TY - JOUR
T1 - Detection of the embryonic ζ chain in blood from newborn babies by reversed-phase high-perfomance liquid chromatogrphy
AU - Kutlar, F.
AU - Fei, Y. J.
AU - Wilson, J. B.
AU - Kutlar, A.
AU - Huisman, T. H.J.
N1 - Funding Information:
Fraction Number Fig. 3. Separationo f Hb componentsb y DEAE-cellulose macrochromatographyA.b out 60 mg Hb was applied on a 30 x 2 cm I.D. column. The samplew as from baby No. 9279w ith an a-thal-2h omo-zygosity( -x+x) and 9.1% Hb Bart’s. ZonesA , FO, No. 1, No. 2, and No. 3 werei solated,c oncentrated, and analyzedb y starchg el electrophoresi(si nsert),a nd by RP-HPLC (Fig. 4).
PY - 1987
Y1 - 1987
N2 - Reversed-phase high-performance liquid chromatography (RP-HPLC) using the large-pore Vydac C4 column has been used to detect and quantitate the embryonic ζ chain in blood samples of normal babies and of newborns with varying degrees of α chain deficiencies. The ζ 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 ζ chain isolated from a blood sample of a fetus without active α globin genes, i.e. with hydrops fetalis (- -/- -). The quantity of ζ in normal babies is less than 0.7% [% of (α + ζ)] and is dependent upon the maturity of the baby as it was only present in babies with low levels of β chain or hemoglobin (Hb) A. The presence of a ζ globin gene deletion [A. E. Felice, Hum. Genet., 73 (1986) 221; and P. Winichagoon, Nucleic Acids Res.,10 (1982) 5853] did not affect the level of ζ in the newborn. All babies with an α-thalassemia-2 heterozygosity, i.e. with three active α globin genes or -α/αα, had ζ in a range of 0.1-0.9%; again the level showed a negative correlation with that of the β chain. Newborns with an α-thalassemia-2 homozygosity or -α/-α had a varying level of ζ of 0.3-2.3%, which did not correlate with the level of β, suggesting that ζ chain production persists after birth in this condition. Macrochromatographic analyses in combination with RP-HPLC indicated that the ζ chain is present as ζ2γ2 or Hb Portland-1, as expected.
AB - Reversed-phase high-performance liquid chromatography (RP-HPLC) using the large-pore Vydac C4 column has been used to detect and quantitate the embryonic ζ chain in blood samples of normal babies and of newborns with varying degrees of α chain deficiencies. The ζ 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 ζ chain isolated from a blood sample of a fetus without active α globin genes, i.e. with hydrops fetalis (- -/- -). The quantity of ζ in normal babies is less than 0.7% [% of (α + ζ)] and is dependent upon the maturity of the baby as it was only present in babies with low levels of β chain or hemoglobin (Hb) A. The presence of a ζ globin gene deletion [A. E. Felice, Hum. Genet., 73 (1986) 221; and P. Winichagoon, Nucleic Acids Res.,10 (1982) 5853] did not affect the level of ζ in the newborn. All babies with an α-thalassemia-2 heterozygosity, i.e. with three active α globin genes or -α/αα, had ζ in a range of 0.1-0.9%; again the level showed a negative correlation with that of the β chain. Newborns with an α-thalassemia-2 homozygosity or -α/-α had a varying level of ζ of 0.3-2.3%, which did not correlate with the level of β, suggesting that ζ chain production persists after birth in this condition. Macrochromatographic analyses in combination with RP-HPLC indicated that the ζ chain is present as ζ2γ2 or Hb Portland-1, as expected.
UR - http://www.scopus.com/inward/record.url?scp=0023219875&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0023219875&partnerID=8YFLogxK
U2 - 10.1016/S0021-9673(01)94185-8
DO - 10.1016/S0021-9673(01)94185-8
M3 - Article
C2 - 2442182
AN - SCOPUS:0023219875
SN - 0021-9673
VL - 394
SP - 333
EP - 343
JO - Journal of Chromatography A
JF - Journal of Chromatography A
IS - 2
ER -