Method for proliferating avian influenza virus on MDCK full suspension cells and application thereof
An avian influenza virus, full suspension technology, applied in the direction of viruses, animal cells, viruses/phages, etc., can solve the problems of large amount of seed virus access, long domestication time, long virus domestication cycle, etc., to shorten the production cycle, seed virus The effect of variant risk reduction
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[0034] The preparation method of the culture medium described in any one of the foregoing includes: mixing all the culture medium components together to form dry culture medium powder and then dispersing in a solvent.
[0035] Preferably, at the time of preparation, the major components of the medium are refined and mixed together to form a dry medium powder, and the minor components are separately formulated into a minor component liquid.
[0036] Preferably, in the above-mentioned preparation method, the method further includes pH adjustment and filter sterilization.
[0037] Preferably, the method as described above, the method also includes:
[0038] Viral antigens are isolated from the virions and purified and / or inactivated.
[0039] According to one aspect of the present invention, the present invention also relates to the application of the aforementioned method in the preparation of avian influenza virus vaccine.
[0040] Preferably, in the above-mentioned applicati...
Embodiment 1
[0043] A serum-free medium adapted to the full suspension culture of MDCK cells, calculated as 1 liter of solution after dissolving with ultrapure water, said medium includes the following components:
[0044] Biotin 1 x 10 -8 M, Calcium Chloride 5×10 -3 M, copper sulfate 2.8×10 -9 M, Cyanocobalamin 1×10 -7 M, D-calcium pantothenate 2×10 -5 M, D-glucose 2.0×10 -2 M, ferrous sulfate 2×10 -6 M, folic acid 0.5×10 -4 M. Glutathione 9.5×10 -7 M. Hydrocortisone 3×10 -8 M, Hypoxanthine 1×10 -5 M. Inositol 10×10 -5 M, ferric nitrate 0.3×10 -7 M, L-alanine 20×10 -5 M, L-arginine 20×10 -4 M, L-asparagine 1×10 -5 M, L-aspartic acid 20×10 -5 M, L-cysteine 0.1×10 - 4 M, L-cystine 10×10 -4 M, L-glutamic acid 1×10 -5 M, L-Glutamine 10×10 -3 M, Glycine 1×10 -4 M, L-histidine 10×10 -4 M, L-isoleucine 0.5×10 -4 M, L-Leucine 10×10 -4 M, L-lysine 1×10 -4 M, L-methionine 10×10 -4 M, L-phenylalanine 1×10 -4M, L-proline 10×10 -4 M, L-serine 1×10 -4 M, L-Threonine 20×10 ...
Embodiment 2
[0047] A serum-free medium adapted to the full suspension culture of MDCK cells, calculated as 1 liter of solution after dissolving with ultrapure water, said medium includes the following components:
[0048] Biotin 3 x 10 -8 M. Calcium chloride 2×10 -3 M, copper sulfate 7.8×10 -9 M, Cyanocobalamin 3×10 -7 M, D-calcium pantothenate 5×10 -5 M, D-glucose 1.8×10 -2 M, ferrous sulfate 5×10 -6 M, folic acid 1×10 -4 M. Glutathione 6.5×10 -7 M. Hydrocortisone 5×10 -8 M, Hypoxanthine 3×10 -5 M. Inositol 7×10 -5 M, ferric nitrate 1.2×10 -7 M, L-alanine 5×10 -5 M, L-arginine 7×10 -4 M, L-asparagine 5×10 -5 M, L-aspartic acid 5×10 -5 M, L-cysteine 1×10 -4 M, L-cystine 1×10 -4 M, L-glutamic acid 5×10 -5 M, L-Glutamine 2.5×10 -3 M. Glycine 2.5×10 -4 M, L-histidine 1.5×10 -4 M, L-isoleucine 4.2×10 -4 M, L-leucine 4.5×10 -4 M, L-lysine 5×10 -4 M, L-methionine 1.2×10 - 4 M, L-phenylalanine 2.2×10 -4 M, L-proline 1.5×10 -4 M, L-serine 2.5×10 -4 M, L-threonine 4.5...
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