Method for producing vaccine by virtue of culturing animal cells
A technology of animal cells and cultured cells, which is applied in the field of bioengineering and can solve problems such as low degree of automation, large differences in product batches, unstable quality of cells and vaccines, etc.
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Embodiment 1(10
[0027] Example 1 (Preparation of PRRS vaccine by culturing Marc-145 cells in a 10L bioreactor)
[0028] Porcine reproductive and respiratory syndrome virus: CH-1R strain.
[0029] After the sheet-shaped carrier is sterilized by gamma rays at 24 kGy, it is first soaked in PBS, then rinsed with PBS, soaked in culture medium overnight, and set aside.
[0030] Inoculate 1 × 10 on the sheet carrier in the bioreactor 7 A Marc-145 cell was cultured and grown at 30-60% DO, pH 7.0-7.2, and 37°C for 5-7 days, during which glucose digestion was detected, and the medium was changed according to the sugar consumption. , so that the amount of sugar can be maintained at a daily sugar consumption of more than 1g / L. With the increase of cell density, the daily sugar consumption will gradually increase until it reaches more than 2.5g / L.
[0031] Drain the culture medium, wash it twice with sterile PBS, and inoculate porcine reproductive and respiratory syndrome virus according to the MOI of v...
Embodiment 2(10
[0036] Example 2 (Preparation of avian influenza vaccine by culturing MDCK cells in a 10L bioreactor)
[0037] After the sheet-shaped carrier is sterilized by gamma rays at 24 kGy, it is first soaked in PBS, then rinsed with PBS, and then soaked in culture medium overnight, and set aside.
[0038] Inoculate 0.5 × 10 on the sheet carrier in the bioreactor 7 For MDCK cells, the DO was controlled at 30-60%, the pH was maintained at 7.2-7.4, and the temperature was 37°C. Detect glucose digestion, and perform fluid replacement or rehydration according to the sugar consumption to keep the sugar level above 1g / L. Basically, the cell density reaches the maximum within 5-7 days.
[0039] Empty the culture medium, wash twice with sterile PBS, and inoculate H5N1 virus according to the MOI of virus infection at 0.01-0.1. All strains that are generally adaptable and sensitive to MDCK cells or other susceptible cells are applicable to the method of the present invention.
[0040] Continu...
Embodiment 3(10
[0044] Embodiment 3 (10L bioreactor cultivates ST cell to prepare swine fever vaccine)
[0045] After the sheet-shaped carrier is sterilized by gamma rays at 24 kGy, it is first soaked in PBS, then rinsed with PBS, soaked in culture medium overnight, and set aside.
[0046] Inoculate 2 × 10 on the sheet carrier in the bioreactor 7 For ST cells, the DO was controlled at 30-60%, the pH was maintained at 7.0-7.2, and the temperature was 37°C. Glucose digestion was detected, and the medium was changed every 2-3 days according to the sugar consumption, so that the sugar content was maintained above 1g / L, and the maximum cell density was reached in 5-7 days.
[0047] Drain all the culture medium, wash the sheet carrier and cells twice with sterile PBS, and inoculate ST cells with a concentration of 2-3% spleen toxin. The methods of the present invention are applicable to all strains that are generally adaptive and sensitive to ST cells or other susceptible cells.
[0048] The ST ...
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