This work reviews our recent progress in development of novel optical methods of detection of mycotoxins in direct assay with either specific antibodies or aptamers. The main method in this work was the total internal reflection ellipsometry (TIRE) combined with LSPR transducers based on gold nano-structures produced by annealing of thin gold films. The gold nano-islands produced were characterised with SEM, AFM, UV-visible absorption spectroscopy, and spectroscopic ellipsometry. The combination of TIRE and LSPR offers superior refractive index sensitivity as compared to traditional UV-vis absorption spectroscopy. The limitations of LSPR related to a short evanescent field decay length can be overcome using small-size bio-receptors, such as halfantibodies and aptamers. The achieved sensitivity of detection of mycotoxins in 0.01ppb level of concentration is sufficient for the use of this method for analysis of agriculture products, food and feed on the presence of mycotoxins. Even higher sensitivity in sub-ppt level was achieved with another optical biosensor developed recently; it is based on optical planar waveguide operating as polarization interferometer (PI). This method is promising for development of portable, highly sensitive, and simple to use biosensors suitable for point-of-need detection of mycotoxins.
The research aim of this work is to develop a simple and highly sensitive optical biosensor for detection of mycotoxins. This sensor is built on a planar waveguide operating on the polarization interferometry principle, i.e., detecting a phase shift between p- and s-components of polarized light developed during the binding of analyte molecules. The operation of the proposed sensor is similar to that of a Mach–Zehnder interferometer, while its design is much simpler and it does not require splitting the waveguide into two arms. The refractive index sensitivity of the polarization interferometer sensor was in the range of 5200 radians per refractive index unit (RIU). Several tests were conducted to detect ochratoxin A (OTA) at different concentrations in direct immunoassay with specific antibodies immobilized in the sensing window. The lowest concentration of OTA of 0.01 ng/mL caused a phase shift of nearly one period. The results obtained prove high sensitivity of the sensors, which are capable of detecting even lower concentrations of mycotoxins at the ppt (part-per-trillion) level.
A planar waveguide (PW) immunosensor working as a polarisation interferometer was developed for the detection of mycotoxin zearalenone (ZON). The main element of the sensor is an optical waveguide consisting of a thin silicon nitride layer between two thicker silicon dioxide layers. A combination of a narrow waveguiding core made by photolithography with an advanced optical set-up providing a coupling of circular polarised light into the PW via its slanted edge allowed the realization of a novel sensing principle by detection of the phase shift between the p- and s-components of polarised light propagating through the PW. As the p-component is sensitive to refractive index changes at the waveguide interface, molecular events between the sensor surface and the contacting sample solution can be detected. To detect ZON concentrations in the sample solution, ZON-specific antibodies were immobilised on the waveguide via an electrostatically deposited polyelectrolyte layer, and protein A was adsorbed on it. Refractive index changes on the surface due to the binding of ZON molecules to the anchored antibodies were detected in a concentration-dependent manner up to 1000 ng/mL of ZON, allowing a limit of detection of 0.01 ng/mL. Structurally unrelated mycotoxins such as aflatoxin B1 or ochratoxin A did not exert observable cross-reactivity.
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