Abstract
In this paper, a novel dual-label time-resolved chemiluminescent multiplexed immunoassay (DLTRC-MIA) based on the distinction of the kinetic characteristics of horseradish peroxidase (HRP) and alkaline phosphatase (ALP) with approximate estimation approach for simultaneous determination of 20 fluoroquinolones (FQs), 15 β-lactams, 15 sulfonamides (SAs), and chloramphenicol (CAP) in milk was developed. The strategy integrated a single-chain variable fragment–alkaline phosphatase fusion protein (scFv-ALP), a recombinant penicillin-binding protein (PBP) 2×*, a monoclonal antibody (MAb), and a polyclonal antibody (PAb) in one immunoassay and in a single well together to fulfill the simultaneous detection of 51 low-molecular weight contaminants (20 FQs, 15 β-lactams, 15 SAs, and CAP). The limits of detection for FQs, β-lactams, SAs, and CAP range from 0.29 μg L−1 for ciprofloxacin (CIP) to 81.6 μg L−1 for trovafloxacin (TRO), 0.27 μg L−1 for ceftiofur (CEF) to 44.1 μg L−1 for cephalexin (CEL), 0.089 μg L−1 for sulfadimethoxine (SDM) to 2.7 μg L−1 for sulfadiazine (SDZ), and 0.028 μg L−1 for CAP, respectively. The results demonstrated that the detection limits of DLTRC-MIA meet the requirement of detection levels for 51 drug residues in milk, suitable for high-throughput screening of low-molecular weight contaminants.
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Acknowledgments
The PAb, MAb, PBP 2×*, and scFv-ALP were donated by the Department of Veterinary Pharmacology and Toxicology, College of Veterinary Medicine, China Agricultural University.
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We acknowledge financial support National Natural Science Foundation of China (Grant No. 31672605) (Grant No. 31671787), China Postdoctoral Science Foundation (Grant No. 2016 M590855) and Fundamental Research Funds for the Central Universities (Grant No. XDJK2015C040) (Grant No. SWU114043) (Grant No. XDJK2016C164) and Chongqing Science and Technology Commission (cstc2014pt-gc8001).
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Xiaoqi Tao declares that he has no conflict of interest. Jia Wang declares that he has no conflict of interest. Yuejie Xie declares that he has no conflict of interest. Xiaowei Zuo declares that he has no conflict of interest. Fang Mo declares that he has no conflict of interest. Song Zhou declares that he has no conflict of interest. Hongjun Li declares that he has no conflict of interest.
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Tao, X., Wang, J., Xie, Y. et al. Dual-Label Chemiluminescence Strategy for Multiplexed Immunoassay of 20 Fluoroquinolones, 15 β-Lactams, 15 Sulfonamides, and CAP in Milk. Food Anal. Methods 10, 3009–3022 (2017). https://doi.org/10.1007/s12161-017-0865-7
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DOI: https://doi.org/10.1007/s12161-017-0865-7