Production and characterization of monoclonal antibodies against urea derivatives |
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Authors: | Manuela Rohde Jörg A. Schenk Stephan Heymann Olaf Behrsing Gudrun Scharte Gerhard Kempter Jochen Woller Wolfgang E. Höhne Axel Warsinke Burkhard Micheel |
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Affiliation: | (1) Institute of Biochemistry and Molecular Physiology, Potsdam University, Germany;(2) Max Delbrück Center for Molecular Medicine (MDC) Berlin-Buch, Germany;(3) Institute of Organic Chemistry, Potsdam University, Germany;(4) Institute of Biochemistry, Humboldt University, Charité, Berlin |
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Abstract: | A panel of monoclonal antibodies was generated against the ureabased haptenN-(2-N-chloroacetylaminobenzyl)-N′-4-chlorophenylurea as a tool for building up sensitive immune assays to detect urea derivatives and to screen them for catalytic antibodies (Abs). Eleven hybridomas were obtained that produced Abs reactive to the hapten. All Abs were of IgG class. Crossreactivities of the Abs to different haptens were examined, especially to a possible transition-state analog. Only four of the hybridomas (R2-DA10/F7, R2-GE7/H2, R2-HC2/A5, R2-HD6/F7) produced Abs crossreactive with the transition-state analog. From the 11 hybridomas, hybridoma B76-BF5 was chosen for further characterization. Compared to the other Abs, B76-BF5 showed the strongest binding and had a rather restricted specificity. These Abs could be used to build up a sensitive enzyme immunoassay for the detection of the hapten. All Abs were screened for crossreactivity with the pesticides monuron and diuron. No reactivity could be detected. In addition, the nucleotide sequences of the variable light and heavy chain genes of the similarly reactive Abs B76-BF5, B76-BB3, R2-DA10/F7, and R2-GA6/G3 were determined to clarify whether structure and binding specificity of these Abs showed any correlation. |
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Keywords: | N-(2-N-chloroacetylaminobenzyl)-N′ -4-chlorophenylurea pesticides catalytic antibodies hybridomas KABAT database |
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