title: Preparation of antibodies and development of an enzyme-linked immunosorbent assay for determination of dealkylated hydroxytriazines.
abstract: The development of an indirect competitive enzyme-linked immunosorbent assay (ELISA) for dealkylated hydroxytriazines is reported here for the first time. The assay uses polyclonal antibodies raised against N-(4-amine-6-hydroxy-[1,3,5]triazin-2-yl)-4-aminobutanoic acid (hapten 2g) conjugated to keyhole limpet hemocyanin by the active ester method. The immunizing hapten was synthesized by first introducing the amino group to the triazine ring in a protected form in order to increase its solubility in organic media. Subsequent steps consisted of reacting this compound with an appropriate spacer arm, followed by removal of the protecting group in acidic media. The resulting assay uses a homologous competitor hapten coupled to conalbumin by the mixed anhydride method. Coating antigens prepared using a homologous covalent coupling procedure failed to produce competitive immunoassays. The assay tolerates media with high ionic strength (up to 70 mS cm(-)(1)) and basic pH values (7.5-9.5 units). Under the optimized conditions, this ELISA is specific for dealkylated hydroxytriazines, reaching suitable limits of detection.
authors.affiliation: Department of Biological Organic Chemistry, IIQAB-CSIC, Jordi Girona, 18-26, 08034-Barcelona, Spain.
authors.forename: Nuria
authors.initials: N
authors.lastname: Sanvicens
authors.forename: Valerie
authors.initials: V
authors.lastname: Pichon
authors.forename: Marie-Claire
authors.initials: MC
authors.lastname: Hennion
authors.forename: M-Pilar
authors.initials: MP
authors.lastname: Marco
mesh_terms: D000818:Animals; D000906:Antibodies / Q000276:immunology; D000941:Antigens / Q000276:immunology; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D005260:Female; D006241:Haptens / Q000737:chemistry / Q000276:immunology; D006433:Hemocyanins / Q000276:immunology; D006540:Herbicides / Q000032:analysis*; D006863:Hydrogen-Ion Concentration; D007114:Immunization; D016014:Linear Models; D009994:Osmolar Concentration; D010573:Pesticide Residues / Q000032:analysis*; D011817:Rabbits; D012680:Sensitivity and Specificity; D014227:Triazines
chemical_list: D000906:Antibodies; D000941:Antigens; D006241:Haptens; D006540:Herbicides; D010573:Pesticide Residues; D014227:Triazines; D006433:Hemocyanins; C032808:keyhole-limpet hemocyanin