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
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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 for Pr...
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 working seed batch (hemagglutination titer 240), which are used for monovalent vaccines preparation.
[0048] Conclusion: B / Malaysia / 2506 / 2004E 4 C 0...
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