METHOD OF EXTRACTING REFOLDED RECOMBINANT PROTEIN FROM PROKARYOTIC CELL CULTURE (VERSIONS) Russian patent published in 2012 - IPC C07K1/14 

FIELD: chemistry.

SUBSTANCE: step is performed for extracting recombinant protein VEGF from a cell culture and dissolving in a first buffer solution, pH higher than 9, which contains the following in the final concentration: 1 M urea, 300 mM arginine, 10 mM CHES, 5 mM EDTA, pH 11, or 1 M urea, 300 mM arginine, 10 mM TRIS, 5 mM EDTA, pH 11. The dissolved protein is then refolded in a second buffer solution with addition of air, pH>9, but ≤11, containing the following in the final concentration: 1 M urea, 15 mM cysteine, 2 mM DTT, 100 mM arginine, 10 mM CHES, 5 mM EDTA, pH 10, or 1 M urea, 15 mM cysteine, 0.5-2 mM DTT, 100 mM arginine, 10 mM TRIS, 5 mM EDTA, pH 10, and the refolded recombinant protein is then extracted. As another version, dissolution and refolding of said protein can be carried out in a combined buffer solution, pH>9, but ≤11, with addition of air or oxygen, where the combined buffer solution contains the following in the final concentration: 1 M urea, 15 mM cysteine, 2 mM DTT, 100 mM arginine, 10 mM CHES, 5 mM EDTA, pH 10, or 1 M urea, 15 mM cysteine, 0.5-2 mM DTT, 100 mM arginine, 10 mM TRIS, 5 mM EDTA, pH 10.

EFFECT: invention enables to extract refolded recombinant protein VEGF from as prokaryotic cell culture.

16 cl, 10 dwg, 6 ex