Isolation of the four forms of transferrin with respect to iron by high-performance liquid chromatography: comparison of three mammalian species.
A method, based on anion exchange combined with high-performance liquid chromatography, is presented for the separation of the four different forms of transferrin with respect to iron, i.e. diferric, monoferric with Fe in the C-terminal lobe, monoferric with Fe in the N-terminal lobe, and apo (iron-free). Depending on the size of column applied, these forms can be obtained on a preparative scale amounting to milligrams. The procedure was also found to free transferrin from loosely bound iron and protein aggregates. Comparative studies with human, murine and rat transferrins showed that optimal resolution of these proteins depended on establishing a distinct elution programme for each species. The order in which the four forms referred to above were displaced from the exchanger differed from species to species.
Department of Pathology, McMaster University Health Sciences Centre, Hamilton, Ont., Canada.
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Regoeczi
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Bolyos
D000818:Animals; D001797:Blood Protein Electrophoresis; D002851:Chromatography, High Pressure Liquid; D003676:Deferoxamine / Q000737:chemistry; D006801:Humans; D007501:Iron / Q000737:chemistry*; D007502:Iron Chelating Agents / Q000737:chemistry; D051379:Mice; D051381:Rats; D013045:Species Specificity; D014168:Transferrin / Q000737:chemistry / Q000302:isolation & purification*
D007502:Iron Chelating Agents; D014168:Transferrin; D007501:Iron; D003676:Deferoxamine