A rabbit monoclonal antibody-based sensitive competitive indirect enzyme-linked immunoassay for rapid detection of chloramphenicol residue

Abstract

A sensitive competitive indirect enzyme-linked immunoassay (ciELISA) based on a rabbit monoclonal antibody (RabMAb) against chloramphenicol (CAP) has been developed and validated in this study. After optimisation of several key physicochemical factors, such as Tween-20 percentage, pH value and ionic strength, the immunoassay showed excellent performance within the linear range of 0.18-6.37 ng mL −1 , with the 50% inhibition concentration (IC 50 ) of 1.06 ng mL −1 and the limit of detection (LOD) of 0.1 ng mL −1 . In addition, the cross-reactivities of RabMAb towards chloramphenicol succinate, florfenicol and thiamphenicol were 2.09, 12.45 and 18.10%, respectively. Finally, the developed method was applied in spiked swine urine, milk and honey samples, with recoveries ranging from 71.03 to 109.62%. The result demonstrated that the developed immunoassay is a valuable method for screening and quantitation of CAP residues in real samples.

Development of a quantitative fluorescence-based lateral flow immunoassay for determination of chloramphenicol, thiamphenicol and florfenicol in milk

“…Considering that, it is necessary to establish a rapid, sensitive assay for the determination of CAP, TAP and FF residues in food samples. A number of analytical methods have been developed to detect CAP, TAP and FF; these include high-performance liquid chromatography (Xie et al, 2011), gas chromatographymass spectrometry (GC-MS) (Shen et al, 2009), liquid chromatography tandem mass spectrometry (LC-MS/MS) (Barreto, Ribeiro, Hoff, & Dalla Costa, 2016;Li, Hu, Huo, & Xu, 2006;Rezende, Filho, & Rocha, 2012), enzyme-linked immunosorbent assay (Fodey, Murilla, Cannavan, & Elliott, 2007;Liu et al, 2014; and lateral flow immunoassay (LFA) (Bai et al, 2013;Guo et al, 2015;Samsonova, Cannavan, & Elliott, 2012;Xu, Xu, Ma, Liu, et al, 2015). Compared with chromatographic methods, immunoassays are simple, rapid and do not require expensive equipment and professional operator.…”

Comparsion of an immunochromatographic strip with ELISA for simultaneous detection of thiamphenicol, florfenicol and chloramphenicol in food samples

“…Several immunological methods have also been developed for screening purposes. These methods are based on antibody preparation and the specific binding reaction of the antibody and antigen (Wang et al ., ; Luo et al ., ; Tao et al ., , , ; Yi et al ., ; Zhang et al ., ; Fodey et al ., ; Liu et al ., ). In recent years, immunoaffinity chromatography cleanup has been used for sample pretreatment in instrument analyses, which can save organic solvents and improve efficiency (Luo et al ., ; Zhang et al ., ; Wang et al ., ).…”

Development of an ultrasensitive ic-ELISA and immunochromatographic strip assay for the simultaneous detection of florfenicol and thiamphenicol in eggs

“…Female BALB/c mice (aged 6-8 weeks) were immunized with 0.1 mL of immunogen per mouse by subcutaneous multisite injections (Liu, Song, et al, 2014;Xing et al, 2015). The mice was immunized firstly using a 1:1 mixture emulsion of FF-HS-MA-BSA (0.1 mg) and complete Freund adjuvant, and then booster immunizations were given over a period of 3 weeks by injecting a 1:1 mixture emulsion of FF-HS-MA-BSA (0.05 mg) and incomplete Freund adjuvant (Liu, Kuang, Peng, Wang, & Xu, 2014).…”