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Highly sensitive detection of Smoothened based on the drug binding and rolling cycle amplification

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Abstract

Metastases are the leading causes of death in cancer patients. Due to intimate connection with metastasis, Smoothened (Smo) has become a therapeutic target for antimetastatic drugs and can provide early warning of metastasis in breast cancer. Thus, we have developed an electrochemical method in Smo analysis based on small-molecule drugs. Smo on the metastatic cell surface can be internalized after combination with the small-molecule drug. The surplus small-molecule drug and rolling circle amplification (RCA) primer are competitively binding with capture probe on the electrode surface through the click chemical reaction. After RCA reaction, methylene blue is used to label the RCA product. In this process, the more Smo on the metastatic cell surface, the more RCA primer is bound with peptide on the electrode. Therefore, the obtained signal response is positively correlated to Smo on the cancer cells. Moreover, the RCA provides sufficiently high sensitivity, enabling the limit of detection of Smo to be calculated as 0.1 pM (S/N = 3). Owing to its desirable sensitivity, excellent reproducibility, and high selectivity, the proposed method may hold great potential in clinical practice in the future.

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Funding

This work is financially supported by the National Natural Science Foundation of China (Grant No. 81703088).

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  1. Jinlong Li and Kai Hu contributed equally to this work.

Authors and Affiliations

  1. Department of Laboratory Medicine, The Second Hospital of Nanjing, Nanjing University of Chinese Medicine, Nanjing, 210003, Jiangsu, China

    Jinlong Li, Yongchen Zhang & Zhaoli Zhang

  2. Department of Ophthalmology, The Nanjing Drum Tower Hospital, The Affiliated Hospital of Nanjing University Medical School, Nanjing, 210004, Jiangsu, China

    Kai Hu

  3. Institute for Advanced Interdisciplinary Research (IAIR), University of Jinan, Jinan, 250022, Shandong, China

    Hao Li

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  1. Jinlong Li
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  2. Kai Hu
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Correspondence to Jinlong Li or Hao Li.

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Li, J., Hu, K., Zhang, Y. et al. Highly sensitive detection of Smoothened based on the drug binding and rolling cycle amplification. Anal Bioanal Chem 411, 5721–5727 (2019). https://doi.org/10.1007/s00216-019-01950-8

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  • DOI: https://doi.org/10.1007/s00216-019-01950-8

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