Citation: Xiaohui Liu, Ruoyu Ba, Fen Wan, Yang Liu, Feng Lu, Jing-Jing Zhang, Peidong Chen, Yi Zhang, Jun-Jie Zhu, Li Zhang, Fang-Fang Cheng. Ultrasensitive upconversion nanoprobe-based biosensor for dual-modal detection of thrombin and hirudin[J]. Chinese Chemical Letters, ;2026, 37(9): 112407. doi: 10.1016/j.cclet.2026.112407 shu

Ultrasensitive upconversion nanoprobe-based biosensor for dual-modal detection of thrombin and hirudin

Xiaohui Liu ^a ,
Ruoyu Ba ^a ,
Fen Wan ^a ,
Yang Liu ^a ,
Feng Lu ^c ,
Jing-Jing Zhang ^b ,
Peidong Chen ^a ,
Yi Zhang ^a ,
Jun-Jie Zhu ^{b, *,} ,
Li Zhang ^{a, *,} ,
Fang-Fang Cheng ^{a, *,}

a.
Jiangsu Collaborative Innovation Center of Chinese Medicinal Resources Industrialization, National and Local Collaborative Engineering Center of Chinese Medicinal Resources Industrialization and Formulae Innovative Medicine, Jiangsu Key Laboratory for High Technology Research of TCM Formulae, Nanjing University of Chinese Medicine, Nanjing 210023, China
b.
State Key Laboratory of Analytical Chemistry for Life Science, School of Chemistry and Chemical Engineering, Nanjing University, Nanjing 210023, China
c.
State Key Laboratory for Organic Electronics and Information Displays and Institute of Advanced Materials IAM, Nanjing University of Posts and Telecommunications, Nanjing 210023, China

jjzhu@nju.edu.cn

zhangli@njucm.edu.cn

ffcheng@njucm.edu.cn

Received Date: 28 May 2025
Revised Date: 6 November 2025
Accepted Date: 12 January 2026
Available Online: 12 January 2026

Figures(4)

Thrombin can participate in many physiological and pathological processes so that its sensitive determination is important in diagnostic and pharmaceutical fields. The content of hirudin in leech is a key criterion for determining the quality of leech. In this work, a dual-modal biosensor was constructed based on fluorescence resonance energy transfer between UCNPs and Cy5 via a thrombin-responsive peptide as a linker. Thrombin cleaved the peptide and Cy5 released, leading to the fluorescence recovery of UCNPs at a specific wavelength, which enabled the successful ratiometric fluorescence detection of thrombin. Simultaneously, Cy5 in the supernatant allowed a colorimetric detection. Hirudin inhibited the activity of thrombin, causing the intensity reduction of the corresponding fluorescence and absorbance signals. The linear ranges of the fluorescence and colorimetric detection for thrombin were 10⁻⁹ ~ 10⁻⁴ U/mL with a detection limit of 1.15 × 10⁻¹⁰ U/mL and 0.0025–5 U/mL with a detection limit of 1.12 × 10⁻³ U/mL, respectively. The linear ranges of the fluorescence and colorimetric detection for hirudin were 10⁻⁸–10⁻⁴ U/mL with a detection limit of 6.026 × 10⁻⁹ U/mL and 0.005–5 U/mL with a detection limit of 2.82 × 10⁻³ U/mL, respectively. This developed biosensor was further applied to the detection of anti-thrombin components in leech extract and the results showed that it had an excellent analytical performance, indicating this dual-modal biosensor provided a new specific and sensitive method for the detection of anti-thrombin components in practical sample, which is significant in quality evaluation of traditional Chinese medicine.
- Thrombin,
- Hirudin,
- Dual-modal,
- Upconversion nanoprobe,
- Chinese medicine
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