| Literature DB >> 29101055 |
Felipe Marques Souza de Oliveira1, Stefan Mereiter2, Peter Lönn1, Benjamin Siart3, Qiujin Shen1, Johan Heldin4, Doroteya Raykova4, Niclas G Karlsson5, Karol Polom6, Franco Roviello7, Celso A Reis8, Masood Kamali-Moghaddam9.
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.Entities:
Keywords: CD44; E-cadherin; EGFR; Gastric cancer; Glycosylation; Phosphorylation; Post-translational modifications; Protein detection; Solid-phase proximity ligation assay (SP-PLA); p53
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Year: 2017 PMID： 29101055 DOI： 10.1016/j.nbt.2017.10.005
Source DB: PubMed Journal: N Biotechnol ISSN： 1871-6784 Impact factor: 5.079