METHOD FOR PREPARING HIGH-PURITY CRYSTALLINE INSULIN OF ANY ORIGIN Russian patent published in 2012 - IPC A61K38/28 C07K14/62 A61P3/10 

FIELD: medicine.

SUBSTANCE: method for preparing high-purity crystalline insulin of any origin involves at least one column common-phase chromatographic purification of raw insulin in water-organic eluates with ionic modifiers on non-polar sorbents with the column purification preceded by primary purification by crystalline modification of raw insulin that is followed by filtration of dissolved crystalline raw insulin in apyrogenic water acidified with 1.5 M muriatic acid to pH 1.5-4.0, at first through an organic filter, and then through a sterile filter; thereafter the filtered solution volume is reduced with muriatic acid to the preset value; and the purified solution is supplied to the column purification performed at flow rate 1-3 cm/min^-1 with a mobile phase for separation presented by a mixture containing orthophosphoric acid (85%) 0.045 M, sodium sulphate 0.010 M and related as 60-70%:40-30% (by volume), apyrogenic water and ethanol, while a mobile phase for washing is presented by a mixture of apyrogenic water and ethanol related as 40-60%: 60-40% (by volume) respectively; thereafter the eluate prepared by the chromatographic purification is placed in a sterile cooled steriliser with keeping pH at 3.3-4.9 and mixed up with cooling to temperature 2-15°C with prepared crystalline insulin dried with ethanol. A cellulose filter is used as said organic filter.

EFFECT: invention provides simplified and reduced technological process ensured by complete single-stage purification of raw insulin during high-effective liquid chromatography.

4 cl, 3 ex