caa a22 4500 467933235 CHVBK 20180406152949.0 cr unu---uuuuu 170328e20060401xx s 000 0 eng 10.1007/s10545-006-0265-4 doi (NATIONALLICENCE)springer-10.1007/s10545-006-0265-4 Direct multiplex assay of enzymes in dried blood spots by tandem mass spectrometry for the newborn screening of lysosomal storage disorders [Elektronische Daten] [Michael Gelb, Frantisek Turecek, C. Scott, Nestor Chamoles] Summary: Tandem mass spectrometry is currently used in newborn screening programmes to quantify the level of amino acids and acylcarnitines in dried blood spots for detection of metabolites associated with treatable diseases. We have developed assays for lysosomal enzymes in rehydrated dried blood spots in which a set of substrates is added and the set of corresponding enzymatic products are quantified using tandem mass spectrometry with the aid of mass-differentiated internal standards. We have developed a multiplex assay of the set of enzymes that, when deficient, cause the lysosomal storage disorders Fabry, Gaucher, Hurler, Krabbe, Niemann-Pick A/B and Pompe diseases. These diseases were selected because treatments are now available or expected to emerge shortly. The discovery that acarbose is a selective inhibitor of maltase glucoamylase allows the Pompe disease enzyme, acid α-glucosidase, to be selectively assayed in white blood cells and dried blood spots. When tested with dried blood spots from 40 unaffected individuals and 10-12 individuals with the lysosomal storage disorder, the tandem mass spectrometry assay led to the correct identification of the affected individuals with 100% sensitivity. Many of the reagents needed for the new assays are commercially available, and those that are not are being prepared under Good Manufacturing Procedures for approval by the FDA. Our newborn screening assay for Krabbe disease is currently being put in place at the Wadsworth Center in New York State for the analysis of ∼1000 dried blood spots per day. Summary We have developed tandem mass spectrometry for the direct assay of lysosomal enzymes in rehydrated dried blood spots that can be implemented for newborn screening of lysosomal storage disorders. Several enzymes can be analysed by a single method (multiplex analysis) and in a high-throughput manner appropriate for newborn screening laboratories. SSIEM and Springer, 2006 Gelb Michael Department of Chemistry, University of Washington, Seattle, Washington, USA aut Turecek Frantisek Department of Chemistry, University of Washington, Seattle, Washington, USA aut Scott C. Department of Pediatrics, University of Washington, Seattle, Washington, USA aut Chamoles Nestor Laboratory of Neurochemistry, Uriarte 2383, 1425, Buenos Aires, Argentina aut Journal of Inherited Metabolic Disease Kluwer Academic Publishers 29/2-3(2006-04-01), 397-404 0141-8955 29:2-3<397 2006 29 10545 https://doi.org/10.1007/s10545-006-0265-4 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s10545-006-0265-4 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Gelb Michael Department of Chemistry, University of Washington, Seattle, Washington, USA aut NATIONALLICENCE 700 1- Turecek Frantisek Department of Chemistry, University of Washington, Seattle, Washington, USA aut NATIONALLICENCE 700 1- Scott C. Department of Pediatrics, University of Washington, Seattle, Washington, USA aut NATIONALLICENCE 700 1- Chamoles Nestor Laboratory of Neurochemistry, Uriarte 2383, 1425, Buenos Aires, Argentina aut NATIONALLICENCE 773 0- Journal of Inherited Metabolic Disease Kluwer Academic Publishers 29/2-3(2006-04-01), 397-404 0141-8955 29:2-3<397 2006 29 10545 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer