The Separation of Globin Chains by High Pressure Cation Exchange Chromatography

Abstract

A high performance cation exchange system is described for isolating globin chains. The method is rapid and inexpensive, using total lysate, a 14 min gradient, and a 3 cm column. The running buffers contain 6 M urea to dissociate and unfold the globin, and a NaCl gradient is employed for elution. The procedure provides a convenient way of detecting mutant globins with an intrinsic alteration in their charge.