Analytical, thermodynamical and kinetic characteristics of photoluminescence immunosensor for the determination of Ochratoxin A

Biosens Bioelectron. 2018 Jan 15:99:237-243. doi: 10.1016/j.bios.2017.07.056. Epub 2017 Jul 22.

Abstract

Ochratoxin A (OTA) is one of the most widespread and dangerous food contaminants. Therefore, rapid, label-free and precise detection of low OTA concentrations requires novel sensing elements with advanced bio-analytical properties. In the present paper we report photoluminescence (PL) based immunosensor for the detection of OTA. During the development of immunosensor photoluminescent ZnO nanorods (ZnO-NRs) were deposited on glass substrate. Then the ZnO-NRs were silanized and covalently modified by Protein-A (Glass/ZnO-NRs/Protein-A). The latest structure was modified by antibodies against OTA (Anti-OTA) in order to form OTA-selective layer (Glass/ZnO-NRs/Protein-A/Anti-OTA). In order to improve immunosensors selectivity the surface of Glass/ZnO-NRs/Protein-A/Anti-OTA was additionally blocked by BSA. Formed Glass/ZnO-NRs/Protein-A/BSA&Anti-OTA structures were integrated within portable fiber optic detection system, what is important for the development of low cost and portable immunosensors. The immunosensor has been tested in a wide range of OTA concentrations from 10-4ng/ml until 20ng/ml. Interaction isotherms were derived from analytical signals of immunosensor. Association constant and Gibbs free energy for the interaction of Glass/ZnO-NRs/Protein-A/Anti-OTA with OTA were calculated, analyzed and compared with some other related results. Sensitivity range and limit of detection were determined as 0.1-1ng/ml and 10-2ng/ml, respectively. Interaction kinetics of ZnO-NRs with OTA was evaluated. Response time of the immunosensor toward OTA was in the range of 500-800s. Some insights related to the mechanism of PL-signal generation are proposed and discussed.

Keywords: Immobilization of antibodies; Interaction kinetics and thermodynamics; Ochratoxin A; Photoluminescence; ZnO-based immunosensor.

MeSH terms

  • Antibodies / chemistry*
  • Biosensing Techniques*
  • Food Analysis*
  • Food Contamination / analysis
  • Gold / chemistry
  • Humans
  • Limit of Detection
  • Nanotubes / chemistry
  • Ochratoxins / isolation & purification*
  • Ochratoxins / toxicity
  • Zinc Oxide / chemistry

Substances

  • Antibodies
  • Ochratoxins
  • ochratoxin A
  • Gold
  • Zinc Oxide