Method for amplifying porcine circovirus type 2 by applying bioreactor and flaky vector

A bioreactor, porcine circovirus technology, applied in the direction of microorganism-based methods, biochemical equipment and methods, microorganisms, etc., can solve the problems of high infection rate of breeding pigs, serious mixed infection, economic loss of pig industry, etc., to achieve The effects of reducing labor intensity, precise cultivation, and increasing virus yield and titer

Inactive Publication Date: 2014-03-05
扬州优邦生物药品有限公司
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Problems solved by technology

At present, circovirus disease has commonly occurred in our country, which is characterized by wide epidemic range, high positive rate, ser

Method used

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Experimental program
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Embodiment 1

[0028] A method for large-scale amplification of porcine circovirus type 2 using bioreactors and sheet-like carriers, PK-15 cell culture circovirus bioreactor cultivation process research The method specifically consists of the following steps:

[0029] 1 material

[0030] 1.1 Cells PK-15 cells (without PCV-1 pollution) were expanded and preserved by Yangzhou Youbang Biopharmaceutical Co., Ltd.

[0031] 1.2 The F15 generation of virus seed PCV-2 (YZ strain) was amplified and preserved by Yangzhou Youbang Biopharmaceutical Co., Ltd.

[0032] 1.3 Medium and reagents DMEM was purchased from Sigma; fetal bovine serum was purchased from Hyclone; D-glucosamine was purchased from BBI; PCV-2 positive serum was purchased from VMRD; goat anti-pig fluorescent secondary antibody was purchased from SouthernBiotech; others Reagents are domestic analytical grade.

[0033] 1.4 Instruments The 7.5L bioreactor and supporting sheet carrier were purchased from NBS Company; the inverted fluoresc...

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Abstract

The invention relates to a method for amplifying porcine circovirus type 2 by applying a bioreactor and a flaky vector, and belongs to the technical field of biological products. The method comprises the following steps: (1) seed cell preparation; (2) bioreactor preparation; (3) PK-15 cell suspension culture; (4) PK-15 cell suspension virus culture; (5) cell suspension virus culture and continuous harvesting; and (6) virus liquid culture period and harvest yield. The method disclosed by the invention exceeds multiple technical bottlenecks of a spinner bottle production process and provides a process which obtains high-titer porcine circovirus type 2 (PCV2) virus liquid with high efficiency, sustainability and large-scale production by carrying out suspension culture on a PK15 cell by utilizing the flaky vector.

Description

technical field [0001] The invention relates to a method for amplifying porcine circovirus type 2 using a bioreactor and a sheet carrier, and belongs to the technical field of biological products. Background technique [0002] Circovirus infection is a relatively difficult infectious disease discovered in recent years. The disease mainly affects suckling and finishing pigs, as well as pregnant sows. Its characteristic clinical symptoms are poor constitution and pale skin in pigs. Diseases caused include porcine post-weaning multisystemic wasting syndrome (PMWS), proliferative necrotizing pneumonia (PNP), porcine dermatitis and nephrotic syndrome (PDNS), porcine respiratory disease syndrome (PRDC), reproductive failure, congenital Tremor and enteritis etc. At the same time, the immunosuppression produced by PCV-2 infection can easily lead to mixed infection or secondary infection of other pathogens. In 2006 and 2007, porcine circovirus played an important role in the occu...

Claims

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Application Information

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IPC IPC(8): C12N7/00C12R1/93
Inventor 钱钟潘杰宋庆庆李群徐萍
Owner 扬州优邦生物药品有限公司
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