The Use of Globin Chain Electrophoresis in Polyacrylamide Gels for the Quantitation of the ^Gγ to ^Aγ Ratio in fetal Hemoglobih

Abstract

Polyacrylamide gel electrophoresis (PAGE) in the presence of urea, acid, and Triton X-100 was used for determination of the ^Gγ to ^Aγ ratio in human Hb F. The data compared most favourably with results obtained by a HPLC procedure and by a chemical procedure. Moreover, its accuracy and reproducibility was determined. The presence of Hb A₂ in a sample with Hb F level below 10% interferes with the determination because of the nearly identical electro-phoretic mobilities of the δ and ^Gγ chains. Thus, the removal of Hb A₂ is required. Samples free of Hb A₂ but with a Hb F level as low as 2% can be analyzed with an accuracy of about 5%. The PAGE method has been applied to the Hb F of 63 subjects with β thalassemia and related conditions, and the results of these analyses are included in this communication.