Ultrasensitive detection of NSE employing a novel electrochemical immunosensor based on a conjugated copolymer

Abstract

In the current study a simple and highly specific label-free impedimetric neuron specific enolase (NSE) immunosensor based on a copolymer matrix-coated disposable electrode was designed and tested. The copolymer matrix was prepared using a very conductive EDOT monomer and semi-conductive thiophene-bearing epoxy groups (ThEp), and the combination of the two monomers enhanced the conductivity and protein loading capacity of the electrode surface. The P(ThEp-co-EDOT) copolymer matrix was prepared via a drop-casting process and anti-NSE recognition biomolecules were immobilized directly on the epoxy groups of the copolymer. After the coupling of NSE molecules on the P(ThEp-co-EDOT) copolymer matrix-coated electrode surface, the charge transfer resistance (R_ct) of the biosensor changed dramatically. These changes in R_ct were proportional to the NSE molecule amounts captured by anti-NSE molecules. Under optimized experimental conditions, the increment in the R_ct value was proportional to the NSE concentration over a range of 0.01 to 25 pg mL⁻¹ with a detection limit (LOD) of 2.98 × 10⁻³ pg mL⁻¹. This copolymer-coated electrode provided a lower LOD than the other biosensors. In addition, the suggested electrochemical immuno-platform showed good selectivity, superior reproducibility, long-term stability, and high recovery of NSE in real serum (95.64–102.20%) and saliva (95.28–105.35%) samples. These results showed that the present system had great potential for electrochemical biosensing applications.