The imidazolinone compounds are a new class of herbicides which are environmentally safe and effective at very low application rates. Because of the shared imidazolinone ring structure amongst several herbicides, generic antibodies were prepared which recognized all the compounds in this class (1). These antibodies are useful not only for developing immunoassays for the detection of low levels of residues in soil or in plant extracts, they are also useful for preparing antibody affinity columns used for sample clean-up in metabolism and residue studies. Imidazolinone-compound containing extracts from wheat plant, corn grain, corn fodder, goat urine, and goat kidney tissue were tested on the antibody columns. It was found that the analytes were bound to the antibody column after some simple sample preparation procedures. The bound analytes were easily eluted with a solution of 30% methanol in water. It was further demonstrated that metabolites of imidazolinone compounds which retain the imidazolinone ring structure can be purified through a simple antibody affinity chromatography procedure and be identified thereafter by mass spectrometry. A comparison of mono-and poly-clonal antibody columns indicates that the monoclonal antibody with its uniform affinity and more restricted binding epitope is better for antibody affinity chromatographies. A lower amount of matrix is bound to the monoclonal antibody column, thus producing a cleaner sample than a polyclonal antibody column.The specificity of antibodies has been utilized extensively in immunoassays where analytes can be quantified in relatively crude matrices. This property can be further harnessed in affinity chromatography. Preparation of monoclonal antibodies allows an unlimited supply of mono-specific antibodies for large scale production of affinity 0097-6156/95/0586-0235$12.00/0
A set of haptens structurally resembling the herbicide imazethapyr (PURSUIT) was synthesized and used to derive monoclonal antibodies (MAbs) and direct and indirect competition enzyme immunoassays (EIAs) which could detect imazethapyr, imazaquin (SCEPTER), imazapic (CADRE), and imazamox (RAPTOR) in the 3-30 ng/mL (parts per billion) range, and imazapyr (ARSENAL) and imazamethabenz-methyl (ASSERT) in the 300-500 ppb range. Two MAbs, 3A2 and 3A5, had affinities of 10-75 nM for imazethapyr. MAbs 1A5, 1D2, and 3A5 were specific for the S isomers of the herbicides. Some MAbs were stable in solutions containing up to 15% methanol and 5% acetonitrile in indirect EIAs. Plates coated with hapten conjugates for indirect EIA could be stored frozen. Selectivity for the imidazolinones by some MAbs varied with different coating conjugates. These MAbs and haptens should prove useful in immunochemical analysis and residue recovery methods for imazethapyr and other imidazolinone herbicides.
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