METHOD OF CLEANING INTENDED TO PRODUCE CLEANED VIRUS OF VESICULAR STOMATITIS FROM CELLULAR CULTURE

摘要

FIELD: biotechnologies.;SUBSTANCE: method includes clarification of a cellular culture liquid by low-speed centrifugation at the speed of lower than 10000 rpm or with the help of a module of volume filtration, the pore size of which makes from 1.0 mcm to 4.5 mcm, to produce a vesicular stomatitis virus in a supernatant. The supernatant is filtered via a filter with holes of the diameter of 0.2-0.45 mcm, and the vesicular stomatitis virus is produced in the filtered solution. The filtered solution, which contains the vesicular stomatitis virus, is loaded to an anion-exchange membrane adsorber balanced with the first buffer salt solution. The first buffer salt solution has ion force from 100 mM to 400 mM. The vesicular stomatitis virus is eluted from the anion-exchange membrane adsorber with the second buffer salt solution. The vesicular stomatitis virus is eluted from the membrane adsorber, when the second buffer salt solution is added, in one stage. Concentration of salt in the single-stage elution makes from 500 mM to 750 mM. Eluted fractions containing the vesicular stomatitis virus are collected. The vesicular stomatitis virus is cleaned with running filtration along the flow direction (press filter) using the membrane, the transmission limit of which is from 300 kDa to 1000 kDa, and the concentrate of the vesicular stomatitis virus is produced. The concentrate containing the vesicular stomatitis virus is filtered via a filter with holes of the diameter of 0.2-0.22 mcm, and the vesicular stomatitis virus is produced in the filtered solution. The method of cleaning does not include use of human embryos.;EFFECT: invention makes it possible to produce a vesicular stomatitis virus of high extent of cleaning with high yield.;13 cl, 8 dwg, 33 tbl, 12 ex