Dual-factor Synergistically Activated ESIPT-based Probe: Differential Fluorescence Signals to Simultaneously Detect α-Naphthyl Acetate and Acid α-Naphthyl Acetate Esterase

Kui Wang, Beidou Feng, Yonggang Yang, Yuehua Chen, Yuzhu Wang, Yafu Wang, Lin Yang, Kai Jiang, Tony D. James, Hua Zhang

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α-Naphthyl acetate esterase (α-NAE) and acid α-naphthyl acetate esterase (ANAE), a class of special esterases, are important for lymphocyte typing and immunocompetence-monitoring. As such, the simultaneous detection of α-NAE and ANAE has become a target to effectively improve the accuracy in lymphocyte typing. Therefore, we developed a dual-factor synergistically activated ESIPT-based probe (HBT-NA) to detect α-NAE and ANAE sensitively, rapidly, and simultaneously in a differential manner. HBT-NA exhibits differential fluorescence signal outputs toward small changes of α-NAE and ANAE activities. HBT-NA displays a weak fluorescence signal at 392 nm over a pH range from 6.0 to 7.4. However, when it interacts with α-NAE (0-25 U) at pH = 7.4, the fluorescence intensity at 392 nm enhanced linearly within 60 s (F392 »nm/F0392 »nm = 0.042 Cα-NAE + 1.1, R2 = 0.99). Furthermore, HBT-NA emits ratiometric fluorescence signals (F505 »nm/F392 »nm) for ANAE (0-25 U) at pH = 6.0 within 2.0 min, exhibiting a good linear relationship (F505 »nm/F392 »nm = 0.83CANAE - 1.75, R2 = 0.99). The differential fluorescence signals can be used to simultaneously detect the activities of α-NAE and ANAE in solutions and complex living organisms. More importantly, based on the differential fluorescence signals toward α-NAE and ANAE, T lymphocytes and B lymphocytes could be successfully typed and differentiated among nontyped lymphocytes, facilitating the real-time evaluation of their immune functions using flow cytometry. Hence, HBT-NA could be used for the ultrasensitive detection of the enzyme activities of α-NAE and ANAE, the real-time precise typing of lymphocytes, and the monitoring of immunocompetence.

Original languageEnglish
Pages (from-to)14471-14480
Number of pages10
JournalAnalytical Chemistry
Issue number43
Early online date25 Oct 2021
Publication statusPublished - 2 Nov 2021

Bibliographical note

Funding Information:
This work was supported by the National Natural Science Foundation of China (21722501, 21803018, 11974103), Henan Special Support for High-Level Talents Central Plains Science and Technology Innovation Leading Talents (204200510006), Key Project of Science and Technology of Henan Province (212102311071, 202102310139), Key Project of Science and Technology of Xinxiang City (GG2020001), Program for Science Technology Innovation Talents in Universities of Henan Province (21HASTIT019), and the High Performance Computing Center of Henan Normal University. T.D.J. wishes to thank the Royal Society for a Wolfson Research Merit Award and the Open Research Fund of the School of Chemistry and Chemical Engineering, Henan Normal University, for support (2020ZD01).

Publisher Copyright:
© Author(s).

ASJC Scopus subject areas

  • Analytical Chemistry


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