Enzyme Immunoassay for 2,4,6-Trichloroanisole Based on Monoclonal Antibodies

Monoclonal antibodies (mabs) are of great interest according to their homologous abilities, high specificity, and unlimited possibility of production. Mabs against different substances have been developed. They are used in many fields of application (e.g., medicine and pesticide analysis). Immunoassays performed with mabs have the advantages of easy handling and high sensitivity. The goal of this study was to develop mabs against 2,4,6-trichloroanisole (TCA). TCA has been identified as the most potent contaminant in wine causing a mouldy taste. Several mice have been immunized with two different TCA haptens. Spleen cells have been gained and prepared following the technique developed by Köhler and Milstein (Köhler, G.; Milstein, C. Continuous cultures of fused cells secreting antibody of predefined specificity. Nature 1975, 256, 495–497). After a screening period with a competitive immunoassay two cells lines have been detected that produce TCA-specific antibodies. Both different antibodies have been used to develop ELISAs. One ELISA with the hapten B mab (Hbab) and one ELISAs with the hapten C mab (Rami). The ELISA with the hapten C mab was afterwards optimized with signal amplification. Each ELISA shows a remarkable detection limit for TCA. The ELISA with signal amplification performed with the mab Rami detects 20 ng/L and the ELISA with the mab Hbab 10 ng/L TCA. Both mabs have been used to measure spiked wine samples. Only with the mab Hbab were spiked cork samples also determined.