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Detection of post-translational modifications using solid-phase proximity ligation assay.

Journal article
Authors Felipe Marques Souza de Oliveira
Stefan Mereiter
Peter Lönn
Benjamin Siart
Qiujin Shen
Johan Heldin
Doroteya Raykova
Niclas G. Karlsson
Karol Polom
Franco Roviello
Celso A Reis
Masood Kamali-Moghaddam
Published in New biotechnology
Volume 45
Issue October
Pages 51-59
ISSN 1876-4347
Publication year 2018
Published at Institute of Biomedicine, Department of Medical Biochemistry and Cell Biology
Pages 51-59
Language en
Links dx.doi.org/10.1016/j.nbt.2017.10.00...
www.ncbi.nlm.nih.gov/entrez/query.f...
Subject categories Medical Biotechnology, Biochemistry and Molecular Biology

Abstract

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.

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