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Capillary zone electrophoresis for determination of carbohydrate‐deficient transferrin in human serum
Author(s) -
Fermo Isabella,
Germagnoli Luca,
Soldarini Armando,
Dorigatti Fernanda,
Paroni Rita
Publication year - 2004
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.200305694
Subject(s) - carbohydrate deficient transferrin , capillary electrophoresis , sialic acid , transferrin , chemistry , neuraminidase , gene isoform , chromatography , carbohydrate , n acetylneuraminic acid , enzyme , microbiology and biotechnology , biochemistry , biology , alcohol , alcohol consumption , gene
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 μm ID, 57 cm in length), applying a voltage of 25 kV and a temperature of 40°C. 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.