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An indirect ELISA-inspired dual-channel fluorescent immunoassay based on MPA-capped CdTe/ZnS QDs

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Abstract

To meet the need for high-throughput immunoassays, many multiplex fluorescent immunoassays have been proposed. Most of them need different kinds of fluorescent label indicators during the test. In this work, a novel indirect ELISA-inspired dual-channel fluorescent immunoassay based on 3-mercaptopropionic acid capped CdTe/ZnS quantum dots (QDs) was constructed. The ELISA wells were coated with two kinds of antigen–QD complex. When the primary antibodies were present in a sample, they mediated the binding of a secondary antibody–DNA–gold nanoparticle complex to the antigen–QD complex. Then the gold nanoparticles quenched the fluorescence of the QDs and a decrease in fluorescence intensity was observed. Thus, the amount of primary antibody could be estimated from the decrease of fluorescence intensity. Owing to the wide absorption range and the relatively narrow emission band of the QDs, the dual-channel fluorescent immunoassay system could work at the same excitation wavelength and the emission wavelengths of each channel had no interference. As a result, two different kinds of primary antibody could be detected at the same time in one ELISA well, which simplified the operation and greatly improved the efficiency. Besides, only one type of secondary antibody needs to be added to the prepared microtiter plates, which further simplified the operation during the detection procedure. This dual-channel fluorescent immunoassay system will provide new insights into high-throughput immunodetection.

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Acknowledgements

The authors thank the National Key Beijing Science Technology New Star Cross Subject (2018019), the Beijing Natural Science Foundation (2172039), National Key Research and Development Program of China (2018YFD0401302), the Fundamental Research Funds for the Central Universities and University of Science and Technology Beijing (USTB), the Open Research Fund Program of the Key Laboratory of Cosmetic(Beijing Technology and Business University), and China National Light Industry (KLC-2017-YB5).

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  1. Xiangyu Jiao and Yabin Zhou contributed equally to this work.

Authors and Affiliations

  1. School of Chemistry and Biological Engineering, University of Science and Technology Beijing, Beijing, 100083, China

    Xiangyu Jiao, Yabin Zhou, Duo Pang, Hongwu Du, Yongqiang Wen & Xueji Zhang

  2. Key Laboratory of Cosmetic, China National Light Industry, Beijing Technology and Business University, Beijing, 100048, China

    Dan Zhao & Changtao Wang

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  1. Xiangyu Jiao
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  2. Yabin Zhou
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Correspondence to Hongwu Du or Yongqiang Wen.

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Jiao, X., Zhou, Y., Zhao, D. et al. An indirect ELISA-inspired dual-channel fluorescent immunoassay based on MPA-capped CdTe/ZnS QDs. Anal Bioanal Chem 411, 5437–5444 (2019). https://doi.org/10.1007/s00216-019-01917-9

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  • DOI: https://doi.org/10.1007/s00216-019-01917-9

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