Suspension culture production method for porcine circovirus 2-type cells

A porcine circovirus, cell suspension technology, applied in microorganism-based methods, biochemical equipment and methods, viruses/phages, etc., can solve the problem of high labor, site and raw material costs, difficulty in meeting vaccine requirements, and difficulty in expanding production, etc. problem, to achieve the effect of simple and stable production process, expansion of production scale, and short production cycle

Active Publication Date: 2012-09-26
成都史纪生物制药有限公司
View PDF4 Cites 11 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0002] At present, the production of porcine circovirus type 2 inactivated vaccine in China is mainly realized by the method of cell spinner bottle culture. The semi-finished product antigen titer produced by this traditional process is low, only 10 4.5-5.5 TCID 50 /ml, it is difficult to meet the requirements of vaccine matching (antigen content per ml should be ≥10 5.5 TICD 50 ), different times of concentration are required
And this method is labor-intensive, requiring 100-200 spinner bottles to produce a batch; time-consuming, 6-7 days; low efficiency, only one harvest per cultivation; high

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

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Suspension culture production method for porcine circovirus 2-type cells
  • Suspension culture production method for porcine circovirus 2-type cells

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] A kind of porcine circovirus type 2 cell suspension culture production method, comprises the steps:

[0036] (1), Subculture and cultivation of healthy cells

[0037] The pK-15 clone cells were digested and passaged with trypsin digestion solution, cultured in cell growth medium at a temperature of 37.0°C (±0.2)°C; when a good cell monolayer was formed, it was used for further passage or inoculated in a bioreactor microcarrier culture;

[0038] (2), preparation and passage of toxic cells

[0039] Digest and passage the healthy pK-15 clone cells with good cell condition and grow into a single layer, then inoculate PCV2 seed virus according to 5% of the cell culture volume, culture for 48 hours, and after the cells grow to more than 50%, change the maintenance solution containing MEM and continue to maintain for 48 hours. After the cells are full, the F1 generation of toxic cells can be obtained; gently wash the F1 generation of toxic cells with preheated sterile PBS bu...

Embodiment 2

[0061] A kind of porcine circovirus type 2 cell suspension culture production method, comprises the steps:

[0062] (1), Subculture and cultivation of healthy cells

[0063] The pK-15 clone cells were digested and passaged with trypsin digestion solution, cultured in cell growth medium at a temperature of 37.0°C (±0.2)°C; when a good cell monolayer was formed, it was used for further passage or inoculated in a bioreactor microcarrier culture;

[0064] (2), preparation and passage of toxic cells

[0065] Digest and passage the healthy pK-15 clone cells with good cell condition and grow into a monolayer, then inoculate PCV2 seed virus according to 1% of the cell culture volume, culture for 24 hours, and after the cells grow to more than 50%, change the maintenance solution containing MEM and continue to maintain for 48 hours. After the cells are full, the F1 generation of toxic cells can be obtained; gently wash the F1 generation of toxic cells with preheated sterile PBS buffe...

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 suspension culture production method for porcine circovirus 2-type cells; the method provided by the invention comprises the production technology of preparing porcine circovirus 2-type inactivated vaccines by suspension culture for poisoned cells through porcine circovirus 2-type virus biological reactor micro carriers. The method provided by the invention can greatly reduce production cost and increase output-input ratio by 5-10 times; and the method shortens production period by 2-3 days and occupies small places, wherein occupied area is only 1/5 of the occupied area in the traditional roller bottle process under the same yield; scale of production is enlarged fast and easily; pollution is little and processes are performed easily under the totally-enclosed tank production condition; degree of automation is high; employee is less; quality stability is realized easily; and the method can significantly reduce production cost and increase yield and quality of vaccines.

Description

technical field [0001] The invention relates to a cell culture production method, in particular to a method for producing porcine circovirus type 2 vaccine antigen using bioreactor microcarrier cell culture technology. Background technique [0002] At present, the production of porcine circovirus type 2 inactivated vaccine in China is mainly realized by the method of cell spinner bottle culture. The semi-finished product antigen titer produced by this traditional process is low, only 10 4.5-5.5 TCID 50 / ml, it is difficult to meet the requirements of vaccine matching (antigen content per ml should be ≥10 5.5 TICD 50 ), different times of concentration are required. And this method is labor-intensive, requiring 100-200 spinner bottles to produce a batch; time-consuming, 6-7 days; low efficiency, only one harvest per cultivation; high production cost, large labor, site and raw material costs; Polluted by the environment, the toxic waste water produced by bottle washing nee...

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
IPC IPC(8): C12N7/00C12R1/93
Inventor 徐宏军丁光星任丽赵英杰胡来根刘玉才
Owner 成都史纪生物制药有限公司
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