An innovative immunosensor for detection of tumor suppressor protein p53 in unprocessed human plasma and cancer cell lysates

Abstract

An innovative mediator-free electrochemical immunosensor for quantitation of p53 tumor suppressor protein based on signal amplification strategy was fabricated. In this work, biotin conjugated p53-antibody (anti-p53) was immobilized onto a green and biocompatible nanocomposite containing poly l-cysteine (P-Cys) as conductive matrix and 3D gold nanoparticles (GNPs) as signal amplification element. Therefore, a novel nanocomposite film based on P-Cys and GNPs was exploited to develop a highly sensitive immunosensor for detection of p53 protein. Importantly, GNPs prepared by sonoelectrodeposition method which lead to compact morphology. Fully electrochemical methodology was used to prepare a new transducer on a gold surface which provided a high surface area to immobilize a high amount of the anti-p53. The surface morphology of electrode was characterized by high-resolution field emission scanning electron microscope (FE-SEM) and energy dispersive spectroscopy (EDX). The immunosensor was employed for the detection of p53 in physiological pH using square wav voltammetry and differential pulse voltammetry (DPVs) techniques. Under optimized condition the calibration curve for p53 concentration by SWV and DPV was linear in 0.0369–50pM and 0.018–2.5pM with lower limit of quantification of 48fM and 18fM, respectively. The method was successfully applied assay of the p53 in unprocessed human plasma samples. Also, the method was applied to the assay of p53 in human plasma sample and normal and malignant cell line lysates such as (L929 normal cell Line from mouse C3H (L929), colon cancer cell-HCT, prostate cancer cell line PC-3, and human breast adenocarcinoma cell line-MCF7).