TABLETED LIVE RECOMBINANT BIVACCINE 'REVAKS VKT' AGAINST NATURAL SMALLPOX AND HEPATITIS B AND METHOD FOR ITS PREPARING

摘要

FIELD: biotechnology, medicine, virology, pharmacy.;SUBSTANCE: invention proposes bivaccine comprising in a single tablet lyophilized viral material produced on the basis of the recombinant smallpox vaccine virus strain GKV № 2131 with typical properties of smallpox virus expressing preS2-Ag and HbsAg of hepatitis B virus with activity 6.5-8.0 lg CPD₅₀ per a tablet, lactose, sucrose, calcium stearate, vanillin, peptone, gelatose and buffer. Smallpox virus is produced in transplanted mammalian cell cultures in the following ratio of components per one tablet, wt.-%: smallpox lyophilized viral material with activity 6.5-8.0 lg CPD₅₀ per a tablet, 1.04-31.3; peptone, 0.12-3.8; gelatose, 0.06-1.9; sucrose, 10.1-11.55; buffer, 0.001-0.027; calcium stearate, 1.94-2.0; vanillin, 0.17-0.2; lactose, the balance, up to 100%. Method for preparing bivaccine involves passaging the strain GKV № 2131 of recombinant smallpox virus in biological system used for culturing viruses, preparing virus-containing material with titer 8.0 lg CPD₅₀/ml, not less, its lyophilization with stabilizing additives, milling, mixing with filling agent and pressing. The transplanted mammalian cell culture 4647 or VERO are used as a biological system with the infection dose with the recombinant strain of smallpox virus 1.0 CPD₅₀ per a cell, not above. Virus is produced in monolayer of indicated mammalian cells in rollers on nutrient medium up to 50% value of cytopathic effect (CPE) followed by exchange of this medium for buffer solution with less volume as compared with volume of the nutrient medium. Preparing virus-containing material is carried out after exchange of the nutrient medium for buffer solution by extraction of viruses in three-fold freezing and thawing the cellular monolayer. As stabilizing additives in lyophilic drying peptone, sucrose and gelatose are used in their final concentration in mixture, wt.-%: 2.0-4.0; 2.0-3.0, and 1.0-3.0, respectively. Vaccine retains activity during processes of drying and tableting and comprises lesser amounts of foreign impurities. Method provides the improved technology in preparing vaccine.;EFFECT: improved preparing method.;5 cl, 4 tbl, 7 ex