Suspension culture and continuous virus collection production method for porcine epidemic diarrhea viruses

A porcine epidemic diarrhea and suspension culture technology, which is applied in the field of porcine epidemic diarrhea virus suspension culture continuous collection production, can solve the problems of high labor intensity, few times of collection, complicated processes, etc., and achieves low labor intensity and good culture conditions. , the effect of simple process

Inactive Publication Date: 2018-11-23
安徽东方帝维生物制品股份有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0002] At present, most of the proliferation of porcine epidemic diarrhea virus (PEDV) adopts the spinner bottle production process. However, the traditional spinner bottle production process has the following disadvantages: labor-intensive, complicated process, slow cell proliferation and small number, unable to provide Optimum conditions such as pH, DO, etc., can only be harvested once, and the virus content of the harvest is low
[0004] However, the existing porcine epidemic diarrhea virus (PEDV) suspension culture production process, although the virus content is improved, but the number of virus collection is less (can only be harvested once), and the production cost is higher

Method used

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  • Suspension culture and continuous virus collection production method for porcine epidemic diarrhea viruses
  • Suspension culture and continuous virus collection production method for porcine epidemic diarrhea viruses

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Experimental program
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Effect test

Embodiment 1

[0044] Such as figure 1 As shown, a kind of porcine epidemic diarrhea virus suspension culture continuous poisoning production method of the present embodiment comprises the following steps:

[0045] (1) Cell recovery and expansion

[0046] The cryopreserved Vero cells (African green monkey kidney passage cells, purchased from ATCC) were resuscitated into T75 cell culture flasks, and cultured at a temperature of 36°C. After the dispersion liquid is digested, add DMEM cell growth medium containing 8% newborn bovine serum for subculture, and then subculture step by step according to the ratio of 1:3 to culture in a 3L spinner bottle, and the spinner bottle is expanded according to the subculture ratio of 1:3;

[0047] (2) Sheet carrier processing

[0048] According to the ratio of 20g / L sheet-shaped carrier, weigh the required sheet-shaped carrier (purchased from ESCO company) into the carrier frame of the 7L suspension culture tank, assemble the suspension culture tank, injec...

Embodiment 2

[0065] Such as figure 1 As shown, a kind of porcine epidemic diarrhea virus suspension culture continuous poisoning production method of the present embodiment comprises the following steps:

[0066] (1) Cell recovery and expansion

[0067] The cryopreserved Vero cells (African green monkey kidney passage cells, purchased from ATCC) were resuscitated into T75 cell culture flasks, and cultured at a temperature of 37°C. After the dispersion liquid is digested, add DMEM cell growth medium containing 10% newborn bovine serum for subculture, and subculture step by step according to the ratio of 1:6 to culture in a 15L spinner bottle, and the spinner bottle is expanded according to the subculture ratio of 1:5;

[0068] (2) Sheet carrier processing

[0069] According to the ratio of 40g / L sheet carrier, weigh the required sheet carrier (purchased from ESCO company) and put it into the carrier frame of the 50L suspension culture tank, assemble the suspension culture tank, inject PBS...

Embodiment 3

[0086] Such as figure 1 As shown, a kind of porcine epidemic diarrhea virus suspension culture continuous poisoning production method of the present embodiment comprises the following steps:

[0087] (1) Cell recovery and expansion

[0088] The cryopreserved Vero cells (African green monkey kidney passage cells, purchased from ATCC) were resuscitated into T75 cell culture flasks, and cultured at a temperature of 36.5°C. After the dispersion liquid is digested, add DMEM cell growth medium containing 9% newborn bovine serum for subculture, and then subculture step by step according to the ratio of 1:4 to a 10L spinner bottle for culture, and the spinner bottle is expanded according to the subculture ratio of 1:4;

[0089] (2) Sheet carrier processing

[0090] According to the ratio of 30g / L sheet carrier, weigh the required sheet carrier (purchased from ESCO company) and put it into the carrier frame of the 40L suspension culture tank, assemble the suspension culture tank, inj...

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Abstract

The invention relates to the technical field of biological products for veterinary use and provides suspension culture and continuous virus collection production method for porcine epidemic diarrhea viruses. The suspension culture and continuous virus collection production method comprises the following steps: carrying out cell resuscitation and amplification, treating a sheet-shaped carrier, sterilizing a suspension culture pot and the sheet-shaped carrier, filling cells into the pot, carrying out cell suspension culture, preparing virus-containing maintenance liquid, receiving the viruses and collecting the viruses. The suspension culture and continuous virus collection production method provided by the invention has the advantages that the sheet-shaped carrier is used for carrying out the suspension culture, the labor intensity is small, a technology is simple, culture conditions are excellent and the cost is low; the quantity of the cells obtained by the suspension culture of the sheet-shaped carrier is very great; the maintenance liquid containing a small amount of pancreatin is added and one part of cells have pathological changes; then the maintenance liquid containing new pancreatin is added to promote one part of the cells to have the pathological changes; the method is repeated so that the cells gradually have the pathological changes and multi-time harvesting can berealized; the content of antigen viruses produced by the method is 10 to 30 times as much as that of a traditional production technology.

Description

technical field [0001] The invention relates to the technical field of veterinary biological products, in particular to a production method for porcine epidemic diarrhea virus suspension culture and continuous poison collection. Background technique [0002] At present, most of the proliferation of porcine epidemic diarrhea virus (PEDV) adopts the spinner bottle production process. However, the traditional spinner bottle production process has the following disadvantages: labor-intensive, complicated process, slow cell proliferation and small number, unable to provide Optimum pH, DO and other conditions can only be harvested once, and the virus content of the harvest is low. Therefore, if want to proliferate in a large amount, then can only increase the method for turning bottle quantity, result in the workshop production required equipment, personnel etc. demand bigger again as a result. [0003] At present, suspension culture technology has become a hot spot for veterinar...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N7/00C12N7/02C12N5/02C12N5/071
CPCC12N5/0686C12N7/00C12N2531/00C12N2770/20051
Inventor 张星曹锋许冬赵文艳魏磊李媛媚池贤凤商俊苏明慧虞慎义张路范龙祥
Owner 安徽东方帝维生物制品股份有限公司
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