高纯胸腺肽α1的制备

doi:10.3969/j.issn.0438-1157.2013.07.026

化工学报 ›› 2013, Vol. 64 ›› Issue (7): 2497-2502.DOI: 10.3969/j.issn.0438-1157.2013.07.026

高纯胸腺肽α1的制备

刘标, 汤斌

安徽工程大学生物与化学工程学院, 安徽芜湖 241000

收稿日期:2012-11-07 修回日期:2013-03-01 出版日期:2013-07-05 发布日期:2013-07-05
通讯作者: 汤斌
作者简介:刘标(1981- ),男,硕士研究生。

Preparation of high-purity thymosin α1

LIU Biao, TANG Bin

College of Biological and Chemical Engineering, Anhui Polytechnic University, Wuhu 241000, Anhui, China

Received:2012-11-07 Revised:2013-03-01 Online:2013-07-05 Published:2013-07-05

摘要/Abstract

摘要： 为了获得高纯产品,在制备胸腺肽α1(Tα1)工艺中分别对合成、纯化两步骤进行控制。在固相多肽合成过程中,以TBTU/HOBt/DIEA体系作为缩合试剂,在易形成α螺旋和β转角的肽序位置,通过采用HOBt/DIC补投的方法促使偶联反应完全。在分离纯化阶段,结合反相色谱和离子色谱的优势,以反相离子对液相色谱法为主要分离手段,通过多步分离实现纯化。获得的粗肽纯度为67.2%,胸腺肽α1产品纯度达99.5%以上,总收率为17.2%。此法可作为制备高纯多肽药物的有效手段。

关键词: 胸腺肽α1, 固相多肽合成, 反相离子对液相色谱

Abstract: The synthesis and purification steps were controlled in preparation of thymosin α1 to obtain the product with high-purity.In the solid-phase peptide synthesis (SPPS)process,TBTU/HOBt/DIEA was employed as the coupling reagent.In the sites where α-helix or β-turn structure formed easily,re-coupling reagent of HOBt/DIC was used to ensure a complete reaction.In the separation and purification process,a multi-step method was employed,in which reversed-phase ion-pair liquid chromatography was used as the main means,combining the advantages of reversed-phase chromatography and ion chromatography.The results showed that through the re-coupling the crude peptide had a purity of 67.2% with the use of TBTU/HOBt/DIEA reagent in SPPS.The purity of the refined thymosin α1 was above 99.5% with a total yield of 17.2% with the reversed-phase ion-pair liquid chromatography.This separation process avoids complex chromatography separation process.The method may be an efficient way for preparation of peptides with high-purity.

Key words: thymosin α1, solid-phase peptide synthesis, reversed-phase ion-pair liquid chromatography

中图分类号:

TQ464.7

刘标, 汤斌. 高纯胸腺肽α1的制备[J]. 化工学报, 2013, 64(7): 2497-2502.

LIU Biao, TANG Bin. Preparation of high-purity thymosin α1[J]. CIESC Journal, 2013, 64(7): 2497-2502.

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高纯胸腺肽α1的制备

Preparation of high-purity thymosin α1

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