Monoclonal antibody for the development of specific immunoassays to detect Enrofloxacin in foods of animal origin

Abstract

The carbodiimide active ester method was employed to synthesise the antigen of 1-cyclopropyl-7-(4-ethylpiperazin-1-yl)-6-fluoro-4-oxo-1,4-dihydroquinoline-3-carboxylic acid [Enrofloxacin (ENFX)], and male Balb/c mice were used to produce anti-ENFX monoclonal antibody. Based on the checkerboard titration, an indirect competitive enzyme-linked immunosorbent assay standard curve was established. This assay was sensitive and highly specific to ENFX with the half maximal inhibitory concentration and limit of detection values of 0.15 ng mL −1 and 0.028ng mL −1 , respectively. Similarly, the antibody had a high affinity as seen in A max values. Spiked cow milk beef liver and fish samples analysed using this kit had recoveries in the range of 90.2-110.2%, 70.7-81.5% and 89.1-101%, respectively, showing satisfactory results. The results suggest that this ELISA kit could be applied as a screening method to detect and control the illegal content of ENFX in food products. ARTICLE HISTORY

Development of a monoclonal antibody assay and a lateral flow strip test for the detection of paromomycin residues in food matrices

“…After preliminary checkerboard titration experiments (results not shown), the optimum concentrations of coating antigen (PR-EDC-OVA) and antibody (2C4) were determined to be 0.8 and 0.6 μg/ml, respectively. The effects of pH and salt concentration on antibody performance were then assessed because, as observed in previous studies (Isanga et al, 2016;Tochi et al, 2015), these two factors vary in different foods and contribute to matrix interference. The interaction between paromomycin and the antibody is also readily affected by assay pH and ionic strength because the drug is water soluble and has many hydroxyl and amino groups in its chemical structure, as shown in Figure 2(a).…”

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

“…Nevertheless, these methods are not suitable for on-site analysis (Wang et al, 2016;Xu et al, 2006); moreover, they are expensive and require more skilled personnel to operate them. Therefore, rapid, sensitive, inexpensive and accurate simple and time-saving methods such as Lateral immunochromatographic strips and indirect competitive enzyme-linked immunosorbent assay (ic-ELISA), which require less knowledge for operation, must be developed complementary to conventional instrumental techniques Tochi et al, 2015;Xu et al, 2015). In this work, we reported the production of a high-affinity monoclonal antibody against HES, the optimization of an Ic-ELISA and immunochromatographic assay to detect HES and DES residues in cow's milk.…”

Development of an ic-ELISA and immunochromatographic strip for detection of sparfloxacin in honey

“…Furthermore, long-term exposure to SPFX can result in the emergence of drug-resistant pathogens which increases the risk of death from common infections (Martín-Galiano & de la Campa, 2003;Minnick, Wilson, Smitherman, & Samuels, 2003;Tlili et al, 2016;Weigel, Anderson, & Tenover, 2002). Due to these concerns, many countries have set maximum residue limits (MRLs) for several quinolones to protect consumers (Tochi, Khaemba, et al, 2016;Zhi et al, 2010). China and the USA do not permit use of SPFX in honey or honey products.…”