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Development and validations of s-triazine and pyrethroid-specific immunosensors


Disposable immunosensors based on non-commercial, monoclonal atrazine-, terbuthylazine- and allethrin-specific antibodies will be developed and validated. They are hoped to find practical application in the rapid, sensitive, specific and inexpensive monitoring of pesticides in water, soil and aerosol samples. An optical and an electrochemical transducer system, based on thick-film electrodes, will be investigated. An important part of this work will focus on the extensive validation of our proposed immunosensors over a wide range of different areas of agricultural production (Germany and Italy) and on the comparison of our results with conventional GC-MS and HPLC-MS analyses.

Monoclonal antibodies (mab) of uniform quality will be provided to all participants concerning the mass production of mab. Using established cell lines, monoclonal antibodies against the herbicides terbuthylazine and atrazine as well as the insecticide allethrin, a pyrethroid, will be produced in cell culture. A roller bottle system allows the production of mab in quantities up to several hundred mg. The purification of the mab required for various applications will be carried out by chromatographic methods. Some applications demand Fab fragments of the antibodies. For this purpose the mab will be cleaved by papain digestion. Further mab directed against other pyrethroids will be produced.

A sensor based on an uniform standardised immunoassay, optimised for atrazine, terbuthylazine and allethrin analysis will be developed. As a first step to immunosensor development, a test strip assay with reflectometric detection will be developed. Starting with the flow injection immunoanalysis (FIIA) for the analysis of several pesticides, the features of the automated flow injection system will be simplified and incorporated into a portable sensing device for field use.

The results will be a novel disposable immunosensing device (based on non-commercial atrazine-, terbuthylazine- and allethrin-specific antibodies) to find practical application in the rapid, sensitive, specific and inexpensive monitoring of pesticides in water and soil samples. Compared to conventional physicochemical methods of pesticide analysis, the immunosensors developed with an optical and an electrochemical transducer system will provide an increase in sensitivity and specificity. Sample preparation will not be required for the measurement of water samples. Other than in standard procedures, no false-negative results are produced. The assay formats chosen for our system will allow for assays on place ("field tests") and thus an extensive validation of various environmental samples in line with the pertinent European regulations, e.g. the European Drinking Water Directives.

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Allmandring 31 Zentrum für Bioverfahrenstechnik
70569 Stuttgart

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