Schartmann B, Gattner H G, Danho W, Zahn H
Hoppe Seylers Z Physiol Chem. 1983 Feb;364(2):179-86.
The yield of insulin by the combination of reduced A and B chains is increased from 15% to 33-34% if the non-insulin-containing fractions of the gel filtration are isolated and used for further combinations (recycling). Crude insulin recovered by gel filtration can be highly purified by preparative high-performance liquid chromatography. Lower yields of insulin were obtained on recombination of chains which had been recycled five times or more. Sulfur analyses show a loss of cystine sulfur. The occurrance of lanthionine and lysinoalanine indicates an alkaline destruction of cystine/cysteine during the repeated combination steps.
如果将凝胶过滤中不含胰岛素的部分分离出来并用于进一步组合（循环利用），还原的A链和B链组合产生胰岛素的产率会从15%提高到33%-34%。通过凝胶过滤回收的粗胰岛素可通过制备型高效液相色谱法进行高度纯化。经过五次或更多次循环的链重新组合时，胰岛素的产率较低。硫分析显示胱氨酸硫有损失。羊毛硫氨酸和赖氨酰丙氨酸的出现表明在重复组合步骤中胱氨酸/半胱氨酸发生了碱性破坏。
