Immunochemical determination of oxytetracycline in fish: Comparison between enzymatic and time-resolved fluorometric assays

Multiresidue determination of antibiotics in feed and fish samples for food safety evaluation. Comparison of immunoassay vs LC-MS-MS

“…An optimized time-resolved fluoroimmunoassay (TR-FIA) previously developed (Cháfer-Pericás et al, 2010b), was applied to determine tetracyclines in fish samples medicated with OTC in a fish farm. Briefly, the procedure consisted of the use of 96-well black and high-binding microtiter plates coated with 100 mL/well of the appropriate concentration (0.0156 mg L À1 ) of OVA-hapten conjugate solution in carbonate buffer, and incubated overnight at 4 C. The competition procedure with serum (1/8000 containing 1.5% (w/v) Hb, in plate) was the same as that described above.…”

“…An immunoassay based on a competitive indirect ELISA format previously optimized (Cháfer-Pericás et al, 2010b) was applied to determine tetracyclines in real samples. The used immunoreagents allowed determining the total tetracycline concentrations (OTC, TC, CTC).…”

Production of a monoclonal antibody against oxytetracycline and its application for oxytetracycline residue detection in shrimp

“…Most of the reported antibodies specific for TC group antibiotics are PAbs (Zhao et al, 2008;Le et al, 2009;Cháfer-Pericása et al, 2010). Because the properties of PAbs vary among batches according to the immune responses of different immunized animals, PAbs are less suitable for long-term application than MAbs (Hock et al, 1995).…”

“…MAb 2-4F showed high crossreactivity with rolitetracycline (RTC) (388%) but low cross-reactivity with other TCs and tested chemicals. A PAb produced against OTC had previously been reported, which also showed high cross-reactivity with TC (1028%) and RTC (449%) (Cháfer-Pericása et al, 2010). The assay was carried out in triplicate (mean±SD) using 2 µg/ml OTC-OVA as the coating antigen and 0.03 µg/ml MAb 2-4F …”

“…Importantly, an immunoassay does not require a complicated step to clean up samples (Cháfer-Pericása et al, 2010). Previously, established ELISA methods have been based on a polyclonal antibody (PAb) for doxycycline (DC) detection (Le et al, 2009) and a monoclonal antibody (MAb) for chlortetracycline (CTC) detection (Le et al, 2011b).…”