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An ultrasensitive ratiometric immunosensor based on the ratios of conjugated distyrylbenzene derivative nanosheets with AIECL properties and electrochemical signal for CYFRA21-1 detection

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Abstract

Aggregation-induced electrochemiluminescence reagent, a distyrylbenzene derivative with donor–acceptor conjugated nanosheet structure, namely TPAPCN, was used as a trace label and modified on the electrode through the formation of classical sandwich complex of antibody-antigen–antibody in this work. In aggregate state, TPAPCN with twisted structure was limited in nanometer space through intermolecular π − π stacking interactions, which not only restricts the intramolecular motions but also combines a large number of singlet excitons to greatly trigger electrochemiluminescence (ECL). The ECL signal of this system enhanced with more captured cytokeratin 19 fragment 21–1 (CYFRA21-1) on the modified electrode. Three-dimensional graphene/platinum nanoparticles with large specific surface, and excellent electroconductivity and biocompatibility were prepared and acted as excellent carriers for thionine handling (3D-GN/PtNPs/Th), which was employed for improving the loading of antibodies and generating internal electrochemical signal. Consequently, a novel ratiometric sandwich immunosensor for CYFRA21-1 detection was fabricated based on TPAPCN and 3D-GN/PtNPs/Th, that is, a rapid and reliable detection was achieved through the ratio between ECL and electrochemical signals. The prepared sensor performed good linearity in the range of 50 fg/mL to 1 ng/mL with a detection limit as low as 16 fg/mL. Moreover, the detection results revealed well in the analysis of human serum samples, demonstrating a significant application for clinical monitoring and biomolecules detection.

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Funding

We received support from the National Natural Science Foundation of China (No. 21705084), and the Natural Science Foundation of Shandong Province of China (No. ZR2017BB074, No. ZR2018PB015), National Training Program of Innovation and Entrepreneurship for Undergraduates (No. S202010431027), Qilu University of Technology of Training Program of Innovation and Entrepreneurship for Undergraduates (No. xj201910431125), the Innovation Team of Jinan City (2018GXRC004), and Special Funds for Taishan Scholars Project.

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  1. State Key Laboratory of Biobased Material and Green Papermaking, School of Food Science and Engineering, Qilu University of Technology, Shandong Academy of Sciences, Jinan, 250353, Shandong, China

    Xiaoyi Lv, Xiaoyun Xu, Yanping Li, Chao Geng, Bo Cui & Yishan Fang

  2. Juye County People’s Hospital, Heze, 274900, Shandong, People’s Republic of China

    Mengmeng Bi

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  1. Xiaoyi Lv
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Lv, X., Bi, M., Xu, X. et al. An ultrasensitive ratiometric immunosensor based on the ratios of conjugated distyrylbenzene derivative nanosheets with AIECL properties and electrochemical signal for CYFRA21-1 detection. Anal Bioanal Chem 414, 1389–1402 (2022). https://doi.org/10.1007/s00216-021-03764-z

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