Validation by isoelectric focusing of the anion-exchange isotransferrin fractionation step involved in determination of carbohydrate-deficient transferrin by the CDTect assay.
Serum concentration of carbohydrate-deficient transferrin (CDT) is used for laboratory diagnosis of chronic alcohol abuse. Using isoelectric focusing for validation of the initial isotransferrin fractionation step involved in the determination of CDT by the CDTect assay, we found a complete in vitro iron saturation of transferrin and sufficient stability of the transferrin iron load during column passage; effective separation of non-CDT-isotransferrins and CDT-isotransferrins at the microcolumns; partial coelution of trisialo-Fe2-transferrin, which did not significantly affect CDT measurement; partial retention of CDT-isotransferrins, especially disialo-Fe2-transferrin, which may cause falsely negative results for CDT at the upper reference limits; good precision of the isotransferrin fractionation step; and no significant effects of low concentrations of serum protein and transferrin. We strongly urge standardization of CDT analysis and suggest isoelectric focusing for validation of CDT analysis methods and verification of odd results.
Arndt
T
T
Abteilung für Klinische Chemie und Zentrallaboratorium, Philipps-Universität, Marburg, Germany. bioscientia@t-online.de
Hackler
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R
Kleine
T O
TO
Gressner
A M
AM
D000437:Alcoholism / Q000097:blood; D000837:Anion Exchange Resins; D001798:Blood Proteins / Q000032:analysis; D002241:Carbohydrates / Q000737:chemistry*; D004355:Drug Stability; D006801:Humans; D007525:Isoelectric Focusing / Q000379:methods; D019158:N-Acetylneuraminic Acid / Q000737:chemistry; D011933:Reagent Kits, Diagnostic; D015203:Reproducibility of Results; D014168:Transferrin / Q000032:analysis* / Q000737:chemistry
D000837:Anion Exchange Resins; D001798:Blood Proteins; D002241:Carbohydrates; D011933:Reagent Kits, Diagnostic; D014168:Transferrin; D019158:N-Acetylneuraminic Acid