Immunodetection by quartz crystal microbalance. A new approach for direct detection of rabbit IgG and peroxidase.

Biodetection is one of the most important challenges for the twenty-first century: many fields are concerned, mainly environmental and medical. The quartz crystal microbalance (QCM) may offer great possibilities for this purpose: a direct response signal, which characterizes the binding event between a sensitive layer, immobilized onto the surface transducer, and the analyte to be detected, can be obtained. However, for the detection of small biomolecules such as antigens, it is quite difficult to obtain an observable signal that corresponds directly to the binding event. In general, this is owing to the lack of mass sensitivity of the commonly used QCM, with 5- to 10-MHz quartz crystals. For improving this mass sensitivity, a 27-MHz quartz resonator was developed and incorporated in a flow-through microcell. Two biospecies, IgG rabbit and peroxidase enzyme, were studied with this ultra-sensitive QCM in terms of specificity, detection limit, and calibration curve.

Validation of antibody-based recognition by piezoelectric transducers through electroacoustic admittance analysis.

The development of immunosensors based on piezoelectric transducers is widely investigated due to their attractive potentialities. The quartz crystal microbalance (QCM) may give a direct response signal which characterizes the binding event between a sensitive layer, immobilized onto the surface transducer, and the analysis to be detected. However, for small biomolecules, such as some antigens, it is quite difficult to obtain an observable signal. This is mainly due to the lack of sensitivity of the commonly used QCM (5 to 10 MHz quartz crystal). Moreover, the mass estimated with the QCM response through the Sauerbrey equation and the mass which can be measured thanks to other analytical techniques, in our case an enzymatic assay, are different: the deposited mass is generally overestimated by the QCM. To validate QCM mass measurements and, therefore antigens recognition, the interactions of acoustic shear waves with a biolayer were investigated during enzyme adsorption onto the microbalance gold electrode or during the antibody/antigen binding. Electroacoustic admittance was measured around the resonance frequency of a 27 MHz quartz resonator in parallel with microbalance measurements. The parameters which characterize the quartz microbalance equivalent circuit were compared with the classical microbalance frequency. The mass overestimation, given by the microbalance, could be explained either by modification of the rheological properties of the sensitive layers and/or by an inadequacy of the assay performed.