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Nanostructured poly(thiophene acetic acid)/Au/poly(methylene blue) interface for electrochemical immunosensing of p53 protein

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Abstract

A poly(thiophene acetic acid)/Au/poly(methylene blue) nanostructured interface was electrochemically assembled step-by-step on screen-printed carbon electrodes (SPCE) for label-free detection of p53 protein. The initial electrical conductive properties of the polymeric interface were increased with an additional layer of poly(methylene blue) electropolymerized in the presence of gold nanoparticles. The nano-immunosensing architecture was prepared by covalent immobilization of anti-p53 antibodies as bioreceptors through the poly(thiophene acetic acid) moieties. The nano-immunosensor assembly was extensively characterized by ultraviolet–visible spectrophotometry, dynamic and electrophoretic light scattering, scanning electron microscopy, X-ray photoelectron spectroscopy, Raman spectroscopy, atomic force microscopy, cyclic voltammetry, and electrochemical impedance spectroscopy. Under optimal conditions, p53 was specifically and selectively detected by square wave voltammetry in a linear range between 1 and 100 ng mL−1 with a limit of detection of 0.65 ng mL−1. In addition, the electrochemical nano-immunosensor detected p53 in spiked human serum samples and colorectal cancer cell lysates, and the results were validated with a standard spectrophotometric method using a paired samples t test, which did not exhibit significant differences between both methods. The resultant p53 nano-immunosensor is simple to assemble, robust, and has the potential for point-of-care biomarker detection applications.

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Acknowledgements

We thank the Ruta N Complex and EPM for hosting the Max Planck Tandem Groups. Finally, we thank Dr. Ernesto Moreno and Dr. Marcela Rubio from the University of Medellín for donating cell lines.

Funding

The work has been funded by Minciencias, Mineducación, MINCIT, and ICETEX through the Program Ecosistema Científico Cod. FP44842-211–2018, project number 58536. J.O. thanks support from the University of Antioquia and the Max Planck Society through the cooperation agreement 566–1, 2014.

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  1. Max Planck Tandem Group in Nanobioengineering, Institute of Chemistry, Faculty of Natural and Exact Sciences, University of Antioquia, Complejo Ruta N, Calle 67 No. 52-20, 050010, Medellín, Colombia

    Andrés F. Cruz-Pacheco, Jennifer Quinchia & Jahir Orozco

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Andrés F. Cruz-Pacheco, conceptualization, methodology, formal analysis, investigation, data curation, and writing, original draft. Jennifer Quinchia, investigation, data curation, and writing, original draft. Jahir Orozco, conceptualization; formal analysis; writing, review and editing; supervision, project administration; and funding acquisition.

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Correspondence to Jahir Orozco.

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Cruz-Pacheco, A.F., Quinchia, J. & Orozco, J. Nanostructured poly(thiophene acetic acid)/Au/poly(methylene blue) interface for electrochemical immunosensing of p53 protein. Microchim Acta 190, 136 (2023). https://doi.org/10.1007/s00604-023-05683-5

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