Determination of p53 biomarker using an electrochemical immunoassay based on layer-by-layer films with NiFe 2 O 4 nanopa
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ORIGINAL PAPER
Determination of p53 biomarker using an electrochemical immunoassay based on layer-by-layer films with NiFe2O4 nanoparticles Gisela Ibáñez-Redín 1 & Nirav Joshi 1 & Gustavo Freitas do Nascimento 1 & Deivy Wilson 1 & Matias E. Melendez 2,3 & André L. Carvalho 3 & Rui Manuel Reis 3,4,5 & Débora Gonçalves 1 & Osvaldo N. Oliveira Jr 1 Received: 1 April 2020 / Accepted: 7 October 2020 # Springer-Verlag GmbH Austria, part of Springer Nature 2020
Abstract A disposable electrochemical immunosensors is presented suitable to detect cancer biomarker p53 using screen-printed carbon electrodes modified with a layer-by-layer (LbL) matrix of carboxylated NiFe2O4 nanoparticles and polyethyleneimine, onto which anti-p53 antibodies were adsorbed. Under optimized conditions, the immunosensors exhibited high surface coverage and high concentration of immobilized antibodies, which allowed for detection of p53 in a wide dynamic range from 1.0 to 10 × 103 pg mL−1, with a limit of detection of 5.0 fg mL−1 at a working potential of 100 mV vs. Ag/AgCl. The immunosensors also exhibited good selectivity with negligible interference upon incubation in complex matrices containing high concentrations of proteins (i.e., fetal bovine serum and cell lysate). The immunosensor performance is among the best reported in the literature for determination of p53, with the additional advantage of being disposable and operating with low-volume solutions. Keywords Electrochemical immunosensors . NiFe2O4 nanoparticles . p53 . Polyethyleneimine . Cancer biomarkers
Introduction The high incidence and morbidity from cancer have created a demand for novel, reliable, and robust techniques for detection
Electronic supplementary material The online version of this article (https://doi.org/10.1007/s00604-020-04594-z) contains supplementary material, which is available to authorized users. * Nirav Joshi [email protected] * Osvaldo N. Oliveira, Jr [email protected] 1
São Carlos Institute of Physics, University of São Paulo, SP 13560-970 São Carlos, Brazil
2
Pelé Little Prince Research Institute, Little Prince Complex, PR 80250-060 Curitiba, Brazil
3
Molecular Oncology Research Center, Barretos Cancer Hospital, SP 14784-400 Barretos, Brazil
4
Life and eHealth Sciences Research Institute (ICVS), Medical School, University ofMinho, Braga, Portugal
5
ICVS/3B’s-PT Government Associate Laboratory, Braga/ Guimarães, Portugal
and quantification of biomarkers, preferably using low-cost and easily deployable devices [1]. The p53 tumor suppressor has an important role in cell growth regulation, genetic stability, and for controlling cell proliferation [2]. Mutations in the p53 gene are noted in many types of cancer, in almost 50% of human tumors [3]; they are related to accumulation of mutated protein in the nucleus of tumor cells, leading to increased concentrations in extracellular fluids such as blood, urine, and saliva [4, 5]. Therefore, the quantification of p53 protein in different matrices can be useful for clinical analysis and for monit
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