Combined assay methodology for the analysis of enzyme activities and biomarker concentrations for Fabry, Gaucher, Krabbe, Niemann Pick types A/B, and Pompe disease

Abstract

To overcome major limitations of the FIA-MS/MS approach used for diagnostics and newborn screening (NBS) for lysosomal storage disorders (LSDs), a combined UHPLC - MS/MS based assay has been developed by utilizing dried blood spots and an Analytical Quality by Design approach. The assay is capable to additionally provide the concentrations of three corresponding biomarkers of five LSDs covered by the NeoLSD Kit (Perkin Elmer). The assay utilizes just a short (4 hours) incubation period, only needs simple precipitation steps and is able to quantify lyso- Gb3, lyso-Gb1/Psychosine and lyso-SM simultaneously in an injection-to- injection time of ~3.2min. The assay has been technical validated and a pre-pilot comparison study with 500 DBS was conducted