A method for fully suspension culture of duck plague virus

A duck plague virus and culture method technology, applied in the field of veterinary biological products, can solve the problems of inability to meet the needs of vaccines, consumption of large SPF chicken embryos, long production cycle, etc., and achieve huge application prospects, social and economic benefits, immunogens The effect of good performance and high production efficiency

Inactive Publication Date: 2019-01-15
ZHAOQING INST OF BIOTECHNOLOGY CO LTD +3
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] At present, chicken embryo attenuated virus strains are often used to inoculate SPF chicken embryo or chicken embryo fibroblasts in the prevention and treatment of this disease, and the infected chicken embryo fluid, fetus and chorioallantoic membrane are mixed and ground or cell culture medium is harvested, and a suitable stabilizer is added. Freeze-drying to make a live vaccine, because the production of this vaccine mainly adopts the traditional method of inoculating chicken embryos to harvest allantoic fluid, but the production of vaccines from chicken embryos needs to consume a large amount of SPF chicken embryos, the production cycle is long, easy to be polluted, and each SPF chicken Embryos can only be injected with one virus strain at a time, especially when various poultry diseases break out, this method cannot meet the poultry industry's demand for vaccines

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Example 1: Effects of different inoculation doses and culture time on the proliferation of duck plague virus in the passaged cell line derived from duck embryo cells

[0030] The sources of materials used in Example 1 of the present invention are as follows:

[0031] 1. Virus: GD strain of duck plague virus, identified, kept and supplied by Zhaoqing Dahuanong Biopharmaceutical Co., Ltd.

[0032] 2. Cells: passaged cells derived from whole suspension duck embryo cells.

[0033]3. The first medium contains 10mg / L alanine, 10mg / L arginine, 10mg / L cysteine, 10mg / L tyrosine, 10mg / L tryptophan, 10mg / L Valine / L, Leucine 10mg / L, Vitamin C 1mg / L, Biotin 1mg / L, Folic Acid 1mg / L, Choline Chloride 1mg / L, Inositol 1mg / L, Cigarette 1mg / L Amide, 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, 0.5mg / ...

Embodiment 2

[0045] Embodiment 2: the influence of different virus medium on the proliferation of duck plague virus

[0046] The source of materials used in Embodiment 2 of the present invention is as follows:

[0047] 1. Virus: the TCID harvested in Example 1 50 Duck plague virus at highest titer.

[0048] 2. Cells: Passage cells derived from fully suspended duck embryo cells.

[0049] 3. The first medium contains 10mg / L alanine, 10mg / L arginine, 10mg / L cysteine, 10mg / L tyrosine, 10mg / L tryptophan, 10mg / L Valine / L, Leucine 10mg / L, Vitamin C 1mg / L, Biotin 1mg / L, Folic Acid 1mg / L, Choline Chloride 1mg / L, Inositol 1mg / L, Cigarette 1mg / L Amide, 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, 0.5mg / L EGF, 1mg / LbFGF.

[0050] 4. The second medium contains 10mg / L alanine, 10mg / L arginine, 10mg / L cysteine, 10...

Embodiment 3

[0058] Example 3: TCID of Duck Plague Virus at Different Harvest Times in Subcultured Cell Cultures Derived from Duck Embryo Cells 50 Compare

[0059] The source of materials used in Embodiment 3 of the present invention is as follows:

[0060] 1. Virus: Adapted strain of duck plague virus in passage cells derived from duck embryo cells.

[0061] 2. Cells: Passage cells derived from duck embryo cells of the whole suspension passage cell line.

[0062] 3. The first medium contains 10mg / L alanine, 10mg / L arginine, 10mg / L cysteine, 10mg / L tyrosine, 10mg / L tryptophan, 10mg / L Valine / L, Leucine 10mg / L, Vitamin C 1mg / L, Biotin 1mg / L, Folic Acid 1mg / L, Choline Chloride 1mg / L, Inositol 1mg / L, Cigarette 1mg / L Amide, 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, 0.5mg / L EGF, 1mg / LbFGF.

[0063] 4. ...

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PUM

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Abstract

The invention discloses a full suspension culture method of a duck plague virus, comprising the following steps: step 1: resuscitation and passage culture of passage cell lines derived from duck embryo cells; 2, inoculation of the duck plague virus into a passage cell line derived from a fully suspend culture duck embryo cell by adopting a second-order culture method to realize large-scale cultureof that duck plague virus. 3, after that virus is inoculated, the TCID50 of the virus is sampled and detected every 12 hours, the virus is harvested when the TCID50 of the virus reach the maximum, and the virus is stored to obtain the cultured duck plague virus. The full suspension culture method of the invention improves virus culture scale and efficiency, accords with the trend of vaccine production in the future, and has wide application prospect and good economic benefit.

Description

technical field [0001] The invention relates to a method for culturing duck plague virus in full suspension, in particular to a method for cultivating duck plague virus in full suspension with passaged cell lines, and belongs to the technical field of veterinary biological products. Background technique [0002] Duck Plague (DP) is an acute contact infectious disease of ducks, geese and swans caused by Duck Plague Virus (DPV), which is characterized by widespread prevalence, rapid spread and high morbidity. and high mortality. In foreign countries, the disease occurs in the Netherlands, France, Belgium, India, the United States and the United Kingdom. In the south of the Yangtze River in my country, especially in the Pearl River Delta where the duck industry is relatively developed, the disease is widespread, and the morbidity and mortality are high. Development of the duck industry. [0003] At present, chicken embryo weakened virus strains are used to inoculate SPF chicke...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N7/00C12R1/93
CPCC12N7/00C12N2710/16051
Inventor 叶俊贤陈瑞爱李延鹏蔡仕君罗琼
Owner ZHAOQING INST OF BIOTECHNOLOGY CO LTD
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