Detection of the embryonic ζ chain in blood from newborn babies by reversed-phase high-perfomance liquid chromatogrphy

F. Kutlar, Y. J. Fei, J. B. Wilson, A. Kutlar, T. H.J. Huisman

Research output: Contribution to journalArticle

18 Citations (Scopus)

Abstract

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.

Original languageEnglish (US)
Pages (from-to)333-343
Number of pages11
JournalJournal of Chromatography A
Volume394
Issue number2
DOIs
StatePublished - 1987

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Globins
Blood
Thalassemia
Genes
High performance liquid chromatography
Reverse-Phase Chromatography
Liquids
High Pressure Liquid Chromatography
Hemoglobin A
Hydrops Fetalis
Viverridae
Gene Deletion
Structural analysis
Nucleic Acids
Fetus
Parturition

ASJC Scopus subject areas

  • Analytical Chemistry
  • Biochemistry
  • Organic Chemistry

Cite this

Detection of the embryonic ζ chain in blood from newborn babies by reversed-phase high-perfomance liquid chromatogrphy. / Kutlar, F.; Fei, Y. J.; Wilson, J. B.; Kutlar, A.; Huisman, T. H.J.

In: Journal of Chromatography A, Vol. 394, No. 2, 1987, p. 333-343.

Research output: Contribution to journalArticle

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title = "Detection of the embryonic ζ chain in blood from newborn babies by reversed-phase high-perfomance liquid chromatogrphy",
abstract = "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.",
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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.

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