METHOD TO PRODUCE ESTROGEN-BINDING PROTEIN ASSOCIATED WITH MALIGNANT TUMORS Russian patent published in 2013 - IPC C07K1/14 C07K1/16 C07K14/00 C07K14/47 A61K35/14 A61K38/00 

FIELD: biotechnologies.

SUBSTANCE: residue A₀ (waste produced during industrial production of a gamma-globulin fraction from abortive blood at the first stage of fractioning) is extracted by 0.05 M sodium-acetate buffer. The produced extract is centrifuged to produce a supernatant. The produced supernatant is chromatographed, and rechromatography is carried out on DEAE cellulose. At the same time the ballast proteins are eluated with 0.05 M sodium-acetate buffer with the following eluation of ESB-containing fraction with a linear gradient of molarity of sodium-acetate buffer from 0.05 M to 0.25 M. Collection of fractions eluated in the range of 0.075 M - 0.15 M of sodium acetate with further concentration, dialysis against distilled water and lyophilisation. The dried fraction is dissolved in 0.015 M sodium-acetate buffer and applied onto KM-cellulose balanced with the same buffer (0.015 M sodium-acetate buffer) with the following performance of the stepped eluation, concentration of the fraction, containing ESB and a-1-acid glycoprotein (α- 1- AGP) and produced with eluation of 0.15 M sodium-acetate buffer. Concentration of this fraction. Dialysis against 0.01 M tris HCl-buffer containing 0.15 M NaCl with subsequent application of the produced fraction onto a column with an immunosorbent IgG anti - α- 1- AGP-sepharose. At the same time the fraction, which is not bound with immunosorbent, is dialysed and lyophilised with subsequent performance of gel-penetrant HPLC on the column with T8K-gel in 0.1 M potassium-phosphate buffer, containing 0.1 M potassium chloride and 5 mM of trilon B. The fraction containing highly pure ESB is dialysed against distilled water and lyophilised.

EFFECT: invention makes it possible to expand arsenal of facilities used in diagnostics of malignant tumors.

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