Method for producing porcine circovirus type 2 antigens in large scale with high density

A porcine circovirus, high-density technology, used in viral antigen components, biochemical equipment and methods, antiviral agents, etc.

Active Publication Date: 2014-08-27
JIANGSU NANNONG HI TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] The object of the invention is to overcome the deficiencies of the existing spinner bottle culture method and the traditional cell suspension microcarrier culture meth

Method used

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  • Method for producing porcine circovirus type 2 antigens in large scale with high density
  • Method for producing porcine circovirus type 2 antigens in large scale with high density
  • Method for producing porcine circovirus type 2 antigens in large scale with high density

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Embodiment Construction

[0012] The bioreactor is used as the culture equipment, the microcarrier is used as the carrier to support cell growth, the cloned cell PK-15W (formerly known as PK-15B1, CCTCC No.C200936) is used as the seedling cell, and PCV2 is used as the seedling poison; The low-sugar DMEM of globulin A and D-glucosamine is used as the seedling cell growth liquid; the serum-free low-sugar DMEM containing concanavalin A and hydrolyzed casein is used as the seedling cell maintenance liquid; and comprises the following steps:

[0013] (1) Seed cell culture

[0014] Wash the overgrown seed cell monolayer once with preheated sterile saline, add 20-40ml of trypsin digestion solution to digest, keep rotating the spinner bottle until all the cells fall off, add cell growth solution to stop digestion, mix well, follow 1 : 3 to 1:5 passage ratio bottle;

[0015] (2) Preparation of seed cells for inoculation

[0016] 1) Preparation of the bioreactor Select a suitable bioreactor according to the vo...

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Abstract

The invention relates to a method for producing porcine circovirus type 2 antigens in large scale with high density. A bioreactor microcarrier suspension culture technology used for replacing an existing spinner bottle culture technology for producing the porcine circovirus type 2 antigens. The method can greatly reduce the production cost and improve the yield. Compared with the spinner bottle technology, the unit antigen cost is reduced by 80% to 90%, the production period is reduced by 7 days, and the yield is improved by 3 times to 10 times; due to the fact that the antigens can be obtained repeatedly, compared with a traditional reactor culture technology, the unit antigen cost is reduced by 40% to 50%, and the yield is improved by 3 times to 5 times. The antigens produced through the method has no serum residues, the produced vaccine is higher in safety and small in batch difference, the quality is stable and easy to control, and the yield and quality of the produced vaccine can be obviously improved.

Description

technical field [0001] The invention relates to a method for large-scale and high-density production of porcine circovirus type 2 vaccine antigen by applying bioreactor microcarrier suspension culture technology, and belongs to the field of veterinary biological products. Background technique [0002] At present, the production of porcine circovirus type 2 (Porcine circovirus2, PCV2) vaccine antigen mainly adopts the method of cell spinner bottle culture, and a few adopts the method of cell suspension culture. 4.5～6.0 TCID 50 / ml, unable to provide stable qualified antigen (qualified antigen antigen content per milliliter should be ≥ 10 5.5 TCID 50 ), requiring high concentration. Moreover, the spin bottle process is labor-intensive. It takes 100-200 spin bottles to produce a batch, and multiple people are required to carry out long-term digestion and passage operations at the same time, which increases the risk of pollution; the production cycle is long, requiring 10 day...

Claims

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Application Information

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IPC IPC(8): C12N7/00C12N7/02A61K39/12A61P31/20
Inventor 董彦鹏孙石静何叶峰缪芬芳何海蓉张志华胡芳
Owner JIANGSU NANNONG HI TECH
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