High-Risk Screening of Fabry Disease: Analysis of Fifteen Urinary Methylated and Non-Methylated Gb3 Isoforms Using Tandem Mass Spectrometry.

High-Risk Screening of Fabry Disease: Analysis of Fifteen Urinary Methylated and Non-Methylated Gb3 Isoforms Using Tandem Mass Spectrometry.
Curr Protoc Hum Genet. 2016 Oct 11;91:17.24.1-17.24.11
Authors: Abaoui M, Boutin M, Lavoie P, Auray-Blais C
Abstract
Fabry disease is a multisystemic, X-linked lysosomal storage disorder caused by mutations in the GLA gene, leading to α-galactosidase A deficiency and resulting in the accumulation of glycosphingolipids in different tissues and biological fluids. Glycosphingolipid biomarkers, such as globotriaosylceramide (Gb3 ) isoforms, globotriaosylsphingosine (lyso-Gb3 ) and related analogs, and galabiosylceramide (Ga2 ) isoforms and analogs, are found to be abnormally increased in urine and in plasma of Fabry patients and have the potential to be used as specific biomarkers of the disease. This unit presents a protocol for the relative quantification of fifteen urinary isoforms of Gb3 analyzed simultaneously with creatinine by ultra-performance liquid chromatography (UPLC) coupled to tandem mass spectrometry (MS/MS). In order to purify urine samples, a liquid-liquid extraction is performed and samples are analyzed by MS/MS in positive electrospray ionization mode. These biomarkers are useful for screening, diagnosis, and long-term monitoring of Fabry disease patients. We have shown that the methylated Gb3 isoforms are particularly useful for screening Fabry patients who present with late-ons...

Source: Current Protocols in Human Genetics - October 13, 2016 Category: Genetics & Stem Cells Tags: Curr Protoc Hum Genet Source Type: research