Synthesis of Haptens and Derivation of Monoclonal Antibodies for Immunoassay of the Phenylurea Herbicide Diuron

Diuron and related phenylurea herbicides and their metabolites are important candidates for sensitive and specific immunodetection. This paper describes a scheme for the synthesis of two different types of phenylurea haptens for immunization and use as detecting conjugates in enzyme immunoassays (EIAs). The haptens were used to develop indirect and direct EIAs and to derive a panel of monoclonal antibodies (MAbs) with different specificities for diuron and its analogs. One of six possible combinations of hapten-spacer arm conjugates tested as immunizing and screening antigens resulted in an indirect competition EIA that was 100-2000-fold more sensitive than the others. The eight most sensitive MAbs had 160 values of 2-20 ppb for diuron. These MAbs gave two different patterns of cross-reactivities with monuron and linuron and negligible recognition of other arylurea herbicides. These MAbs and EIAs are potentially suitable for identification as well as detection of diuron, monuron, and linuron.

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