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A fluorometric clenbuterol immunoassay based on the use of organic/inorganic hybrid nanoflowers modified with gold nanoclusters and artificial antigen

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Abstract

Organic/inorganic hybrid nanoflowers were synthesized from calcium phosphate and protein modified fluorescent gold nanoclusters and antigens. These nanoflowers are shown to be well suited labels for bioassay because they fulfill the functions of biological recognition and signal output. A fluorometric immunoassay was developed that was combined with immunomagnetic separation. In the detection system, the red fluorescence of the supernatant (measured at excitation/emission wavelengths of 360/640 nm) is found to be proportional to the clenbuterol (Clen) concentration after two immunomagnetic separations. The assay has a linear response in the 0.5 μg L−1 to 40 μg L−1 Clen concentration range, and 0.167 μg L−1 limit of detection. This makes it well suited for food safety monitoring. The average recoveries from spiked samples range from 92.7 to 109.1% (intra-assay) and 101.2 to 125.7% (inter-assay) with relative standard deviations of <11.6%. Spiked swine urine samples were analyzed by this method, and the results correlated well with data obtained by LC-MS/MS.

Fluorescent hybrid nanoflowers were fabricated with gold nanoclusters (BSA-AuNCs) and antigens. A fluorometric immunoassay based on the use of such nanoflowers and based on immunomagnetic separation was developed to detect clenbuterol residues in swine urine with satisfactory recoveries and acceptable accuracy.

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Acknowledgments

This work was supported by the National Natural Science Foundation of China (31672600) and Sanming Project of Medicine in Shenzhen (SZSM201611068). And the authors appreciate the cooperation of other faculty members in the Department of Pharmacology and Toxicology of the College of Veterinary Medicine at China Agricultural University.

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Authors and Affiliations

  1. Beijing Advanced Innovation Center for Food Nutrition and Human Health, College of Veterinary Medicine, China Agricultural University, Beijing Key Laboratory of Detection Technology for Animal-Derived Food Safety, Beijing Laboratory for Food Quality and Safety, Beijing, 100193, People’s Republic of China

    Tao Peng, Jianyi Wang, Sanlei Xie, Kai Yao, Pimiao Zheng, Sihan Wang & Haiyang Jiang

  2. China Animal Health and Epidemiology Center, Qingdao, 266032, People’s Republic of China

    Sijun Zhao

  3. Shenzhen Center for Disease Control and Prevention, Shenzhen, 518055, People’s Republic of China

    Yuebin Ke

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  1. Tao Peng
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  2. Jianyi Wang
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  4. Sanlei Xie
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Correspondence to Haiyang Jiang.

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This article does not contain any studies with human participants or animals performed by any of the authors. The author(s) declare that they have no competing interests.

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Peng, T., Wang, J., Zhao, S. et al. A fluorometric clenbuterol immunoassay based on the use of organic/inorganic hybrid nanoflowers modified with gold nanoclusters and artificial antigen. Microchim Acta 185, 366 (2018). https://doi.org/10.1007/s00604-018-2889-0

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