Adaption method of influenza virus vaccine strains on Vero cells

An adaptation method, a technology for influenza vaccine, applied in the direction of microorganism-based methods, animal cells, viruses/phages, etc., which can solve the problems of conventional methods such as troublesome

Active Publication Date: 2013-06-05
吉林亚泰生物药业股份有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The main problems of chicken embryo vaccines: on the one hand, there are strict requirements on eggs, not every egg can be used to produce vaccines, and only certified chicken farms can provide such eggs; on the other hand, it involves the production of embryonated eggs Conventional methods for influenza vaccines are cumbersome, involving the handling of thousands of eggs per week, and large-scale purification of virus suspensions obtained from allantoic fluid to ensure they are free of egg proteins
However, the experimental results of our study did not show that this method can change the fundamental problem that influenza virus is difficult to adapt to in Vero cells

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Type A virus strain, approved by Who, Solomon strain, A / Solomon / 3 / 06 (H1N1)-, IVR-145, from NIBSC, UK, batch number: Ivr-145, 06 / 234, generation E 7 (The passage of the virus in the chicken embryo is represented by E and the number, and the passage of the virus on the cell is represented by C and the number)

[0028] SPF eggs: Beijing Meria Verton Laboratory Animal Technology Co., Ltd.

[0029] Incubate in a biochemical incubator at 37°C for 10 days.

[0030] Vero monolayer cells, passage 140-145;

[0031] MDCK cells, passage 99-100;

[0032] A / Solomon / 3 / 06 (H1N1)E 7 C 0 Infected MDCK cells and chicken embryos with NIBSC virus seeds for subculture, after passage of chicken embryos, the generation of virus seeds is E 8 C 0 , the hemagglutination titer was 640 and the passage of MDCK cells was E 7 C 1 , The hemagglutination titer is 20-40. E 8 C 0 The Vero cells were infected with the virus species and continued to be subcultured, harvested when the cytopathic...

Embodiment 2

[0035] A type 3 virus, A / wisconsin / 67 / 2005 (H3N2)-, NYMC-161, from NIBSC, batch number: 06 / 112, generation E 9

[0036] SPF eggs: Beijing Meria Verton Laboratory Animal Technology Co., Ltd.

[0037] Incubate in a biochemical incubator at 37°C for 10 days.

[0038] Vero monolayer cells, passage 140-145;

[0039] MDCK cells, passage 99-100;

[0040] A / wisconsin / 67 / 2005 (H3N2) E9C0 derived from NIBSC virus seed was infected with MDCK cells and chicken embryos for passage, and the passage of virus seed after passage of chicken embryo was E 10 C 0 , the hemagglutination titer was 480 and the passage of MDCK cells was E 9 C 1 , The hemagglutination titer was 160. Passaging MDCK to E 9 C 1 The Vero cells were infected with the virus species and continued to be subcultured, harvested when the cytopathic changes reached +++-++++, and the main generation seed batch (hemagglutination titer 240) and working seed batch (hemagglutination titer 480) were established and used fo...

Embodiment 3

[0043] Type B virus, B / Malaysia / 2506 / 2004, from NIBSC, UK, batch number: 06 / 104, generation E 4

[0044] SPF eggs: Beijing Meria Verton Laboratory Animal Technology Co., Ltd.

[0045] Incubate in a biochemical incubator at 37°C for 10 days.

[0046] Vero monolayer cells, passage 140-145;

[0047] Will B / Malaysia / 2506 / 2004E 4 C 0 Originated from NIBSC virus species infected chicken embryos and Vero cells, the hemagglutination titer reached 240, and the virus generation after passage was E 5 C 0 , while Vero cells were passaged to E 4 C 1 , with a hemagglutination titer of 10. Will B / Malaysia / 2506 / 2004E 4 C 1 Virus seeds are returned to chicken embryos, and the hemagglutination titer can reach 480, and then continue to be passed on Vero cells to establish the main generation seed batch (hemagglutination titer 160) and the working seed batch (hemagglutination titer 240), which are used for monovalent vaccines preparation.

[0048] Conclusion: B / Malaysia / 2506 / 20...

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Abstract

The invention provides an adaption method of influenza virus vaccine strains on Vero cells. The influenza virus vaccine strains are rapidly adapted on the Vero cells; and various effective virus adaption methods are established, so that influenza virus seeds can be adapted on the Vero cells, and virus progenies used as vaccines are produced under the condition of minimizing virus surface antigen variation caused by virus adaptation selection. The hemagglutination titer of the virus seeds prepared by vaccine strains adapted by the method is more than or equal to 160, and meets the requirementsof three parts of the Chinese pharmacopoeia, 2010 edition.

Description

technical field [0001] The invention relates to a method for adapting multiple influenza virus vaccine strains on Vero cells, and obtains vaccine-adapted strains and virus seed batches to prepare Vero cell influenza vaccines, belonging to the field of biotechnology. Background technique [0002] The traditional method of preparing influenza vaccine is to use chicken embryos, which have been used in China for more than 50 years to cultivate vaccines. The currently used influenza vaccines are mainly inactivated influenza trivalent split vaccines prepared from chicken embryos, including influenza A strains, H 3 N 2 and influenza B strains. There are also influenza vaccines (including H 5 N 1 wait). The source of the vaccine strain is mainly determined by the surface antigen of the influenza virus epidemic strain recommended by the WHO according to the annual change of the global influenza virus for the next year's influenza vaccine production. strain (A / PR / 8 / 34 (H 1 N 1...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N7/00C12N5/071C12R1/93
Inventor 于洪涛李光谱张莹周庆玲董唤岳振齐胡晓明郭岩李淑兰
Owner 吉林亚泰生物药业股份有限公司
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