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Anal Chem. 2018 Feb 6;90(3):2216-2223. doi: 10.1021/acs.analchem.7b04590. Epub 2018 Jan 11.

Liquid Chromatography-High Resolution Mass Spectrometry Analysis of Platelet Frataxin as a Protein Biomarker for the Rare Disease Friedreich's Ataxia.

Author information

1
Penn SRP Center and Center of Excellence in Environmental Toxicology Center, Department of Systems Pharmacology and Translational Therapeutics, Perelman School of Medicine, University of Pennsylvania , Philadelphia, Pennsylvania 19104, United States.
2
Penn/CHOP Center of Excellence in Friedreich's Ataxia , Philadelphia, Pennsylvania 19104, United States.
3
Departments of Pediatrics and Neurology, Children's Hospital of Philadelphia , Philadelphia, Pennsylvania 19104, United States.
4
Departments of Pediatrics and Neurology, Perelman School of Medicine, University of Pennsylvania , Philadelphia, Pennsylvania 19104, United States.
5
Department of Medicine, Division of Hematology-Oncology, Perelman School of Medicine, University of Pennsylvania , Philadelphia, Pennsylvania 19104, United States.

Abstract

Friedreich's ataxia (FA) is an autosomal recessive disease caused by an intronic GAA triplet expansion in the FXN gene, leading to reduced expression of the mitochondrial protein frataxin. FA is estimated to affect 1 in 50 000 with a mean age of death in the fourth decade of life. There are no approved treatments for FA, although experimental approaches, which involve up-regulation or replacement of frataxin protein, are being tested. Frataxin is undetectable in serum or plasma, and whole blood cannot be used because it is present in long-lived erythrocytes. Therefore, an assay was developed for analyzing frataxin in platelets, which have a half-life of 10 days. The assay is based on stable isotope dilution immunopurification two-dimensional nano-ultra high performance liquid chromatography/parallel reaction monitoring/mass spectrometry. The lower limit of quantification was 0.078 pg frataxin/μg protein, and the assay had 100% sensitivity and specificity for discriminating between controls and FA cases. The mean levels of control and FA platelet frataxin were 9.4 ± 2.6 and 2.4 ± 0.6 pg/μg protein, respectively. The assay should make it possible to rigorously monitor the effects of therapeutic interventions on frataxin expression in this devastating disease.

PMID:
29272104
PMCID:
PMC5817373
DOI:
10.1021/acs.analchem.7b04590
[Indexed for MEDLINE]
Free PMC Article

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