Preparation of anti-fragrant monoclonal antibodies by the rat lymph node method and their characterization using enzyme-linked immunosorbent assay

In the food and beverage industries there is a great demand for a sensitive and easy detection system of fragrant compounds in order to ensure product quality control and to avoid contamination. Benzaldehyde (BZ) and furfural (FF) are typical fragrant compounds for peach-flavored beverages, and methyl anthranilate (MA) is a typical flavor of grape beverages. For sensitive detection of these flavors by immunoreaction, we prepared monoclonal antibodies against BZ, FF and MA by using the rat lymph node method. The antibody for BZ had high selectivity toward BZ compared with other fragrant compounds and high affinity with an affinity constant of 1.9 x 10E6/M. The limit of detection (LOD) of BZ by indirect competitive ELISA (ic-ELISA) was 0.6 ng/mL. For FF, anti-FF mAb had high selectivity toward FF and high affinity constant of 2.5 x 10E4/M. The LOD of FF by ic-ELISA was 100 ng/mL. For MA, anti-MA mAb had high selectivity toward MA and high affinity constant of 1.3 x 10E5/M. The LOD of MA by ic-ELISA was 200 ng/ml.These monoclonal antibodies were suitable for detection of very small amounts of fragrant compounds in rinsed water of beverage industries.