A semi-perfusion culture method for duck Tembusu virus

A technology of duck Tembusu virus and its cultivation method, which is applied in the direction of virus, virus/bacteriophage, positive-sense single-stranded RNA virus, etc., can solve the problems of difficult vaccine quality assurance, high labor intensity, and low preparation efficiency, and promote continuous Divide, improve production efficiency, increase the effect of cell number

Active Publication Date: 2018-05-18
GUANGDONG HAID GROUP +1
View PDF3 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

For virus propagation, most of them use the spinner bottle production process. Due to the limitation of the spinner bottle surface area and culture conditions, the cell density is low, the labor intensity is high, the preparation efficiency is low, the operation process is easy to be polluted, and the quality of the vaccine is difficult to be guaranteed.
There are few reports about the perfusion culture method used in vaccine production

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] A semi-perfusion culture method for duck Tembusu virus, comprising the following steps:

[0030] 1) Use double-arm cell culture device for virus proliferation: Autoclave the double-arm cell culture device with the carrier, add culture medium, culture at 4°C for 36 hours, then place in 37°C water bath for 30 minutes; sterile Access the vero cells at 37 °C, 5% CO 2 Cell expansion culture under the condition of 36h;

[0031] 2) Replacement of the culture solution: measure the glucose content and pH value in the culture solution every day, and replace the culture solution when the sugar content is 1 mmol and the pH value is 6.5;

[0032] 3) Virus inoculation: when the number of African green monkey kidney cells reaches 10 8 At 37°C, 5% CO 2 Carry out virus amplification culture under the condition;

[0033] 4) Supplement culture medium: monitor the glucose content and pH value every day after inoculation, when the sugar content in the culture medium is 1 mmol, and the p...

Embodiment 2

[0037] A semi-perfusion culture method for duck Tembusu virus, comprising the following steps:

[0038] 1) Use double-arm cell culture device for virus proliferation: Autoclave the double-arm cell culture device with the carrier, add culture medium, culture at 20°C for 24 hours, then place in 37°C water bath for 30 minutes; sterile Access the vero cells at 37 °C, 5% CO 2 Cell expansion and culture under the condition of 100h;

[0039] 2) Replacement of the culture medium: measure the glucose content and pH value in the culture medium every day, and replace the culture medium when the sugar content is 10 mmol and the pH value is 7;

[0040] 3) Virus inoculation: when the number of African green monkey kidney cells reaches 10 8 At 37°C, 5% CO 2 Carry out virus amplification culture under the condition;

[0041] 4) Supplement culture fluid: Monitor the glucose content and pH value every day after inoculation. When the sugar content in the culture fluid is 10 mmol and the pH v...

Embodiment 3

[0045] 1) Virus multiplication using a dual-arm cell culture device: Autoclave the dual-arm cell culture device with the carrier, add culture medium, culture at 25°C for 36 hours, then place in a 37°C water bath for 30 minutes; sterile Access the vero cells at 37 °C, 5% CO 2 Cell expansion and culture under the condition of 120h;

[0046] 2) Replacement of the culture medium: measure the glucose content and pH value in the culture medium every day, and replace the culture medium when the sugar content is 10 mmol and the pH value is 7.5;

[0047] 3) Virus inoculation: when the number of African green monkey kidney cells reaches 10 8At 37°C, 5% CO 2 Carry out virus amplification culture under the condition;

[0048] 4) Supplementary culture medium: Monitor the glucose content and pH value every day after inoculation. When the sugar content in the culture medium is 10 mmol and the pH value is 7.5, collect 90% of the crude virus liquid, and then add an equal amount of fresh cul...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

No PUM Login to view more

Abstract

The invention discloses a semi-pouring flow type culture method for the duck Tembusu virus. The culture method comprises the steps that firstly, a cell culture device is used for carrying out virus reproduction, wherein the cell culture device containing a carrier is subjected to autoclaved sterilization, a culture solution is added, and African green monkey kidney cells are inoculated in a sterile mode; secondly, the culture solution is replaced, wherein the glucose content in the culture solution and the pH value are measured every day, and the culture solution is replaced; thirdly, virus inoculation is carried out, wherein allantoic fluid containing the duck Tembusu virus is inoculated, and virus multiplication culture is carried out; fourthly, the culture solution is supplemented, wherein crude virus liquor is collected, and then the fresh culture solution with the equal amount is supplemented to continue culture; fifthly, preservation is carried out, wherein the crude virus liquor collected in the fourth step is subjected to multigelation 1-5 times at minus 100-minus 20 DEG C, cell fragments are extracted through centrifuging after multigelation, filtering is carried out, and virus liquor is obtained and preserved at minus 20 DEG C. By means of the semi-pouring flow type culture method, the production efficiency of the virus can be improved, and crude virus liquor can be continuously gained multiple times every time each batch of cells are cultured.

Description

technical field [0001] The invention relates to a semi-perfusion culture method for duck Tembusu virus, which belongs to the field of biotechnology. Background technique [0002] In the prior art, virus culture cells were used for virus propagation in the early stage, and viruses were used in vaccines, which has become a common means in vaccine preparation technology. For virus propagation, most of them adopt the spinner bottle production process. Due to the limitation of the spinner bottle surface area and culture conditions, the cell density is low, the labor intensity is high, the preparation efficiency is low, the operation process is easy to be polluted, and the quality of the vaccine is difficult to be guaranteed. There are few reports about the perfusion culture method used in vaccine production. Perfusion culture is that on the one hand, fresh medium is continuously added to the reactor, and on the other hand, the reactor liquid is continuously taken out, so that th...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
Patent Type & Authority Patents(China)
IPC IPC(8): C12N7/00C12N7/02
CPCC12N7/00C12N2770/24051
Inventor 王伟冯晓声罗梦萍王贵平龚凤平
Owner GUANGDONG HAID GROUP
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap