Gamma chain heterogeneity: determination of Hb F composition by perfusion chromatography

The Ggamma:Agamma ratio is around 70:30 at the time of birth and usually 40:60 in the trace amounts of Hb F found in the adult. Changes in this ratio are observed in several hemoglobin disorders providing insights on the genetics and molecular pathophysiology of these diseases. Several techniques have been proposed to measure the Ggamma:Agamma ratio. We here describe perfusion chromatography which is now in routine use in one of our laboratories. The method involves a high velocity flow of the mobile phase through a porous reversed phase chromatographic stationary bed and allows us to determine this parameter one order of magnitude faster than with conventional high performance liquid chromatography.