Separation of hemoglobin variants in single human erythrocytes by capillary electrophoresis with laser-induced native fluorescence detection.
Single human red blood cells, in which the hemoglobin (Hb) molecules exist in their native, tetrameric states, were analyzed. Upon injection and lysis of a cell, the tetramers were dissociated on-column into their respective polypeptide chains, separated, and detected by laser-induced native fluorescence detection with 275-nm excitation. This technique was applied to the determination of hemoglobin variants as found in adult (normal and elevated Hb A1) and fetal erythrocytes. Normal adult cells contained 9.6% and 4.8% glycated beta- and alpha-chains, respectively. Cells with elevated Hb A1 gave 30% and 12%, respectively. The amounts of glycated Hb and total Hb in a given cell were found to be uncorrelated.
Lillard
S J
SJ
Department of Chemistry, Iowa State University, Ames 50011, USA.
Yeung
E S
ES
Lautamo
R M
RM
Mao
D T
DT
D000328:Adult; D019075:Electrophoresis, Capillary / Q000379:methods*; D004912:Erythrocytes / Q000737:chemistry*; D005319:Fetal Hemoglobin / Q000032:analysis / Q000302:isolation & purification; D005453:Fluorescence; D006442:Glycated Hemoglobin / Q000032:analysis / Q000302:isolation & purification*; D006418:Heme; D006441:Hemoglobin A / Q000032:analysis / Q000302:isolation & purification; D006454:Hemoglobins / Q000032:analysis / Q000235:genetics / Q000302:isolation & purification*; D006801:Humans; D007834:Lasers; D008706:Methemoglobin / Q000032:analysis / Q000302:isolation & purification; D010084:Oxidation-Reduction; D010455:Peptides / Q000032:analysis / Q000737:chemistry; D011487:Protein Conformation; D011489:Protein Denaturation; D013501:Surface-Active Agents / Q000494:pharmacology
D005319:Fetal Hemoglobin; D006442:Glycated Hemoglobin; D006418:Heme; D006441:Hemoglobin A; D006454:Hemoglobins; D008706:Methemoglobin; D010455:Peptides; D013501:Surface-Active Agents