title: 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.affiliation: Department of Chemistry, University of Washington, Seattle, Washington 98195-1700, USA.
authors.forename: Brian J
authors.initials: BJ
authors.lastname: Wolfe
authors.forename: Sophie
authors.initials: S
authors.lastname: Blanchard
authors.forename: Martin
authors.initials: M
authors.lastname: Sadilek
authors.forename: C Ronald
authors.initials: CR
authors.lastname: Scott
authors.forename: Frantisek
authors.initials: F
authors.lastname: Turecek
authors.forename: Michael H
authors.initials: MH
authors.lastname: Gelb
mesh_terms: D001800:Blood Specimen Collection; D006801:Humans; D007231:Infant, Newborn; D008247:Lysosomes / Q000201:enzymology*; D015394:Molecular Structure; D016532:Mucopolysaccharidosis II / Q000097:blood* / Q000175:diagnosis* / Q000201:enzymology; D015997:Neonatal Screening / Q000379:methods*; D053719:Tandem Mass Spectrometry / Q000379:methods*