Mass Spectrometry-based Identification of Transferrin Glycosylation Variants for Diagnosis of Congenital Disorders of Glycosylation
DOI:
https://doi.org/10.52783/jns.v14.1757Keywords:
Transferrin, Congenital Disorders of Glycosylation, Iso-Electric Focusing, MALDI-TOFAbstract
Congenital disorders of glycosylation (CDG) are a group of metabolic conditions resulting from impaired glycosylation of proteins and/or lipids. Transferrin, a vital glycoprotein present in blood plasma, plays a significant role in our research. We aim to tackle the diagnostic challenges associated with CDG by focusing on transferrin as a key protein for study. Several techniques are used for the quantification of human transferrin, each offering valuable insights into protein levels. While these methods can be highly sensitive, they often fall short in terms of specificity and reproducibility. In contrast, Liquid Chromatography-Mass Spectrometry (LC-MS/MS) offers a more reliable and precise approach, even at low concentrations, making it a superior tool for transferrin analysis in CDG research.
In this study, we present: (i) an efficient and rapid method for purifying transferrin from human serum using rivanol, (ii) a comparative analysis of various techniques for screening CDG samples, including Isoelectric Focusing (IEF), 2D gel electrophoresis, MALDI, and LC-MS/MS, with LC-MS/MS proving to be the most reliable method, and (iii) an LC-MS/MS-based proteomics assay to quantify transferrin isoform levels in human serum, utilizing specific glycosylated peptides. We identified significant biomarker peptides with glycosylation variants, and their relative levels were used for confirmatory diagnosis of CDG. Early and accurate diagnosis of CDG is crucial for the timely initiation of appropriate therapies, which can greatly enhance clinical outcomes. We aim to validate these findings further in our ongoing research using a larger cohort of samples to develop a fast and reliable diagnostic test for CDG.
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