单组分胰岛素的制备及质量分析

Preparation and Analysis of Monocomponent Insulin

单峰胰岛素经DEAE 琼脂糖快流速离子交换层析后用醋酸锌结晶,得到结晶单组分胰岛素.层析和结晶的胰岛素回收率分别为93.8%±1.0%(n=3)和94.7%±1.5%(n=3).产品经PAGE分析为单带,HPLC检测高分子量杂蛋白和脱酰胺胰岛素的含量分别为0.42%±0.06%(n=3)和1.29%±0.06%(n=3),锌的残留量为0.187%±0.006%(n=3),产品效价为27.13±0.40U/mg(n=3).所有指标均达到USP24的质量标准.该工艺具有回收率高、产品质量好的特点.

Single peak insulin was purified by using DEAE-Sepharose fast flow ion exchange chromatography and crystallizing with zinc component. The insulin recovery of the chromatography and zinc crystallization was 93.8%±1.0%(n=3)and 94.7%±1.5%(n=3)respectively.The products showed single band on PAGE analysis. The contents of high molecular weight protein and monodesamido-insulin determined by HPLC were 0.42%±0.06 %(n=3)and 1.29%±0.06 %(n=3).Zinc content was 0.187%±0.006 %(n=3). Potency was 27.13±0.40 U/mg(n=3).All conform to the relative standard of USP24. This technology could prepare monocomponent insulin with high insulin recovery.