Development of a Biosensor-based Immunoassay for Screening of Chloramphenicol Residues in Milk

Abstract

Biosensor immunoassays were developed recently for antibiotics with an established maximum residue limit (MRL). In this study, according to the regulatory banning of chloramphenicol (CAP) use for food producing animals, the main objectives were: the specificity of the biosensor assay and the lowest detection limit possible. The assay was based on the inhibition of the binding of polyclonal antibodies against CAP to immobilized CAP on a sensor chip by CAP in solution. The response varied inversely with the antibiotic concentration in the sample. Two different antibodies and two immobilization protocols were tested. As in ELISA tests the antibody influenced the assay performances. Moreover, we showed that particular care should be concentrated on the immobilization step because it is a critical point in the assay development. Three different protocols were developed in milk. The best assay was obtained with antibody 1 in milk on the CAP base surface because of its very low detection limit (0.1 m g l -1 ) and the decreased consumption of antibody (four times less than on the CAP surface). This assay is rapid (3 min/run), sensitive, and specific for CAP and CAP glucuronide. It could be integrated in a multi-residue screening test and applied to other matrices (bile, urine, meat).

A label-free and portable multichannel surface plasmon resonance immunosensor for on site analysis of antibiotics in milk samples

“…In this context, surface plasmon resonance (SPR) allows for qualitative and quantitative measurements of biomolecular interactions in real-time without requiring a labeling procedure. In the framework of food safety, SPR has been used for the detection of antibiotics in honey (Ashwin et al, 2005;Ferguson et al, 2005;Yuan et al, 2009), shrimps (Dumont et al, 2006), milk (Ashwin et al, 2005;Ferguson et al, 2005;Gaudin and Maris, 2001;Laurentie and Gaudin, 2009;Mellgren and Sternesjo, 1998;Sternesjo et al, 1995), porcine muscle (McGrath et al, 2005), eggs, fish, poultry meat Huet et al, 2008), prawns , kidney (Ashwin et al, 2005) or broiler (Haasnoot et al, 2007), achieving good detectabilities. Specifically for milk samples, detectabilities reaching sub and low g L −1 range for CAP (Ashwin et al, 2005;Gaudin and Maris, 2001), fluoroquinolones (Mellgren and Sternesjo, 1998) and sulfonamides (Sternesjo et al, 1995), have been reported using laboratory equipment devices such as Biacore.…”

Improved screening method for the detection of a range of nitroimidazoles in various matrices by optical biosensor

“…doi:10.1016/j.aca.2008.09.040 of many laboratories worldwide [9][10][11][12][13][14][15]. In this study, an optical biosensor-screening assay was initially developed for the detection of nitroimidazoles in porcine, bovine and ovine kidney tissue, as this is the matrix specified by the United Kingdom National Surveillance Scheme (UKNSS).…”

Biosensors for environmental pollutants and food contaminants

“…Vibrio cholerae detection via its toxin was suggested by Swanson et al by combining the natural receptor for the toxin, the glycolipid GM1, with a fluorescence energy transfer optical transducer [168]. Antibiotic and other drug residue analysis in food samples have been published, such as the detection of penicillin in milk using photometric immunosensors [169], chloramphenicol was quantified in bile, urine, and meat [170], a variety of veterinary drugs, such as clenbuterol and ethinyl estradiol in urine, and enrofloxacin in milk were detected using an eight-channel surface plasmon resonance immunosensor [171], Hassnoot et al described the use of the Biacore system for the identification of gentamicin in milk samples [172], and compare direct versus competitive strategies for (dihydro)streptomycin immunoassays [173].…”