ISOELECTRIC-FOCUSING OF HUMAN HEMOGLOBIN - ITS APPLICATION TO SCREENING, TO CHARACTERIZATION OF 70 VARIANTS, AND TO STUDY OF MODIFIED FRACTIONS OF NORMAL HEMOGLOBINS
Isoelectric focusing on slabs of acrylamide gel was adapted for the screening of abnormal Hb, the characterization of 70 human variants and the study of minor fractions of normal Hb. The screening method was as fast and inexpensive as conventional techniques, allowed the simultaneous analysis of some 50 samples of whole blood and yielded resolution superior to that obtained by other methods with hemolysates. Among the 70 variants, 31 mutants could not be separated from HbS by cellulose acetate electrophoresis. The characterization technique of electrofocusing allowed discrimination between most variants. Only 1 mutant, Hb Galveston, could be confused with HbS. Hb Koln, the most frequent unstable mutant, exhibited a special pattern. HbA1C was separated from HbA. Quantitation of HbA1C by gel scanning is feasible.