BACKGROUND: We expanded the use of tandem mass spectrometry combined with liquid chromatography (LC-MS/MS) for multiplex newborn screening of seven lysosomal enzymes in dried blood spots (DBS). The new assays are for enzymes responsible for the mucopolysaccharidoses (MPS-I, -II, -IIIB, -IVA, -VI, and -VII) and type 2 neuronal ceroid lipofuscinosis (LINCL). METHODS: New substrates were prepared and characterized for tripeptidyl peptidase 1 (TPP1), α-N-acetylglucosaminidase (NAGLU), and lysosomal β-glucuronidase (GUSB). These assays were combined with previously developed assays to provide a multiplex LC-MS/MS assay of 7 lysosomal storage diseases. Multiple reaction monitoring of ion dissociations for enzyme products and deuterium-labeled internal standards was used to quantify the enzyme activities. RESULTS: Deidentified DBS samples from 62 nonaffected newborns were analyzed to simultaneously determine (run time 2 min per DBS) the activities of TPP1, NAGLU, and GUSB, along with those for α-iduronidase (IDUA), iduronate-2-sulfatase (I2S), N-acetylgalactosamine-6-sulfatase (GALNS), and N-acetylgalactosamine-4-sulfatase (ARSB). The activities measured in the 7-plex format showed assay response-to-blank-activity ratios (analytical ranges) of 102-909 that clearly separated healthy infants from affected children. CONCLUSIONS: The new multiplex assay provides a robust comprehensive newborn screening assay for the mucopolysaccharidoses. The method has been expanded to include additional lysosomal storage diseases.
BACKGROUND: We expanded the use of tandem mass spectrometry combined with liquid chromatography (LC-MS/MS) for multiplex newborn screening of seven lysosomal enzymes in dried blood spots (DBS). The new assays are for enzymes responsible for the mucopolysaccharidoses (MPS-I, -II, -IIIB, -IVA, -VI, and -VII) and type 2 neuronal ceroid lipofuscinosis (LINCL). METHODS: New substrates were prepared and characterized for tripeptidyl peptidase 1 (TPP1), α-N-acetylglucosaminidase (NAGLU), and lysosomal β-glucuronidase (GUSB). These assays were combined with previously developed assays to provide a multiplex LC-MS/MS assay of 7 lysosomal storage diseases. Multiple reaction monitoring of ion dissociations for enzyme products and deuterium-labeled internal standards was used to quantify the enzyme activities. RESULTS: Deidentified DBS samples from 62 nonaffected newborns were analyzed to simultaneously determine (run time 2 min per DBS) the activities of TPP1, NAGLU, and GUSB, along with those for α-iduronidase (IDUA), iduronate-2-sulfatase (I2S), N-acetylgalactosamine-6-sulfatase (GALNS), and N-acetylgalactosamine-4-sulfatase (ARSB). The activities measured in the 7-plex format showed assay response-to-blank-activity ratios (analytical ranges) of 102-909 that clearly separated healthy infants from affected children. CONCLUSIONS: The new multiplex assay provides a robust comprehensive newborn screening assay for the mucopolysaccharidoses. The method has been expanded to include additional lysosomal storage diseases.
Authors: Zdeněk Spáčil; Susan Elliott; Steven L Reeber; Michael H Gelb; C Ronald Scott; František Tureček Journal: Anal Chem Date: 2011-05-17 Impact factor: 6.986
Authors: Naveen Kumar Chennamaneni; Arun Babu Kumar; Mariana Barcenas; Zdeněk Spáčil; C Ronald Scott; František Tureček; Michael H Gelb Journal: Anal Chem Date: 2014-04-21 Impact factor: 6.986
Authors: Xinying Hong; Arun Babu Kumar; Jessica Daiker; Fan Yi; Martin Sadilek; Fabiola De Mattia; Francesca Fumagalli; Valeria Calbi; Roberta Damiano; Maria Della Bona; Giancarlo la Marca; Adeline L Vanderver; Amy T Waldman; Laura Adang; Omar Sherbini; Sarah Woidill; Teryn Suhr; Joanne Kurtzberg; Maria L Beltran-Quintero; Maria Escolar; Alessandro Aiuti; Alan Finglas; Amber Olsen; Michael H Gelb Journal: Anal Chem Date: 2020-04-16 Impact factor: 6.986
Authors: Fan Yi; Xinying Hong; Arun Babu Kumar; Chengli Zong; Geert-Jan Boons; C Ronald Scott; Frantisek Turecek; Bruce H Robinson; Michael H Gelb Journal: Mol Genet Metab Date: 2018-05-23 Impact factor: 4.797