Production of Monoclonal Antibodies for the Determination of s-Triazines with Enzyme Immunoassays

Two triazine derivatives, ametryn sulphoxide (2-ethylamino-4-isopropylamino-6-methylsulphoxide-l,3,5-triazine) and dichloroatrazine (2,6-dichloro-4-isopropylamino-1,3,5-triazine) were conjugated to bovine serum albumin (BSA) and used for the immunization of BALB/c mice. Hybridomas were produced by cell fusion of immune spleen and mouse myeloma cells (PAI-B3AG8J). After screening with a competitive enzyme-linked immunosorbent assay (ELISA), four anti-triazine monoclonal antibodies from permanent hybridoma cell lines were selected for further characterization. Cross-reaction studies with the antibodies developed against the ametryn sulphoxide conjugate showed strong affinities to terbutryn and prometryn. ELISAs with antibodies from clone AS-K1F4 reached a detection limit of approximately 0.1μg/l for terbutryn, and with AS-K1A11 antibodies a limit of 0–3 μg/l for prometryn was attained. Cell lines from mice immunized with the dichloroatrazine-conjugate produced antibodies with the highest affinities to aziprotryn. The detection limit of the corresponding ELISA was 1μg/l (clone DC-C3K5). The scope of the monoclonal antibodies for use in pesticide residue analysis is discussed with respect to polyclonal antibodies.