Monoclonal antibody based enzyme-linked immunosorbent assay for the specific detection of ciprofloxacin and enrofloxacin residues in fishery products

5-sulfosalicylic Acid Dihydrate-Based Pretreatment for the Modification of Enzyme-Linked Immunoassay of Fluoroquinolones in Fishery Products

“…The mean recovery for three fishery samples at lower spike concentration was relatively higher than those at higher concentration. Several previous studies also reported that the recovery for samples contained higher concentrations of fluoroquinolones was lower than those contained lower concentrations [11,12,23] and the reason may be attributed the overall affluence of many factors. The most likely explanation is that the extraction efficiency of FQs at different concentrations is not consistent.…”

“…Hu et al [12] used solvent extraction in sample preparation for determination of fluoroquinolone residues. Andrzej Posyniak et al [18] introduced solid phase extraction as a further clean-up step.…”

“…In some reports defatting by hexane and some other clean-up steps were also required. [11,12] Such complicated and time consuming pretreatments make the ELISA and other immunoassays not "fast and simple" enough for routine monitoring of large numbers of food samples, even in comparison to some HPLC-based methods, especially those exploiting automatic samplers. To a large extent such a problem could be attributed to the significant interference from the complex food matrix to the immunoassays, which has been confirmed by many previous studies.…”

Influence of hapten density on immunogenicity for anti-ciprofloxacin antibody production in mice

“…In our previous studies, a monoclonal antibody (mAb) against small hapten-CPFX with high affinity and specificity was produced and was used for rapid CPFX immunoassays in food stuff of animal origin (9). Furthermore, with the CPFX-specific mAbs, an indirect competitive enzyme-linked immunosorbent assay (ELISA) was developed for the sensitive and specific detection of CPFX residues in fishery products (10). In this study, CPFX as a hapten was covalently attached to BSA by a modified carbodiimide method using 1-ethl-3-carbodiimide methiodide (EDC) (1,8).…”

“…A hundred μL of CPFX-conjugated ovalbumin, which was prepared in our laboratory (10), with a concentration of 400 ng/mL was added into wells of an ELISA plate (Dingguo Biotechnology Co., Ltd.) and incubated at 37°C for 1 h. After coating, the wells were washed 3 times (3 min each) with phosphate buffer containing 0.5% Tween-20 (washing solution). The wells were blocked with 200 μL 5% milk and stored at 4°C overnight.…”