Peptide purification by affinity chromatography based on α‐ketoacyl group chemistry |
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Authors: | Toshiaki Hara Akira Tainosho Ken'ichiroh Nakamura Takeshi Sato Toru Kawakami Saburo Aimoto |
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Affiliation: | Institute for Protein Research, Osaka University, 3‐2 Yamadaoka, Suita, Osaka 565‐0871, Japan |
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Abstract: | 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 α‐ketoacyl groups in peptides. The α‐ketoacyl peptide was generated from Nε‐acyl‐lysyl‐peptide in the solid phase via a transamination reaction using glyoxylic acid and nickel(II) ion. Cleavage of the α‐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 © 2009 European Peptide Society and John Wiley & Sons, Ltd. |
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Keywords: | peptide purification α ‐ketoacyl group transamination amide‐bond scission |
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