| Tandem mass spectrometry for the direct assay of lysosomal enzymes in dried blood spots: application to screening newborns for mucopolysaccharidosis II (Hunter Syndrome). | |
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MedLine Citation:
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PMID: 21192662 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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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%. |
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Authors:
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Brian J Wolfe; Sophie Blanchard; Martin Sadilek; C Ronald Scott; Frantisek Turecek; Michael H Gelb |
Publication Detail:
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Type: Journal Article; Research Support, N.I.H., Extramural Date: 2010-12-30 |
Journal Detail:
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Title: Analytical chemistry Volume: 83 ISSN: 1520-6882 ISO Abbreviation: Anal. Chem. Publication Date: 2011 Feb |
Date Detail:
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Created Date: 2011-01-31 Completed Date: 2011-05-17 Revised Date: 2012-05-30 |
Medline Journal Info:
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Nlm Unique ID: 0370536 Medline TA: Anal Chem Country: United States |
Other Details:
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Languages: eng Pagination: 1152-6 Citation Subset: IM |
Affiliation:
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Department of Chemistry, University of Washington, Seattle, Washington 98195-1700, USA. |
Export Citation:
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APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
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Blood Specimen Collection Humans Infant, Newborn Lysosomes / enzymology* Molecular Structure Mucopolysaccharidosis II / blood*, diagnosis*, enzymology Neonatal Screening / methods* Tandem Mass Spectrometry / methods* |
| Grant Support | |
ID/Acronym/Agency:
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R01 DK067859/DK/NIDDK NIH HHS; R01 DK067859/DK/NIDDK NIH HHS |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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