Porous silicon based photoluminescence immunosensor for rapid and highly-sensitive detection of Ochratoxin A

Rok publikacji: 2018
Wydawca:  Biosensors and Bioelectronics, 2018, 99, 661-667.
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V. Myndrul, R. Viter, M. Savchuk, N. Shpyrka, D. Erts, D. Jevdokimovs, V. Silamiķelis, V. Smyntyna, A. Ramanavicius, I. Iatsunskyi
A rapid and low cost photoluminescence (PL) immunosensor for the determination of low concentrations of Ochratoxin A (OTA) has been developed. This immunosensor was based on porous silicon (PSi) and modified by antibodies against OTA (anti-OTA). PSi layer was fabricated by metal-assisted chemical etching (MACE) procedure. Main structural parameters (pore size, layer thickness, morphology and nanograins size) and composition of PSi were investigated by means of X-Ray diffraction (XRD), scanning electron microscopy (SEM) and Raman spectroscopy. PL-spectroscopy of PSi was performed at room temperature and showed a wide emission band centered at 680 ± 20nm. Protein A was covalently immobilized on the surface of PSi, which in next steps was modified by anti-OTA and BSA in this way a anti-OTA/Protein-A/PSi structure sensitive towards OTA was designed. The anti-OTA/Protein-A/PSi-based immunosensors were tested in a wide range of OTA concentrations from 0.001 upto 100ng/ml. Interaction of OTA with anti-OTA/Protein-A/PSi surface resulted in the quenching of photoluminescence in comparison to bare PSi. The limit of detection (LOD) and the sensitivity range of anti-OTA/Protein-A/PSi immunosensors were estimated. Association constant and Gibbs free energy for the interaction of anti-OTA/Protein-A/PSi with OTA were calculated and analyzed using the interaction isotherms. Response time of the anti-OTA/Protein-A/PSi-based immunosensor toward OTA was in the range of 500-700s. These findings are very promising for the development of highly sensitive, and potentially portable immunosensors suitable for fast determination of OTA in food and beverages.

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