Production of monoclonal antibodies for the determination of s‐triazines with enzyme immunoassays

Abstract

Two triazine derivatives, ametryn sulphoxide (2‐ethylamino‐4‐isopropylamino‐6‐methyl‐sulphoxide‐1,3,5‐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‐B3AG8.I). 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 0.1 μg/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 1 μg/l (clone DC‐C3K5). The scope of the monoclonal antibodies for use in pesticide residue analysis is discussed with respect to polyclonal antibodies.