著者

AL-MASHIKHI S.A. NAKAI Shuryo

出版者

社団法人日本農芸化学会

雑誌

Agricultural and Biological Chemistry (ISSN:00021369)

巻号頁・発行日

vol.51, no.11, pp.2881-2887, 1987-11-23

被引用文献数

19

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Immobilized metal affinity chromatography (IMAC) was used to separate ovotransferrin (OVT) by a single chromatographic step. Ovotransferrin was bound strongly to immobilized copper ions, while other components of egg white passed through the column. Using acidic pH and/or a strong competing solute, imidazole, ovotransferrin was recovered in 94〜98% pure form as indicated by SDS-polyacrylamide gel electrophoresis and immunoelectrophoresis. The elution profiles from the IMAC column indicated the presence of two forms of ovotransferrin. The capacity of the IMAC column was 20mg OVT/ml copper loaded gel. Saturation of the metal binding sites of ovotransferrin with Fe^<2+> and Cu^<2+> did not interfere with the binding of ovotransferrin to the IMAC column. However, modification of the histidine residues in ovotransferrin with diethyl pyrocarbonate almost completely destroyed the binding to the IMAC column.