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Time-resolved fluorescence microspheres-antibody-penicillin-binding protein assisted construction of immunochromatographic assay for sensitive detection of 22 β-lactams in milk

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Abstract

A sensitive immunochromatographic assay (ICA) using time-resolved fluorescence microspheres (TRFMs) coupled with an indirect-labeling mode was developed for simultaneously determining 22 kinds of β-lactams in milk samples. The TRFMs labeled anti-receptor monoclonal antibodies (mAbs) conjugated to penicillin-binding proteins (PBPs) as ternary TRFMs-mAb-PBPs (TMP) nanoscaffolds provide excellent solubility, brightness, and stability. Thanks to the fact that they not only fully expose the binding sites of PBPs, thereby enhancing the biological affinity of PBPs towards the target, but also generated superb fluorescence signals, the versatile TMP manifested unique possibilities as efficient probes for ICA with remarkable enhancement in sensitivity in β-lactams screening. The results showed that the standard curves of the 22 varying β-lactams displayed linearity in their respective concentration ranges (R2 > 0.98), with the cutoff values of 1–100 ng/mL. The constructed TMP-ICA was successfully applied to the analysis of real milk, with consistent results compared with liquid chromatography-tandem mass spectrometry (LC-MS), providing an effective method for sensing β-lactams in food matrices.

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Funding

This work was supported by the Key Scientific and Technological Project of Henan Provincial Education Department of China (222102310162), the Henan Postgraduate Joint Training Base Project (YJS2022JD16), and the Program for Innovative Research Team (in Science and Technology), University of Henan Province (No. 23IRTSTHN023).

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  1. Yawei Sun and Xiangjie Deng contributed equally to this work.

Authors and Affiliations

  1. College of Animal Science and Veterinary Medicine, Henan Institute of Science and Technology, Xinxiang, 453003, Henan, People’s Republic of China

    Yawei Sun & Xiangjie Deng

  2. Henan Engineering Technology Research Center of Food Processing and Circulation Safety Control, College of Food Science and Technology, Henan Agricultural University, 63 Nongye Road, Zhengzhou, 450002, Henan, China

    Changwei Luo, Wentao Ma, Wanjing Liu, Jinkui Wang, Qingyue Li, Tong Bu & Xiya Zhang

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  1. Yawei Sun
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Sun, Y., Deng, X., Luo, C. et al. Time-resolved fluorescence microspheres-antibody-penicillin-binding protein assisted construction of immunochromatographic assay for sensitive detection of 22 β-lactams in milk. Microchim Acta 191, 50 (2024). https://doi.org/10.1007/s00604-023-06106-1

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