METHOD FOR OBTAINING RECOMBINANT PROTEIN SAV-RGD, SPECIFICALLY RECOGNISING MELANOMA CELLS Russian patent published in 2015 - IPC C07K1/18 C07K14/00 C12N1/21 A61P35/00 

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention relates to biotechnology. A method of obtaining the recombinant protein SAV-RGD, specifically recognising human melanoma cells, includes cultivation in a nutrient medium of the strain Escherichia coli MG1655/pSAV-RGD, obtaining a periplasmatic fraction of the said strain cells, purification of the recombinant protein by ion-exchange chromatography with the further ultrafiltration and lyophilisation. The strain E. coli MG1655/pSAV-RGD is obtained by the transformation of the strain E. coli MG1655 with the plasmid pSAV-RGD, obtained on the basis of the vector pUC18 and including a sequence, coding the fused pro-protein SAV-RGD (pro-sav-rgd) under the control of a promoter of uridine phosphorylase (Pudp) E. coli. In the process of cultivation feeding with a 40% solution of glucose is realised for the first 10 hours at the feeding rate of 25 ml/hour and supporting glucose at the level of 1 g/l, sterile peptone solution 10 g/l, yeast extract 5 g/l and ampicilline 10 mg/l are fractionally added with the continuation of cultivation for 8 hours. The periplasmatic fraction of cells is obtained by the addition to the cell suspension of a lysing buffer, containing lysozyme in the concentration of 1 g/l, with the ratio of volumes of the lysing buffer and the cell suspension of 1:20.

EFFECT: invention makes it possible to obtain the target protein with high purity, constituting not less than 97%.

2 cl, 15 dwg, 2 tbl, 8 ex