Direct capillary electrophoretic detection of carbohydrate-deficient transferrin in neat serum.
Transferrin, an iron transport protein found in serum and cerebrospinal fluid, is known to be microheterogeneous with respect to its carbohydrate and sialic acid content. The forms of transferrin deficient in sialic acid and/or carbohydrate, termed carbohydrate-deficient transferrin (CDT), have been of clinical interest for almost two decades as a result of the initial finding that elevated CDT concentrations are associated with chronic, excessive alcohol abuse. We demonstrate the utility of capillary electrophoresis for examining the CDT sialoform profile via the direct electrophoresis of serum. The need for negligible preelectrophoresis sample preparation and absence of postelectrophoresis processing dramatically decreases analysis time compared to slab gel-based separations. Using a fluorocarbon-coated capillary containing a hydroxyethyl cellulose/borate buffer, the high resolution separation of serum components is effected in less than 30 min. Under these conditions, the beta region proteins (including transferrin) are well resolved from the alpha-2 and gamma zone proteins in a window where the individual transferrin sialoforms can be detected. The usefulness of this method is demonstrated with the electrophoresis of serum from subjects known to be either non-alcoholic and alcoholic.
Department of Chemistry, University of Pittsburgh, PA, USA.
A L
AL
Trout
R
R
Prasad
D
D
Coffin
A
A
DiMartini
T
T
Lane
C
C
Blessum
N
N
Khatter
J P
JP
Landers
D000428:Alcohol Drinking / Q000097:blood*; D019075:Electrophoresis, Capillary / Q000295:instrumentation / Q000379:methods; D006801:Humans; D008104:Liver Cirrhosis, Alcoholic / Q000097:blood; D008105:Liver Cirrhosis, Biliary / Q000097:blood; D008113:Liver Neoplasms / Q000097:blood; D020033:Protein Isoforms / Q000032:analysis; D012794:Sialic Acids / Q000378:metabolism; D014168:Transferrin / Q000031:analogs & derivatives* / Q000032:analysis
D020033:Protein Isoforms; D012794:Sialic Acids; D014168:Transferrin; C056759:carbohydrate-deficient transferrin