Atypical serum transferrin isoform distribution in liver cirrhosis studied by HPLC, capillary electrophoresis and transferrin genotyping
Introduction
Analysing human serum transferrin by HPLC or capillary electrophoresis (CE) yields transferrin isoform patterns which can be classified into normal or abnormal [1]. Normal human serum transferrin isoform patterns show a major fraction of tetrasialotransferrin of approximately 80% and minor fractions of disialo-, trisialo- and pentasialotransferrin [1], [2]. Atypical transferrin isoform patterns can be categorized into (a) patterns with twin bands, most clearly visible for tetrasialotransferrin, caused by genetic transferrin variants like BC or CD [3], (b) patterns with increased amounts of asialo- and disialotransferrin caused by alcohol abuse [1] or rarely by congenital disorders of glycosylation [4], (c) patterns with increased fractions of trisialotransferrin e. g. caused by primary biliary cirrhosis [5] or anorexia nervosa [6] and (d) patterns with combinations of these characteristics.
We report about transferrin isoform patterns which can neither be categorized as normal nor as abnormal according to categories a–d and which were seen quite frequently in large routine CDT analysis series (50–60 analysis/night) using the Recipe CDT on-line HPLC assay [7]. These transferrin isoform patterns attracted attention by a significantly incomplete separation of the disialotransferrin and trisialotransferrin peaks which we like to call the di-tri-bridging phenomenon or shortly di-tri-bridge. We investigated this phenomenon by different HPLC methods, capillary electrophoresis and transferrin genotyping as well as by assessment of the clinical background of the patients.
Section snippets
Materials and methods
All procedures in this retrospective study were performed in accordance with the current Helsinki Declaration.
Results and discussion
Fig. 1 shows (for comparison with Fig. 2) a transferrin isoform pattern obtained by the routine CDT HPLC analysis method (Recipe, Munic) of a healthy control with normal CDT. Fig. 2 demonstrates the variability of the di-tri-bridging phenomenon between patients with alcohol-induced liver cirrhosis. Fig. 2A and B shows chromatograms obtained with the routine Recipe HPLC method, Fig. 2C and D the corresponding chromatograms obtained with the candidate reference HPLC method, and Fig. 2 E and F the
Conclusion
A distinctly incomplete separation by HPLC or CE of the disialotransferrin and trisialotransferrin fractions with formation of a disialotransferrin-trisialotransferrin-bridge (di-tri-bridge) was in 19 out of 21 cases related to patients with liver cirrhosis. It could not be explained by the presence of an increased trisialotransferrin fraction or genetic transferrin variants. Our data provide some evidence that this di-tri-bridging phenomenon might be related to (alcohol-induced) liver
Acknowledgement
The skilful analytical assistance of Mrs. Riekie te Stoet is gratefully acknowledged.
References (11)
- et al.
Primary biliary cirrhosis is not a clinical condition for increased carbohydrate-deficient transferrin: experience with 4 independent CDT analysis methods
Clin Chim Acta
(2006) - et al.
High prevalence of increased trisialotransferrin concentrations in patients with anorexia nervosa: implications for determination of carbohydrate-deficient transferrin
Clin Chim Acta
(2007) - et al.
Forensic analysis of carbohydrate-deficient transferrin by HPLC — statistics and extreme CDT values
Forens Sci Int
(2008) - et al.
Analytical evaluation of a new capillary electrophoresis method for carbohydrate-deficient transferrin measurement
Clin Chim Acta
(2007) - et al.
Molecular characterization of a case of atransferrinemia
Blood
(2000)
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Reprint of Standardisation and use of the alcohol biomarker carbohydrate-deficient transferrin (CDT)
2017, Clinica Chimica ActaIFCC approved HPLC reference measurement procedure for the alcohol consumption biomarker carbohydrate-deficient transferrin (CDT): Its validation and use
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Standardisation and use of the alcohol biomarker carbohydrate-deficient transferrin (CDT)
2016, Clinica Chimica ActaFirst evaluation of a multi-capillary electrophoresis CDT assay on Helena Biosciences' V8 analyser
2014, Clinical BiochemistryCitation Excerpt :The measurement of CDT is mainly performed for identification and follow-up of individuals suffering from alcohol consumption disorders, and also for medico-legal decisions, such as in traffic medicine, occupational healthcare and social services. However, it is important to recognize other clinical conditions that can lead to an abnormal Tf glycosylation pattern including polymorphisms within the Tf gene [4,5], rare hereditable deficiency in the glycosylation enzymes [6–8], hereditary fructose intolerance and galactosemia [9] and some other clinical conditions that are apparently unrelated to excessive alcohol consumption [10–12]. In the routine clinical laboratory, automation, high throughput instruments and robustness are always a critical issue.