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Electrochemical detection of different p53 conformations by using nanostructured surfaces
Sarah Tonello1, Francesca Stradolini2, Giulia Abate3
1Department of Information Engineering, University of Brescia, Brescia, Italy. s.tonello@unibs.it.
Scientific Reports
|November 24, 2019
Summary
This study introduces novel electrochemical methods to analyze the metalloprotein p53 and its redox forms. These techniques offer a cost-effective alternative to traditional assays for protein analysis.
Area of Science:
- Electrochemistry
- Biochemistry
- Protein Science
Background:
- Protein electrochemistry is vital for studying protein structure and function.
- Existing biochemical assays have limitations in assessing protein conformational states and are costly.
Purpose of the Study:
- To explore electrochemical methods for investigating the metalloprotein p53 and its redox products.
- To compare label-free and label-based electrochemical approaches for p53 analysis.
- To evaluate different electrode materials for p53 detection.
Main Methods:
- Utilized label-free direct electrochemistry and label-based antibody-specific assays.
- Qualitatively analyzed p53 redox products on carbon electrodes.
- Quantitatively analyzed denatured p53 using various carbon and platinum electrodes (bulk and nanostructured).
Main Results:
- Successfully discriminated four p53 forms (wild type to denatured).
- Label-free analysis indicated single electron exchange with electron transfer rate constants around 1 s-1.
- Label-based analysis revealed reduced pAb240 affinity for denatured, oxidized, and nitrated p53.
- Platinum nanostructured electrodes enhanced the limit of detection to 100 ng/ml.
Conclusions:
- Developed and validated electrochemical methods for p53 analysis.
- Demonstrated the potential of label-free and label-based approaches for discriminating p53 redox states.
- Highlighted platinum nanostructured electrodes for improved sensitivity in p53 quantification.
- Paved the way for integrated devices for clinical protein analysis beyond simple quantification.
