690636-76-1Relevant articles and documents
Enzyme-Linked Immunosorbent Assays based on Rabbit Polyclonal and Rat Monoclonal Antibodies against Isoproturon
Kraemer, Petra M.,Goodrow, Marvin H.,Kremmer, Elisabeth
, p. 2462 - 2471 (2007/10/03)
This work describes the production and characterization of rabbit polyclonal antisera (pAb) and rat monoclonal antibodies (mAb) against isoproturon. Coating antigen and enzyme-tracer formats were developed. Standard curves for isoproturon were conducted either in 40 mM phosphate buffered saline (PBS) or in Milli-Q water. PAb 352 together with the best enzyme tracer revealed in the optimized ELISA (enzyme tracer format) a test midpoint of 1.06 ± 0.34 μg/L (n = 19, standard set up in Milli-Q water) with a detection limit of about 0.1 μg/L. The comparable ELISA with mAb IOC 7E1 had test midpoints of 0.07 ± 0.04 μg/L (n = 7, standards in Milli-Q water) and 0.11 ± 0.08 μg/L (n = 33; standards in 40 mM PBS). The limits of detection were about 0.003 and 0.01 μg/L in Milli-Q water and PBS, respectively. Noticeable cross reactivities (CRs) were seen with the major metabolites, namely 4-isopropylaniline, 4-isopropylphenylurea, and 1-(4-isopropylphenyl)-3-methylurea. With pAb 352, these CRs were 5%, 7%, and 31%, respectively, and with mAb IOC 7E1, they were 3%, 5%, and ca. 19%, respectively. All arylurea herbicides had only minor CRs, which ranged from no CR (e.g., chlorosulfuron) to a maximum of 3.3% (chlortoluron). Influences of organic solvents (methanol, ethanol, acetonitrile, and acetone) were evaluated. Both pAb- and mAb-based immunoassays showed the highest tolerance for methanol, up to 5%. Ethanol and acetonitrile could not be used above 2% without an influence on the assays. The same was true for acetone, although tested only in the mAb-based assay. Water samples of different origins and matrices were spiked and analyzed with these pAb and mAb ELISAs. The results demonstrated that these immunoassays are useful screening tools.
