A monoclonal antibody-based piezoelectric immunosensor for the analysis of carbaryl

Authors: C. March, A. Abad, A. Arnau, Y. Jiménez, T. Sogorb, A. Montoya

Event: 4th MGPR International Symposium of pesticides in food and the environment in Mediterranean Countries. Pine Bay Holiday Resort Kusadasi-Aydin. Turkey (2005)

 

Immunosensors combine immunoassay selectivity with the high sensitivity often provided by electronic signal transducers. In this communication, the development of a monoclonal antibody-based Piezoelectric Immunosensor for the analysis of carbaryl is described and coupled to BSA were covalently attached to gold coated AT- cut 9 MHz quartz crystals, previously functionalized with a self-assembled alkanethiol monolayer. The crystal was inserted in a homemade Arnite cell that allowed only one face of the crystal to be in contact with the reagents. The cell was integrated in a flow-through system and pesticide detection was accomplished by competitive immunoassay in the conjugate-coated format. The assay consisted of the incubation of the analyte with a specific monoclonal antibody, followed by the immunoreaction of the mixture with the carbaryl derivative immobilized on the sensor surface. The frequency variations of the piezoelectric crystal were measured with a commercial Research Quartz Crystal Microbalance (RQCM) and correlated with the carbaryl concentration in the standards. The total time required for a complete assay cycle, including regeneration, was 20 min. As it corresponds to binding inhibition immunoassays, the frequency signal provided by the sensor was inversely proportional to the analyte concentration. A sigmoidal calibration curve was obtained by fitting experimental points to a four-parameter equation. The immunoassay sensitivity, expressed as the carbaryl concentration that reduced the assay signal by 50% (I50), was around 25 µg/l. The inhibition curve would allow the detection of carbaryl in fruits and vegetables at European regulatory levels.