METHOD FOR PREPARING CARBOHYDRATE-ENRICHED ALPHA-FETOPROTEIN

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Patent number: RU2232770
Publication date: 2004-07-20
Inventor: KHOPERSKAJA O A (RU); MAL DOV D G (RU); TATARINOV JU S (RU); ZARAJSKIJ E I (RU)
Applicant:
Classification:
- international: C07K14/435; C07K1/36
- european:
Application number: RU20020120579 20020805
Priority number(s): RU20020120579 20020805
Abstract of RU2232770

FIELD: biotechnology. ^ SUBSTANCE: invention can be used for preparing alpha-fetoprotein of high purity enriched with carbohydrates. Method for preparing involves diluting the parent raw that is carried out by two-fold with 0.1 M phosphate-saline buffet, pH 7.2-7.4, its centrifugation and chromatography of the end product on affinity column with low-avidity highly specific monoclonal antibodies cross-linked covalently with cyanobrominated Sepharose. Then column is washed out with 10-fold volume of 2 M NaCl in 0.15 M carbonate buffer, pH 9.3, and elution is carried out with 2.5 M carbonate buffer, pH 11.2, followed by neutralization of eluate with 1 M hydrochloric acid. Product is dialyzed against phosphate-saline buffer and product is centrifuged. Before sterilization product is sorbed in affinity columns and saline elution is carried out on DEAE-Sepharose. Then sucrose or starch is added, ready product is sterilized by filtration and the following lyophilization of ready product or its storing as a solution. Invention provides carrying out the industrial technology for preparing the preparation of high purity degree with the retained carbohydrate fraction that can be used in curative practice. ^ EFFECT: improved preparing method. ^ 1 ex