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Human globin chain separation by capillary electrophoresis in acidic isoelectric buffers
Author(s) -
Righetti Pier Giorgio,
Saccomani Arianna,
Stoyanov Alexander V.,
Gelfi Cecilia
Publication year - 1998
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.1150191034
Subject(s) - capillary electrophoresis , chromatography , chemistry , isoelectric focusing , electrophoresis , iminodiacetic acid , electrolyte , urea , isoelectric point , globin , hemoglobin , biochemistry , inorganic chemistry , chelation , electrode , enzyme
A simple and reliable method, utilizing capillary electrophoresis in uncoated capillaries in acidic isoelectric buffers, is reported for screening for thalassemia and other defects on the synthesis of human globin chains. A solution of 50 mM iminodiacetic acid (p i 2.23), containing 7 M urea and 0.5% hydroxyethyl‐cellulose (apparent pH 3.2) is used as background electrolyte for fast separation of heme‐free, denatured globin (α, β and γ) chains. Due to the low conductivity of such a buffer, high voltage gradients (600 V/cm) can be applied, thus reducing the separation time to only a few minutes. It is additionally shown that inclusion of 2% surfactant (Tween 20) in the background electrolyte induces the splitting of the γ chains into two zones, called Aγ and Gγ, which represent the products of two genes coding for Ala or Gly as residue 136 of the chain.