Monoclonal-Based ELISA for the Identification of Herbicidal Cyclohexanedione Analogues That Inhibit Graminaceous Acetyl Coenzyme-A Carboxylase

Abstract

Cyclohexanediones are one of four known structural classes of herbicides that inhibit graminaceous acetyl coenzyme-A carboxylase (ACCase; EC 6.4.1.2). Five monoclonal antibodies were raised against cyclohexanediones conjugated to bovine serum albumin. Cross-reactivity studies using a homologous competitive indirect enzyme-linked immunosorbent assay (ciELISA) against 24 cyclohexanedione analogues revealed that two monoclonal antibodies (mAb A and mAb B) could segregate the analogues into active and inactive ACCase inhibitors on the basis of the analogue concentration required to inhibit 50% of antibody binding to the coating conjugate (IC(50)). Both mAb A and mAb B were also found to cross-react with various members of the indolizidinedione structural class of ACCase inhibitors in ciELISA, suggesting that both cyclohexanediones and indolizidinediones possess features recognized by monoclonal antibodies important for the inhibition of ACCase activity. In conclusion, pharmacophore-specific antibodies may be potentially valuable screening tools for the identification of new lead chemistries in a pesticide discovery program.

Development and Evaluation of an Immunological Approach for the Identification of Novel Acetyl Coenzyme-A Carboxylase Inhibitors: Assay Optimization and Pilot Screen Results

“…For example, none of the active aryloxyphenoxypropionates tested inhibited mAb A binding to CCA by 50%. These results are consistent with earlier studies (Webb and Hall, 2000). However, active analogues 17-20 (i.e., 67% of all actives) ( Figure 3) inhibited both mAb A and mAb B binding by 50% when coating conjugate CCE was used (Table 2).…”

Synthesis of Ligand-Specific Phage-Display ScFv against the Herbicide Picloram by Direct Cloning from Hyperimmunized Mouse

“…The versatility and flexibility of antibody (Ab) technology can be used in agriculture in numerous ways. Antibodies can be used in immunoassays to monitor agrochemicals in the environment for registration and stewardship purposes ( − ), in affinity chromatography for extraction and purification of ligands of interest (), and to screen potential lead chemistry for development of new drugs and pesticides ( − ). Production of recombinant Abs using phage-display technology has many advantages over conventional polyclonal and monoclonal Ab production ( 2 , 8 , 9 ).…”

Immunomodulation confers herbicide resistance in plants

“…As exemplified by the diverse array of antibodies available for binding herbicides with unique chemical structures and modes of action, antibodies can be raised against or selected for specific binding to a seemingly unlimited repertoire of haptens (< 1000 Da) (Meulenberg et al ., 1995; Parnell and Hall, 1998; Clegg et al ., 1999, 2001; Webb and Hall, 2000). In this study, we tested the hypothesis that antibodies with specific xenobiotic affinities are able to generate xenobiotic resistance when expressed in plants.…”

“…As exemplified by the diverse array of antibodies available for binding herbicides with unique chemical structures and modes of action, antibodies can be raised against or selected for specific binding to a seemingly unlimited repertoire of haptens (< 1000 Da) (Meulenberg et al, 1995;Parnell and Hall, 1998;Clegg et al, 1999Clegg et al, , 2001Webb and Hall, 2000).…”