High-Throughput Assay of 9 Lysosomal Enzymes for Newborn
Screening

J 2013

High-Throughput Assay of 9 Lysosomal Enzymes for Newborn Screening

SPÁČIL, Zdeněk, H TATIP2AKA, M BARCENAS, CR SCOTT, F TURECEK et. al.

Základní údaje

Originální název

High-Throughput Assay of 9 Lysosomal Enzymes for Newborn Screening

Autoři

SPÁČIL, Zdeněk, H TATIP2AKA, M BARCENAS, CR SCOTT, F TURECEK a MH GELB

Vydání

Clinical Chemistry, Washington, USA, Amer. Assoc. Clinical Chemistry, 2013, 0009-9147

Další údaje

Jazyk

angličtina

Typ výsledku

Článek v odborném periodiku

Utajení

není předmětem státního či obchodního tajemství

Impakt faktor

Impact factor: 7.768

DOI

http://dx.doi.org/10.1373/clinchem.2012.189936

UT WoS

000317337600010

Změněno: 29. 9. 2017 13:51, PharmDr. Zdeněk Spáčil, Ph.D.

Anotace

V originále

BACKGROUND: There is interest in newborn screening of lysosomal storage diseases (LSDs) because of the availability of treatments. Pilot studies have used tandem mass spectrometry with flow injection of samples to achieve multiplex detection of enzyme products. We report a multiplexing method of 9 enzymatic assays that uses HPLC-tandem mass spectrometry (MS/MS). METHODS: The assay of 9 enzymes was carried out in 1 or 2 buffers with a cassette of substrates and internal standards and 1 or 2 punches of a dried blood spot (DBS) from a newborn screening card as the source of enzymes. The pre-HPLC-MS/MS sample preparation required only 4 liquid transfers before injection into a dual-column HPLC equipped with switching valves to direct the flow to separation and column equilibration. Product-specific and internal standard-specific ion fragmentations were used for MS/MS quantification in the selected reaction monitoring mode. RESULTS: Analysis of blood spots from 58 random newborns and lysosomal storage disease-affected patients showed that the assay readily distinguished affected from nonaffected individuals. The time per 9-plex analysis (1.8 min) was sufficiently short to be compatible with the workflow of newborn screening laboratories. CONCLUSIONS: HPLC-MS/MS provides a viable alternative to flow-injection MS/MS for the quantification of lysosomal enzyme activities. It is possible to assay 9 lysosomal enzymes using 1 or 2 reaction buffers, thus minimizing the number of separate incubations necessary. (C) 2012 American Association for Clinical Chemistry

Citovat

SPÁČIL, Zdeněk, H TATIP2AKA, M BARCENAS, CR SCOTT, F TURECEK a MH GELB. High-Throughput Assay of 9 Lysosomal Enzymes for Newborn Screening. Clinical Chemistry. Washington, USA: Amer. Assoc. Clinical Chemistry, 2013, roč. 59, č. 3, s. 502-511. ISSN 0009-9147. Dostupné z: https://dx.doi.org/10.1373/clinchem.2012.189936.

@article{1391010,
   author = {Spáčil, Zdeněk and Tatip2aka, H and Barcenas, M and Scott, CR and Turecek, F and Gelb, MH},
   article_location = {Washington, USA},
   article_number = {3},
   doi = {http://dx.doi.org/10.1373/clinchem.2012.189936},
   language = {eng},
   issn = {0009-9147},
   journal = {Clinical Chemistry},
   title = {High-Throughput Assay of 9 Lysosomal Enzymes for Newborn Screening},
   volume = {59},
   year = {2013}
}

TY  - JOUR
ID  - 1391010
AU  - Spáčil, Zdeněk - Tatip2aka, H - Barcenas, M - Scott, CR - Turecek, F - Gelb, MH
PY  - 2013
TI  - High-Throughput Assay of 9 Lysosomal Enzymes for Newborn Screening
JF  - Clinical Chemistry
VL  - 59
IS  - 3
SP  - 502-511
EP  - 502-511
PB  - Amer. Assoc. Clinical Chemistry
SN  - 00099147
N2  - BACKGROUND: There is interest in newborn screening of lysosomal storage diseases (LSDs) because of the availability of treatments. Pilot studies have used tandem mass spectrometry with flow injection of samples to achieve multiplex detection of enzyme products. We report a multiplexing method of 9 enzymatic assays that uses HPLC-tandem mass spectrometry (MS/MS). METHODS: The assay of 9 enzymes was carried out in 1 or 2 buffers with a cassette of substrates and internal standards and 1 or 2 punches of a dried blood spot (DBS) from a newborn screening card as the source of enzymes. The pre-HPLC-MS/MS sample preparation required only 4 liquid transfers before injection into a dual-column HPLC equipped with switching valves to direct the flow to separation and column equilibration. Product-specific and internal standard-specific ion fragmentations were used for MS/MS quantification in the selected reaction monitoring mode. RESULTS: Analysis of blood spots from 58 random newborns and lysosomal storage disease-affected patients showed that the assay readily distinguished affected from nonaffected individuals. The time per 9-plex analysis (1.8 min) was sufficiently short to be compatible with the workflow of newborn screening laboratories. CONCLUSIONS: HPLC-MS/MS provides a viable alternative to flow-injection MS/MS for the quantification of lysosomal enzyme activities. It is possible to assay 9 lysosomal enzymes using 1 or 2 reaction buffers, thus minimizing the number of separate incubations necessary. (C) 2012 American Association for Clinical Chemistry
ER  -

SPÁČIL, Zdeněk, H TATIP2AKA, M BARCENAS, CR SCOTT, F TURECEK a MH GELB. High-Throughput Assay of 9 Lysosomal Enzymes for Newborn Screening. \textit{Clinical Chemistry}. Washington, USA: Amer. Assoc. Clinical Chemistry, 2013, roč.~59, č.~3, s.~502-511. ISSN~0009-9147. Dostupné z: https://dx.doi.org/10.1373/clinchem.2012.189936.

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