Isolation of high-purity peptide Val-Val-Tyr-Pro from Globin Peptide using MCI gel column combined with high-speed counter-current chromatography
Isolation of high-purity peptide Val-Val-Tyr-Pro from Globin Peptide using MCI gel column combined with high-speed counter-current chromatography
复制标题
使用 MCI 凝胶柱结合高速逆流色谱从球蛋白肽中分离高纯度肽 Val–Val–Tyr–Pro
DOI：
10.1002/jssc.201800972
复制
发表时间：
2018-12-01
影响因子：
3.1
通讯作者：
Di, Duo-Long
中科院分区：
文献类型：
--
作者：
Pei, Dong;Xi, Xing-Jun;Di, Duo-Long
在过去的几十年里，肽由于其治疗作用而获得了越来越多的关注。本研究采用MCI凝胶柱层析和高速逆流色谱相结合的方法从珠蛋白肽中分离出高纯度的Val-Val-Tyr-Pro肽。首先，通过使用MCI凝胶柱以乙醇/水（20：80，v/v/v）的混合物获得纯度为15.8%的Val-Val-Tyr-Pro混合物的级分。采用乙醇/乙腈/异丙醇/（NH_4）（2）SO_4饱和溶液/H_2O（0.5：0.5：0.25：1.5：0.7，v/v）的双水相系统，高速逆流色谱分离得到高纯度的Val-Val-Tyr-Pro。通过MCI凝胶柱层析，使用乙醇/水以分步洗脱模式从目标化合物中除去高速逆流色谱馏分中的硫酸铵。从30 g粗珠蛋白肽中成功纯化出78 mg Val-Val-Tyr-Pro，纯度为98.80%。通过电喷雾电离高分辨串联质谱法测定Val-Val-Tyr-Pro的氨基酸序列。该方法为大规模分离珠蛋白肽中的纯肽Val-Val-Tyr-Pro提供了一种实用策略，也为从其他多肽混合物中获得高纯度肽提供了参考方法。
Peptides have gained increased interest over the past several decades because of their therapeutics. In this research, a strategy combining MCI gel column chromatography and high-speed countercurrent chromatography was developed for the separation of high-purity peptide Val-Val-Tyr-Pro from Globin Peptide. First, the fraction of Val-Val-Tyr-Pro mixtures with a purity of 15.8% was obtained by using MCI gel column with a mixture of ethanol/water (20: 80, v/v/v). Then, the high-purity Val-Val-Tyr-Pro was separated by high-speed countercurrent chromatography with a aqueous two phase systems of ethanol/acetonitrile/iso-propyl alcohol/(NH4)(2)SO4Saturated solution/H2O (0.5:0.5:0.25:1.5:0.7, v/v). The ammonium sulfate from high-speed countercurrent chromatography fractions was removed from target compound by MCI gel column chromatography using ethanol/water in stepwise elution mode. A 78 mg of Val-Val-Tyr-Pro was successfully purified with the purities of 98.80% from 30 g crude Globin Peptide. The amino acid sequence of the Val-Val-Tyr-Pro was determined by electrospray ionization high resolution tandem mass spectrometry. The method presents a practical strategy for the large-scale separation of pure peptide Val-Val-Tyr-Pro from Globin Peptide, and provides a reference method for obtaining high-purity peptide from other polypeptide mixtures.