FIELD: veterinary science; medicine; biotechnology.
SUBSTANCE: disclosed is a vaccine for immunoprophylaxis of tissue forms of echinococcosis of farm animals and humans, containing recombinant oncosphere protective Echinococcus antigen EG95tr, encoded by nucleotide sequence SEQ ID NO: 1 or SEQ ID NO: 3, and auxiliary components. Disclosed is a method of producing a vaccine for immunoprophylaxis of tissue forms of echinococcosis of farm animals and humans. Method provides for obtaining DNA coding recombinant Echinococcus EG95 antigen, having nucleotide sequence SEQ ID NO: 1 or SEQ ID NO: 3, cloning said DNA into a shuttle vector and delivering the genetic construct into Pichia pastoris yeast cells to obtain a genetically engineered microorganism Pichia pastoris carrying the EG95 protective antigen gene, fermentation on YPD, MMY, MGY, BMM, BMG, BMGY media, definitive media, purification of the obtained recombinant Echinococcus antigen EG95tr from the culture medium by centrifugation or decantation of the fermentation medium, followed by dilution of the culture medium, followed by ultrafiltration through filters with a pore size of 50 kDa and subsequent ultrafiltration through filters with a pore size of 3–5 kDa, with subsequent purification of the desired recombinant antigen from the culture medium or filtrate, preservation of the purified recombinant antigen by instantaneous freezing in liquid nitrogen or lyophilic drying, addition of auxiliary components represented by adjuvants, excipients, isotonic agents, stabilizers and/or preservatives. Disclosed is use of vaccine for immunoprophylaxis of tissue forms of echinococcosis of farm animals and humans in therapeutic doses based on protective antigen for subcutaneous 40 mcg/kg and 100 mcg/kg for intranasal immunization, in combination with an immunostimulating carrier, at least twice with interval of 10–14 days for immunoprophylaxis of tissue forms of echinococcosis of farm animals and humans.
EFFECT: oncosphere antigen EG95tr has high protective activity, which provides stable production of antibodies to the recombinant antigen in the blood serum of immunized animals for at least 4 months, as well as reduction of infection of target animals with tissue helminths by not less than 78% for immunoprophylaxis of cystic hydatidosis.
3 cl, 13 dwg, 7 ex
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Authors
Dates
2024-11-26—Published
2023-08-09—Filed