Transfer factor specific for SARS virus and method for preparing same
A transfer factor and specific technology, applied in antiviral agents, antiviral immunoglobulins, pharmaceutical formulations, etc., can solve problems such as drugs that have not been developed for SARS
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Embodiment 1
[0033] The preparation of embodiment 1 SARS inactivated vaccine
[0034] The bottom area is 275cm 2 Cultivate VeroE6 (National Institute of Virology, China) in cell flasks. After the cells grow into a dense monolayer, wash the cells with serum-free medium for 3 times, add 100ml of serum-free medium, and inoculate F69 and Z 2 -Z 3 SARS-Cov virus strain, when cytopathic to 75%-100%, titrate virus, TCID 50 is log10×10 7 / ml. Freeze and thaw the culture flask 3 times in a -20°C refrigerator, shake well and place in a 2-8°C refrigerator overnight, inactivate with 0.4% formaldehyde for 24 hours, 5000rpm×30min, discard the precipitate, and use the supernatant as a SARS inactivated vaccine.
Embodiment 2
[0035] Embodiment 2 immunization program (SARS inactivated vaccine immunization alone)
[0036]17 healthy castrated boars (body weight 50Kg, fine breed pig farm in Guangzhou City) use embodiment SARS inactivated vaccine (TCID50Log107 / ml). A total of 4 times, the first time was Freund's complete adjuvant + SARS inactivated vaccine; the second time was Freund's incomplete adjuvant + SARS inactivated vaccine, and the third and fourth times were simple SARS inactivated vaccine immunization; Dosage 3×TCID50; Inoculation method Multi-point intramuscular injection on the back. The time interval of immunization is 0d, 15d, 21d, 40d. 1-3ml of whole blood was collected from the ear vein, and the intervals of serum collection were 0d, 5d, 7d, 10d, 15d, 21d, 28d, 35d and 42d. The serum was then centrifuged and stored at -20°C for future use.
[0037] Embodiment 2. Immunization program (SARS inactivated vaccine and influenza vaccine combined immunization)
[0038] 13 healthy cast...
Embodiment 3
[0039] Example 3. Test of immune effect Detection of neutralizing antibody
[0040] The immunized pigs in Examples 1 and 2 were carried out according to the method for evaluating and detecting the immune effect of inactivated vaccines.
[0041] 1. Virus dilution Dilute the titrated virus to 100TCID 50 / 25ul.
[0042] 2. Animal serum The serum of pigs immunized with SARS virus in different periods was collected in the
[0043] Start from 1:10 on a sterile 96-well plate, and serially dilute to 1:
[0044] 10240, 2 wells for each dilution, and collect the pre-immunization at the same time
[0045] Various animals were used as controls.
[0046] 3. Neutralization test Add 25ul 100TCID to the above serum dilution well 50 / 25ul
[0047] For Z2-Y3 virus application solution, shake gently and place at 36°C 5% CO 2
[0048] Culture in an incubator, and set normal cell control and virus titer back at the same t...
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