IMPROVED METHOD FOR PREPARING INSULIN PRECURSOR WITH CORRECTLY LINKED CYSTINE BRIDGES

摘要

FIELD: biochemistry, chemistry of proteins, hormones. SUBSTANCE: invention relates to method for preparing precursor of insulin or insulin derivatives with correctly linked cystine bridges in the presence of cysteine or cysteine hydrochloride and chaotropic accessory substance characterizing by that the following stages are carried one after another: (a) mixing and aqueous suspension of precursor of insulins or insulin derivatives with amount of cysteine or cysteine hydrochloride resulting up to 15 SH-residues of cysteine or cysteine hydrochloride per one cysteine residue of precursor; (b) addition of cysteine- or cysteine hydrochloride-containing suspension of precursor to 4-9 M solution of chaotropic accessory substance at pH value from 8 to 11.5 and temperature from 15 to 55 C followed by keeping the prepared mixture at this temperature for 10-60 min; (c) addition of mixture at pH value from 8 to 11.5 and temperature from 5 to 30 C to water amount that provides dilution of cysteine or cysteine hydrochloride concentration in the mixture by 1-5 mM and a chaotropic substance by 0.2-1.0 M. Method provides high yield of correctly stacked precursors of insulin or insulin derivatives and low reaction time for the stacking process. EFFECT: improved preparing method. 13 cl, 4 ex