METHOD OF PRODUCING BACTERIOPHAGE Russian patent published in 2016 - IPC C12N7/00 A61K35/76 

FIELD: biotechnology.

SUBSTANCE: invention relates to production of a bacteriophage. Disclosed method involves inoculation of bacterial cell suspension in the titre of 10⁸-10¹² CFU/ml on a dense nutrient medium with the layer thickness from 3 to 25 mm and cultivation with no foreign microflora, when the air layer thickness over the surface of the dense nutrient medium is from 5 to 50 mm and at optimum temperature for a host strain culture growth. In 30-120 minutes after the beginning of culturing every 30-60 minutes smears are taken from the surface of the dense nutrient medium, coloured with an intercalating dye and microscoped. Cultivation is stopped when reaching at least 10 % in relation to the total amount of cells of the host cell fraction strain with heterogeneous fluorescent cytoplasm staining. Then the obtained lawn of the host strain culture is inoculated with a mother bacteriophage in the titre of 10⁷-10¹⁰ BFU/ml. Cultivated for 13-15 hours with no foreign microflora, at the air layer thickness over the surface of the dense nutrient medium from 5 to 50 mm and at optimum temperature for the bacteriophage strain culture growth. Obtained a phage lysate when suspending, sucked off the phage lysate into a sterile container and cleaned. Invention provides reaching a stable high titre of bacteriophage (10¹²-10¹⁴ BFU/ml) when producing phage lysates both at varying formulations of nutrient media and increasing the span variation of values of culturing the host strain and the bacteriophage.

EFFECT: invention can be used in biotechnology for obtaining products containing bacteriophages.

9 cl