Preparation method of avian influenza virus and fowl adenovirus combined inactivated vaccine
A dual-inactivated vaccine and avian influenza virus technology, applied in biochemical equipment and methods, vaccines, viruses, etc., can solve the problems of immutable downstream technologies, low virus titers, and easy contamination, and achieve increased virus content, Improve the effect of vaccines and increase the effect of proliferation
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Embodiment 1
[0043] Embodiment 1, a kind of preparation method of avian influenza virus, avian adenovirus dual inactivated vaccine
[0044] Preparation of S1 cell carrier: Digest and disperse LMH cells with trypsin, and use DMEM medium containing 5% newborn bovine serum and 1000 units / mL penicillin-streptomycin double antibody at 37°C, 5% CO 2 LMH cells were cultured for 2 days under certain conditions, and then the cells were washed twice with serum-free DMEM medium to obtain cell carriers; the LMH cells were cultured in spinner bottles or placed in microcarrier reactors.
[0045] Inoculation of S2 virus: inoculate the virus liquid of avian influenza virus subtype H9 and avian adenovirus type 4 into the cell carrier obtained in step S1 at a final volume of 1:50, place it at 37°C for 30 minutes, and then suck out the virus liquid , followed by medium A, at 37°C, 5% CO 2 48h under the condition of culturing, 80% cytopathy occurs, and diseased cells are obtained;
[0046] Described medium ...
Embodiment 2
[0054] Embodiment 2, a kind of preparation method of avian influenza virus, avian adenovirus dual inactivated vaccine
[0055] Preparation of S1 cell carrier: Digest and disperse LMH cells with trypsin, and use DMEM medium containing 8% newborn bovine serum and 1500 units / mL penicillin-streptomycin double antibody at 37°C, 5% CO 2 The conditions were cultivated for 2 days, and then the cells were washed 3 times with serum-free DMEM culture medium to obtain cell carriers; the LMH cells were cultured in a microcarrier reactor, and the amount of microcarriers used was 6 g / L.
[0056] Inoculation of S2 virus: inoculate the virus liquid of avian influenza virus subtype H9 and avian adenovirus type 4 into the cell carrier obtained in step S1 at a final volume of 1:100, place it at 37°C for 40 minutes, and then suck out the virus liquid , followed by medium A, at 37°C, 5% CO 2 Under the conditions of 56 hours, 80% of the cells were damaged, and the diseased cells were obtained;
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Embodiment 3
[0065] Embodiment 3, a kind of preparation method of avian influenza virus, avian adenovirus dual inactivated vaccine
[0066] Preparation of S1 cell carrier: Digest and disperse LMH cells with trypsin, and incubate at 37°C, 5% CO with DMEM medium containing 10% newborn bovine serum and 2000 units / mL penicillin-streptomycin double antibody 2 The conditions were cultivated for 3 days, and then the cells were washed 3 times with serum-free DMEM culture medium to obtain cell carriers; the LMH cells were cultured in a microcarrier reactor, and the amount of microcarriers used was 8 grams per liter.
[0067] Inoculation of S2 virus: inoculate the virus liquid of avian influenza virus subtype H9 and avian adenovirus type 4 into the cell carrier obtained in step S1 at a final volume of 1:200, place it at 37°C for 60 minutes, and then suck out the virus liquid , followed by medium A, at 37°C, 5% CO 2 72h under the condition of culture, 80% cytopathy occurs, and diseased cells are obt...
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