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Application of an immunosensor for the detection of the β-lactam antibiotic, cephalexin
Authors:Paul P. Dillon   Stephen J. Daly  John G. Browne  Bernadette M. Manning  Elma Loomans  Aart Van Amerongen  Richard O'Kennedy
Affiliation: a School of Biotechnology, Dublin City University, Dublin, Republic of Irelandb NCSR (National Centre for Sensor Research), Dublin City University, Dublin, Republic of Irelandc ATO BV Agrotechnological Research Institute, Wageningen, Bornsesteeg, The Netherlands
Abstract:Public concern surrounding antibiotic contamination in food and food products has made it imperative to develop analytical methods for their detection. Polyclonal antibodies were used in the development of a surface plasmon resonance (SPR)-based inhibition immunoassay for cephalexin. A conjugate consisting of cephalexin-bovine serum albumin (BSA) was immobilized on the dextran gel surface of the sensor chip. Binding/regeneration studies of antibody to immobilized cephalexin were studied and dissociation of the antibody from the immobilized cephalexin was easily achieved with 10 mmol l-1 NaOH. Forty surface regeneration cycles were carried out and found to be reproducible with only a 7.4% decrease in binding over this number of regenerations. Model inhibition immunoassays for cephalexin were developed in PBS and spiked milk samples with detection ranges of 4.88 to 2,500 ng ml-1 and 244 to 3,906 pg ml-1, respectively.
Keywords:Cephalexin  antibiotics  surface plasmon resonance  regeneration  inhibitive immunoassay
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