Screening of umbilical cord blood hemoglobins by isoelectric focusing in capillaries

M. Conti, C. Gelfi, P. G. Righetti

Research output: Contribution to journalArticle

47 Citations (Scopus)

Abstract

Separation and quantitation of the three main hemoglobin components of umbilical cord blood (fetal, acetylated fetal and adult hemoglobins; Hb F, F(ac), A) by capillary isoelectric focusing (IEF) in a pH 6-8 gradient is reported. Even in coated capillaries (with covalently bound chains of linear acrylamido derivatives, notably N-acryloylaminoethoxyethanol), no base line separation is obtained between Hb F and A, although this is routinely achieved in gel slab IEE. However, when the carrier ampholyte buffers were added to 3% short-chain liquid linear polyacrylamide, base line resolution and stabilization of peak transit times were obtained. This suggests that even in the best coating procedures, patches of the inner capillary surface could still be naked, so that the static coating is complemented by a dynamic coating on the unoccupied sites. An additional improvement in separation occurs if the above mixture, comprising 5% carrier ampholytes in the pH 6-8 range and 3% soluble polyacrylamide, is made to contain 50 mM β-alanine, a 'separator' known to flatten the pH gradient around pH 7. In the normal newborns analyzed (n = 30), the following average values were obtained: Hb F, 70.1% (range 65-75%); Hb A, 20.2% (range 15-25%); and HbF(ac), 9.5% (range 7-11%).

Original languageEnglish
Pages (from-to)1485-1491
Number of pages7
JournalElectrophoresis
Volume16
Issue number8
Publication statusPublished - 1995

Fingerprint

Isoelectric Focusing
Ampholyte Mixtures
Fetal Blood
Screening
Hemoglobins
Blood
Coatings
Fetal Hemoglobin
Proton-Motive Force
Separators
Alanine
Buffers
Stabilization
Gels
Derivatives
Liquids
polyacrylamide

Keywords

  • Hemoglobin
  • Isoelectric focusing
  • Thalassemia
  • Umbilical blood

ASJC Scopus subject areas

  • Clinical Biochemistry

Cite this

Screening of umbilical cord blood hemoglobins by isoelectric focusing in capillaries. / Conti, M.; Gelfi, C.; Righetti, P. G.

In: Electrophoresis, Vol. 16, No. 8, 1995, p. 1485-1491.

Research output: Contribution to journalArticle

Conti, M. ; Gelfi, C. ; Righetti, P. G. / Screening of umbilical cord blood hemoglobins by isoelectric focusing in capillaries. In: Electrophoresis. 1995 ; Vol. 16, No. 8. pp. 1485-1491.
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