Peptide purification by affinity chromatography based on α-ketoacyl group chemistry
Peptide purification by affinity chromatography based on α-ketoacyl group chemistry
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DOI：
10.1002/psc.1127
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发表时间：
2009-05-01
影响因子：
2.1
通讯作者：
Aimoto, Saburo
中科院分区：
文献类型：
--
作者：
Hara, Toshiaki;Tainosho, Akira;Aimoto, Saburo
在肽/蛋白质合成领域已经取得了重大进展，允许制备大的复杂分子。然而，在肽纯化领域中仍然存在缺点。本文详细介绍了我们进行的一些研究，以开发一种新的策略，肽纯化的基础上的反应性的α-酮酰基的肽。通过使用乙醛酸和镍（II）离子的转氨反应，在固相中从N-乙酰基-赖氨酰-肽生成α-酮酰基肽。用邻苯二胺裂解α-酮酰基，以可接受的产率和纯度得到目标肽。我们首先使用模型肽对纯化条件进行了仔细的逐步优化。然后，该策略用于纯化跨膜肽，该跨膜肽由于肽的强疏水性和其高聚集倾向而不能使用常规RP-HPLC系统有效纯化。版权所有（C）2009欧洲肽协会和约翰威利父子有限公司。
Significant advances have been achieved in the fields of peptide/protein synthesis, permitting the preparation of large, complex molecules. Shortcomings, however, continue to exist in the area of peptide purification. This paper details some studies we undertook to develop a new strategy for peptide purification based on a reactivity of alpha-ketoacyl groups in peptides. The alpha-ketoacyl peptide was generated from N-epsilon-acyl-lysyl-peptide in the solid phase via a transamination reaction using glyoxylic acid and nickel(II) ion. Cleavage of the alpha-ketoacyl group with o-phenylenediamine gave the target peptide in an acceptable yield and purity. We first carried out a careful step-by-step optimization of the purification conditions using a model peptide. The strategy was then used in the purification of a transmembrane peptide that could not be effectively purified using a conventional RP-HPLC system due to the strong hydrophobicity of the peptide and its high tendency to aggregate. Copyright (C) 2009 European Peptide Society and John Wiley & Sons, Ltd.