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Target-triggered “signal-off” electrochemical aptasensor assisted by Au nanoparticle–modified sensing platform for high-sensitivity determination of circulating tumor cells

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Abstract

In this study, we fabricated a high-sensitivity “signal-off” electrochemical aptasensing platform for quantifying circulating tumor cells (CTCs) based on target-triggered signal readout of methylene blue (MB). Au nanoparticles (AuNPs) were introduced to enlarge the specific surface area of the gold electrode (GE), which would immobilize homogeneous and more MB-aptamers. MB-modified and stem-loop-like aptamers were assigned as a recognition element with K562 cells. Thiolated complementary strands hybridized with MB-aptamers to form double-stranded DNA (dsDNA) conformation which were further self-assembled on the surface of AuNP-modified GE, leading to a marked current peak of MB signal. In the presence of K562 cells, the MB-aptamers preferred to recognize and bind with the cells, causing the disassembly of MB-aptamers from the GE surface. Therefore, the reduced value of MB signal was related to the number of K562 cells. With the proposed aptasensor, a dynamic linear range from 1 × 102 to 1 × 106 cells mL−1 was obtained with a detection limit of 23 cells mL−1. Moreover, the aptasensor showed good selectivity, stability, and reproducibility as well as potential use in the clinical setting. Meanwhile, characterization techniques such as field-emission scanning electron microscopy, energy-dispersive X-ray spectroscopy, atomic force microscopy, cyclic voltammetry, and electrochemical impedance spectroscopy were performed to analyze the evolution of the morphology and each fabricated step of the constructed aptasensor. Our proposed aptasensor could be designed as a universal platform for CTC determination by replacing tumor cell–specific aptamers, which is a promising strategy for basic research and clinical applications.

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Funding

This study was supported by the National Natural Science Foundation of China (Grant Nos. 81430053, 81401751, 81972027), Chongqing Health Commission (2018QNXM049, 2019ZDXM025), and Medical pre-research project of the Army Medical University (2018XYY04).

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Authors and Affiliations

  1. Department of Clinical Laboratory Medicine, Southwest Hospital, Third Military Medical University (Army Medical University), 30 Gaotanyan, Shapingba District, Chongqing, 400038, China

    Yang Wang, Wenqing Zhang, Xiaoqi Tang, Yunxia Wang, Weiling Fu, Kai Chang & Ming Chen

  2. College of Pharmacy and Laboratory Medicine, Third Military Medical University (Army Medical University), 30 Gaotanyan, Shapingba District, Chongqing, 400038, China

    Yang Wang & Ming Chen

  3. State Key Laboratory of Trauma, Burn and Combined Injury, Third Military Medical University (Army Medical University), 30 Gaotanyan, Shapingba District, Chongqing, 400038, China

    Ming Chen

Authors
  1. Yang Wang
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  2. Wenqing Zhang
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  3. Xiaoqi Tang
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  4. Yunxia Wang
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  5. Weiling Fu
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  6. Kai Chang
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  7. Ming Chen
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Correspondence to Kai Chang or Ming Chen.

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The experiments were approved by the ethics committee of Southwest Hospital (China).

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Wang, Y., Zhang, W., Tang, X. et al. Target-triggered “signal-off” electrochemical aptasensor assisted by Au nanoparticle–modified sensing platform for high-sensitivity determination of circulating tumor cells. Anal Bioanal Chem 412, 8107–8115 (2020). https://doi.org/10.1007/s00216-020-02940-x

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