Development of an ELISA and Immunochromatographic Assay for Tetracycline, Oxytetracycline, and Chlortetracycline Residues in Milk and Honey Based on the Class-Specific Monoclonal Antibody

Abstract

A sensitive class-specific monoclonal antibody against tetracyclines (TCs) was generated and used to develop an enzyme-linked immunosorbent assay (ELISA) and an immunochromatographic assay for TC, oxytetracycline (OTC), and chlortetracycline (CTC) detection in milk and honey samples. The dynamic range of detection for TC in ELISA was 0.26-2.00 μg L −1 with an IC 50 of 0.72 μg L −1 . The IC 50 value of OTC and CTC was 3.2 and 6.4 μg L −1 , respectively. The recovery of TC, OTC, and CTC in milk samples was 82-102, 91-105, and 90-101 %, respectively, and 88-101, 89-101, and 89-95 % in honey samples, respectively. In the immunochromatographic assay, the cutoff values for TC, OTC, and CTC were 15, 15, and 50 μg L −1 in milk, respectively, and 40, 40, and 40 μg L −1 in honey, respectively. The results revealed that ELISA and the immunochromatographic assay can be applied for the rapid and sensitive detection of TC, OTC, and CTC in milk and honey samples.

Establishment of a monoclonal antibody-based indirect enzyme-linked immunosorbent assay for the detection of trimethoprim residues in milk, honey, and fish samples

“…The enzyme-linked immunosorbent assay (ELISA) is a sensitive, simple, and cost-effective method, which has been widely used in the analyses of large compounds such as microbes , proteins , and allergens and small compounds such as antibiotics (Chen et al, 2016;Suryoprabowo, Liu, Peng, Kuang, & Xu, 2014), pesticides (Chen, Liu, Kuang, Song, & Xu, 2013;, hormones Liu, Peng, Jin, & Xu, 2007), and toxins . There are also some papers which have reported the detection of TMP in food-based antibodies.…”

Development of an immunochromatographic assay for hexestrol and diethylstilbestrol residues in milk

“…The production of monoclonal antibody was done according to the method as described by Chen et al (2015). Murine myeloma cells (SP20-Ag14) and mouse spleen cells were fused at a ratio of 1:1 in the presence of PEG-1500 resulting in hybridoma cells.…”

“…The cross-reactivity (CR) values were calculated according to the following equation (Chen et al, 2015): CR% = (IC 50 value of HES)/(IC 50 value of Competitor) × 100.…”

Development of a monoclonal antibody assay and immunochromatographic test strip for the detection of amikacin residues in milk and eggs

“…However, these methods are costly, time-consuming, require sophisticated laboratory equipment and technically trained personnel (Chen, Kong, et al, 2016;Li et al, 2016;Lourenco, Barbosa, & Pinto, 2011;Xu et al, 2016;Xu et al, 2011). In addition, the amikacin molecule has no chromophores that can give a reliable signal in the UV region, lacks volatility and is very hydrophilic (Farouk, Azzazy, & Niessen, 2015).…”

“…Enzyme-linked immunosorbent assay (ELISA) detection methods are semi-quantitative, simple, highly specific, sensitive, and cost effective, hence they are convenient for routine monitoring and screening of drug residues in food samples (Chen, Kong, et al, 2016;Liu, Yan, Zhang, Kuang, & Xu, 2015;Tian, Chen, Guo, Guo, & Mei, 2015). Previously, commercially available polyclonal antibodies were used to develop a surface plasmon resonance immunoassay for therapeutic drug monitoring of amikacin (Losoya-Leal et al, 2015) and a fluorescence polarization immunoassay for detection of amikacin (SanchezMartinez, Aguilar-Caballos, & Gomez-Hens, 2007), but neither studies provided detailed information on antibody preparation and cross-reactivity analysis.…”