Preparation method for duck tembusu virus liquid and product of duck tembusu virus liquid
A technology of duck Tembusu virus and virus liquid, applied in biochemical equipment and methods, viruses, viruses/phages, etc., can solve the problem of increasing foreign virus and mycoplasma contamination, increasing the difficulty of downstream separation and purification, and difficult product quality. Control and other issues to achieve the effect of improving equipment utilization, shortening production time, and expanding production scale
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Embodiment 1
[0026] Example 1 Acclimatization of DF-1 cells adapted to the serum-free medium and growing in a suspension state The DF-1 cells were acclimatized by gradually lowering the serum to adapt to the culture at low concentration of serum. When DF-1 cells growing stably in MEM medium containing 10% newborn bovine serum grow to the mid-logarithmic growth phase, replace them with MEM medium containing 5% newborn bovine serum until the cells grow to 80%-90% At confluence, trypsinize the cells to a cell density of 2.0 x 10 5 The cells / ml were subcultured in MEM medium containing 5% newborn bovine serum. After several generations, the survival rate of DF-1 cells in the MEM medium containing 5% newborn bovine serum was maintained at more than 90%, and maintained a relatively fast growth rate, which was used to further reduce serum acclimation culture. In the same way, DF-1 cells were gradually adapted to the culture condition of MEM containing 1% newborn bovine serum. The DF-1 cells ada...
Embodiment 2
[0028] The preparation of embodiment 2 duck Tembusu virus liquid
[0029] (1) The DF-1-XF cells obtained in Example 1 were domesticated and cultured according to 0.5×10 6 cells / ml is the initial inoculation density of the cells and inoculated them into a 7L bioreactor for cell suspension culture, and the culture time is 36 hours; other suspension culture conditions are as follows: pH control is 7.2, dissolved oxygen (DO) is controlled at 50%, and temperature is controlled at 37°C, the stirring speed is controlled at 100r / min;
[0030] (2) Inoculate Duck Tembusu virus (SD strain) to the DF-1-XF cells that were cultured in suspension for 48 hours according to the inoculation dose of 5% (V / V) to carry out virus proliferation and culture; harvest the virus liquid after 72 hours of proliferation and culture ; Other suspension culture conditions are as follows: pH control is 7.2, dissolved oxygen (DO) control is 50%, temperature control is 37 ℃, stirring speed control is 100r / min; ...
Embodiment 3
[0031] The preparation of embodiment 3 duck Tembusu virus liquid
[0032] (1) The DF-1-XF cells obtained in Example 1 were domesticated and cultured according to 0.3×10 6 cells / ml is the initial inoculation density of the cells and inoculate them into a 7L bioreactor for cell suspension proliferation culture; other suspension culture conditions are as follows: pH control is 7.2, dissolved oxygen (DO) is controlled at 50%, temperature is controlled at 37°C, stirring speed The control is 100r / min;
[0033] (2) Inoculate duck Tembusu virus (SD strain) to the DF-1-XF cells that were cultured in suspension for 48 hours according to the inoculation dose of 7.5% (V / V) to carry out virus proliferation and culture; harvest the virus liquid after 48 hours of proliferation and culture ; After testing, the virus price is 10 5.05 EID 50 / 0.2ml.
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