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

Oliveira FMS, Mereiter S, Lönn P, Siart B, Shen Q, Heldin J, Raykova D, Karlsson NG, Polom K, Roviello F, Reis CA, Kamali-Moghaddam M

N Biotechnol 45 (-) 51-59 [2018-10-25; online 2017-10-31]

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.

Affinity Proteomics Uppsala [Technology development]

Glycoproteomics and MS Proteomics [Technology development]

PLA and Single Cell Proteomics [Technology development]

PubMed 29101055

DOI 10.1016/j.nbt.2017.10.005

Crossref 10.1016/j.nbt.2017.10.005

pii: S1871-6784(17)30423-5

Publications 9.5.0