Tandem mass spectrometry for the direct assay of lysosomal enzymes in dried blood spots: application to screening newborns for mucopolysaccharidosis II (Hunter Syndrome).

Abstract

We have developed a tandem mass spectrometry based assay of iduronate-2-sulfatase (IdS) activity for the neonatal detection of mucopolysaccharidosis II (MPS-II, Hunter Syndrome). The assay uses a newly designed synthetic substrate (IdS-S) consisting of α-L-iduronate-2-sulfate, which is glycosidically conjugated to a coumarin and a linker containing a tert-butyloxycarbamido group. A short synthesis of the substrate has been developed that has the potential of being scaled to multigram quantities. Sulfate hydrolysis of IdS-S by IdS found within a 3 mm dried blood spot specifically produces a nonsulfated product (IdS-P) which is detected by electrospray tandem mass spectrometry and quantified using a deuterium-labeled internal standard, both carried out in positive ion mode. Analysis of DBS from 75 random human newborns showed IdS activities in the range of 4.8-16.2 (mean 9.1) μmol/(h L of blood), which were clearly distinguished from the activities measured for 14 MPS-II patients at 0.17-0.52 (mean 0.29) μmol/(h L of blood). The assay shows low blank activity, 0.15 ± 0.03 μmol/(h L of blood). The within-assay coefficient of variation (CV) was 3.1% while the interassay CV was 15%.

• Authors: Brian J Wolfe, Sophie Blanchard, Martin Sadilek, C Ronald Scott, Frantisek Turecek, Michael H Gelb
• Journal: Analytical chemistry (Anal Chem) Vol. 83 Issue 3 Pg. 1152-6 (Feb 01 2011) ISSN: 1520-6882 [Electronic] United States
• PMID: 21192662 (Publication Type: Journal Article, Research Support, N.I.H., Extramural)

Topics

• Blood Specimen Collection
• Humans
• Infant, Newborn
• Lysosomes (enzymology)
• Molecular Structure
• Mucopolysaccharidosis II (blood, diagnosis, enzymology)
• Neonatal Screening (methods)
• Tandem Mass Spectrometry (methods)