Characterisation of antibodies to chloramphenicol, produced in different species by enzyme-linked immunosorbent assay and biosensor technologies |
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Authors: | Fodey Terence Murilla Grace Cannavan Andrew Elliott Christopher |
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Affiliation: | a Agri-Food and Biosciences Institute, Veterinary Sciences Division, Stoney Road, Belfast BT4 3SD, UK b Kenya Agricultural Research Institute, Residue Analysis Section, Kikuyu, Kenya c International Atomic Energy Agency, Agriculture and Biotechnology Laboratory, A-2444 Seibersdorf, Austria d Agri-Food and Land Use, Queens University Belfast, David Keir Building, Stranmillis Road, Belfast BT9 5AG, UK |
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Abstract: | Six polyclonal antisera to chloramphenicol (CAP) were successfully raised in camels, donkeys and goats. As a comparison of sensitivity, IC50 values ranged from 0.3 ng mL−1 to 5.5 ng mL−1 by enzyme-linked immunosorbent assay (ELISA) and from 0.7 ng mL−1 to 1.7 ng mL−1 by biosensor assay. The introduction of bovine milk extract improved the sensitivity of four of the antisera by ELISA and two by biosensor assay; a reduction in sensitivity of the remaining antisera ranged by a factor of 1.1-2.6. Porcine kidney extract reduced the sensitivity of all the antisera by a factor ranging from 1.1 to 7 by ELISA and a factor of 1.5 to 4 by biosensor. A low cross-reactivity with thiamphenicol (TAP) and florfenicol (FF) was displayed by antiserum G2 (1.2% and 18%, respectively) when a homologous ELISA assay format was employed. No cross-reactivity was displayed by any of the antisera when a homologous biosensor assay format was employed. Switching to a heterologous ELISA format prompted three of the antisera to display more significant cross-reactivity with TAP and FF (53% and 82%, respectively, using D1). The heterologous biosensor assay also increased the cross-reactivity of D1 for TAP and FF (56% and 129%, respectively) and of one other antiserum (G1) to a lesser degree. However, unlike the ELISA, the heterologous biosensor assay produced a substantial reduction in sensitivity (by a factor of 6 for D1). |
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Keywords: | CAP, chloramphenicol TAP, thiamphenicol FF, florfenicol ELISA, enzyme-linked immunosorbent assay HRP, horseradish peroxidase HSA, human serum albumin SPE, solid-phase extraction SPR, surface plasmon resonance NHS, N-hydroxysuccinimide EDC, 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride |
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