A sandwich-type electrochemical aptasensor for the carcinoembryonic antigen via biocatalytic precipitation amplification and by using gold nanoparticle composites.

Abstract

A sandwich-type electrochemical aptasensor is described for detecting the carcinoembryonic antigen (CEA) with high sensitivity and accuracy. Two kinds of nanomaterials are used. The first was obtained by modifying gold nanoparticles with reduced graphene oxide and hemin (Hemin-rGO-AuNPs). The second consists of horseradish peroxidase-modified organic-inorganic hybrid nanoflowers linked to gold nanoparticles to obtain an architecture of type HRP-Cu3(PO4)2-HNF-AuNPs). These serve as carriers for two aptamers (apt1 and apt2) against CEA. Simultaneously, they were used to catalyze the precipitation reaction between 4-chloro-1-naphthol(4-CN) and H2O2. A sandwich-type assay linked to enzyme inhibition amplification was established for electrochemical determination of CEA. Under optimal experimental conditions and by using differential pulse voltammetry, the response peak currents (best measured at -0.34V vs. Ag/AgCl) increases linearly with the logarithm of the CEA concentration in the range between 100fgmL-1 and 100ngmL-1. The detection limit is as low as 29fgmL-1. Graphical abstract Schematic representation of the sandwich-type electrochemical aptasensor based on signal inhibition amplification from biocatalytic precipitation reaction. (HRP-Cu3(PO4)2 hybrid nanoflowers: Horseradish Peroxidase-Cu3(PO4)2 hybrid nanoflowers; AuNPs: Gold Nanoparticles; Hemin-rGO-AuNPs: Hemin-Reduced Graphene Oxide-Gold Nanoparticles; BSA: Bovine Serum Albumin; CEA: Carcinoembryonic Antigen; CEAapt1: 5'-SH-(CH2)6-ATA CCA GCT TAT TCA ATT-3'; CEAapt2: 5'-NH2-(CH2)6-AGG GGG TGA AGG GAT ACC C-3'; GCE: Glassy carbon electrode; 4-CN: 4-Chloro-1-naphthol; DPV: Differential pulse voltammetry).