Method for the complete separation of radioactively labeled human γ-globin chains from pre- βA chains on CM-cellulose chromatography: Use of cold carrier globin chains

Abstract

Carboxymethyl (CM)-cellulose chromatography of globins is the technique used most frequently in analysis of hemoglobin synthesis. However, if this method is to be reliable in cases where only small amounts of fetal hemoglobin ( α 2 γ 2) compared to adult hemoglobin ( α 2 β 2 A) are synthesized, it is important to obtain a clean separation of γ chains from pre- β A chains. In the past, it has been found that small amounts of pre- β A chains tend to elute with the γ chains. Radioactively labeled γ chains can be completely and reproducibly separated from small amounts of labeled pre- β A chains by the addition of unlabeled β J chains (Hb J Baltimore = β16 Gly → Asp) which elute at the same position as the pre β A chains, thus increasing the quantity of this peak and allowing a clean separation from the γ chains.