发明名称 METHOD TO PRODUCE ESTROGEN-BINDING PROTEIN ASSOCIATED WITH MALIGNANT TUMORS
摘要 FIELD: biotechnologies.SUBSTANCE: residue A(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 (?- 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 - ?- 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.1 tbl, 1 ex
申请公布号 RU2489440(C1) 申请公布日期 2013.08.10
申请号 RU20120126721 申请日期 2012.06.26
申请人 FEDERAL'NOE GOSUDARSTVENNOE BJUDZHETNOE UCHREZHDENIE NAUKI TIKHOOKEANSKIJ INSTITUT BIOORGANICHESKOJKHIMII IM. G.B. ELJAKOVA DAL'NEVOSTOCHNOGO OTDELENIJA ROSSIJSKOJ AKADEMII NAUK (TIBOKH DVO RAN) 发明人 BULGAKOV ALEKSANDR ALEKSANDROVICH;PETROVA IRINA JUR'EVNA
分类号 C07K1/14;A61K35/14;A61K38/00;C07K1/16;C07K14/00;C07K14/47 主分类号 C07K1/14
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