Human globin chain separation by isolectric focusing

Pier Giorgio Righetti, Elisabetta Gianazza, Alessandro M. Gianni, Paola Comi, Barbara Giglioni, Sergio Ottolenghi, Camillo Secchi, Luigi Rossi-Bernardi

Research output: Contribution to journalArticle

Abstract

Human globin chain separation, a key procedure in the study of hemoglobinopathies, is routinely performed by chromatography on carboxymethylcellulose. This method, though relatively easy and highly reliable, is expensive and time consuming. A new procedure, based on isoelectric focusing, is presented which allows the simultaneous separation of globin chains from multiple samples (at least 20 per gal slab). The method is rapid, inexpensive and can be easily carried out in clinical laboratories, and its high sensitivity allows the identification of radioactive bands even with minute amounts of labelled material. A new phenomenon, called the 'Nonidet P-40 effect', which greatly enhances the separation between γ and β chainss by binding to these two chains and shifting their pI values in opposite directions, is described.

Original languageEnglish
Pages (from-to)45-57
Number of pages13
JournalJournal of Biochemical and Biophysical Methods
Volume1
Issue number1
DOIs
Publication statusPublished - 1979

Fingerprint

Globins
Clinical laboratories
Carboxymethylcellulose Sodium
Chromatography
Hemoglobinopathies
Isoelectric Focusing

Keywords

  • detergent binding
  • globin chains
  • hemoglobinopathies
  • isoelectric focusing

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics

Cite this

Human globin chain separation by isolectric focusing. / Righetti, Pier Giorgio; Gianazza, Elisabetta; Gianni, Alessandro M.; Comi, Paola; Giglioni, Barbara; Ottolenghi, Sergio; Secchi, Camillo; Rossi-Bernardi, Luigi.

In: Journal of Biochemical and Biophysical Methods, Vol. 1, No. 1, 1979, p. 45-57.

Research output: Contribution to journalArticle

Righetti, PG, Gianazza, E, Gianni, AM, Comi, P, Giglioni, B, Ottolenghi, S, Secchi, C & Rossi-Bernardi, L 1979, 'Human globin chain separation by isolectric focusing', Journal of Biochemical and Biophysical Methods, vol. 1, no. 1, pp. 45-57. https://doi.org/10.1016/0165-022X(79)90045-9
Righetti PG, Gianazza E, Gianni AM, Comi P, Giglioni B, Ottolenghi S et al. Human globin chain separation by isolectric focusing. Journal of Biochemical and Biophysical Methods. 1979;1(1):45-57. https://doi.org/10.1016/0165-022X(79)90045-9
Righetti, Pier Giorgio ; Gianazza, Elisabetta ; Gianni, Alessandro M. ; Comi, Paola ; Giglioni, Barbara ; Ottolenghi, Sergio ; Secchi, Camillo ; Rossi-Bernardi, Luigi. / Human globin chain separation by isolectric focusing. In: Journal of Biochemical and Biophysical Methods. 1979 ; Vol. 1, No. 1. pp. 45-57.
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