Production of polyclonal antibodies towards the immunodetection of insecticide phosalone
| Autor: | B. Caporiccio, V. Issert, P. Grenier, V. Rollande, V. Bellon-Maurel, René Lazaro, P. Besançon, Frédéric Lamaty |
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| Přispěvatelé: | Institut des Biomolécules Max Mousseron [Pôle Chimie Balard] (IBMM), Centre National de la Recherche Scientifique (CNRS)-Institut de Chimie du CNRS (INC)-Université de Montpellier (UM)-Ecole Nationale Supérieure de Chimie de Montpellier (ENSCM), division giqual, Centre national du machinisme agricole, du génie rural, des eaux et forêts (CEMAGREF), Lab. Génie Biologique et Sciences des aliments, Unité de nutrition- département agroressources et procédés biologiques, Université Montpellier 2 - Sciences et Techniques (UM2) |
| Rok vydání: | 2000 |
| Předmět: |
0106 biological sciences
Male Insecticides Magnetic Resonance Spectroscopy hapten synthesis medicine.medical_treatment Clinical Biochemistry Enzyme-Linked Immunosorbent Assay 01 natural sciences Biochemistry Horseradish peroxidase Sensitivity and Specificity Antibodies chemistry.chemical_compound Antibody Specificity 010608 biotechnology medicine Animals Phosalone [SDV.BBM.BC]Life Sciences [q-bio]/Biochemistry Molecular Biology/Biochemistry [q-bio.BM] Bovine serum albumin Antiserum biology [CHIM.ORGA]Chemical Sciences/Organic chemistry Immune Sera 010401 analytical chemistry Organic Chemistry polyclonal Organothiophosphorus Compounds phosalone 0104 chemical sciences immunodetection chemistry Polyclonal antibodies biology.protein Amino acids Thyroglobulin Rabbits Hapten Haptens Conjugate |
| Zdroj: | Amino Acids Amino Acids, Springer Verlag, 1999, 17 (4), pp.377-389. ⟨10.1007/BF01361663⟩ Scopus-Elsevier |
| ISSN: | 0939-4451 1438-2199 |
| DOI: | 10.1007/BF01361663⟩ |
| Popis: | 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. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
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