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Testing the feasibility of fully automated chip‐based nanoelectrospray ionization mass spectrometry as a novel tool for rapid diagnosis of Fabry disease

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2013

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Flangea, Corina
Mosoarca, Cristina
Galusca, Mirela
Zamfir, Alina D.

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Electrophoresis. 2013, 34(11), pp. 1572-1580. ISSN 0173-0835. eISSN 1522-2683. Available under: doi: 10.1002/elps.201200665

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Fabry condition, a lysosomal storage disease (LSD) is characterized by the absence or reduction of the α-galactosidase A activity. Recently, a new diagnostic method for detection of α-galactosidase activity from dried blood spots (DBS) using a chemical substrate and quantification of reaction mixture was developed. To improve this method in the terms of automation, reproducibility, sensitivity, and data reliability, we introduce here an innovative analytical approach based on chip-nanoESI MS. The α-galactosidase assay products derived from DBS of 11 healthy donors and 11 Fabry disease patients were analyzed by NanoMate robot coupled to a high-capacity ion trap MS. Confirmation and structural analysis of the reaction products was achieved by CID and electron transfer dissociation (ETD) MS/MS. The cleavage of a substrate GLA-S generated a product, GLA-P, which was quantified related to an internal standard GLA-IS. Comparative patient versus control analysis indicated a 13-fold reduction in GLA-P/GLA-IS ratio in the case of the patients. Moreover, our method provided direct data on the enzyme, from which it was for the first time possible to discriminate between the patients lacking the enzyme and those presenting a less active one. GLA-IS and GLA-P were confirmed by CID/ETD, which applied together, increased considerably the sequence coverage and provided complementary information for unambiguous product identification. The present chip-nanoESI CID and ETD MS(n) strategy introduced here for first time in LSD diagnosis, provided a maximum confidence in assay product identification, a high sensitivity, speed of analysis, and result reproducibility.

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ISO 690FLANGEA, Corina, Cristina MOSOARCA, Claudia COZMA, Mirela GALUSCA, Michael PRZYBYLSKI, Alina D. ZAMFIR, 2013. Testing the feasibility of fully automated chip‐based nanoelectrospray ionization mass spectrometry as a novel tool for rapid diagnosis of Fabry disease. In: Electrophoresis. 2013, 34(11), pp. 1572-1580. ISSN 0173-0835. eISSN 1522-2683. Available under: doi: 10.1002/elps.201200665
BibTex
@article{Flangea2013-06Testi-42254,
  year={2013},
  doi={10.1002/elps.201200665},
  title={Testing the feasibility of fully automated chip‐based nanoelectrospray ionization mass spectrometry as a novel tool for rapid diagnosis of Fabry disease},
  number={11},
  volume={34},
  issn={0173-0835},
  journal={Electrophoresis},
  pages={1572--1580},
  author={Flangea, Corina and Mosoarca, Cristina and Cozma, Claudia and Galusca, Mirela and Przybylski, Michael and Zamfir, Alina D.}
}
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