Diploid cell rabies vaccine and method for preparing purified rabies vaccine
A technology of diploid cells and vaccines is applied in the field of preparation of purified rabies vaccines, which can solve the problems that exogenous factors cannot be guaranteed, that cells cannot be carried, and the tumorigenicity of cell matrix and cell DNA cannot be guaranteed, so as to achieve good antibodies. Level, stability improvement, effect of increasing persistence
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Embodiment 1
[0055] Diploid cells come from the Chinese Center for Disease Control and Prevention, and the serum-free medium of the cell nutrient solution neutralizes 25IU / ml gentamicin or 25IU / ml kanamycin, and adjusts the pH to 7.2. After the dense monolayer was amplified according to the seeding ratio of 1:2, when it was amplified to the production batch, after about 4 days, when the cells grew into a dense monolayer, the cell nutrient solution was discarded, and the SNK-CTN virus was inoculated with a virus seed concentration of MOI 0.1~0.01, after 3 days of cultivation, start to harvest the virus, harvest once every 3-4 days, and collect 3 times in total, each batch of virus liquid is sampled for virus titration and sterility test, and it is required to harvest the virus drops of the virus liquid for the virus liquid degrees up to 10 5.0 LD50 / ml or above; add formaldehyde solution or β-propiolactone (final concentration 1 / 4000) or other suitable inactivating agents to the combined vir...
Embodiment 2
[0059] Diploid cells come from the Chinese Center for Disease Control and Prevention, and the cell nutrient solution is serum-free medium neutralized with 25IU / ml gentamicin or 25IU / ml kanamycin, and adjust the pH to 7.2, culture in a 3L spinner bottle at 37°C After forming a dense monolayer, amplify according to the seeding ratio of 1:2. When the amplified to the production batch, after about 4 days, when the cells grow into a dense monolayer, discard the cell nutrient solution, inoculate diploid cell rabies virus, and use Rabies virus SNK-CTN. The second-generation working virus seed of the strain passed on diploid cells, the concentration of the virus seed is MOI 0.05, and the cell maintenance medium is 199 culture medium containing 0.4-0.5% (W / W) human albumin, pH7.6, 35°C After 24 hours, replace with fresh cell maintenance medium and continue to cultivate for 2 days. Harvest the virus every 3.4 days, a total of 3 times. Each batch of virus liquid is sampled for virus titr...
Embodiment 3
[0063] With the inactivated rabies vaccine prepared by the method of claim 1, after 14 days of inoculating the purified rabies vaccine to those who have not been vaccinated against rabies, the positive conversion rate of neutralizing antibodies is higher than 95.0%, and no side effects occur.
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