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Heat-resisting protective agent for freeze-dried vaccine and preparing method and application thereof

A heat-resistant protective agent and freeze-drying technology, which can be used in freeze-dried transportation, antiviral agents, and pharmaceutical formulations, can solve the problems of less research and inconvenient production for vaccine companies, and achieve easy preparation, suitable for large-scale production, and formulation simple effect

Active Publication Date: 2016-12-21
成都史纪生物制药有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] However, there is less research on a heat-resistant protective agent suitable for different viruses
Different viruses require different heat-resistant protective agents and different freeze-drying processes, which brings great inconvenience to the production of vaccine companies

Method used

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  • Heat-resisting protective agent for freeze-dried vaccine and preparing method and application thereof
  • Heat-resisting protective agent for freeze-dried vaccine and preparing method and application thereof
  • Heat-resisting protective agent for freeze-dried vaccine and preparing method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] 1. Cultivation and Harvest of Porcine Pseudomonas Virus

[0036] Passage and culture of Vero cells: When the Vero cells in the culture flask are covered with a single layer, discard the cell culture medium, and digest the cells with cell digestion solution containing 0.25% trypsin, and the digested cells in the culture flask are observed under an inverted microscope. When it becomes round and shrunk, immediately add cell growth solution to the culture flask, shake the culture flask to digest and disperse the cells into single cells, and culture the digested cells at a ratio of 1:3 for the next generation of cells.

[0037] Proliferation, culture and harvest of the virus: take the wet virus of PRV Bartha-k61 strain from the ultra-low temperature refrigerator at -70°C, inoculate the Vero cell culture bottle that has grown to a single layer at a ratio of 1% (V / V), and inoculate and continue to cultivate Observe and harvest when the cells have more than 80% lesions, freeze ...

Embodiment 2

[0062] 1. Culture and Harvest of Porcine Reproductive and Respiratory Syndrome Virus

[0063] Subculture and culture of cells: When the Marc-145 cells in the cell culture flask are covered with a monolayer, discard the growth medium, and digest the cells with 0.25% trypsin. The digested cells in the culture flask are round and shrunk when observed under an inverted microscope When it is in shape, immediately add cell growth solution to the culture flask, shake the culture flask to digest and disperse the cells into single cells, and expand the digested cells at a ratio of 1:4.

[0064] Proliferation, cultivation and harvest of the virus: Get the seed virus of PRRSV JXA1-R strain from the refrigerator at -70°C, and inoculate the Marc-145 cell culture bottle that has overgrown a single layer by a 0.5% (V / V) inoculation ratio. Continue to cultivate and observe, and harvest when the lesions of the cells reach more than 75%. Freeze and thaw the culture flask once at -20°C to collec...

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Abstract

The invention discloses a heat-resisting protective agent for freeze-dried vaccine and a preparing method and application thereof. The heat-resisting protective agent is prepared from, by mass, 0.3-0.6% of L-sodium glutamate, 8-14% of saccharose, 2-5% of D-sorbitol, 1-3% of bovine serum albumin, 0.1-0.3% of polyvinylpyrrolidone, 0.2-1% of thiourea, 3-5% of NZ-Amine AS, 0.8-1.5% of hydrolyzed gelatin and the balance PBS with PH of 7.2. The heat-resisting protective agent for freeze-dried vaccine can be applied to freeze-drying of porcine pseudorabies live vaccine and porcine reproductive and respiratory syndrome live vaccine after being subjected to asepsis treatment. By adopting the same heat-resisting protective agent and freeze-drying technology for the two viruses, freeze-drying loss is lower than 0.2 titer, virus content titer is lower than one titer after the viruses are stored at 37 DEG C for seven days, and the viruses can be stored for 24 months at 2-8 DEG C; convenience is brought for production of freeze-dried vaccine, and product quality and competitiveness are improved.

Description

technical field [0001] The invention relates to the field of biological products, in particular to a freeze-dried vaccine heat-resistant protective agent and its preparation method and application. Background technique [0002] Lyoprotectants may affect the quality, potency and stability of biological products. Heat-resistant lyoprotectants have immunological activity but no pharmacological activity. It not only protects the biological activity of products, but also has excipients and antioxidants. The role of the vaccine is directly related to the titer and stability of the vaccine. It can maintain the stability of the vaccine during freeze-drying and storage, and ensure that the product can be stored at 2-8°C for up to 24 months. Even at 37°C, it can be stored for more than ten days, thus solving the problem of heat resistance, low storage temperature, short storage period and harsh freezing conditions caused by conventional vaccines during long-distance transportation, lo...

Claims

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Application Information

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IPC IPC(8): A61K47/42A61K47/18A61K47/26A61K47/32A61K47/20A61K47/02A61K39/245A61K39/12A61K9/19A61P31/22A61P31/14
CPCA61K9/19A61K39/12A61K39/245A61K47/02A61K47/18A61K47/183A61K47/20A61K47/26A61K47/32A61K47/42
Inventor 徐宏军胡来根高鹏岳丰雄黄杰
Owner 成都史纪生物制药有限公司
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