화학공학소재연구정보센터
Electrophoresis, Vol.25, No.3, 469-475, 2004
Capillary zone electrophoresis for determination of carbohydrate-deficient transferrin in human serum
Carbohydrate-deficient transferrin (CDT) is the most specific marker for diagnosis of chronic excessive alcohol consumption and includes the serum transferrin (T-f) isoforms with two or less sialic acid residues (di-, mono-, and asialo-T-f). To monitor serum CDT, we developed a capillary zone electrophoresis (CZE) method based on the dynamic capillary coating with diethylenetriamine (DETA). The separation was performed in a bare fused-silica capillary (50 mum ID, 57 cm in length), applying a voltage of 25 kV and a temperature of 40degreesC. Using a 100 mmol/L borate buffer, pH 8.4 with 3 mmol/L DETA, the T-f isoforms (asialo- to pentasialo-T-f) were resolved within 16 min. Enzymatic cleavage of sialic acid residues with neuraminidase and immunosubtraction were used to identify CDT isoforms. The relative amount of CDT expressed as area % of disialo-T-f isoform related to the area of tetrasialo-T-f in 50 healthy donors (24 males and 26 females; aged 25-50 years) was 3.15 +/- 0.76% (mean +/- SD). The comparison between CDT values obtained by this CZE procedure and the "Axis-Shield %CDT" kit gave r = 0.644, p < 0.001 (n = 290). This easy to use and inexpensive CZE procedure could be an ideal tool to investigate CDT proteins for clinical or forensic purposes.