Liquid chromatography-tandem mass spectrometry assay of leukocyte acid α-glucosidase for post-newborn screening evaluation of pompe disease

Na Lin, Jingyu Huang, Sara Violante, Joseph J. Orsini, Michele Caggana, Erin E. Hughes, Colleen Stevens, Lisa DiAntonio, Hsuan Chieh Liao, Xinying Hong, Farideh Ghomashchi, Arun Babu Kumar, Hui Zhou, Ruth Kornreich, Melissa P. Wasserstein, Michael H. Gelb, Chunli Yu

Research output: Contribution to journalArticle

7 Citations (Scopus)

Abstract

BACKGROUND: Pompe disease (PD) is the first lysosomal storage disorder to be added to the Recommended Uniform Screening Panel for newborn screening. This condition has a broad phenotypic spectrum, ranging from an infantile form (IOPD), with severe morbidity and mortality in infancy, to a late-onset form (LOPD) with variable onset and progressive weakness and respiratory failure. Because the prognosis and treatment options are different for IOPD and LOPD, it is important to accurately determine an individual's phenotype. To date, no enzyme assay of acid α-glucosidase (GAA) has been described that can differentiate IOPD vs LOPD using blood samples. METHODS: We incubated 10 μL leukocyte lysate and 25 μL GAA substrate and internal standard (IS) assay cocktail for 1 h. The reaction was purified by a liquid-liquid extraction. The extracts were evaporated and reconstituted in 200 μL methanol and analyzed by LC-MS/MS for GAA activity. RESULTS: A 700-fold higher analytical range was observed with the LC-MS/MS assay compared to the fluorometric method. When GAA-null and GAA-containing fibroblast lysates were mixed, GAA activity could be measured accurately even in the range of 0%-1% of normal. The leukocyte GAA activity in IOPD (n = 4) and LOPD (n = 19) was 0.44 -1.75 nmol · h-1 · mg-1 and 2.0-6.5 nmol · h-1 · mg-1, respectively, with no overlap. The GAA activity of pseudodeficiency patients ranged from 3.0 -28.1 nmol · h-1 · mg-1, showing substantial but incomplete separation from the LOPD group. CONCLUSIONS: This assay allows determination of low residual GAA activity in leukocytes. IOPD, LOPD, and pseudodeficiency patients can be partially differentiated by measuring GAA using blood samples.

Original languageEnglish (US)
Pages (from-to)842-851
Number of pages10
JournalClinical Chemistry
Volume63
Issue number4
DOIs
StatePublished - Apr 1 2017
Externally publishedYes

Fingerprint

Glycogen Storage Disease Type II
Glucosidases
Liquid chromatography
Tandem Mass Spectrometry
Liquid Chromatography
Mass spectrometry
Assays
Screening
Leukocytes
Acids
Liquid-Liquid Extraction
Blood
Enzyme Assays
Respiratory Insufficiency
Methanol
Liquids
Fibroblasts
Newborn Infant
Morbidity
Phenotype

ASJC Scopus subject areas

  • Clinical Biochemistry
  • Biochemistry, medical

Cite this

Liquid chromatography-tandem mass spectrometry assay of leukocyte acid α-glucosidase for post-newborn screening evaluation of pompe disease. / Lin, Na; Huang, Jingyu; Violante, Sara; Orsini, Joseph J.; Caggana, Michele; Hughes, Erin E.; Stevens, Colleen; DiAntonio, Lisa; Liao, Hsuan Chieh; Hong, Xinying; Ghomashchi, Farideh; Kumar, Arun Babu; Zhou, Hui; Kornreich, Ruth; Wasserstein, Melissa P.; Gelb, Michael H.; Yu, Chunli.

In: Clinical Chemistry, Vol. 63, No. 4, 01.04.2017, p. 842-851.

