Graphene-based immunosensor for electrochemical quantification of phosphorylated p53 (S15)
We reported a graphene-based immunosensor for electrochemical quantification of phosphorylated p53 on serine 15 (phospho-p5315), a potential biomarker of gamma-radiation exposure. The principle is based on sandwich immunoassay and the resulting immunocomplex is formed among phospho-p53 capture antibody, phospho-p5315 antigen, biotinylated phospho-p5315 detection antibody and horseradish peroxidase (HRP)-labeled streptavidin. The introduced HRP results in an electrocatalytic response to reduction of hydrogen peroxide in the presence of thionine. Graphene served as sensor platform not only promotes electron transfer, but also increases the surface area to introduce a large amount of capture antibody, thus increasing the detection sensitivity. The experimental conditions including blocking agent, immunoreaction time and substrate concentration have been optimized. Under the optimum conditions, the increase of response current is proportional to the phospho-p5315 concentration in the range of 0.2–10 ng mL-1, with the detection limit of 0.1 ng mL-1. The developed immunosensor exhibits acceptable stability and reproducibility and the assay results for phospho-p5315 are in good correlation with the known values. This easily fabricated immunosensor provides a new promising tool for analysis of phospho-p5315 and other phosphorylated proteins.
- Research Organization:
- Pacific Northwest National Lab. (PNNL), Richland, WA (United States)
- Sponsoring Organization:
- USDOE
- DOE Contract Number:
- AC05-76RL01830
- OSTI ID:
- 1021833
- Report Number(s):
- PNNL-SA-80745; ACACAM; 400412000; TRN: US201117%%406
- Journal Information:
- Analytica Chimica Acta, Vol. 699, Issue 1; ISSN 0003-2670
- Publisher:
- Elsevier
- Country of Publication:
- United States
- Language:
- English
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