[Characterization of genetic variants of human albumin by isoelectric focusing].
Normal human serum albumin and bisalbuminic fractions from genetic variants of european origin have been studied by ultrathin-layer isoelectric focusing performed on whole sera or after purification of albumin fractions by affinity chromatography on Blue-Trisacryl. Narrow range ampholytes giving pH gradient between 5 and 8, together with the use of 8 M urea, provided suitable patterns allowing to discriminate the various allotypes. In these conditions, both normal albumin and heterozygous variants were microheterogeneous with several main bands; in the case of normal albumin, four major bands were found, while variants exhibited additional bands differing in number and pI. The position of additional bands comparatively to that of normal albumin was consistent with the electrophoretic behavior of variants. Fast moving allotypes exhibited additional bands with more anodal pIs, whereas slow moving variants were characterized by cathodal additional bands. The number and position of these bands allowed to characterize some variants, when they failed to distinguish between some others, identical patterns being correlated to identical mutations arising at different locations on the albumin molecule. These observations indicate that isoelectric focusing could allow a clear distinction of albumin variants, provided that their mutation were different, or could confirm the occurrence of identical mutation when IEF patterns are indistinguishable. In addition to electrophoretic mobilities at various pH, analytical isoelectric focusing could be a useful technique employed as a second step in the identification of allotypes prior to the determination of the structural change characterizing the variant.
Rochu
D
D
Laboratoire d'Immunochimie Analytique, Institut National de Transfusion Sanguine, Paris.
Crespeau
H
H
Fine
J M
JM
D015153:Blotting, Western; D002846:Chromatography, Affinity; D014644:Genetic Variation / Q000235:genetics*; D006579:Heterozygote; D006801:Humans; D007525:Isoelectric Focusing; D012709:Serum Albumin / Q000737:chemistry / Q000235:genetics*
D012709:Serum Albumin