The invention discloses a method for large scale cultivation in a
bioreactor, comprising the following steps: (1) making a recombinant H5N1
avian influenza virus adapt to MDCK cells; (2) preparing a seed lot by using the adapted
virus strain as a reactor large-scale culture
virus, wherein the seed lot
virus generation is no less than F6, the
hemagglutination titer is no less than 28, and EID50 is no less than 107.5; (3) culturing the MDCK cells by using
polyester sheets as a carrier and the reactor as a basket
fixed bed, and pouring the cells on every
gram of the carrier, wherein the culture medium is newborn calf serum DMEM; (4) inoculating the virus on the cultured MDCK cells, washing the cells by using a virus
diluent, conducting
virus inoculation, after absorption, and replacing a
cell maintenance media; (5) maintaining the MDCK cells and conducting virus propagation, stirring, and replacing the
cell maintenance media; and (6) monitoring the
hemagglutination titer and harvesting the virus, and harvesting the reactor to culture a volume virus liquid. According to the invention, the method is easy to operate and the operation is simple, the HA
titer of the virus after adaption is higher than 2<7>, and the problem that the culture virus titer cannot satisfy the demand of produce vaccine in the existing culture of the recombinant H5 subtype
avian influenza virus on the MDCK cells is solved.