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Detection of post-translational modifications using solid-phase proximity ligation assay
Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Immunology, Genetics and Pathology, Molecular tools.
i3S – Instituto de Investigação e Inovação em Saúde and IPATIMUP – Institute of Molecular Pathology and Immunology of the University of Porto, Portugal.
Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Immunology, Genetics and Pathology, Molecular tools.
Department of Anthropology, University of Vienna, Austria; Department of Behavioral Biology, University of Vienna, Austria .
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2017 (English)In: New Biotechnology, ISSN 1871-6784, E-ISSN 1876-4347Article in journal (Refereed) Epub ahead of print
Abstract [en]

Post-translational modifications (PTMs) regulate protein activities to help orchestrate and fine-tune cellular processes. Dysregulation of PTMs is often related with disorders and malignancies, and may serve as a precise biomarker of disease. Developing sensitive tools to measure and monitor low-abundant PTMs in tissue lysates or serum will be instrumental for opening up new PTM-based diagnostic avenues. Here, we investigate the use of solid-phase proximity ligation assay (SP-PLA) for detection of different PTMs. The assay depends on the recognition of the target protein molecule and its modification by three affinity binders. Using antibodies and lectins, we applied the method for detection of glycosylated CD44 and E-Cadherin, and phosphorylated p53 and EGFR. The assay was found to have superior dynamic range and limit of detection compared to standard ELISAs. In summary, we have established the use of SP-PLA as an appropriate method for sensitive detection of PTMs in lysates and sera, which may provide a basis for future PTM-based diagnostic and prognostic biomarkers

Place, publisher, year, edition, pages
2017.
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Other Medical Sciences Medical Biotechnology (with a focus on Cell Biology (including Stem Cell Biology), Molecular Biology, Microbiology, Biochemistry or Biopharmacy)
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URN: urn:nbn:se:uu:diva-334520DOI: 10.1016/j.nbt.2017.10.005OAI: oai:DiVA.org:uu-334520DiVA: diva2:1159731
Available from: 2017-11-23 Created: 2017-11-23 Last updated: 2017-11-24

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de Oliveira, Felipe
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Molecular toolsDepartment of Pharmaceutical Biosciences
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