title: Erythrocyte membrane protein analysis by sodium dodecyl sulphate-capillary gel electrophoresis in the diagnosis of hereditary spherocytosis.
abstract: BACKGROUND
  Sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) is currently the reference method for detecting protein deficiencies related to hereditary spherocytosis. The aim of the study was to evaluate an automated capillary gel electrophoresis system, the Experion instrument from BioRad, for its ability to separate and quantify the erythrocyte membrane proteins.
  
  
  METHODS
  The major erythrocyte membrane proteins (actin, protein 4.2, protein 4.1, band 3, ankyrin, α- and β-spectrin) were extracted and purified from membrane ghosts by centrifugation, immunoprecipitation and electroelution. Analyses were performed using SDS-PAGE and sodium dodecyl sulphate capillary gel electrophoresis (SDS-CGE) to establish a separation profile of the total ghosts. Then, the samples from patients received for investigations of erythrocyte membrane defects were analysed.
  
  
  RESULTS
  Five of the seven expected erythrocyte membrane proteins were finally separated and identified. In the 20 studied cases, taking into account the screening test results and the clinical and family histories, the SDS-CGE method allowed us to achieve the same conclusion as with SDS-PAGE, except for the patient with elliptocytosis.
  
  
  CONCLUSIONS
  The new SDS-CGE method presents interesting features that could make this instrument a powerful diagnostic tool for detection of erythrocyte membrane protein abnormalities, and can be proposed as an automated alternative method to the labour intensive SDS-PAGE analysis.
authors.affiliation: Department of Clinical Chemistry, Hôpital Erasme, Université Libre de Bruxelles (ULB), Brussels, Belgium.
authors.forename: France
authors.initials: F
authors.lastname: Debaugnies
authors.forename: Frédéric
authors.initials: F
authors.lastname: Cotton
authors.forename: Charles
authors.initials: C
authors.lastname: Boutique
authors.forename: Béatrice
authors.initials: B
authors.lastname: Gulbis
mesh_terms: D004591:Electrophoresis, Polyacrylamide Gel; D004910:Erythrocyte Membrane / Q000737:chemistry*; D004912:Erythrocytes / Q000737:chemistry*; D006801:Humans; D008565:Membrane Proteins / Q000032:analysis*; D013103:Spherocytosis, Hereditary / Q000175:diagnosis*
chemical_list: D008565:Membrane Proteins