New hepatitis A inactivated vaccine virus strain and method for culturing same
A technology of hepatitis A virus and hepatitis A, applied in the direction of antiviral agent, inactivation/attenuation, and resistance to vector-borne diseases, etc., can solve the problem that hepatitis A vaccine cannot meet the requirements of the domestic market, and achieve the goal of being suitable for large Large-scale industrial production, the effect of improving yield and quality
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Embodiment 1
[0017] Embodiment 1 The culture method of hepatitis A virus strain JS-4 strain
[0018] The feces collected from clinical hepatitis A patients in the acute phase were prepared into 20% suspension, treated with 5% plant activated carbon, centrifuged at 10000rpm for 30min, the supernatant was absorbed and appropriate amount of antibiotics were added, and placed at 4°C for later use.
[0019] After the vero cells grow into a single layer in the small square bottle, wash the cell surface once, inoculate each bottle with 2ml of feces suspension, absorb at 37°C for 2 hours, supplement the cell maintenance solution and culture at 36°C for 28 days to collect the virus, and change the medium every seven days during this period .
[0020] The samples separated and passaged in step (b) continue to be adapted for Vero cell culture. The difference is that each bottle of cells was inoculated with 1mL sample, adsorbed at 37°C for 2 hours, and cultured at 36°C for 28 days. After the 10th pas...
Embodiment 2
[0035] Example 2 The method for preparing hepatitis A inactivated vaccine virus antigen
[0036] Take out the cell cryopreservation tube from the Vero cell working seed bank, put it into 40°C water to dissolve quickly, then add it to the 199 culture medium pre-warmed at 37°C, plant it into the cell bottle, culture it at 37°C, and grow into single cells after 3-4 days After layering, amplify for several generations until the specified generation for production.
[0037] Add the above-mentioned cells to the hepatitis A virus strain JS-4 strain obtained in Example 1 at 0.2 MOI, inoculate them in a 3L three-layer flask after adsorbing the suspended cells at 37°C for 40 minutes, and culture them at 36°C, during which time, replace them every seven days. The virus-cell culture suspension was digested with digestive juice after the culture period expired, and the culture medium was broken and extracted, concentrated and purified. After inactivation, the hepatitis A inactivated vacci...
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