Abstract
A novel lateral flow immunoassay biosensor, combining the receptor-ligand reaction and antigen-antibody reaction was developed for the detection of β-lactam antibiotics in milk. The receptor protein, the carboxy-terminal domain of β-lactam sensor-transducer mutant from Bacillus. licheniformis, served as the recognition element. Colloidal gold (CG)-labelled receptor antibody-receptor ternary complex was applied as a signal transducer probe. The presence of β-lactams was identified through a color change in the test zone, where the reaction between the complex of CG/anti-receptor antibody/receptor and β-lactam antigen on the test line could be inhibited by β-lactam residues in the sample, thus leading to a reduction in color signal. Based on the indirect labeling of the receptor with an antibody as a linker, the ability to detect 33 β-lactams was below or near the corresponding maximum residue limit. The proposed lateral flow immunoassay biosensor appears to be an excellent field-based screening tool for the qualitative screening of β-lactams in milk.
摘要
的侧流免疫传感器, 用于牛奶中β-内酰胺类抗生素的残留检测. 该传感器以地衣芽孢杆菌来源的突变型β-内酰胺类抗生素传感受体蛋白(BlaR-CTD-M)为识别元件, 以胶体金(CG)标记的受体抗体-受体复合物为信号传递探针. 基于样品中游离的β-内酰胺类抗生素竞争抑制CG/受体抗体-受体复合物与检测线处固定的β-内酰胺类抗生素抗原的结合, 根据检测线的颜色变化可实现β-内酰胺类抗生素的快速定性检测. 通过受体抗体作为连接臂, 以CG间接标记受体, 使该传感器能检测牛奶中33种β-内酰胺类抗生素, 检测限符合最大残留限量. 本文所建的传感器检测方法是一种现场快速定性筛查β-内酰胺类抗生素残留的有效工具.
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Acknowledgements
This work was financially supported by the National Key R&D Program of China (2019YFC1606603).
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Author contributions Kuang H and Xu C conceived and designed the experiments; Li Y performed the experiments; Sun L, Liu L and Kuang H performed the data analysis. All the authors drafted the manuscript, and read and approved the manuscript prior to submission.
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Yue Li is a PhD candidate under the supervision of Prof. Chuanlai Xu at Jiangnan University. Her research interests focus on developing rapid and sensitive methods to determine analytes using immunochromatography strip test.
Chuanlai Xu is the Director of the International Lab of Biointerface & Biodetection, Jiangnan University, leader of the Food Safety Innovation Team in Priority Areas Accredited by the Ministry of Science and Technology and fellow of the Royal Society of Chemistry. His research interests include: (1) fabrication of nano-devices and their applications in biotechnology; (2) preparation of antibodies against hazards (such as heavy metals, pesticides, and veterinary drug) and their applications in rapid detection.
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40843_2020_1578_MOESM1_ESM.pdf
Integration of Antibody-Antigen and Receptor-Ligand Reactions to Establish a Gold Strip Biosensor for Detection of 33 β-Lactam Antibiotics
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Li, Y., Liu, L., Xu, C. et al. Integration of antibody-antigen and receptor-ligand reactions to establish a gold strip biosensor for detection of 33 β-lactam antibiotics. Sci. China Mater. 64, 2056–2066 (2021). https://doi.org/10.1007/s40843-020-1578-0
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DOI: https://doi.org/10.1007/s40843-020-1578-0