FIELD: veterinary medicine.
SUBSTANCE: cultivation of a culture of continuous BHK cells is carried out for 48-72 hours to the concentration of (2.5-3.0)×106 cells/ml, followed by infecting the cells with rabies virus at a dose of 0.01-0.1 MMLD50/cells. It is maintained at a temperature of 38.5-39.5°C for 60-90 minutes, followed by dilution with growth medium to the concentration of the cell subline of (0.5-0.6)×106 cells/ml and continuation of culturing the resulting suspension for 48-72 hours at a temperature of 37-38°C and pH 7.1-7.3. After which the virus-containing suspension is cooled to a temperature of 4-8°C at pH 7.8-8.0 and tris (hydroxymethyl)aminomethane and disodium salt of ethylenediaminetetraacetic acid is added, taken in the final concentrations of 0.7-0.9% and 0.006-0.01%, respectively. To inactivate the virus, β-propiolactone is applied in the bioreactor.
EFFECT: use of this method enables to improve the quality of the desired product by increasing the yield of viral antigen, the accumulation of virus particles, glycoprotein which is responsible for production of virus-neutralising antibodies, and enables to improve stabilisation of the antigenic properties of the viral glycoprotein.
4 cl, 1 tbl, 9 ex
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Authors
Dates
2015-01-10—Published
2013-04-11—Filed