Method for producing avian encephalomyelitis virus inactivated vaccine

A technology for avian encephalomyelitis and virus inactivation, applied in biochemical equipment and methods, antiviral agents, viruses/phages, etc., can solve the problems of difficulty in improving vaccine yield and quality, large differences between product batches, and high production costs , to achieve the effect of low probability of foreign virus contamination, small quality difference between batches and low cost

Inactive Publication Date: 2015-06-10
RINGPU (BAODING) BIOLOGICAL PHARMACEUTICAL CO LTD
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  • Summary
  • Abstract
  • Description
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  • Application Information

AI Technical Summary

Problems solved by technology

Using chicken embryos to cultivate avian encephalomyelitis virus requires a large number of SPF chicken embryos. The production cost is high, the labor

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Example 1 Cloning, Purification and Screening of Baby Hamster Kidney Passage Cells (BHK21) by Limiting Dilution

[0030] Young hamster kidney passage cells (BHK21) and avian encephalomyelitis virus Van Roekel strain (AEV VR strain) were purchased from China Veterinary Drug Administration.

[0031] The formula of the cell growth solution is 92% low-sugar DMEM solution by volume, 8% newborn bovine serum, and the pH value is adjusted to 7.2;

[0032] A. Preparation of Monoclonal Cell Lines

[0033] a. Limiting dilution of cells

[0034] Prepare BHK21 cell suspension, after cell counting, use cell growth medium to carry out gradient dilution to 10 1 cells / ml, that is, 1 cell per 0.1ml.

[0035] b. Clonal culture of cells

[0036] Add the above-mentioned diluted cell suspension into a 96-well cell culture plate, 0.1ml / well, observe under a microscope, discard the cell wells with non-single cells and poor cell state, and place at 37°C, 5% CO 2 Cultivate, observe the grow...

Embodiment 2

[0051] Example 2 Production of Inactivated Avian Encephalomyelitis Virus Vaccine Using Baby Hamster Kidney Passage Cells (BHK21)

[0052] The cells are the same as the baby hamster kidney passage cells (BHK21) preserved in the strain in Example 1;

[0053] The seed virus is the same as the Van Roekel strain of avian encephalomyelitis virus (AEV VR strain) in Example 1;

[0054] The formula of the cell growth solution is 92% by volume low-sugar DMEM solution, 8% newborn bovine serum, and the pH value is adjusted to 7.3;

[0055] The formula of the cell maintenance solution is 98% by volume low-sugar DMEM solution, 2% newborn bovine serum, and the pH value is adjusted to 7.4;

[0056] a. Select the cell line of young hamster kidney passage cells (BHK21) as the cells for making seedlings;

[0057] b. Subculture and cultivation of cells for seedling production

[0058] When the young hamster kidney passaged cells (BHK21) form a good monolayer, after digested with EDTA-trypsin, ...

Embodiment 3

[0070] Example 3 Production of Avian Encephalomyelitis Virus Inactivated Vaccine Using Chicken Embryo Fibroblast Cell Line (DF-1)

[0071] Chicken embryo fibroblast cell line (DF-1) and avian encephalomyelitis virus Van Roekel strain (AEV VR strain) were purchased from China Veterinary Drug Administration.

[0072] The formula of the cell growth solution is 90% DMEM / F12 solution by volume, 10% newborn bovine serum, and the pH value is adjusted to 7.0;

[0073] The formula of the cell maintenance solution is 98% DMEM / F12 solution by volume, 2% newborn bovine serum, and the pH value is adjusted to 7.3;

[0074] a. Select the chicken embryo fibroblast cell line (DF-1) as the cells for making seedlings;

[0075] b. Subculture and cultivation of cells for seedling production

[0076] When chicken embryo fibroblasts (DF-1) form a good monolayer, after digested with EDTA-trypsin, add cell growth medium to prepare DF-1 cell suspension, which is used for continuous passage or virus i...

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Abstract

The invention discloses a method for producing an avian encephalomyelitis virus inactivated vaccine. The method comprises the following steps: a. selecting cell lines to serve as cells for preparing the vaccine; b. passing and culturing the cells for preparing the vaccine; c. breeding a virus seed for production; d. producing venom for preparing the vaccine; and e. preparing the vaccine. According to the method disclosed by the invention, continuous cell lines are used as the cells for preparing the vaccine, and sensitive cell lines are screened to reinforce the match of the virus and the cells. The method disclosed by the invention has the advantages of simple and stable production process, easiness in operation, small batch difference and low cost, and the safety and immunogenicity of the produced avian encephalomyelitis virus inactivated vaccine are good.

Description

technical field [0001] The invention relates to a method for producing an inactivated vaccine of avian encephalomyelitis virus by using a cell line, and belongs to the field of biotechnology. Background technique [0002] Avian Encephalomyelitis (AE), also known as epidemic tremor, is caused by avian encephalomyelitis virus (AEV), which mainly harms chicks under 4 weeks of age, in order to infringe the central nervous system of chicks, causing non-suppurative Encephalitis is a viral infectious disease with the main pathological features. The main manifestations of the disease are neurological symptoms such as ataxia, head and neck tremor, and numbness of the hind toes. It has the characteristics of sudden appearance and unpredictable duration, which can cause the loss of chicks and the decline of egg production rate of hens. It has become one of the main diseases that endanger the development of poultry industry in the world. Since the report of this disease occurred in our...

Claims

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Application Information

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IPC IPC(8): A61K39/125A61P31/14C12N7/00
Inventor 韩佳丽邱贞娜梁武刘涛柳珊朱秀同郁宏伟杨保收何平有徐倩倩郑朝朝
Owner RINGPU (BAODING) BIOLOGICAL PHARMACEUTICAL CO LTD
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