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Clinical Chemistry, Vol 40, 2288-2295, Copyright © 1994 by American Association for Clinical Chemistry
JM Hempe and RD Craver
Department of Pediatrics, LSU School of Medicine.
Capillary isoelectric focusing (cIEF) was used to identify and quantify major and minor hemoglobin (Hb) variants. Whole blood (approximately 10 microL required) hemolysate was analyzed with a commercial instrument equipped with a 50 microns (i.d.) x 27 cm coated capillary filled with 20 g/L ampholytes (pH 6-8) in 4 g/L methylcellulose (MC). Cathode and anode solutions were 20 mol/L NaOH and 100 mol/L H3PO4 in MC, respectively. Samples (approximately 40 nL) were applied via autosampler by low-pressure injection, focused for 3 min at 30 kV, and mobilized by simultaneous voltage and low pressure past the detector, where absorbance at 415 nm was analyzed by an automated data acquisition system. Blood from subjects with sickle cell trait, Hb S/C disease, and various beta-thalassemias were analyzed by cIEF in < 15 min. cIEF was used to separate Hb S from Hb D-Los Angeles. Assay precision determined with commercial controls gave CV < 2% for Hb A and S, and 1-11% for minor Hb variants A2, F, and A1c. Results obtained by cIEF for patients' samples agreed well with values determined by conventional assays (r2 > 0.95). The results demonstrate that cIEF is a rapid, sensitive, high-resolution automated method for routine quantitative clinical analysis of Hb variants.
The following articles in journals at HighWire Press have cited this article:
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  J. M. Hempe, J. Ory-Ascani, and D. Hsia Genetic Variation in Mouse Beta Globin Cysteine Content Modifies Glutathione Metabolism: Implications for the Use of Mouse Models Experimental Biology and Medicine, March 1, 2007; 232(3): 437 - 444. [Abstract] [Full Text] [PDF]
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 G. M. Clarke and T. N. Higgins Laboratory Investigation of Hemoglobinopathies and Thalassemias: Review and Update Clin. Chem., August 1, 2000; 46(8): 1284 - 1290. [Abstract] [Full Text] [PDF]
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N. Mario, B. Baudin, A. Bruneel, J. Janssens, and M. Vaubourdolle Capillary Zone Electrophoresis for the Diagnosis of Congenital Hemoglobinopathies Clin. Chem., February 1, 1999; 45(2): 285 - 288. [Full Text] [PDF]
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Y. Henskens, J. de Winter, M. Pekelharing, and G. Ponjee Detection and identification of monoclonal gammopathies by capillary electrophoresis Clin. Chem., June 1, 1998; 44(6): 1184 - 1190. [Abstract] [Full Text] [PDF]
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N. Mario, B. Baudin, C. Aussel, and J. Giboudeau Capillary isoelectric focusing and high-performance cation-exchange chromatography compared for qualitative and quantitative analysis of hemoglobin variants Clin. Chem., November 1, 1997; 43(11): 2137 - 2142. [Abstract] [Full Text] [PDF]
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A. A. Mohammad, A. O. Okorodudu, M. G. Bissell, P. Dow, G. Reger, A. Meier, P. Guodagno, and J. R. Petersen Clinical Application of Capillary Isoelectric Focusing on Fused Silica Capillary for Determination of Hemoglobin Variants Clin. Chem., September 1, 1997; 43(9): 1798 - 1799. [Full Text]
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C. J. A. Doelman, C. W. M. Siebelder, W. A. Nijhof, C. W. Weykamp, J. Janssens, and T. J. Penders Capillary electrophoresis system for hemoglobin A1c determinations evaluated Clin. Chem., April 1, 1997; 43(4): 644 - 648. [Abstract] [Full Text] [PDF]
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