Research output: Contribution to journalArticle

Lin, N, Huang, J, Violante, S, Orsini, JJ, Caggana, M, Hughes, EE, Stevens, C, DiAntonio, L, Liao, HC, Hong, X, Ghomashchi, F, Kumar, AB, Zhou, H, Kornreich, R, Wasserstein, MP, Gelb, MH & Yu, C 2017, 'Liquid chromatography-tandem mass spectrometry assay of leukocyte acid α-glucosidase for post-newborn screening evaluation of pompe disease', Clinical Chemistry, vol. 63, no. 4, pp. 842-851. https://doi.org/10.1373/clinchem.2016.259036
Lin, Na ; Huang, Jingyu ; Violante, Sara ; Orsini, Joseph J. ; Caggana, Michele ; Hughes, Erin E. ; Stevens, Colleen ; DiAntonio, Lisa ; Liao, Hsuan Chieh ; Hong, Xinying ; Ghomashchi, Farideh ; Kumar, Arun Babu ; Zhou, Hui ; Kornreich, Ruth ; Wasserstein, Melissa P. ; Gelb, Michael H. ; Yu, Chunli. / Liquid chromatography-tandem mass spectrometry assay of leukocyte acid α-glucosidase for post-newborn screening evaluation of pompe disease. In: Clinical Chemistry. 2017 ; Vol. 63, No. 4. pp. 842-851.
@article{7626236e44a147fe8cf943d66fb110f5,
title = "Liquid chromatography-tandem mass spectrometry assay of leukocyte acid α-glucosidase for post-newborn screening evaluation of pompe disease",
abstract = "BACKGROUND: Pompe disease (PD) is the first lysosomal storage disorder to be added to the Recommended Uniform Screening Panel for newborn screening. This condition has a broad phenotypic spectrum, ranging from an infantile form (IOPD), with severe morbidity and mortality in infancy, to a late-onset form (LOPD) with variable onset and progressive weakness and respiratory failure. Because the prognosis and treatment options are different for IOPD and LOPD, it is important to accurately determine an individual's phenotype. To date, no enzyme assay of acid α-glucosidase (GAA) has been described that can differentiate IOPD vs LOPD using blood samples. METHODS: We incubated 10 μL leukocyte lysate and 25 μL GAA substrate and internal standard (IS) assay cocktail for 1 h. The reaction was purified by a liquid-liquid extraction. The extracts were evaporated and reconstituted in 200 μL methanol and analyzed by LC-MS/MS for GAA activity. RESULTS: A 700-fold higher analytical range was observed with the LC-MS/MS assay compared to the fluorometric method. When GAA-null and GAA-containing fibroblast lysates were mixed, GAA activity could be measured accurately even in the range of 0{\%}-1{\%} of normal. The leukocyte GAA activity in IOPD (n = 4) and LOPD (n = 19) was 0.44 -1.75 nmol · h-1 · mg-1 and 2.0-6.5 nmol · h-1 · mg-1, respectively, with no overlap. The GAA activity of pseudodeficiency patients ranged from 3.0 -28.1 nmol · h-1 · mg-1, showing substantial but incomplete separation from the LOPD group. CONCLUSIONS: This assay allows determination of low residual GAA activity in leukocytes. IOPD, LOPD, and pseudodeficiency patients can be partially differentiated by measuring GAA using blood samples.",
author = "Na Lin and Jingyu Huang and Sara Violante and Orsini, {Joseph J.} and Michele Caggana and Hughes, {Erin E.} and Colleen Stevens and Lisa DiAntonio and Liao, {Hsuan Chieh} and Xinying Hong and Farideh Ghomashchi and Kumar, {Arun Babu} and Hui Zhou and Ruth Kornreich and Wasserstein, {Melissa P.} and Gelb, {Michael H.} and Chunli Yu",
year = "2017",
month = "4",
day = "1",
doi = "10.1373/clinchem.2016.259036",
language = "English (US)",
volume = "63",
pages = "842--851",
journal = "Clinical Chemistry",
issn = "0009-9147",
publisher = "American Association for Clinical Chemistry Inc.",
number = "4",

}

TY - JOUR

T1 - Liquid chromatography-tandem mass spectrometry assay of leukocyte acid α-glucosidase for post-newborn screening evaluation of pompe disease

AU - Lin, Na

AU - Huang, Jingyu

AU - Violante, Sara

AU - Orsini, Joseph J.

AU - Caggana, Michele

AU - Hughes, Erin E.

