COMPLETELY SCALABLE METHOD FOR PRODUCING rAAV BASED ON COLUMNS Russian patent published in 2021 - IPC A61K38/46 C12N15/86 

FIELD: biochemistry.

SUBSTANCE: method for purification of particles of a recombinant vector of an adeno-associated virus (hereinafter – rAAV) is described, including stages of: (a) collecting cells and cell culture supernatant containing particles of the rAAV vector to obtain a collection; (b) optionally, concentrating the specified collection obtained at stage (a) to obtain a concentrated collection; (c) lysing the specified collection received at stage (a), or the specified concentrated collection received at stage (b), to obtain lysate; (d) processing lysate obtained at stage (c) to reduce the content of contaminating nucleic acid in lysate and, thus, to obtain lysate with the reduced content of nucleic acid; (e) filtering the specified lysate with the reduced content of nucleic acid obtained at stage (d) to obtain purified lysate and, optionally, dilute the specified purified lysate to obtain diluted purified lysate; (f) subjecting the specified purified lysate or diluted purified lysate obtained at stage (e) to anion exchange chromatography on columns to obtain column eluate consisting of rAAV vector particles, and, optionally, concentrating the specified column eluate to obtain concentrated column eluate; (g) subjecting the specified column eluate or the specified concentrated column eluate obtained at stage (f) to exclusion column chromatography to obtain the second column eluate consisting of rAAV vector particles, and, thus, separating rAAV vector particles and protein impurities and, optionally, diluting the specified second column eluate to obtain diluted second column eluate; (h) subjecting the specified second column eluate or the specified diluted second column eluate obtained at stage (g) to cation exchange chromatography on columns to obtain the third column eluate consisting of rAAV vector particles, and, thus, separating rAAV vector particles and protein or other industrial impurities and, optionally, concentrating the specified third column eluate to obtain concentrated third column eluate; and (i) filtering the specified third column eluate or the specified concentrated third column eluate obtained at stage (h), and, thus, obtaining purified rAAV vector particles.

EFFECT: invention makes it possible to achieve significant amount of AAV particle yield.

74 cl, 3 dwg