Analytical and Bioanalytical Chemistry

, Volume 410, Issue 20, pp 4991–4999 | Cite as

Molecular switch-modulated fluorescent copper nanoclusters for selective and sensitive detection of histidine and cysteine

  • Zefeng Gu
  • Zhijuan CaoEmail author
Research Paper


A novel assay for histidine and cysteine has been constructed based on modulation of fluorescent copper nanoclusters (CuNCs) by molecular switches. In our previous work, a dumbbell DNA template with a poly-T (thymine) loop has been developed as an excellent template for the formation of strongly fluorescent CuNCs. Herein, for the first time, we established this biosensor for sensing two amino acids by using dumbbell DNA-templated CuNCs as the single probe. Among 20 natural amino acids, only histidine and cysteine can selectively quench fluorescence emission of CuNCs, because of the specific interaction of these compounds with copper ions. Furthermore, by using nickel ions (Ni2+) and N-ethylmaleimide as the masking agents for histidine and cysteine respectively, an integrated logic gate system was designed by coupling with the fluorescent CuNCs and demonstrated selective and sensitive detection of cysteine and histidine. Under optimal conditions, cysteine can be detected in the concentration ranges of 0.01–10.0 μM with the detection limit (DL) of as low as 98 pM, while histidine can be detected in the ranges of 0.05–40.0 μM with DL of 1.6 nM. In addition, histidine and cysteine can be observed with the naked eye under a hand-held UV lamp (DL, 50 nM), which can be easily adapted to automated high-throughput screening. Finally, the strategy has been successfully utilized for biological fluids. The proposed system can be conducted in homogeneous solution, eliminating the need for organic cosolvents, separation processes of nanomaterials, or any chemical modifications. Overall, the assay provides an alternative method for simultaneous detection of cysteine and histidine by taking the advantages of high speed, no label and enzyme requirement, and good sensitivity and specificity, and will satisfy the great demand for determination of amino acids in fields such as food processing, biochemistry, pharmaceuticals, and clinical analysis.

Graphical abstract


Dumbbell DNA template Histidine Cysteine Logic gate Fluorescent copper nanoclusters 


Funding information

The study was financially supported by the National Natural Science Foundation of China (No. 21505023).

Compliance with ethical standards

Conflict of interest

The authors declare that they have no competing interests.

Human participation and rights

This study was approved by the Institutional Review Board at Fudan University, and the experiments were performed in accordance with the ethical standards.

Informed consent

The authors declare that urine samples were collected at the Fudan University and obtained with informed consent.

Supplementary material

216_2018_1149_MOESM1_ESM.pdf (617 kb)
ESM 1 (PDF 616 kb)


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Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  1. 1.School of PharmacyFudan UniversityShanghaiChina

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