METHOD OF PREPARING THE HIGHLY PURIFIED MONOCOMPONENT INSULIN

摘要

FIELD: biotechnology, biochemistry, endocrinology. SUBSTANCE: method involves purification of sodium-insulin by chromatography and involves chromatography on macroporous sulfocationite КУ-2ЗИ, isolation of insulin from eluates by isoelectric precipitation, gel-filtration on Sephadex G-50 and insulin elution with 1 M acetic acid followed by chromatography of insulin in monopeak fraction on column with anion-exchange silica gel PAE-300. Insulin is eluted from the latter column at exponential gradient of NaCl concentration under the following dependence: C = K x e^N where: C - NaCl concentration in eluting solution, M/l; K - coefficient of gradient steepness, K = 0.0041-0.011; e - natural logarithm base, e = 2.71828...; N - ratio of eluent volume to packing column volume. Highly purified insulin is isolated from eluate as crystalline zinc-insulin. Obtained insulin contains 98.5% monofraction (not less), 0.5% deamidoinsulin (not above), 1.0% unidentified impurities (not above, by HPLC method), 10 ppm proinsulin and its derivatives (not above). Yield is 55-65% of the parent sodium-insulin. EFFECT: enhanced quality, increased yield of the end product.