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Heat-proof freeze-dried protective agent and preparation method and application thereof

A heat-resistant freeze-drying and protective agent technology, which is applied in the direction of freeze-drying transportation, antiviral agents, pharmaceutical formulations, etc., to achieve the effects of shortening the freeze-drying time, improving the protection period, and long-lasting antibodies

Active Publication Date: 2014-04-09
哈药集团生物疫苗有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0002] my country's research on heat-resistant freeze-drying protective agents started relatively late, but now the research in this area has achieved certain results. The patent publication number is CN1426816A, and the invention name is heat-resistant freeze-drying protective agent for veterinary virus biological products and its preparation. The patent application for the invention of the process discloses a heat-resistant freeze-drying protectant for veterinary virus products. The components of the protectant include commonly used sucrose, skimmed milk powder, gelatin, and dextrin. The application further discloses a The composition ratio of the protective agent of the attenuated Newcastle disease freeze-dried vaccine: gelatin 1.5-2%, sucrose 5-6%, dextrin 5-10%, PVP (K90) 2-5%, sorbitol 2%, sodium glutamate 1-2%

Method used

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  • Heat-proof freeze-dried protective agent and preparation method and application thereof
  • Heat-proof freeze-dried protective agent and preparation method and application thereof
  • Heat-proof freeze-dried protective agent and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Embodiment 1 Preparation of heat-resistant lyoprotectant

[0024] (1) Weigh each substance according to the following weight:

[0025] Weigh 2 g of trehalose, 3 g of sodium thiosulfate, 1.5 g of enzymatically hydrolyzed casein, 1.5 g of thiourea, GIBCO199 medium (purchased from Invitrogen, Cat. No. 31100-035, Lot. No. ) is 909374, powder) 1g, O-2′-hydroxypropyltrimethylammonium chloride chitosan 0.5g, gelatin 2g, peptone 8g, sucrose 8g;

[0026] (2) Preparation of liquid A:

[0027] The trehalose taken by weighing, sodium thiosulfate, enzymatic casein hydrolysis, thiourea, 199 medium, O-2'-hydroxypropyltrimethylammonium chloride chitosan were added to a small amount of mass percentage concentration of 4% polyvinylpyrrolidone (PVPK 30 ) in aqueous solution, fully dissolved, stirred evenly, and then with 4% polyvinylpyrrolidone (PVPK 30 ) aqueous solution to 100ml, sterilized with a 0.22um filter, and set aside to obtain liquid A;

[0028] (3) Preparation of liquid B...

Embodiment 2

[0031] Embodiment 2 Preparation of heat-resistant lyoprotectant

[0032] (1) Weigh each substance according to the following weight:

[0033] Weigh 1 g of trehalose, 3 g of sodium thiosulfate, 1 g of enzymatically hydrolyzed casein, and 2 g of thiourea, and GIBCO199 culture medium (purchased from Invitrogen, Cat. No. 31100-035, Lot. No.) is 909374, powder) 2g, O-2′-hydroxypropyltrimethylammonium chloride chitosan 0.1g, gelatin 3g, peptone 5g and sucrose 10g.

[0034] (2) Preparation of liquid A:

[0035] The trehalose, sodium thiosulfate, enzymatic hydrolyzed casein, thiourea, 199 medium and O-2'-hydroxypropyltrimethylammonium chloride chitosan were added to a small amount of mass percentage concentration of 2% polyvinylpyrrolidone (PVPK 30 ) in aqueous solution, fully dissolved, stirred evenly, and then with 2% polyvinylpyrrolidone (PVPK 30 ) aqueous solution to 100ml, sterilized with a 0.22um filter, and set aside to obtain liquid A;

[0036] (3) Preparation of liquid B...

Embodiment 3

[0039] Embodiment 3 Preparation of heat-resistant lyoprotectant

[0040] (1) Weigh each substance according to the following weight:

[0041] Weigh 3 g of trehalose, 1 g of sodium thiosulfate, 2 g of enzymatically hydrolyzed casein, 1 g of thiourea, and GIBCO199 medium (purchased from Invitrogen, Cat. No. 31100-035, Lot. No. ) 909374, powder) 0.5g, O-2′-hydroxypropyltrimethylammonium chloride chitosan 1g, gelatin 1g, peptone 10g and sucrose 5g;

[0042] (2) Preparation of liquid A:

[0043] The trehalose, sodium thiosulfate, enzymatic hydrolyzed casein, thiourea, 199 medium and O-2'-hydroxypropyltrimethylammonium chloride chitosan were added to a small amount of mass percentage concentration of 6% polyvinylpyrrolidone (PVPK 30 ) in aqueous solution, fully dissolved, stirred evenly, and then with 6% polyvinylpyrrolidone (PVPK 30 ) aqueous solution to 100ml, sterilized with a 0.22um filter, and set aside to obtain liquid A;

[0044] (3) Preparation of liquid B:

[0045] Ad...

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Abstract

The invention discloses a heat-proof freeze-dried protective agent and a preparation method and application thereof. The heat-proof freeze-dried protective agent is prepared by mixing a solution A and a solution B according to the volume ratio of 1:1, wherein the solution A comprises 0.01-0.03 g / ml of trehalose, 0.01-0.04 g / ml of sodium thiosulfate, 0.01-0.02 g / ml of casein enzymatic hydrolysate, 0.01-0.02 g / ml of thiocarbamide, 0.005-0.02 g / ml of 199 culture medium and 0.001-0.01 g / ml of O-2'-hydroxypropyl trimethyl ammonium chloride chitosan, is dissolved and determined in volume by using a polyvinyl pyrrolidone (PVPK30) water solution with the mass percent concentration of 2%-6%, and is disinfected by using 0.22 micrometer filter; the solution B contains 0.01-0.03 g / ml of gelatin, 0.05-0.1 g / ml of peptone and 0.05-0.1 g / ml of sucrose, is dissolved and determined in volume by using deionized water, and is autoclaved for later use. Compared with the existing product, the improved heat-proof freeze-dried protective agent disclosed by the invention can increase the protection duration of a product, furthest maintain the virus titer and shorten the freeze-drying time, and is long in antibody duration time and favorable to use in industrialized large-scale production.

Description

technical field [0001] The invention relates to a heat-resistant freeze-drying protective agent and its preparation method and application. Belongs to the field of biotechnology. Background technique [0002] my country's research on heat-resistant freeze-drying protective agents started relatively late, but now the research in this area has achieved certain results. The patent publication number is CN1426816A, and the invention name is heat-resistant freeze-drying protective agent for veterinary virus biological products and its preparation. The patent application for the invention of the process discloses a heat-resistant freeze-drying protectant for veterinary virus products. The components of the protectant include commonly used sucrose, skimmed milk powder, gelatin, and dextrin. The application further discloses a The composition ratio of the protective agent of the attenuated Newcastle disease freeze-dried vaccine: gelatin 1.5-2%, sucrose 5-6%, dextrin 5-10%, PVP (K90) ...

Claims

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Application Information

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IPC IPC(8): A61K47/42A61K47/36A61K47/32A61K39/17A61K9/19A61P31/14
Inventor 孙德君丁国杰刘鑫莹武佳斌吴琼孙心孙凯李静王昊
Owner 哈药集团生物疫苗有限公司
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