Detection of the embryonic zeta chain in blood from newborn babies by reversed-phase high-performance liquid chromatography.
Reversed-phase high-performance liquid chromatography (RP-HPLC) using the large-pore Vydac C4 column has been used to detect and quantitate the embryonic zeta chain in blood samples of normal babies and of newborns with varying degrees of alpha chain deficiencies. The zeta chain eluted at the end of the chromatogram at about 130 min using a modified and extended gradient. Its identity was confirmed by structural analysis of zeta chain isolated from a blood sample of a fetus without active alpha globin genes, i.e. with hydrops fetalis (--/--). The quantity of zeta in normal babies is less than 0.7% [% of (alpha + zeta)] and is dependent upon the maturity of the baby as it was only present in babies with low levels of beta chain or hemoglobin (Hb) A. The presence of a zeta globin gene deletion [A. E. Felice et al., Hum. Genet., 73 (1986) 221; and P. Winichagoon et al., Nucleic Acids Res., 10 (1982) 5853] did not affect the level of zeta in the newborn. All babies with an alpha-thalassemia-2 heterozygosity, i.e. with three active alpha globin genes or -alpha/alpha alpha, had zeta in a range of 0.1-0.9%; again the level showed a negative correlation with that of the beta chain. Newborns with an alpha-thalassemia-2 homozygosity or -alpha/-alpha had a varying level of zeta of 0.3-2.3%, which did not correlate with the level of beta, suggesting that zeta chain production persists after birth in this condition. Macrochromatographic analyses in combination with RP-HPLC indicated that the zeta chain is present as zeta 2 gamma 2 or Hb Portland-I, as expected.
D000510:Alpha-Globulins / Q000032:analysis; D002848:Chromatography, DEAE-Cellulose; D002851:Chromatography, High Pressure Liquid; D004247:DNA / Q000032:analysis; D004912:Erythrocytes / Q000032:analysis; D005312:Fetal Blood / Q000032:analysis; D005319:Fetal Hemoglobin / Q000032:analysis*; D006455:Hemoglobins, Abnormal / Q000032:analysis; D006801:Humans; D007231:Infant, Newborn
D000510:Alpha-Globulins; D006455:Hemoglobins, Abnormal; D004247:DNA; D005319:Fetal Hemoglobin; C030448:hemoglobin Bart's