title: Production of polyclonal antibodies towards the immunodetection of insecticide phosalone.
abstract: 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-benzoxazolone 1. Two haptens containing different spacers have been prepared: N-5-carboxypentyl-6-chloro-2-benzoxazolone 7 and N-(2-oxo-3-aza-5-carboxypentyl)-6-chloro-2-benzoxazolone 12. 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 0.05 micrograms/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.
authors.affiliation: Cemagref, Division GIQUAL, Montpellier, France.
authors.forename: V
authors.initials: V
authors.lastname: Issert
authors.forename: R
authors.initials: R
authors.lastname: Lazaro
authors.forename: F
authors.initials: F
authors.lastname: Lamaty
authors.forename: V
authors.initials: V
authors.lastname: Rolland
authors.forename: P
authors.initials: P
authors.lastname: Besançon
authors.forename: B
authors.initials: B
authors.lastname: Caporiccio
authors.forename: P
authors.initials: P
authors.lastname: Grenier
authors.forename: V
authors.initials: V
authors.lastname: Bellon-Maurel
mesh_terms: D000818:Animals; D000906:Antibodies / Q000276:immunology*; D000918:Antibody Specificity; D004797:Enzyme-Linked Immunosorbent Assay; D006241:Haptens / Q000276:immunology; D007106:Immune Sera; D007306:Insecticides / Q000032:analysis* / Q000276:immunology; D009682:Magnetic Resonance Spectroscopy; D008297:Male; D009946:Organothiophosphorus Compounds / Q000032:analysis* / Q000276:immunology; D011817:Rabbits; D012680:Sensitivity and Specificity
chemical_list: D000906:Antibodies; D006241:Haptens; D007106:Immune Sera; D007306:Insecticides; D009946:Organothiophosphorus Compounds; C003117:phosalone