METHOD FOR PURIFICATION OF RECOMBINANT PROTEIN CONTAINING MYELOPEPTIDES SEQUENCES Russian patent published in 2017 - IPC C07K14/47 C12N15/63 C12N15/12 C07K1/18 C07K1/20 C07K1/22 A61K38/17 

FIELD: biotechnology.

SUBSTANCE: invention relates to a method for isolation of a recombinant protein comprising myelopeptides sequences and represented by the sequence of SEQ ID NO:1. The plasmid DNA of pET-32a/REC-MP carrying the gene represented by the sequence of SEQ ID NO:2 is used to transform competent bacterial cells of Escherichia coli BL21 (DE3) strain to obtain a soluble fraction of the thioredoxin/REC-MP fusion protein. Further, the fusion protein is purified on the metal-affinity sorbent using a Ni-Sepharose carrier. The fusion protein is then hydrolized with enterokinase-F enzyme, incubating for 1 hour at 37°C. Impurities are denaturated with heating to 74°C for 60 min. FPLC purification of the target recombinant protein on the SP-Sepharose ion exchange sorbent and reverse phase HPLC purification on a Kromasil 100C18 column are performed.

EFFECT: invention allows to increase the expression of the target protein, improve the protein purification efficiency, resulting in homogeneity and almost 100% chromatographic purity of the product obtained.

10 dwg, 3 tbl, 6 ex