The separation of human globin chains by ion-exchange chromatography on CM-Sepharose CL-6B

A chromatographic procedure for the separation of human globin chains is described. This method uses CM-Sepharose CL-6B as ion-exchanger and NaCl gradients formed in sodium phosphate buffers containing urea and 0.05M mercaptoethanol to elute the chains. One advantage of this system is that a column, once packed, may be used repeatedly for several chain separations, thus reducing cost and practical work. The resolution in our hands was also better than that on CM-Cellulose using phosphate buffer gradients.