DNA hybridization biosensor using chitosan–carbon nanotubes composite film as an immobilization platform and [Cu(bpy)(MBZ)2(H2O)] (bpy = 2,2′-bipyridine, MBZ = p-methylbenzoate) as a novel redox indicator

In this paper, a new DNA hybridization detection strategy was developed based on the immobilization of capture probe DNA on a chitosan (CS)–carbon nanotubes (CNTs) composite modified glassy carbon electrode (CS–CNTs/GCE) and the use of a copper complex, [Cu(bpy)(MBZ)2(H2O)] (bpy = 2,2′-bipyridine, MBZ = p-methylbenzoate), as a new redox hybridization indicator. The electrochemical characterization experiments showed that the nanocomposite film of CS–CNTs could effectively immobilize the capture probe DNA and greatly improve the electron-transfer reactions of the electroactive molecules. Electrochemical and fluorescent spectroscopic analysis revealed that the polypyridyl copper complex of [Cu(bpy)(MBZ)2(H2O)] bound to DNA via a typical intercalation mode. Surface studies further showed that the copper complex can discriminate between double-stranded and single-stranded DNA that immobilized on the surface of CS–CNTs/GCE. When being utilized as a redox indicator for the detection of hybridization for short DNA species related to phosphinothricin acetyltransferase (PAT), the indicator showed good specificity for recognizing the complementary, three-base mismatched and non-complementary DNA. Under the optimized conditions, the oxidation peak currents of the copper complex enhanced linearly with increases in the concentration of the complementary sequence in the range from 5.0 × 10−10 to 1.0 × 10−8 M. A detection limit of 5.0 × 10−10 M was also obtained based on the constructed DNA biosensor.

Research highlights► Chitosan–carbon nanotubes composite was used as a platform for DNA immobilization. ► Discrimination of a synthetic copper complex to ds- and ss-DNA was obtained. ► Hybridization detection was performed using the copper complex as an indicator. ► Good specificity and high sensitivity were achieved on the constructed DNA biosensor.