METHOD OF INSULIN PURIFICATION Russian patent published in 2000 - IPC

FIELD: medicine, endocrinology, biochemistry. SUBSTANCE: invention relates to methods of purification of insulin isolated from animal raw or prepared by biotechnology approach (for example, human genetic engineering insulin). Insulin to be purified is subjected for gel-chromatography using 0.7-1.2 M acetic acid on column filled with hydrophilic gel obtained by polycondensation of polyglucin with epichlorohydrin and equilibrated with 0.7-1.2 M acetic acid solution. Insulin loading on column is 4-7 g/l of equilibrated gel at insulin concentration in applied solution 5-15%. Insulin is eluted from column at 8-15 C at the flow rate 2.0-5.0 ml/cm²/h and the end product is isolated by zinc precipitation and subjected for citrate crystallization. The yield of insulin is 75-85%, content of insulin monofraction is 98.0-98.5%, content of proinsulin is less 2 p.p.m., content of bacterial peptides (in human genetic engineering insulin) - is less 1 p.p.m. (i. e. less 0.0001%), amount of deamidoinsulin and nonidentified impurities (by methods of RIA and HPLC) - 1%, not above, in all cases. EFFECT: improved method of purification, increased yield, high purity of insulin. 5 cl, 3 ex