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Preparation method of porcine epidemic diarrhea virus liposome diluent freeze-dried product

A technology for porcine epidemic diarrhea and freeze-dried products, which is applied in the directions of liposome delivery, virus/phage, biochemical equipment and methods, etc. The effect of increasing viral content and enhancing immunogenicity

Inactive Publication Date: 2017-04-19
浙江美保龙生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] One of the most important indicators for the quality assessment of veterinary biological products is the batch-to-batch stability. However, it is difficult to control the batch-to-batch stability of semi-finished products in the actual production process. The main reason is that the proportion of live organisms inoculated is uncertain , that is to say, the unstable inoculation amount leads to large differences in the production of semi-finished products, which affects the reproduction of the virus and the virus content, so that the vaccination cannot effectively prevent the occurrence of porcine epidemic diarrhea

Method used

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  • Preparation method of porcine epidemic diarrhea virus liposome diluent freeze-dried product
  • Preparation method of porcine epidemic diarrhea virus liposome diluent freeze-dried product
  • Preparation method of porcine epidemic diarrhea virus liposome diluent freeze-dried product

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] (1) Prepare phosphate buffer:

[0025] a. Weigh the following raw material components: sodium chloride 6g, potassium chloride 0.05g, disodium hydrogen phosphate 1g, potassium dihydrogen phosphate 0.05g, calcium chloride 0.05g and magnesium chloride hexahydrate (MgCl 2 ·6H 2 O) 0.05g, mix well and set aside;

[0026] b. Add 1000 mL of double-distilled water to the mixture of raw material components obtained in step a, and after fully dissolving, autoclave at 116°C for 30 minutes to obtain a phosphate buffer solution with pH=7.

[0027] (2) Extraction of Ophiopogon japonicus flavonoids:

[0028] Weigh 5g of Radix Ophiopogon japonicus, add 90% ethanol according to the ratio of solid to liquid 1:14, set the power of microwave oven to 900W, extract in microwave for 4min, reflux in water bath for 30min, and the obtained extract is filtered and evaporated to dryness to obtain Radix Ophiopogon japonicus flavonoids.

[0029] (3) Preparation of liposomes

[0030] A. Weigh ...

Embodiment 2

[0037] (1) Prepare phosphate buffer:

[0038] a. Weigh the following raw material components: sodium chloride 8g, potassium chloride 0.3g, disodium hydrogen phosphate 1.1g, potassium dihydrogen phosphate 0.25g, calcium chloride 0.1g and magnesium chloride hexahydrate (MgCl 2 ·6H 2 O) 0.13g, mix well and set aside;

[0039] b. Add 1000 mL of double-distilled water to the mixture of raw material components obtained in step a, and after fully dissolving, autoclave at 116°C for 30 minutes to obtain a phosphate buffer solution with pH=7.

[0040] (2) Extraction of Ophiopogon japonicus flavonoids:

[0041] Weigh 5g of Radix Ophiopogon japonicus, add 90% ethanol according to the ratio of solid to liquid 1:14, set the power of microwave oven to 900W, extract in microwave for 4min, reflux in water bath for 30min, and the obtained extract is filtered and evaporated to dryness to obtain Radix Ophiopogon japonicus flavonoids.

[0042] (3) Preparation of liposomes

[0043] A. Weigh ...

Embodiment 3

[0050] (1) Prepare phosphate buffer:

[0051] a. Weigh the following raw material components: sodium chloride 10g, potassium chloride 0.5g, disodium hydrogen phosphate 1.2g, potassium dihydrogen phosphate 0.5g, calcium chloride 0.2g and magnesium chloride hexahydrate (MgCl 2 ·6H 2 O) 0.2g, mix well and set aside;

[0052] b. Add 1000 mL of double-distilled water to the mixture of raw material components obtained in step a, and after fully dissolving, autoclave at 116°C for 30 minutes to obtain a phosphate buffer solution with pH=7.

[0053] (2) Extraction of Ophiopogon japonicus flavonoids:

[0054] Weigh 5g of Radix Ophiopogon japonicus, add 90% ethanol according to the ratio of solid to liquid 1:14, set the power of microwave oven to 900W, extract in microwave for 4min, reflux in water bath for 30min, and the obtained extract is filtered and evaporated to dryness to obtain Radix Ophiopogon japonicus flavonoids.

[0055] (3) Preparation of liposomes

[0056] A. Weigh 4...

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Abstract

The invention belongs to the technical field of veterinary biological products, and in particular relates to a preparation method of a porcine epidemic diarrhea virus liposome diluent freeze-dried product, and the method comprises the following steps: weighing raw materials of 5-15 parts by weight of yam polysaccharide, 1-5 parts by weight of ophiopogonone,1-5 parts by weight of rheum emodin, 5-10 parts by weight of liposome and 10-30 parts by weight of a freeze-drying protective additive; mixing evenly the raw material components, adding the raw materials to a phosphate buffer liquid of pH = 7 for oscillation hydration to form a homogeneous solution; placing a filtrate obtained by filtering the homogeneous solution in the environment of -40 DEG C for freezing, then placing the filtrate in a room temperature environment for natural thawing, after repeating for 3 to 6 times, placing the filtrate in the environment of-40 DEG C for prefreezing for 4-8h, and then placing the filtrate in a vacuum freeze drying machine for drying to obtain the porcine epidemic diarrhea virus liposome diluent freeze-dried product. The method can reduce batch difference among semi-finished products, can promote the reproduction of porcine epidemic diarrhea virus, improves the virus content, and enhances the immunogenicity of a vaccine.

Description

technical field [0001] The invention belongs to the technical field of veterinary biological products, and in particular relates to a method for preparing a freeze-dried product of porcine epidemic diarrhea virus liposome dilution. Background technique [0002] Porcine epidemic diarrhea (Porcine epidemic diarrhea, PED) is caused by porcine epidemic diarrhea virus (Porcine epidemic diarrhea, PEDV), characterized by severe enteritis, vomiting, watery diarrhea, dehydration and high mortality. Highly contagious infectious disease. The disease mainly harms healthy suckling piglets at the age of 1-2 weeks, and the mortality rate can be as high as 100%, which has caused huge economic losses to the pig industry, and vaccination is the most effective way to prevent the disease from occurring. [0003] One of the most important indicators for the quality assessment of veterinary biological products is the batch-to-batch stability. However, it is difficult to control the batch-to-batc...

Claims

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Application Information

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IPC IPC(8): A61K9/19A61K9/127A61K47/26A61K39/215A61P31/14A61K31/715A61K31/352A61K31/122
CPCA61K9/19A61K9/127A61K31/122A61K31/352A61K31/715A61K39/12A61K47/26A61K2039/552C12N2770/20034A61K2300/00
Inventor 沈建军张秀文李阳
Owner 浙江美保龙生物技术有限公司
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