Development of a capillary isoelectric focusing immunoassay to measure DJ-1 isoforms in biological samples |
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| Authors: | D. Besong Agbo,H. Klafki,G. Poschmann,K. Seyfarth,J. Genius,C. Janß en,K. Stü hler,W. Wurst,H.E. Meyer,M. Klingenspor,J. Wiltfang |
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| Affiliation: | 1. LVR-Hospital Essen, Department of Psychiatry and Psychotherapy, Faculty of Medicine, University of Duisburg-Essen, 45147 Essen, Germany;2. Molecular Proteomics Laboratory, Biologisch Medizinisches Forschungszentrum (BMFZ), Heinrich Heine Universität Düsseldorf, 40225 Düsseldorf, Germany;3. Chair for Molecular Nutritional Medicine, Technische Universität München, Else Kröner-Fresenius Zentrum (EKFZ) and ZIEL, Research Center for Nutrition and Food Sciences, 85350 Freising, Germany;4. Helmholtz Center Munich, Institute for Developmental Genetics, 85764 Munich, Germany;5. Chair for Developmental Genetics, Technische Universität München, 80333 Munich, Germany;6. Molecular Neurogenetics, Max Planck Institute of Psychiatry, 80804 Munich, Germany;g Deutsches Zentrum für Neurodegenerative Erkrankungen, Standort München, 80336 Munich, Germany;h Medizinisches Proteom Center, Ruhr Universität Bochum, 44801 Bochum, Germany;i Leibniz-Institut für Analytische Wissenschaften (ISAS) e.V., 44227 Bochum, Germany |
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| Abstract: | We report on the development of a novel assay protocol for the separation and detection of charge isoforms of DJ-1 in biological samples by automated capillary isoelectric focusing followed by immunological detection. DJ-1 (PARK7) is considered as a biomarker candidate for Parkinson’s disease and may potentially support the differentiation of clinical subtypes of the disease. The new method allows for separation and subsequent relative quantitative comparison of different isoforms of DJ-1 in biological samples. The assay was successfully applied to the analysis of DJ-1 isoform patterns in brains from mice subjected to normal or high-fat diet and revealed statistically significant group differences. Furthermore, in a pooled and concentrated sample of human cerebrospinal fluid that was depleted of albumin and immunoglobulin G, four different charge variants of DJ-1 could be detected. Taken together, the capillary isoelectric focusing immunoassay for DJ-1 represents a promising tool that may ultimately serve in clinical biomarker studies. |
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| Keywords: | DJ-1 PARK7 Isoelectric focusing Immunoassay CSF Biomarker |
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