METHOD OF PRODUCING ANTIGEN OF ZIKA VIRUS, HAVING IMMUNOGENIC AND ANTIGENIC PROPERTIES Russian patent published in 2020 - IPC C12N7/02 

FIELD: biotechnology.

SUBSTANCE: invention represents a method for producing inactivated, concentrated and purified antigen of Zika virus (ZIKV). Method for preparing the Zika virus antigen involves developing the virus on a monolayer of Vero cell culture, inactivating the virus-containing material with formaldehyde, concentrating and purifying the obtained antigen. Prior to inactivation, all virus-containing biomass of Vero cells is subjected to triple freezing at minus 20 °C and thawing at temperature of +20–22 °C for destruction of cell membranes, inactivation of a virus-containing material in the form of a mixture of cell suspension with broken membranes is carried out with a solution of formaldehyde in a final concentration in range of 0.00016–0.016 % of the volume during incubation for 24 hours at 37 °C. After that, fractionation of the inactivated virus-containing material into Vero cell precipitate and culture growth medium in form of supernatant. Vero cell sediment is sonicated for 30 sec on ice to obtain a cell lysate. Concentration of total proteins from supernatant fraction is performed by 10 % solution of polyethylene glycol with molecular weight 6,000 Da for 5 days at +4 °C. Purification of two fractions of virus-containing material: infected cell lysate and concentrated fraction of growth medium in form of supernatant is performed by gradient centrifugation 30 % glycerine and 40 % potassium/sodium tartrate by Meichi method. Deposits of the purified antigen are dissolved in 1 ml of 0.05M sodium-phosphate buffer solution (pH 8.0). Invention allows to increase output of Zika virus antigen.

EFFECT: high output of the antigen of Zika virus and creation of more sparing conditions for inactivation of the virus.

3 cl, 5 dwg, 5 tbl, 8 ex