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Method for producing swine fever live vaccine with cell line

A live swine fever vaccine and cell line technology, which can be used in pharmaceutical formulations, medical preparations containing active ingredients, antibody medical ingredients, etc., can solve the difficulties in improving vaccine production and efficacy, exogenous virus contamination of cells, and toxin-producing drops It can achieve good economic benefits and application prospects, high virus content, and solve the effect of exogenous pathogen contamination of cattle.

Active Publication Date: 2010-01-06
CHINA INST OF VETERINARY DRUG CONTROL +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Since BVD / MDV disease is relatively common in cattle herds in my country, the use of primary cells of bovine testis to produce vaccines may easily cause cell exogenous virus contamination, and the titer of toxin production is not high, and there are large batch-to-batch differences, which will bring great benefits to the improvement of vaccine production and efficacy. difficulty

Method used

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  • Method for producing swine fever live vaccine with cell line
  • Method for producing swine fever live vaccine with cell line
  • Method for producing swine fever live vaccine with cell line

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] (1) Selection of cells for seedling production: use porcine testis (ST) cell line;

[0028] (2) Subculture and culture of cells for seedling production: ST cell line is digested and subcultured with EDTA-trypsin cell dispersion, then cultured with cell growth solution at 36°C to form a good monolayer, used for continued subculture or virus inoculation;

[0029] (3) Propagation of cytotoxic species: Use cell maintenance solution to prepare a 0.3% virus suspension from fresh spleen venom, inoculate a well-growing ST cell line monolayer, and continue culturing at 36°C. Harvest and change the liquid every 4 days, and take the secondary and tertiary cell culture venom as the production poison;

[0030] Identification of cytotoxic species: The identification of cytotoxic species is in full compliance with the standard of attenuated strains of classical swine fever lapinization and has no side effects on pigs. The cytotoxic species contains more than 500,000 rabbit infections per 1 m...

Embodiment 2

[0038] (1) Selection of cells for seedling production: use pig kidney (PK15) cell line;

[0039] (2) Subculture and culture of cells for seedling production: PK15 cell line is digested and subcultured with EDTA-trypsin cell dispersion, then cultured with cell growth solution at 37°C to form a good monolayer, used for continued subculture or virus inoculation;

[0040] (3) Propagation of cytotoxic species: Using cell maintenance solution, fresh spleen toxin was made into a 0.4% virus suspension, inoculated with a monolayer of well-growing PK15 cell line, and kept at 37°C to continue the culture. Harvest and change the liquid every 5 days, and take the secondary and tertiary cell culture venom as the production poison;

[0041] Identification of cytotoxic species: The identification of cytotoxic species is in full compliance with the standard of attenuated strains of classical swine fever lapinization and has no side effects on pigs. The cytotoxic species contains more than 500,000 rab...

Embodiment 3

[0049] (1) Selection of cells for seedling production: use pig kidney (IBRS-2) cell line;

[0050] (2) Subculture and culture of cells for seedling production: IBRS-2 cell line is digested and subcultured with EDTA-trypsin cell dispersion, then cultured with cell growth medium at 37°C to form a good monolayer, used to continue subculture or inoculation virus;

[0051] (3) Propagation of cytotoxic species: Use cell maintenance solution to make 0.5% virus suspension from fresh spleen venom, inoculate a monolayer of well-growing IBRS-2 cell line, and continue culturing at 37°C. Harvest and change the liquid every 4 days, and take the secondary and tertiary cell culture venom as the production poison;

[0052] Identification of cytotoxic species: The identification of cytotoxic species is in full compliance with the standard of attenuated strains of classical swine fever lapinization and has no side effects on pigs. The cytotoxic species contains more than 500,000 rabbit infections per ...

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Abstract

The invention discloses a method of using clone for producing swine fever living vaccine. The invention comprises the following technical steps: (1) selecting clone as the cell for producing vaccine; (2) the culture transferring and culturing of the cell for producing vaccine; (3) the breeding of the cell virus; (4) the breeding of the virus liquid for producing vaccine; (5) vaccine preparing, split charging and freeze-drying. The invention has the advantages of simple and stable technique, easy operation, high virus content, small inter-batch difference, easy controlled quality, which can markedly improve the vaccine output and quality. The swine fever living vaccine produced by adopting the invention is good in safety and high in immunity effect, which has complete protecting function against the attack of the swine fever strong virus.

Description

Technical field [0001] The invention belongs to the technical field of veterinary biological products, and more specifically relates to a method for producing a swine fever live vaccine using a cell line. Background technique [0002] At present, the cells used in the production of lapinized attenuated swine fever vaccine in my country are primary bovine testicular cells. Because BVD / MDV disease is common in cattle herds in my country, the use of primary cells of bovine testis to produce vaccines is likely to cause foreign virus contamination, and the titer of toxin production is not high, and there are large differences between batches, which will increase the yield and efficacy of vaccines. difficult. Summary of the invention [0003] The purpose of the present invention is to overcome the shortcomings of the prior art and provide a method for producing swine fever live vaccine using cell lines. The method has the advantages of simple and stable production process, easy operati...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K39/187A61P31/14
Inventor 宁宜宝吴文福林旭埜张毓金李嘉爱赖月辉张国丽岑小清游启有王少英司徒剑谋
Owner CHINA INST OF VETERINARY DRUG CONTROL
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