Repetitive Immunoassay with a Surface Acoustic Wave Device and a Highly Stable Protein Monolayer for On-Site Monitoring of Airborne Dust Mite Allergens.

This work describes a sensor to be incorporated into the on-site monitoring system of airborne house dust mite (HDM) allergens. A surface acoustic wave (SAW) device was combined with self-assembled monolayers of a highly stable antibody capture protein on the SAW surface that have high resistance to pH change. A sandwich assay was used to measure a HDM allergen, Der f 1 derived from Dermatophagoides farinae. Capture antibodies were cross-linked to a protein G based capture layer (ORLA85) on the sensor surface, thereby only Der f 1 and detection antibodies were regenerated by changing pH, resulting in fast repetition of the measurement. The sensor was characterized through 10 repetitive measurements of Der f 1, which demonstrated high reproducibility of the sensor with the coefficient of variation of 5.6%. The limit of detection (LOD) of the sensor was 6.1 ng·mL(-1), encompassing the standard (20 ng·mL(-1)) set by the World Health Organization. Negligible sensor outputs were observed for five different major allergens including other HDM allergens which tend to have cross-reactivity to Der f 1 and their mixtures with Der f 1. Finally, the sensor lifetime was evaluated by conducting three measurements per day, and the sensor output did not substantially change for 4 days. These characteristics make the SAW immunosensor a promising candidate for incorporation into on-site allergen monitoring systems.

Repeated immunosensing by a dithiobis(succinimidyl propionate)-modified SAW device.

The possibility of dithiobis(succinimidyl propionate) (DSP) for repeated immunoassay with a surface acoustic wave (SAW) immunosensor was explored. In the sensor, DSP was used to modify a gold-coated quartz sensing area of a SAW device by forming a self-assembled monolayer on the gold surface. In a model sandwich assay using mouse (mIgG) and anti-mouse (a-mIgG) antibodies, the primary antibody, mIgG, firstly reacted with N-hydroxysuccinimide ester groups of DSP and was immobilized on the SAW device to fabricate the SAW immunosensor. Optimization of adsorption time of mIgG revealed that both degrees of adsorption and immobilization of mIgG reached a saturation at 30 min although the immobilization was more dependent on the adsorption time. Through characterization of the DSP-modified SAW immunosensor, a high selectivity, with which no sensor output was observed from various kinds of secondary antibodies except for a-mIgG, along with 8-fold shorter measurement time (15 min) than that of enzyme-linked immunosorbent assay were obtained. Furthermore, 10 repeated measurement of a-mIgG demonstrated a high reproducibility of the sensor output (coefficient of variation of 7.0%). These validate the utility of DSP in the SAW immunosensor for rapid and repeated measurement of antigens.