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Production method of porcine circovirus type 2

A technology of porcine circovirus and its production method, which is applied in the production process field of the spinner bottle culture method, can solve problems such as difficult to meet the requirements of vaccine matching, low antigen titer of semi-finished products, hidden dangers of vaccine quality, etc., achieve easy quality, improve vaccine quality and output , The production process is simple and stable

Inactive Publication Date: 2014-02-05
成都史纪生物制药有限公司
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  • Claims
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Problems solved by technology

[0002] At present, the domestic production of porcine circovirus type 2 inactivated vaccines is mainly realized by the method of cell transfer bottle culture. This traditional process produces semi-finished products with low antigen titers, and it is difficult to meet the requirements for vaccine matching. It requires labor-intensive, time-consuming and low-efficiency. , high production costs; easy to be polluted by the environment; large differences between different production batches or between different bottles of the same production batch; difficult to expand production; prone to hidden dangers of vaccine quality caused by bacteria or other virus contamination, involving biological safety and public health Hygiene issue

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  • Production method of porcine circovirus type 2

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Embodiment Construction

[0038] The present invention will be further described below in conjunction with the accompanying drawings and specific embodiments.

[0039] (1) Choose a bioreactor as a means of cultivation: 3L-3000L bioreactor.

[0040] (2) Select microcarriers as the carrier for cell-attached growth

[0041] Microcarrier cleaning and sterilization methods:

[0042] 1) Weigh 3-5g / L of Cytodex microcarriers and soak them in an appropriate amount of PBS for three hours.

[0043] 2) Wash 3 times with appropriate amount of PBS each time.

[0044] 3) Add an appropriate amount of PBS solution to soak the microcarriers, and steam sterilize at 121°C for 30 minutes.

[0045] (3) Select pK-15A1# cells as cells for seedling production:

[0046] (4) Subculture and cultivation of cells for seedling production:

[0047] The above cells were digested and passaged with trypsin digestion solution (containing 0.25% trypsin (1:250)), and then cultured with growth medium at a temperature of 37.0 (±0.2) °C...

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Abstract

The invention discloses a production process for producing porcine circovirus type 2 by culturing cells pK-15A1# in a bioreactor. The production process comprises the technical steps of (1) selecting pK-15A1# cells for multiplying circovirus type 2; (2) utilizing the bioreactor as a cell culture tool; (3) carrying out a virus culture process of synchronous virus inoculation; (4) culturing the pK-15A1# cells on micro-carriers in the bioreactor; (5) carrying out multiplication culture on viruses of porcine circovirus type 2 in the cells on the micro-carriers in the bioreactor; and (6) harvesting virus culturing materials of porcine circovirus type 2. The production process has the advantages that the production cost can be lowered, and compared with that of a traditional spinner bottle production process, the input-output ratio of the production process is increased by 5-10 times; the production cycle is short, the occupied area is small, the production scale can be easily and rapidly expanded, the environmental pollution is slight and is easy to avoid, the automation degree is high, few workers are required, the quality is stable, the cost can be remarkably lowered, and the yield and quality of vaccines can be remarkably improved.

Description

technical field [0001] The invention relates to a vaccine production method using bioreactor cell culture technology, which can be used for industrial production of porcine circovirus type 2 vaccine and replaces the traditional production process of rotary bottle culture method. Background technique [0002] At present, the domestic production of porcine circovirus type 2 inactivated vaccines is mainly realized by the method of cell spinner bottle culture. This traditional process produces semi-finished products with low antigen titers, and it is difficult to meet the requirements for vaccine matching. It requires labor-intensive, time-consuming and low-efficiency. , high production costs; easy to be polluted by the environment; large differences between different production batches or between different bottles of the same production batch; difficult to expand production; prone to hidden dangers in vaccine quality caused by bacteria or other virus contamination, involving bio...

Claims

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Application Information

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IPC IPC(8): A61K39/12A61P31/20C12N7/04C12R1/93
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
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