Nanofluidics-Based Mass Spectrometry. Applications for Biomarker Discovery in Lysosomal Storage Diseases

2014 ◽  
pp. 137-165 ◽  
Author(s):  
Mirela Sarbu ◽  
Alina D. Zamfir
Keyword(s):  
Mass Spectrometry ◽  
Biomarker Discovery ◽  
Lysosomal Storage Diseases ◽  
Lysosomal Storage ◽  
Storage Diseases

scholarly journals Tandem Mass Spectrometry Has a Larger Analytical Range than Fluorescence Assays of Lysosomal Enzymes: Application to Newborn Screening and Diagnosis of Mucopolysaccharidoses Types II, IVA, and VI

2015 ◽  
Vol 61 (11) ◽  
pp. 1363-1371 ◽  
Author(s):  
Arun Babu Kumar ◽  
Sophia Masi ◽  
Farideh Ghomashchi ◽  
Naveen Kumar Chennamaneni ◽  
Makoto Ito ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Newborn Screening ◽  
Lysosomal Enzymes ◽  
Lysosomal Storage Diseases ◽  
Tandem Mass ◽  
Lysosomal Storage ◽  
Storage Diseases ◽  
Mps Ii ◽  
Analytical Range ◽  
Screening And Diagnosis

Abstract BACKGROUND There is interest in newborn screening and diagnosis of lysosomal storage diseases because of the development of treatment options that improve clinical outcome. Assays of lysosomal enzymes with high analytical range (ratio of assay response from the enzymatic reaction divided by the assay response due to nonenzymatic processes) are desirable because they are predicted to lead to a lower rate of false positives in population screening and to more accurate diagnoses. METHODS We designed new tandem mass spectrometry (MS/MS) assays that give the largest analytical ranges reported to date for the use of dried blood spots (DBS) for detection of mucopolysaccharidoses type II (MPS-II), MPS-IVA, and MPS-VI. For comparison, we carried out fluorometric assays of 6 lysosomal enzymes using 4-methylumbelliferyl (4MU)-substrate conjugates. RESULTS The MS/MS assays for MPS-II, -IVA, and -VI displayed analytical ranges that are 1–2 orders of magnitude higher than those for the corresponding fluorometric assays. The relatively small analytical ranges of the 4MU assays are due to the intrinsic fluorescence of the 4MU substrates, which cause high background in the assay response. CONCLUSIONS These highly reproducible MS/MS assays for MPS-II, -IVA, and -VI can support multiplex newborn screening of these lysosomal storage diseases. MS/MS assays of lysosomal enzymes outperform 4MU fluorometric assays in terms of analytical range. Ongoing pilot studies will allow us to gauge the impact of the increased analytical range on newborn screening performance.

scholarly journals Oligosaccharide Analysis in Urine by MALDI-TOF Mass Spectrometry for the Diagnosis of Lysosomal Storage Diseases

2013 ◽  
Vol 59 (9) ◽  
pp. 1357-1368 ◽  
Author(s):  
Baoyun Xia ◽  
Ghazia Asif ◽  
Leonard Arthur ◽  
Muhammad A Pervaiz ◽  
Xueli Li ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Genetic Diseases ◽  
Age Groups ◽  
Time Of Flight ◽  
Lysosomal Storage Diseases ◽  
Lysosomal Storage ◽  
Storage Diseases ◽  
Maldi Tof ◽  
Specificity And Sensitivity ◽  
Free Oligosaccharides

BACKGROUND There are 45 known genetic diseases that impair the lysosomal degradation of macromolecules. The loss of a single lysosomal hydrolase leads to the accumulation of its undegraded substrates in tissues and increases of related glycoconjugates in urine, some of which can be detected by screening of free oligosaccharides (FOS) in urine. Traditional 1-dimensional TLC for urine oligosaccharide analysis has limited analytical specificity and sensitivity. We developed fast and robust urinary FOS and glycoaminoacid analyses by MALDI–time-of-flight/time-of-flight (MALDI-TOF/TOF) mass spectrometry for the diagnosis of oligosaccharidoses and other lysosomal storage diseases. METHODS The FOS in urine equivalent to 0.09 mg creatinine were purified through sequential passage over a Sep-Pak C18 column and a carbograph column and were then permethylated. MALDI-TOF/TOF was used to analyze the permethylated FOS. We studied urine samples from individuals in 7 different age groups ranging from 0–1 months to ≥17 years as well as urine from known patients with different lysosomal storage diseases. RESULTS We identified diagnostic urinary FOS patterns for α-mannosidosis, galactosialidosis, mucolipidosis type II/III, sialidosis, α-fucosidosis, aspartylglucosaminuria (AGU), Pompe disease, Gaucher disease, and GM1 and GM2 gangliosidosis. Interestingly, the increase in urinary FOS characteristic of lysosomal storage diseases relative to normal FOS appeared to correlate with the disease severity. CONCLUSIONS The analysis of urinary FOS by MALDI-TOF/TOF is a powerful tool for first-tier screening of oligosaccharidoses and lysosomal storage diseases.

scholarly journals Mass Spectrometry-based Protein Profiling to Determine the Cause of Lysosomal Storage Diseases of Unknown Etiology

2009 ◽  
Vol 8 (7) ◽  
pp. 1708-1718 ◽  
Author(s):  
David E. Sleat ◽  
Lin Ding ◽  
Shudan Wang ◽  
Caifeng Zhao ◽  
Yanhong Wang ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Lysosomal Storage Diseases ◽  
Protein Profiling ◽  
Unknown Etiology ◽  
Lysosomal Storage ◽  
Storage Diseases

scholarly journals Pilot study of newborn screening for six lysosomal storage diseases using Tandem Mass Spectrometry

2016 ◽  
Vol 118 (4) ◽  
pp. 304-309 ◽  
Author(s):  
Susan Elliott ◽  
Norman Buroker ◽  
Jason J. Cournoyer ◽  
Anna M. Potier ◽  
Joseph D. Trometer ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Pilot Study ◽  
Tandem Mass Spectrometry ◽  
Newborn Screening ◽  
Lysosomal Storage Diseases ◽  
Tandem Mass ◽  
Lysosomal Storage ◽  
Storage Diseases

scholarly journals Diagnosing lysosomal storage diseases in a Brazilian non-newborn population by tandem mass spectrometry

Clinics ◽  
2013 ◽  
Vol 68 (11) ◽  
pp. 1469-1473 ◽  
Author(s):  
GD Brand ◽  
HC Matos ◽  
GC Cruz ◽  
NC Fontes ◽  
M Buzzi ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Tandem Mass Spectrometry ◽  
Lysosomal Storage Diseases ◽  
Tandem Mass ◽  
Lysosomal Storage ◽  
Storage Diseases

scholarly journals Multiplex Tandem Mass Spectrometry Enzymatic Activity Assay for Newborn Screening of the Mucopolysaccharidoses and Type 2 Neuronal Ceroid Lipofuscinosis

2017 ◽  
Vol 63 (6) ◽  
pp. 1118-1126 ◽  
Author(s):  
Yang Liu ◽  
Fan Yi ◽  
Arun Babu Kumar ◽  
Naveen Kumar Chennamaneni ◽  
Xinying Hong ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Tandem Mass Spectrometry ◽  
Newborn Screening ◽  
Neuronal Ceroid Lipofuscinosis ◽  
Lysosomal Storage Diseases ◽  
Tandem Mass ◽  
Lysosomal Storage ◽  
Storage Diseases ◽  
Ceroid Lipofuscinosis

Abstract 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.

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