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. 2024 Mar 16;14(1):6350.
doi: 10.1038/s41598-024-56784-x.

Novel electrochemical platform based on C3N4-graphene composite for the detection of neuron-specific enolase as a biomarker for lung cancer

Zhang Junping  1 , Wei Zheng  2 , Tang ZhengFang  3 , L I Ji Yue  3 , An PengHang  3 , Zhang Mingli  4 , An Hongzhi  5
Affiliations

Novel electrochemical platform based on C3N4-graphene composite for the detection of neuron-specific enolase as a biomarker for lung cancer

Zhang Junping et al. Sci Rep. .

Abstract

Lung cancer remains the leading cause of cancer mortality worldwide. Small cell lung cancer (SCLC) accounts for 10-15% of cases and has an overall 5-years survival rate of only 15%. Neuron-specific enolase (NSE) has been identified as a useful biomarker for early SCLC diagnosis and therapeutic monitoring. This work reports an electrochemical immunosensing platform based on a graphene-graphitic carbon nitride (g-C3N4) nanocomposite for ultrasensitive NSE detection. The g-C3N4 nanosheets and graphene nanosheets were synthesized via liquid exfoliation and integrated through self-assembly to form the nanocomposite. This nanocomposite was used to modify screen-printed carbon electrodes followed by covalent immobilization of anti-NSE antibodies. The unique properties of the graphene-g-C3N4 composite facilitated efficient antibody loading while also enhancing electron transfer efficiency and electrochemical response. Systematic optimization of experimental parameters was performed. The immunosensor exhibited a wide linear detection range of 10 pg/mL to 100 ng/mL and low limit of detection of 3 pg/mL for NSE along with excellent selectivity against interferences. Real serum matrix analysis validated the applicability of the developed platform for sensitive and accurate NSE quantifica-tion at clinically relevant levels. This novel graphene-g-C3N4 nanocomposite based electro-chemical immunoassay demonstrates great promise for early diagnosis of SCLC.

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Conflict of interest statement

The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.

Figures

Figure 1
Figure 1
Fabrication process of proposed electrochemical sensor.
Figure 2
Figure 2
SEM image of (A) g-C3N4 nanosheets and (B) graphene-g-C3N4 composite.
Figure 3
Figure 3
(A) XRD patterns of g-C3N4 nanosheets and graphene-g-C3N4 composite. (B) Raman spectra of g-C3N4 nanosheets, graphene and graphene-g-C3N4 composite.
Figure 4
Figure 4
FTIR spectra of g-C3N4 nanosheets, graphene and graphene-g-C3N4 composite.
Figure 5
Figure 5
Cyclic voltammograms of bare SPCE, graphene/SPCE, g-C3N4-graphene/SPCE, and anti-NSE/g-C3N4-graphene/SPCE in 0.1 M PBS containing 5 mM [Fe(CN)6]3−/4− at a scan rate of 50 mV/s.
Figure 6
Figure 6
Nyquist plots obtained from electrochemical impedance spectroscopy of bare SPCE, graphene/SPCE, g-C3N4-graphene/SPCE, and anti-NSE/g-C3N4-graphene/SPCE in 0.1 M PBS containing 5 mM [Fe(CN)6]3−/4−. Inset shows the equivalent circuit model used for fitting the EIS data.
Figure 7
Figure 7
Effect of (A) incubation time; (B) incubation temperature; and (C) pH on the response of the immunosensor.
Figure 8
Figure 8
(A) Calibration curve of the immunosensor for different concentrations of NSE; (B) Selectivity of the immunosensor for 5 ng/mL NSE against other proteins.
Figure 9
Figure 9
(A) Correlation of determined and spiked NSE concentrations in serum samples; (B) Comparison of NSE levels in serum samples as measured by the immunosensor and ELISA.
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