Avian adenovirus strain, inactivated vaccine and preparation method
A technology of avian adenovirus and inactivated vaccine, which is applied in the field of bioengineering to achieve the effects of high safety, high protection rate against virus and rapid immune response
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Embodiment 1
[0039] Example 1 Preparation of avian adenovirus inactivated vaccine using chicken embryo kidney cells (CEK cells)
[0040] The formula of the cell growth solution is: M199 solution with a volume percentage of 90%, 10% newborn calf serum, and adjust the pH to 7.2;
[0041] The cell maintenance solution formula is: M199 solution with a volume percentage of 98%, 2% newborn calf serum, 2.4mmol / L arginine, and adjust the pH to 7.4;
[0042] a. Preparation of CEK cells: Take 17-day-old SPF chicken embryos, take out the kidneys in a sterile environment, place them in an appropriate amount of pancreatin, and digest them at 37°C. After the digestion is complete, take the supernatant and add an appropriate amount of serum to stop the digestion, centrifuge at 1500 rpm for 10 minutes, discard the supernatant, resuspend the cell pellet with cell growth fluid and dilute to a final concentration of 1×10 cells 6 Cells / ml, cultured at 37°C for 72 hours, the cells grow into a good monolayer;
[0043] ...
Embodiment 2
[0053] Example 2 Preparation of avian adenovirus inactivated vaccine using chicken embryo fibroblast cell line (DF1 cells)
[0054] The growth solution formula for cells is: DMEM / F12 solution with a volume percentage of 92%, 8% newborn calf serum, and adjust the pH to 7.2;
[0055] The cell maintenance solution formula is: DMEM / F12 solution with a volume percentage of 97%, 3% newborn calf serum, 2.2mmol / L arginine, and adjust the pH to 7.3;
[0056] a. Cell culture:
[0057] The DF1 seed cells were removed from the liquid nitrogen and melted in a 37°C water bath, and resuscitated and cultured with cell growth medium (DMEM / F12 medium). After 3 days of culture, the number of cells was 4.0×10 7.0 Cells / ml, and passage 1:3, culture at 37°C for 2 days;
[0058] b. Virus inoculation: Press 10 for poultry adenovirus 3.0 ELD 50 / 0.1ml inoculate DF1 cells, add cell maintenance medium (DMEM / F12 medium), continue culture at 37°C;
[0059] c. Harvesting the venom: After the virus inoculated cells an...
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