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Heatproof freeze-drying protective agent of live vaccine, live vaccine freeze-drying powder and preparation method and preparation method of live vaccine freeze-drying powder

A technology of heat-resistant freeze-drying and live vaccines, applied in the field of heat-resistant freeze-drying protective agents for live vaccines, can solve the problems of limited heat resistance, low stability, damage, etc., to reduce physical damage and enhance heat resistance , The effect of heat resistance enhancement

Active Publication Date: 2013-04-17
JIANGSU ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In the conventional freeze-drying process, the live vaccine is first frozen at ultra-low temperature (below -40°C), and the ice crystals formed will cause varying degrees of damage to the virus envelope or cell wall, thereby affecting the activity of the dried product after hydration
Therefore, in the conventional freeze-drying process, no matter how much the heat-resistant protective agent formula is improved, the heat-resistant performance of the product is still limited, and the stability at room temperature or even higher temperature is not high

Method used

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  • Heatproof freeze-drying protective agent of live vaccine, live vaccine freeze-drying powder and preparation method and preparation method of live vaccine freeze-drying powder
  • Heatproof freeze-drying protective agent of live vaccine, live vaccine freeze-drying powder and preparation method and preparation method of live vaccine freeze-drying powder

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Example 1 Preparation of heat-resistant lyoprotectant

[0030] Unless otherwise specified, the percentages in this embodiment are percentages by weight.

[0031] Among them, casein hydrolyzate was purchased from ShanHai Richu Bioscience Co..Ltd, batch number 090901; polyvinylpyrrolidone (K-30) was purchased from Sinopharm Chemical Reagent Co., Ltd., batch number F20111009; Pluronic F 68 was purchased from From Shanghai Sangon Biotechnology Co., Ltd., batch number HJ0522B2012.

[0032] Heat-resistant lyoprotectant 1 formula:

[0033] Trehalose 68%,

[0034] Glycine 6%,

[0035] Gelatin 3%,

[0036] Casein Hydrolyzate 6%,

[0037] Polyvinylpyrrolidone 6%,

[0038] Glycerin 5%,

[0039] Plunik 68 6%.

[0040] Heat-resistant lyoprotectant 2 formula:

[0041] Trehalose 38%, Sucrose 20%,

[0042] Arginine 1%, glycine 1%, methionine 1%, glutamic acid 1%, isoleucine 3%,

[0043] Gelatin 8%,

[0044] Casein Hydrolyzate 4%,

[0045] Polyvinylpyrrolidone 9%,

[0046...

Embodiment 2

[0074] Example 2 Preparation and Heat Resistance of Newcastle Disease Live Vaccine Freeze-dried Powder

[0075] (1) Preparation of heat-resistant lyoprotectant

[0076] Heat-resistant lyoprotectant 1-5 was prepared according to the method in Example 1.

[0077] (2) Preparation of chicken Newcastle disease virus liquid (live virus liquid)

[0078] Chicken Newcastle disease virus liquid (live virus liquid) was provided by Nanjing Tianbang Biotechnology Co., Ltd. Specific preparation method: According to the relevant requirements of "Compilation of Quality Standards for Veterinary Biology" (2006-2008), chicken Newcastle disease virus liquid (La Sota strain) was prepared, and the virus content of each virus liquid was 10 7.5 EID 50 (chicken embryo test method).

[0079] (3) Preparation of freeze-dried powder of Newcastle disease live vaccine

[0080] Mix the heat-resistant lyoprotectant 1 with sterile water for injection at a mass ratio of 3:7, and after the heat-resistan...

Embodiment 3

[0097] Example 3 Preparation and heat resistance of porcine reproductive and respiratory syndrome live vaccine freeze-dried powder

[0098] (1) Preparation of heat-resistant lyoprotectant

[0099] Heat-resistant lyoprotectants 1-5 were prepared according to the method in Example 1.

[0100] (2) Preparation of porcine reproductive and respiratory syndrome virus liquid (live virus liquid)

[0101] Porcine reproductive and respiratory syndrome virus liquid (live virus liquid) was provided by Nannong Hi-Tech Biological Products Co., Ltd. Specific preparation method: According to the relevant requirements of the "Compilation of Quality Standards for Veterinary Biology" (2006-2008), the porcine reproductive and respiratory syndrome virus liquid (R98 strain) was prepared, and the virus content of each virus liquid was 10 6.0 TCID 50 (cytometry).

[0102] (3) Preparation of lyophilized powder of porcine reproductive and respiratory syndrome live vaccine

[0103] The correspondi...

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Abstract

The invention provides a heatproof freeze-drying protective agent, live vaccine freeze-drying powder and a preparation method of the live vaccine freeze-drying powder, and relates to the field of microorganism deposit. The heatproof freeze-drying protective agent comprises 46-90% of oligosaccharide, 1-9% of amino acid, 1-9% of gelatin, 1-9% of casein hydrolysate, 1-9% of polyvinyl pyrrolidone, 1-9% of glycerol and 1-9% of pluronic 68. The preparation method of the live vaccine freeze-drying powder comprises the steps of preparing a protective agent solution and the live vaccine, drying, keeping for 2-6h at -10-2 DEG C and atmospheric pressure, keeping 10-26h for 0-20 DEG C at 100-800mbar, and keeping 2-8h at 0.01-50.mbar and 10-28 DEG C. The heatproof freeze-drying protective agent is reasonable in component and content collocation, and allows the live vaccine to be heatproof. According to the preparation method of the live vaccine freeze-drying powder, formation of ice crystals is avoided, and the obtained live vaccine freeze-drying powder can be kept for 3 months at 37 DEG C and 18 months at 20 DEG C.

Description

technical field [0001] The invention relates to the field of preservation of living microorganisms, in particular to a heat-resistant freeze-drying protective agent for live vaccines, live vaccine freeze-dried powder and a preparation method thereof. Background technique [0002] The heat sensitivity of viruses and other biologically active substances and their related products has always been the bottleneck of long-term storage of live vaccines. my country has a vast territory and great differences in economic levels. The phenomenon of short-term exposure of vaccines to high temperature environments (even above 45°C) still exists during transportation and use. Therefore, higher requirements are put forward for the heat resistance of live vaccines. [0003] The heat-resistant lyoprotectant considers the impact of the physical and chemical changes that may occur in the lyophilized substance on the survival of the live vaccine when the live vaccine is stored at a higher temper...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/12A61K39/17A61K47/42A61K47/36A61K47/26A61P31/12A61P31/14
Inventor 吕芳卢宇邓碧华张金秋侯继波
Owner JIANGSU ACAD OF AGRI SCI
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