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Separation of hemoglobin variants with similar charge by capillary isoelectric focusing: Value of isoelectric point for identification of common and uncommon hemoglobin variants
Author(s) -
Hempe James M.,
Craver Randall D.
Publication year - 2000
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/(sici)1522-2683(20000301)21:4<743::aid-elps743>3.0.co;2-1
Subject(s) - isoelectric focusing , hemoglobin variants , isoelectric point , hemoglobin , pi , chemistry , chromatography , microbiology and biotechnology , biochemistry , biology , enzyme
Clinical assays for the primary evaluation of congenital hemoglobin (Hb) disorders must detect and identify a variety of Hb variants. We analyzed hemolysates containing Hb variants with similar charge to evaluate the diagnostic sensitivity and specificity of automated capillary isoelectric focusing (CIEF). Peak separation was observed for each variant in samples containing Hb S, D, and G. The calculated isoelectric points (p I ) of these variants were significantly different such that each could be identified in a single run with p I as the sole criterion of identification. The p I of Hb C was significantly different from that of Hb E, C‐Harlem, and O‐Arab. Hb E, C‐Harlem, and O‐Arab had similar p I and were not readily differentiated. Hb Koln, M‐Saskatoon, Aida, and S/Aida hybrid were readily separated from common Hb variants and detected by CIEF. We conclude that CIEF exhibits both diagnostic sensitivity and specificity, and that p I is an objective and specific criterion of Hb variant identification.