DEAE-Affi-Gel Blue chromatography of human serum: use for purification of native transferrin |
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Authors: | P A Werner R M Galbraith P Arnaud |
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Affiliation: | Department of Basic and Clinical Immunology and Microbiology, and Department of Medicine, Medical University of South Carolina, Charleston, South Carolina 29425 USA |
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Abstract: | Human serum was subjected to chromatography on DEAE-Affi-Gel Blue which combines ion-exchange and pseudo-ligand-affinity chromatography in a 0.02 M phosphate buffer, pH 7.0. All serum proteins were bound with the exception of transferrin, IgG (immunoglobulin G) and trace amounts of IgA. After a second step of Sephadex G-100 gel chromatography, or affinity chromatography against goat anti-human IgG F(ab')2 coupled to AH-Sepharose 4B, IgG and IgA were removed. The transferrin obtained was homogeneous and of high yield (greater than 80%), and was unaltered as judged by analyses of molecular weight, isoelectric point, iron-binding capacity, antigenicity, and ability to bind to high-affinity specific cellular receptors. Thus, DEAE-Affi-Gel Blue chromatography may be used as the basis for a simple, rapid, two-step method for the purification of large amounts of native transferrin from serum. |
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Keywords: | To whom correspondence should be addressed. |
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