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Full suspension culture method of chicken infectious bursal disease virus

A technology for bursal disease and chicken infectivity, which is applied in the direction of virus, culture process, tissue culture, etc. to achieve high production efficiency, improved scale and efficiency, and good immunogenicity.

Inactive Publication Date: 2019-02-01
ZHAOQING INST OF BIOTECHNOLOGY CO LTD +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The use of MDCK cells (canine kidney cells) for full suspension culture of avian influenza virus and the use of BHK-21 cells (milk hamster kidney cells) for full suspension culture of foot-and-mouth disease virus have been verified in production, but there is no full suspension culture method for IBDV in China. According to reports, the production method still has a lot of room for optimization

Method used

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  • Full suspension culture method of chicken infectious bursal disease virus
  • Full suspension culture method of chicken infectious bursal disease virus
  • Full suspension culture method of chicken infectious bursal disease virus

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028]Example 1: Effects of different inoculated doses and culture times on the proliferation of chicken infectious bursal disease virus in passaged cells derived from duck embryo cells

[0029] The sources of materials used in Embodiment 1 of the present invention are as follows:

[0030] 1. Virus: Chicken Infectious Bursal Disease Virus is IBDV (GT strain), identified, kept and supplied by Zhaoqing Dahuanong Biopharmaceutical Co., Ltd.

[0031] 2. Cells: Passage cells derived from fully suspended duck embryo cells, provided by Gansu Jianshun Biotechnology Co., Ltd.

[0032] 3. Culture medium: the name is medium EBLM004, containing 10mg / L alanine, 10mg / L arginine, 10mg / L cysteine, 10mg / L tyrosine, 10mg / L chromosine amino acid, 10mg / L valine, 10mg / L leucine, 1mg / L vitamin C, 1mg / L biotin, 1mg / L folic acid, 1mg / L choline chloride, 1mg / L inositol, 1mg / L Niacinamide, 1mg / L Pyridoxine Hydrochloride, 50mg / L Potassium Chloride, 50mg / L Sodium Chloride, 50mg / L Disodium Hydrogen Phos...

Embodiment 2

[0043] Example 2: Passage acclimation of chicken infectious bursal disease virus in the passage cell line derived from duck embryo cells

[0044] The present invention implements 2 used material source as follows:

[0045] 1. Virus: EID harvested from previous generation 50 The highest IBDV was used as seed poison;

[0046] 2. Cells: Passage cell lines derived from fully suspended duck embryo cells, provided by Gansu Jianshun Biotechnology Co., Ltd.;

[0047] 3. Culture medium: the name is medium EBLM004, containing 10mg / L alanine, 10mg / L arginine, 10mg / L cysteine, 10mg / L tyrosine, 10mg / L chromosine amino acid, 10mg / L valine, 10mg / L leucine, 1mg / L vitamin C, 1mg / L biotin, 1mg / L folic acid, 1mg / L choline chloride, 1mg / L inositol, 1mg / L Niacinamide, 1mg / L Pyridoxine Hydrochloride, 50mg / L Potassium Chloride, 50mg / L Sodium Chloride, 50mg / L Disodium Hydrogen Phosphate, 50mg / L Sodium Bicarbonate, 1000mg / L Glucose, 0.1mg / L manganese sulfate, 0.01mg / L ferric nitrate, 0.1mg / L insu...

Embodiment 3

[0057] Example 3: Comparison of production of IBDV by subcultured cell lines derived from duck embryonic cells in suspension culture in different media

[0058] The source of materials used in Embodiment 3 of the present invention is as follows:

[0059] 1. Virus: IBDV (GT strain), identified, kept and supplied by Zhaoqing Dahuanong Biopharmaceutical Co., Ltd.

[0060] 2. Cells: Passage cell lines derived from whole suspension duck embryo cells

[0061] 3. Culture medium: the name is medium EBLM004, containing 10mg / L alanine, 10mg / L arginine, 10mg / L cysteine, 10mg / L tyrosine, 10mg / L chromosine amino acid, 10mg / L valine, 10mg / L leucine, 1mg / L vitamin C, 1mg / L biotin, 1mg / L folic acid, 1mg / L choline chloride, 1mg / L inositol, 1mg / L Niacinamide, 1mg / L Pyridoxine Hydrochloride, 50mg / L Potassium Chloride, 50mg / L Sodium Chloride, 50mg / L Disodium Hydrogen Phosphate, 50mg / L Sodium Bicarbonate, 1000mg / L Glucose, 0.1mg / L manganese sulfate, 0.01mg / L ferric nitrate, 0.1mg / L insulin, 0....

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Abstract

The invention discloses a full suspension culture method of chicken infectious bursal disease virus, wherein the method comprises the following steps: step 1, carrying out resuscitation and subcultureof subcultured cell lines derived from duck embryo cells; step 2, adopting a second-order culture method, inoculating the subcultured cell lines derived from the duck embryo cells subjected to full suspension culture with the chicken infectious bursal disease virus, and carrying out virus large-scale culture; and step 3, after inoculating with the virus, taking samples every other 12 h and measuring the virus TCID50, harvesting the virus and preserving when the virus TCID50 reaches the maximum, and thus obtaining the chicken infectious bursal disease virus after culture. The full suspension culture method improves the scale and efficiency of virus culture, conforms to the tendency of future vaccine production, has broad application prospects and contains good economic benefits.

Description

technical field [0001] The invention relates to a full-suspension culture method for chicken infectious bursal disease virus, in particular to a method for full-suspension culture of chicken infectious bursal disease virus with a subcultured cell line, and belongs to the technical field of veterinary biological products. Background technique [0002] Chicken infectious bursal disease (Infectious bursal disease, IBD) is an acute, highly contagious infectious disease of chickens caused by chicken infectious bursal disease virus (Infectious bursal disease Virus, IBDV), which is required by the regulations of our country. Class II animal diseases strictly controlled. IBDV mainly invades the chick's humoral immune organ, the bursa of Fabricius, and damages lymphocytes, which leads to immune dysfunction in chickens, which in turn increases the susceptibility to other pathogens, and even reduces the immune response to other vaccines. After being infected with IBDV, a large number ...

Claims

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Application Information

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IPC IPC(8): C12N7/00C12N7/02C12N5/02C12N5/073
CPCC12N5/0603C12N7/00C12N2500/12C12N2500/16C12N2500/20C12N2500/24C12N2500/32C12N2500/34C12N2500/38C12N2501/11C12N2501/115C12N2501/33C12N2720/10051
Inventor 李延鹏叶俊贤陈瑞爱温良海罗琼
Owner ZHAOQING INST OF BIOTECHNOLOGY CO LTD
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