powder, add 15 times its weight in purified water, stir evenly, add 8mol / L HCL to the suspension to adjust the pH value of the solution to 2.0, add 1200U / g of pepsin, enzymolyze at 42°C for 10h, and use 1mol / L HCL or NaOH to keep the pH value of the solution unchanged, inactivate the enzyme at 95°C for 20 minutes after enzymolysis, and centrifuge to take the supernatant after cooling, concentrate under reduced pressure, and dry under reduced pressure to obtain the enzymatic hydrolyzate of wheat protein;

...

Embodiment 3

[0036] Example 3 Heat-resistant lyoprotectant for recombinant pseudorabies virus vaccine of the present invention

[0037] Recombinant pseudorabies virus vaccine heat-resistant lyoprotectant, which is made of the following components by weight percentage: trehalose 4%, sucrose 3%, sorbitol 0.7%, polyvinylpyrrolidone K-30 2%, soybean isolate Protein 5%, wheat protein hydrolyzate 0.8%, and the balance is water for injection.

[0038] The preparation method is the same as in Example 2.

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Abstract

The invention relates to a heatproof freeze-drying protecting agent for recombined pseudorabies virus vaccines. The heatproof freeze-drying protecting agent comprises, by weight, 4-8% of trehalose, 0-3% of cane sugar, 0.2-0.8% of sorbitol, 1-3% of polyvinylpyrrolidone K-30, 4-7% of soybean protein isolate, 0.5-1% of wheat proteolysis product and the balance injection water. The invention further provides a preparation method of the heatproof freeze-drying protecting agent for the recombined pseudorabies virus vaccines. The heatproof freeze-drying protecting agent is simple in formula, easy to prepare and suitable for mass production, has the advantages of high vaccine stability, prolonged vaccine shelf life and the like, and does not contain gelatin and animal source ingredients. Safety of the vaccines is high when the protecting agent is applied to the vaccines.

Description

technical field [0001] The invention relates to a heat-resistant freeze-drying protective agent for vaccines and a preparation method thereof, in particular to a heat-resistant freeze-drying protective agent for recombinant pseudorabies virus vaccines and a preparation method thereof. Background technique [0002] Pseudorabies virus (PRV), also known as porcine herpesvirus type I, is one of the main pathogens causing reproductive disorders in pigs. The PRV genome is huge and contains multiple genes that can be deleted and are not necessary for virus proliferation, which can be inserted into foreign genes. The recombinant virus inserted with foreign genes can stably express foreign genes while proliferating, thus providing protection against pseudorabies virus and other viruses. Corresponding infectious disease resistance, so it can be used as a recombinant virus vector for the development of recombinant vaccines. The research on recombinant pseudorabies virus mainly involve...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K47/26A61K47/32A61K47/42A61K39/245A61K39/12A61P31/22A61P31/20
CPCA61K39/12A61K47/26A61K47/32A61K47/42A61K2039/53A61K2039/70C12N2710/16734C12N2750/10034
Inventor 陈瑞爱徐家华张文炎何芳唐兆新高艳
Owner ZHAOQING DAHUANONG BIOLOGIC PHARMA
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