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Process for preparing avian influenza inactivated vaccine by full suspended culture cells and product

A technology for culturing cells and inactivating vaccines, applied in the field of vaccine preparation, can solve problems such as inability to meet, and achieve the effects of low cost, high pollution rate, and increased culture density

Inactive Publication Date: 2014-08-27
郑州爱科生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, limited by the carrier amplification process, it cannot meet the needs of large-scale industrial production of high-quality avian influenza inactivated vaccines

Method used

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  • Process for preparing avian influenza inactivated vaccine by full suspended culture cells and product
  • Process for preparing avian influenza inactivated vaccine by full suspended culture cells and product
  • Process for preparing avian influenza inactivated vaccine by full suspended culture cells and product

Examples

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preparation example Construction

[0045] The preparation process of the present invention will be described in detail below.

[0046] (1) Full suspension culture of MDCK cells

[0047] (1.1) Shake flask suspension culture of MDCK cells

[0048] Take out two suspensions of MDCK cells frozen in liquid nitrogen, and after thawing quickly, directly add the suspended nutrient solution to 200ml in a 500ml shaker flask, and place them in suspension culture at 36-37°C. After the cells grow for 60-72 hours, the ratio of 1:3 Passage to other shake flasks at a ratio of ~1:5, and amplify to the required volume of the reactor;

[0049] (1.2) Reactor suspension culture of MDCK cells

[0050] (1.2.1) MDCK cell suspension culture in reactors above 20L

[0051] Dilute the MDCK cells cultured in shake flask suspension in step (1) to 0.5×10 6 After each / ml, inoculate into a bioreactor of more than 20L (20L refers to the working volume, all reactor volumes mentioned below are working volumes, and the working volume of the rea...

Embodiment 1

[0066] Full suspension culture cell manufactures avian influenza inactivated vaccine, comprises the following steps:

[0067] (1) Full suspension culture of MDCK cells

[0068] (1.1) Shake flask suspension culture of MDCK cells

[0069] Take out two suspended MDCK cells cryopreserved in liquid nitrogen, and after rapid thawing, directly add the suspension nutrient solution to 200ml in a 500ml shake flask, place it in a suspension culture at 37°C, and after the cells grow for 60 hours, pass to the In other shake flasks, amplify to the required volume of the reactor;

[0070] (1.2) Reactor suspension culture of MDCK cells

[0071] (1.2.1) MDCK cell suspension culture in 20L reactor

[0072] Dilute the MDCK cells cultured in shake flask suspension in step (1.1) to 0.5×10 6 cells / ml and inoculated into a 20L bioreactor, supplemented with suspension nutrient solution, the cell culture temperature was set to 36°C, the pH value was 7.2, and the dissolved oxygen was 40%; the cells...

Embodiment 2

[0086] Full suspension culture cell manufactures avian influenza inactivated vaccine, comprises the following steps:

[0087] (1) Full suspension culture of MDCK cells

[0088] (1.1) Shake flask suspension culture of MDCK cells

[0089] Take out two suspended MDCK cells cryopreserved in liquid nitrogen, and after rapid thawing, directly add the suspension nutrient solution to 200ml in a 500ml shake flask, place it in a suspension culture at 37°C, and after the cells grow for 72 hours, pass passage to In other shake flasks, amplify to the required volume of the reactor;

[0090] (1.2) Reactor suspension culture of MDCK cells

[0091] (1.2.1) MDCK cell suspension culture in 20L reactor

[0092] Dilute the MDCK cells cultured in shake flask suspension in step (1.1) to 0.5×10 6 cells / ml and inoculated into a 20L bioreactor, supplemented with suspension nutrient solution, the cell culture temperature was set at 36°C, the pH value was 7.2, and the dissolved oxygen was 60%; the c...

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Abstract

The invention relates to a process for preparing an avian influenza inactivated vaccine by full suspended culture cells and a product. The avian influenza inactivated vaccine product is prepared by the following steps: inoculating an avian influenza virus by the full suspended culture MDCK cells; clarifying, inactivating, degerming, concentrating and purifying and emulsifying the obtained avian influenza virus liquid. Compared with the prior art, the process provided by the invention is optimal and low in cost. Under the condition of guaranteeing the virus titer, the MDCK cell culture density can be greatly improved by the process provided by the invention, so that amplification on a large scale is easy to realize, and the avian influenza virus is proliferated in a great quantity. Compared with existing carrier suspended culture, the process provided by the invention is free from carrier intervention, so that the risk on pollution is greatly reduced. The process provided by the invention efficiently solves the problems of high pollution rate, low antigen output, long production period, insufficient SPF chicken embryo supply and the like of a chicken embryo method and is suitable for popularization and application of the reactor cell suspension culture technology in the avian influenza industry.

Description

technical field [0001] The invention relates to a preparation method and products of vaccines, in particular to a process for producing inactivated avian influenza vaccines by whole suspension culture cells and products obtained by the process. Background technique [0002] At present, the production of inactivated avian influenza vaccines and other animal influenza vaccines mainly relies on the traditional "chicken embryo method" process. The "chicken embryo method" has a long history of producing vaccines. It was first developed in the 1950s. Workers inject the virus into chicken embryos, allowing the virus to multiply naturally along with the development of the chicken embryos. After the chicken embryos develop for about 3 days, the workers will Open the eggshells one by one and collect the antigens one by one. Generally speaking, tens of thousands of eggs are often needed to produce a batch of vaccine antigens, which is a heavy workload. Although the current process of ...

Claims

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Application Information

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IPC IPC(8): A61K39/145A61P31/16
Inventor 李少英徐树兰吴华伟刘宏斌
Owner 郑州爱科生物科技有限公司
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