技术支持 Top-Peptide

 Cys(tBu)一般在TFA和I2的情况下比较稳定,因此常用来组合生产第三对及以上二硫键的多肽,而不打乱前面的二硫键配对.去除掉tBu的方法通常有醋酸汞和三氯甲基硅烷/二苯基亚砜法.
下面介绍一下醋酸汞法的操作步骤
1. 将带tBu的多肽溶解在冰TFA中(5-10mg/ml)
2. 加入10倍当量的醋酸汞,搅拌反应3小时
3. 减压旋转蒸发掉多余的TFA,再溶解到10%的醋酸水溶液中
4. 加入20倍当量的巯基乙醇,静置5小时
5. 离心去除沉淀再进行脱盐或者HPLC纯化.


1. Dissolve peptide in ice-cold TFA(5-10mg/ml)
2. Add Mercury acetate (10 eq./tBu) and stir the mixture gently at rt for 3 hr under a blanket of N2.
3. Remove the TFA by evaporation under reduced pressure at rt and redissolve the residue in 10% aqueous acetic acid
4. Add β-mercaptoethanol (20eq./tBu) and leave the mixture to stand for 5 hr.
5. Remove the precipitate by centrifugation and desalt the supernatant by HPLC.