METHOD OF OBTAINING BOTULINUM NEUROTOXIN (VERSIONS) Russian patent published in 2015 - IPC C07K1/18 C07K14/33 B01D15/36 C07K1/20 A61K39/08 

FIELD: chemistry.

SUBSTANCE: group of inventions relate to field of biotechnology. Claimed are methods of obtaining biologically active botulinum neurotoxin with application of chromatography, in fact without application of animal-derived products. Cultivation of bacteria Clostridium botulinum in culture medium in fact without application of animal-derived products is realised. After that, bacteria are fermented into fermentation medium in fact without application of animal-derived products. At least one culture medium and fermentation medium includes protein of vegetable origin. Fermentation medium is collected by removal of cell residues. Collected medium is concentrated by filtration. Concentrated medium is diluted with buffer. Anion-exchange chromatography is carried out. Botulinum neurotoxin is eluted from anion-exchange medium with obtaining first eluent. First eluent is passed through cation-exchange medium with obtaining second eluent. In one version of method second eluent is processed with diafiltration. After that, processed eluent is filtered with obtaining complex of biologically active botulinum neurotoxin of A type, possessing activity 2.0×10⁷ - 6.0×10⁷ units/mg. In other version of method second eluent is filtered with obtaining biologically active botulinum neurotoxin, containing 1 ng or less of residual nucleic acid per each mg of obtained botulinum neurotoxin, with realising method within one week or less.

EFFECT: inventions provide fast and effective purification of botulinum neurotoxin within short time period, homogeneity of product characteristics from batch to batch.

8 cl, 1 dwg, 3 ex