A novel electrochemical immunosensor based on Au nanoparticles and horseradish peroxidase signal amplification for ultrasensitive detection of α-fetoprotein

  • Dingqiang Lu
  • Qiuda Xu
  • Guangchang PangEmail author
  • Fuping LuEmail author


An electrochemical double-layer Au nanoparticle membrane immunosensor was developed using an electrochemical biosensing signal amplification system with Au nanoparticles, thionine, chitosan, and horseradish peroxidase, which was fabricated using double self-adsorption of Au nanoparticle sol followed by anti-α-fetoprotein Balb/c mouse monoclonal antibody adsorption. The AuNPs sol was characterized by spectrum scanning and transmission electron microscopy. The immunosensor was characterized by atomic force microscopy, cyclic voltammetry, and alternating-current impedance during each stage of adsorption and assembly. The amperometric I-t curve method was used to measure α-fetoprotein (AFP) diluted in phosphate buffered saline. The result indicated a wide linear range, and the change rate of steady-current before and after immune response had linear correlation within the range 0.1–104 pg/mL AFP. The current change rate equation was △I = 5.82334 lgC + 37.01195 (R2 = 0.9922). The lowest limit of detection was 0.03 pg/mL (S/N = 3), and the reproducibility of the sensor was good. Additionally, the sensor could be stably stored above phosphate buffered saline at 4 °C for more than 24 days. More importantly, the sensor is label-free, reagentless and low fouling, making it capable of assaying AFP in real serum samples without suffering from significant interference or biofouling.


α-fetoprotein Au nanoparticles Amperometric I-t curve method Electrochemical immunosensor Horseradish peroxidase Membrane potential 



This research was financially supported by “The National Natural Science Foundation of China (grant numbers: 31671857, 31371773)”.

Compliance with ethical standards

Conflict of interests

The authors declare that there is no conflict of interests regarding the publication of this paper.


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Authors and Affiliations

  1. 1.College of BiotechnologyTianjin University of Science &TechnologyTianjinChina
  2. 2.Key Laboratory of Industrial Fermentation Microbiology, Ministry of EducationTianjinChina
  3. 3.College of Biotechnology & Food ScienceTianjin University of CommerceTianjinChina
  4. 4.Tianjin Key Laboratory of Food BiotechnologyTianjinChina

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