Analyte comigrating with trisialotransferrin during capillary zone electrophoresis of sera from patients with cancer.
BACKGROUND
Serum concentrations of monoglycosylated isoforms of transferrin are increased by chronic ethanol intake. We investigated transferrin glycosylation in patients with cancer, in which aberrant glycosylation is also induced.


METHODS
We used a P/ACE 5000 series capillary zone electrophoresis (CZE) apparatus and a CZE carbohydrate-deficient transferrin reagent set to study 200 cancer patients who consumed alcohol moderately and 33 who were alcohol abusers; we then compared these patients with 56 healthy teetotalers, 89 moderate, and 112 excessive alcohol drinkers without known malignancies. Transferrin isoforms were identified by immunosubtraction with anti-human transferrin polyclonal antibody.


RESULTS
Seven peaks, P0-P6, were visualized and completely or partly immunosubtracted when CZE separation was performed at pH 8.5. P0 was present in 95% of alcohol abusers with or without cancer. P3 was significantly higher in cancer patients and was only partly immunosubtracted as trisialotransferrin in all groups. The comigrating analyte was not altered by papain, precipitation by ethanol, or extraction by organic solvents, but was sensitive to acid hydrolysis, suggesting a polysaccharide structure. When isolated at pH 8.25, this analyte was higher in cancer patients. ROC curve analysis identified localized malignant neoplasia at P3 values above 5.8% of total transferrin (sensitivity, 0.78; specificity, 0.87), regardless of alcohol consumption. Disseminated cancers were better differentiated above 8% (sensitivity, 0.94; specificity, 0.96).


CONCLUSIONS
Malignant neoplasia, unlike excessive ethanol intake, did not alter the addition of two N-glycans to transferrin. A peak comigrating with trisialotransferrin had characteristics of a polysaccharide in all adults and was increased in sera of patients with cancer.
University Department of Gastroenterology, Centre Hospitalier Universitaire de Charleroi, 92, Boulevard Janson, 6000 Charleroi, Belgium.
Brahim
B
Ramdani
Vincent
V
Nuyens
Thierry
T
Codden
Gael
G
Perpete
Jacques
J
Colicis
Anne
A
Lenaerts
Jean-Pol
JP
Henry
Franz J
FJ
Legros
D000428:Alcohol Drinking / Q000097:blood; D000437:Alcoholism / Q000097:blood / Q000150:complications; D019075:Electrophoresis, Capillary; D005260:Female; D006031:Glycosylation; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007106:Immune Sera; D008297:Male; D008875:Middle Aged; D019158:N-Acetylneuraminic Acid / Q000378:metabolism*; D009369:Neoplasms / Q000097:blood* / Q000150:complications / Q000175:diagnosis; D020033:Protein Isoforms / Q000097:blood; D012372:ROC Curve; D012016:Reference Values; D014168:Transferrin / Q000031:analogs & derivatives* / Q000032:analysis* / Q000276:immunology / Q000378:metabolism
D007106:Immune Sera; D020033:Protein Isoforms; D014168:Transferrin; C056759:carbohydrate-deficient transferrin; D019158:N-Acetylneuraminic Acid