Abstract
In this study, mAb-5A7 and mAb-1D7, two highly specific monoclonal antibodies against tylosin (TYL), were prepared by using hapten TYL-carboxymethoxylamine hemihydrochloride (TYL-CMO), and they showed IC50 values of 13.84 ng/mL and 13.01 ng/mL and LODs of 1.21 and 0.98 ng/mL, respectively, in indirect competitive enzyme-linked immunosorbent assay. An immunochromatographic assay based on quantum dot (QD)-embedded nanobead (QB-ICA) and mAb-1D7 was developed for the detection of TYL in milk and honey. The IC50 values of QB-ICA in PBS, milk, and honey were calculated to be 22.4, 18.5, and 24.41 ng/mL, with LODs of 2.32, 1.84, and 2.69 ng/mL, respectively, satisfying the requirement of the maximum residue limit for TYL in food samples of 100–300 μg/kg in China. The cross-reactivity (CR) values with two related compounds, tilmicosin and abamectin, were 4.48% and 1.87%, respectively, and the CR values with other drugs were all less than 0.2%, which indicated that the developed QB-ICA was highly specific for the determination of TYL. In addition, recovery experiments indicated that the recoveries ranged from 92.16 to 108.5% in milk and from 84.12 to 110.5% in honey. Therefore, the developed QB-ICA is reliable for determining TYL in milk and honey. In conclusion, the QB-ICA we developed can be considered an effective and rapid tool for screening TYL in food products.
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Funding
This work was supported by the National Science Foundation for Young Scientists of China (31800776), Jiangxi Special Fund for Agro-scientific Research in the Collaborative Innovation (JXXTCX201703-1), earmarked fund for Jiangxi Agriculture Research System (JXARS-03), Key Research and Development Program of Jiangxi Province (20192BBF60046), and the free explore issue of State Key Laboratory of Food Science and Technology of Nanchang University(SKLF-ZZB-201719).
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Xiaocui Lai declares that she has no conflict of interest; Xi Lv declares that she has no conflict of interest; Ganggang Zhang declares that he has no conflict of interest; Zhijuan Xiong declares that she has no conflict of interest; Weihua Lai declares that he has no conflict of interest; Juan Peng declares that she has no conflict of interest.
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The protocol for carrying out animal experiments was approved by the Nanchang University Animal Ethics Committee, and all the ethical requirements to conduct the experiment were met. All procedures were conducted in compliance with provided protocols and approved by the Animal Care Review Committee (approval number 0064257) of Nanchang University (Jiangxi, China).
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Lai, X., Lv, X., Zhang, G. et al. Highly Specific Anti-tylosin Monoclonal Antibody and Its Application in the Quantum Dot Bead-Based Immunochromatographic Assay. Food Anal. Methods 13, 2258–2268 (2020). https://doi.org/10.1007/s12161-020-01846-9
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DOI: https://doi.org/10.1007/s12161-020-01846-9