title: A high throughput electrochemiluminescence assay for the quantification of frataxin protein levels.
abstract: Friedreich's ataxia (FRDA) is an autosomal recessive neurodegenerative disease affecting 1 in 50,000 people and is caused by a GAA-trinucleotide expansion in the frataxin gene located on chromosome locus 9q13 which results in a markedly reduced expression of frataxin, a small mitochondrial protein. The exact function of frataxin is still unknown and currently there is no approved treatment available. In the near future there will be a high demand for measuring frataxin protein levels due to the development of therapeutic strategies for FRDA based on manipulating frataxin expression levels in vivo. In this paper we describe the development of an electrochemiluminescence assay (ECLIA) to measure frataxin protein levels in a 96-well plate format. The ECLIA for frataxin is able to measure human and mouse samples and is highly quantitative, accurate and reproducible, with low intra- and inter-assay error throughout a wide working range. The assay has an excellent precision and provides a new tool for the set up of high-throughput screening for basic research and for clinical studies with FRDA patients.
authors.affiliation: Medical University of Vienna, Department of Medical Chemistry, Waehringerstr. 10, 1090 Vienna, Austria. hannes.steinkellner@meduniwien.ac.at
authors.forename: Hannes
authors.initials: H
authors.lastname: Steinkellner
authors.forename: Barbara
authors.initials: B
authors.lastname: Scheiber-Mojdehkar
authors.forename: Hans
authors.initials: H
authors.lastname: Goldenberg
authors.forename: Brigitte
authors.initials: B
authors.lastname: Sturm
mesh_terms: D000818:Animals; D045744:Cell Line, Tumor; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D005621:Friedreich Ataxia / Q000175:diagnosis / Q000378:metabolism; D006367:HeLa Cells; D057166:High-Throughput Screening Assays; D006801:Humans; D033862:Iron-Binding Proteins / Q000032:analysis*; D020014:K562 Cells; D008163:Luminescent Measurements / Q000379:methods*; D051379:Mice; D000096923:Frataxin
chemical_list: D033862:Iron-Binding Proteins; D000096923:Frataxin