Open AccessComparison of transferrin isoform analysis by capillary electrophoresis and HPLC for screening congenital disorders of glycosylation
Author(s) -
Dave Mihika B.,
Dherai Alpa J.,
Udani Vrajesh P.,
Hegde Anaita U.,
Desai Neelu A.,
Ashavaid Tester F.
Publication year - 2018
Publication title -
journal of clinical laboratory analysis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.536
H-Index - 50
eISSN - 1098-2825
pISSN - 0887-8013
DOI - 10.1002/jcla.22167
Subject(s) - transferrin , gene isoform , glycosylation , carbohydrate deficient transferrin , capillary electrophoresis , sialic acid , high performance liquid chromatography , glycoprotein , chemistry , biochemistry , chromatography , microbiology and biotechnology , biology , gene , alcohol , alcohol consumption
Background Transferrin, a major glycoprotein has different isoforms depending on the number of sialic acid residues present on its oligosaccharide chain. Genetic variants of transferrin as well as the primary (CDG) & secondary glycosylation defects lead to an altered transferrin pattern. Isoform analysis methods are based on charge/mass variations. We aimed to compare the performance of commercially available capillary electrophoresis CDT kit for diagnosing congenital disorders of glycosylation with our in‐house optimized HPLC method for transferrin isoform analysis. Methods The isoform pattern of 30 healthy controls & 50 CDG‐suspected patients was determined by CE using a Carbohydrate‐Deficient Transferrin kit. The results were compared with in‐house HPLC‐based assay for transferrin isoforms. Results Transferrin isoform pattern for healthy individuals showed a predominant tetrasialo transferrin fraction followed by pentasialo, trisialo, and disialotransferrin. Two of 50 CDG‐suspected patients showed the presence of asialylated isoforms. The results were comparable with isoform pattern obtained by HPLC. The commercial controls showed a <20% CV for each isoform. Bland Altman plot showed the difference plot to be within +1.96 with no systemic bias in the test results by HPLC & CE. Conclusion The CE method is rapid, reproducible and comparable with HPLC and can be used for screening Glycosylation defects.