Method for preparing enterovirus vaccines
An enterovirus and vaccine technology, applied in the field of enterovirus vaccine preparation, can solve the problems of yield limitation, low efficiency, low reproduction titer and the like, and achieve the effects of saving consumption, simple production process and convenient purification
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Embodiment 1
[0021] Embodiment 1: Preparation of EV71 monovalent vaccine
[0022] (1) Preparation of cell monolayer
[0023] Take gerbils aged 10-15 days, cut off the carotid artery and bleed to death, soak in 0.2% (v / v) new clean and disinfectant solution for 30 minutes, take the muscle cells of the hind legs aseptically, cut them into pieces, and contain 100U / ml card Wash twice with the MEM culture solution of Namycin, remove the washing solution; add 0.2% (w / v) collagenase II and 0.25% (w / v) trypsin digestion solution of 10 times the weight of muscle tissue, and place at 4°C Digestion: Discard the digestive fluid after 16-20 hours, add MEM solution 10 times the mass of muscle tissue to repeatedly blow and disperse the cells, pass through 100, 200 and 400 mesh stainless steel filters in turn, collect the filtrate, centrifuge, discard the supernatant, and precipitate the cells Resuspend with MEM, add newborn bovine serum to a final concentration of 10% (v / v), inoculate cell bottles, and ...
Embodiment 2
[0035] Embodiment 2: Preparation of CoxA16 vaccine
[0036] In Example 1, the inoculated virus was replaced with the CoxA16 virus strain (provided by Hangzhou Sixth People's Hospital), and the other preparation methods were the same, and CoxA16 vaccine semi-finished products and finished vaccines could be prepared.
Embodiment 3
[0037] Embodiment 3: Preparation of EV71 and CoxA16 bivalent vaccine
[0038] The EV71 vaccine semi-finished product prepared in Example 1 and the CoxA16 vaccine semi-finished product prepared in Experimental Example 2 were mixed at a volume ratio of 1:1 and subpackaged to obtain a bivalent enterovirus vaccine.
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