Production of polyclonal antibodies towards the immunodetection of insecticide phosalone |
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Authors: | V Issert R Lazaro F Lamaty V Rollande P Besançon B Caporiccio P Grenier V Bellon-Maurel |
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Institution: | (1) Division GIQUAL, Cemagref, Montpellier, France;(2) LAPP, UMR CNRS 5810, Université Montpellier II, Place Eugène Bataillon, F-34095 Montpellier Cedex 5, France;(3) Laboratoire Génie Biologique et Sciences des Aliments, Unité de Nutrition, Département Agroressources et Procédés Biologiques, Université Montpellier II, Montpellier, France |
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Abstract: | Summary Hapten synthesis for the production of specific insecticide phosalone polyclonal antibodies was carried out starting from an intermediate of the phosalone synthesis, 6-chloro-2-benzoxazolone1. Two haptens containing different spacers have been prepared: N-5-carboxypentyl-6-chloro-2-benzoxazolone7 and N-(2-oxo-3-aza-5-carboxypentyl)-6-chloro-2-benzoxazolone12. Each of these two haptens conjugated to bovine serum albumine (BSA) was used to immunize four rabbits. Immunoassays of phosalone were performed with ELISA using solid-phase bound hapten thyroglobulin conjugate and horseradish peroxidase labelled goat antirabbit IgG. The more sensitive response was observed when the antiserum obtained from the rabbit immunized with the hapten-BSA conjugate containing the N-2-oxo-3-aza-5-carboxypentyl spacer was in competition with the same hapten coupled to thyroglobulin. An identical response was obtained under the same conditions when using benzoxazolone instead of phosalone as competitor, showing a good recognition of the specific aromatic part of the organophosphate insecticide phosalone. Reduction of coating conjugate concentration (from 2 to 0.05g/well) and also the use of heterologous coating protein instead of homologous did improve the sensitivity, resulting in a concentration of phosalone required to inhibit binding by 50% of 2 mg/l and a detection limit of 0.02 mg/l. |
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Keywords: | Amino acids Polyclonal antibodies Phosalone Immunodetection Hapten synthesis |
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