AU - Stevens, Colleen

AU - DiAntonio, Lisa

AU - Liao, Hsuan Chieh

AU - Hong, Xinying

AU - Ghomashchi, Farideh

AU - Kumar, Arun Babu

AU - Zhou, Hui

AU - Kornreich, Ruth

AU - Wasserstein, Melissa P.

AU - Gelb, Michael H.

AU - Yu, Chunli

PY - 2017/4/1

Y1 - 2017/4/1

N2 - BACKGROUND: Pompe disease (PD) is the first lysosomal storage disorder to be added to the Recommended Uniform Screening Panel for newborn screening. This condition has a broad phenotypic spectrum, ranging from an infantile form (IOPD), with severe morbidity and mortality in infancy, to a late-onset form (LOPD) with variable onset and progressive weakness and respiratory failure. Because the prognosis and treatment options are different for IOPD and LOPD, it is important to accurately determine an individual's phenotype. To date, no enzyme assay of acid α-glucosidase (GAA) has been described that can differentiate IOPD vs LOPD using blood samples. METHODS: We incubated 10 μL leukocyte lysate and 25 μL GAA substrate and internal standard (IS) assay cocktail for 1 h. The reaction was purified by a liquid-liquid extraction. The extracts were evaporated and reconstituted in 200 μL methanol and analyzed by LC-MS/MS for GAA activity. RESULTS: A 700-fold higher analytical range was observed with the LC-MS/MS assay compared to the fluorometric method. When GAA-null and GAA-containing fibroblast lysates were mixed, GAA activity could be measured accurately even in the range of 0%-1% of normal. The leukocyte GAA activity in IOPD (n = 4) and LOPD (n = 19) was 0.44 -1.75 nmol · h-1 · mg-1 and 2.0-6.5 nmol · h-1 · mg-1, respectively, with no overlap. The GAA activity of pseudodeficiency patients ranged from 3.0 -28.1 nmol · h-1 · mg-1, showing substantial but incomplete separation from the LOPD group. CONCLUSIONS: This assay allows determination of low residual GAA activity in leukocytes. IOPD, LOPD, and pseudodeficiency patients can be partially differentiated by measuring GAA using blood samples.

AB - BACKGROUND: Pompe disease (PD) is the first lysosomal storage disorder to be added to the Recommended Uniform Screening Panel for newborn screening. This condition has a broad phenotypic spectrum, ranging from an infantile form (IOPD), with severe morbidity and mortality in infancy, to a late-onset form (LOPD) with variable onset and progressive weakness and respiratory failure. Because the prognosis and treatment options are different for IOPD and LOPD, it is important to accurately determine an individual's phenotype. To date, no enzyme assay of acid α-glucosidase (GAA) has been described that can differentiate IOPD vs LOPD using blood samples. METHODS: We incubated 10 μL leukocyte lysate and 25 μL GAA substrate and internal standard (IS) assay cocktail for 1 h. The reaction was purified by a liquid-liquid extraction. The extracts were evaporated and reconstituted in 200 μL methanol and analyzed by LC-MS/MS for GAA activity. RESULTS: A 700-fold higher analytical range was observed with the LC-MS/MS assay compared to the fluorometric method. When GAA-null and GAA-containing fibroblast lysates were mixed, GAA activity could be measured accurately even in the range of 0%-1% of normal. The leukocyte GAA activity in IOPD (n = 4) and LOPD (n = 19) was 0.44 -1.75 nmol · h-1 · mg-1 and 2.0-6.5 nmol · h-1 · mg-1, respectively, with no overlap. The GAA activity of pseudodeficiency patients ranged from 3.0 -28.1 nmol · h-1 · mg-1, showing substantial but incomplete separation from the LOPD group. CONCLUSIONS: This assay allows determination of low residual GAA activity in leukocytes. IOPD, LOPD, and pseudodeficiency patients can be partially differentiated by measuring GAA using blood samples.

UR - http://www.scopus.com/inward/record.url?scp=85017027439&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85017027439&partnerID=8YFLogxK

U2 - 10.1373/clinchem.2016.259036

DO - 10.1373/clinchem.2016.259036

M3 - Article

C2 - 28196920

AN - SCOPUS:85017027439

VL - 63

SP - 842

EP - 851

JO - Clinical Chemistry

JF - Clinical Chemistry

SN - 0009-9147

IS - 4

ER -