Capillary zone electrophoresis of transferrin and EDTA samples in congenital disorders of glycosylation screening: CaNOt do, really?

Background: The Congenital Disorders of Glycosylation (CDG) are an expanding group of metabolic diseases with a broad clinical presentation. We sought to validate a new Capillary Zone Electrophoresis (CZE) method (Sebia CAPILLARYS™ CDT) to screen for CDG. Methods: We analyzed 119 serum samples from children of varying ages and of both sexes to establish a reference range of transferrin glycoforms including CDT (Carbohydrate Deficient Transferrin). We then studied serums from 8 known CDG patients and compared the CZE results to the isoelectric focusing (IEF) profiles. We also analyzed serums after extraction from spotted Guthrie cards. Results: The mean (SD) percentage of transferrin glycoforms is 18.5 (4.4), 78.5 (4.2), 2.5 (1.3) and 0.6 (0.3) for penta-, tetra-, trisialotransferrin and CDT, respectively. There is no statistically significant difference between the different age groups analyzed (0-5, 6-11, 12-15, 16-18, and > 18 years) or between sexes. We observed a good correlation between the CZE and IEF profiles with both fresh serum and serum extracted from Guthrie cards. Conclusions: The Sebia CAPILLARYS™ CDT system is a simple and reliable method to screen for CDG in pediatric and adult patients with an unexplained clinical syndrome, particularly when the nervous system is involved.

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Diagnosis of Congenital Disorders of Glycosylation by Capillary Zone Electrophoresis of Serum Transferrin

Clinical Chemistry ◽

10.1373/clinchem.2003.021568 ◽

2004 ◽

Vol 50 (1) ◽

pp. 101-111 ◽

Cited By ~ 72

Author(s):

Hubert A Carchon ◽

Roland Chevigné ◽

Jean-Bernard Falmagne ◽

Jaak Jaeken

Keyword(s):

Capillary Zone Electrophoresis ◽

Reference Group ◽

Congenital Disorders ◽

Serum Transferrin ◽

Congenital Disorders Of Glycosylation ◽

Zone Electrophoresis ◽

Carbohydrate Deficient Transferrin ◽

Group A ◽

Capillary Zone ◽

Relative Migration

Abstract Background: Congenital disorders of glycosylation (CDG) are usually diagnosed by isoelectric focusing (IEF) of serum transferrin (Tf). The aim of this study was to evaluate capillary zone electrophoresis (CZE) as a diagnostic alternative to IEF. Methods: We performed 792 CZE analyses of Tf, using the CEofixTM-CDT (carbohydrate-deficient transferrin) assay. Peak identification was based on relative migration times (RMTs) to reduce migration variability. Results: Tf profiles comprised three main groups (A–C). Groups A and B were characterized by one or two dominant tetrasialo-Tf peaks, whereas group C showed a widely variable Tf isoform composition. Group A was composed of four subgroups: a major group with a typical Tf profile (considered as reference group), two minor groups with decreased or moderately increased trisialo-Tf isoform, and a group showing the presence of unknown compounds with RMTs similar to mono- and disialo-Tf. However, these compounds were absent on IEF. Group C contained all profiles from patients with confirmed as well as putative CDG. From the reference group, 99% confidence intervals were calculated for the RMTs of the Tf isoforms, and percentiles representing the Tf isoform distributions were defined. Conclusions: All patients with abnormal IEF results and confirmed CDG were identified by CZE; thus, this method can be used as a diagnostic alternative to IEF in a manner suitable for automation. Because whole serum is analyzed, it should be kept in mind that CZE profiles can show substances other than Tf.

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Comparison between high performance liquid chromatography and capillary zone electrophoresis for the diagnosis of congenital disorders of glycosylation

Journal of Chromatography B ◽

10.1016/j.jchromb.2009.06.031 ◽

2009 ◽

Vol 877 (24) ◽

pp. 2513-2518 ◽

Cited By ~ 17

Author(s):

Ester Quintana ◽

Raquel Montero ◽

Mercedes Casado ◽

Aleix Navarro-Sastre ◽

María A. Vilaseca ◽

...

Keyword(s):

High Performance Liquid Chromatography ◽

Liquid Chromatography ◽

Capillary Zone Electrophoresis ◽

High Performance ◽

Congenital Disorders ◽

Congenital Disorders Of Glycosylation ◽

Zone Electrophoresis ◽

Capillary Zone

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Congenital Disorders of Glycosylation: What Clinicians Need to Know?

Frontiers in Pediatrics ◽

10.3389/fped.2021.715151 ◽

2021 ◽

Vol 9 ◽

Author(s):

Patryk Lipiński ◽

Anna Tylki-Szymańska

Keyword(s):

High Performance ◽

Diagnostic Algorithm ◽

Diagnostic Methods ◽

Congenital Disorders ◽

Congenital Disorders Of Glycosylation ◽

Acid Deficiency ◽

Zone Electrophoresis ◽

Capillary Zone ◽

Generation Sequencing

Congenital disorders of glycosylation (CDG) are a group of clinically heterogeneous disorders characterized by defects in the synthesis of glycans and their attachment to proteins and lipids. This manuscript aims to provide a classification of the clinical presentation, diagnostic methods, and treatment of CDG based on the literature review and our own experience (referral center in Poland). A diagnostic algorithm for CDG was also proposed. Isoelectric focusing (IEF) of serum transferrin (Tf) is still the method of choice for diagnosing N-glycosylation disorders associated with sialic acid deficiency. Nowadays, high-performance liquid chromatography, capillary zone electrophoresis, and mass spectrometry techniques are used, although they are not routinely available. Since next-generation sequencing became more widely available, an improvement in diagnostics has been observed, with more patients and novel CDG subtypes being reported. Early and accurate diagnosis of CDG is crucial for timely implementation of appropriate therapies and improving clinical outcomes. However, causative treatment is available only for few CDG types.

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