Generation of antiserum to Irgarol 1051 and development of a sensitive enzyme immunoassay using a new heterologous hapten derivative.
A polyclonal antiserum to Irgarol 1051 was developed in sheep and used to construct an enzyme immunoassay method for the measurement of the antifouling compound in river and seawater samples. The antiserum was generated by a hapten derivative, 2-(tert-butylamino)-4-(cyclopropylamino)-6-(thiopropionic acid)-1,3,5-triazine, coupled to a mixture of keyhole limpet hemocyanin and bovine serum albumin, and the competitive enzyme immunoassay was constructed using a plate-coating antigen made of a heterologous new hapten derivative, 2-(tert-butylamino)-4-(cyclopropylamino)-6-(phenoxybenzoic acid)-1,3,5-triazine, linked to gelatine. The assay showed a sensitivity of about 5 ng L(-1) in river and seawater matrices with reasonable specificity with respect to commonly used triazines such as atrazine (3%), simazine (>0.1%) and desethylatrazine (>0.01%). However, high cross-reactivity levels were found with ametryn (56%) and prometryn (60%). Tests on the effects of organic solvents on assay performance indicated a high tolerance to methanol but much less so to acetonitrile. The assay was found to be highly reproducible and robust owing to the stability of the sheep antibody and the highly optimised competitive assay reagents which included the use of the new triazine-O-phenoxybenzoic acid derivative.
Abuknesha
Ramadan A
RA
School of Health and Life Sciences, Analytical Research Group, Pharmaceutical Sciences Division, King's College London, University of London, 150 Stamford Street, London SE1 9NN, UK. ram.abuknesha@kcl.ac.uk
Griffith
Hannah M T
HM
D006241:Haptens / Q000737:chemistry*; D007106:Immune Sera / Q000276:immunology*; D007124:Immunoenzyme Techniques / Q000379:methods*; D012680:Sensitivity and Specificity; D014227:Triazines / Q000276:immunology*
D006241:Haptens; D007106:Immune Sera; D014227:Triazines; C103341:irgarol 1051