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. 2024 Sep 18;14(9):446.
doi: 10.3390/bios14090446.

Ultrasensitive Electrochemical Biosensors Based on Allosteric Transcription Factors (aTFs) for Pb2+ Detection

Ningkang Yu  1   2 , Chen Zhao  2 , Xiaodan Kang  1   2 , Cheng Zhang  2 , Xi Zhang  2 , Chenyu Li  2 , Shang Wang  2 , Bin Xue  2 , Xiaobo Yang  2 , Chao Li  2 , Zhigang Qiu  2 , Jingfeng Wang  2 , Zhiqiang Shen  1   2
Affiliations

Ultrasensitive Electrochemical Biosensors Based on Allosteric Transcription Factors (aTFs) for Pb2+ Detection

Ningkang Yu et al. Biosensors (Basel). .

Abstract

Exposure to Pb2+ in the environment, especially in water, poses a significant threat to human health and urgently necessitates the development of highly sensitive Pb2+ detection methods. In this study, we have integrated the high sensitivity of electrochemical techniques with allosteric transcription factors (aTFs) to develop an innovative electrochemical biosensing platform. This biosensors leverage the specific binding and dissociation of DNA to the aTFs (PbrR) on electrode surfaces to detect Pb2+. Under the optimal conditions, the platform has a broad linear detection range from 1 pM to 10 nM and an exceptionally low detection threshold of 1 pM, coupled with excellent selectivity for Pb2+. Notably, the biosensor demonstrates regenerative capabilities, enabling up to five effective Pb2+ measurements. After one week of storage at 4 °C, effective lead ion detection was still possible, demonstrating the biosensor's excellent stability, this can effectively save the cost of detection. The biosensor also achieves a recovery rate of 93.3% to 106.6% in real water samples. The biosensor shows its potential as a robust tool for the ultrasensitive detection of Pb2+ in environmental monitoring. Moreover, this research provides new insights into the future applications of aTFs in electrochemical sensing.

Keywords: Pb2+; PbrR; allosteric transcription factors (aTFs); electrochemical.

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Conflict of interest statement

The authors declare no conflicts of interest.

Figures

Figure 1
Figure 1
The principle of electrochemical biosensors based on aTFs.
Figure 2
Figure 2
Characterization of the electrochemical biosensor for Pb2+ detection based on aTFs by (A) Cyclic voltammograms, (B) Nyquist diagrams and (C) Square-wave voltammogram.
Figure 3
Figure 3
Optimization of electrochemical biosensors based on aTFs. (A) Signal response at different DNA concentrations. (B) Electrochemical signals of reactions with different PbrR concentrations. (C) Signal changes for different incubation times.
Figure 4
Figure 4
Quantitative analysis and selectivity of biosensors. (A,B). Signal change values of the biosensor after 10 min of incubation with different concentrations of Pb2+. (C). The linear relationship between the signal change and the concentration of Pb2+. (D). The biosensor signal changes after 10 min of incubation with different heavy metal ions. The concentration of all interfering heavy metal ions was 1 nM, and that of Pb2+ was 1 pM. Error bars; SD, n = 3. (* p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001).
Figure 5
Figure 5
Regeneration and stability of biosensors. (A) Regeneration of biosensors. Where N represents the number of regenerations. (B) Stability of biosensors. Error bars; SD, n = 3. (* p < 0.05, ** p < 0.01, *** p < 0.001).
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Grants and funding

This research received financial support from the China National Key R&D Program (grant no. 2018YFC1603702).

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