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<p> The use of a protein-based electrochemical sensor for rapid and sensitive detection of ochratoxin (OTA) is explored. We have employed a combinatorial approach involving <i>in silico</i> target fishing and molecular docking analysis to identify the protein candidates with high affinity for OTA. Porcine pancreatic α-amylase has emerged as a promising candidate. α-Amylase was immobilized on multi-walled carbon nanotube (MWCNT) and amine modified titanium dioxide nanoparticles (TNP-NH<sub>2</sub>) nanocomposite coated glassy carbon electrode (GCE) (α-amylase/NH<sub>2</sub>-TNP + MWCNT/GCE) <i>via</i> inverse NHS-EDC chemistry within 40&#xa0;min. The successful immobilization of α-amylase was confirmed by Fourier transform Infrared (FT-IR) spectroscopy and bicinchoninic acid (BCA) assay. The biosensor has displayed an excellent limit of detection (LOD) of 0.074 ng/mL, good specificity towards OTA in the presence of other mycotoxins (patulin and aflatoxin B1), and interfering molecules in foods such as starch and some tested polyphenols. This sensor has good reproducibility with relative standard deviation (RSD) of 0.735%, is stable for 11 days to efficiently detect OTA and for single use only. Our sensor has displayed a similar detection performance of OTA as the conventional method i.e. ultra high performance liquid chromatography-ultra-violet detection (UPLC-UV) studies in OTA spiked three food samples (corn, chilli, and rice). The swift preparation time, coupled with high sensitivity and selectivity, positions this sensor as a promising tool for enhancing food safety measures and mitigating the risks associated with OTA contamination.</p> Graphical Abstract <p></p>

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Bioinformatic based protein-ochratoxin A screening studies and fabrication of a protein-based electrochemical sensor for specific detection of ochratoxin A in food samples

  • 期刊论文

  • 发表日期：

    2025年11月12日

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  • Megala Ulaganambi,
  • Kishore K. R. Tetala

摘要

The use of a protein-based electrochemical sensor for rapid and sensitive detection of ochratoxin (OTA) is explored. We have employed a combinatorial approach involving in silico target fishing and molecular docking analysis to identify the protein candidates with high affinity for OTA. Porcine pancreatic α-amylase has emerged as a promising candidate. α-Amylase was immobilized on multi-walled carbon nanotube (MWCNT) and amine modified titanium dioxide nanoparticles (TNP-NH2) nanocomposite coated glassy carbon electrode (GCE) (α-amylase/NH2-TNP + MWCNT/GCE) via inverse NHS-EDC chemistry within 40 min. The successful immobilization of α-amylase was confirmed by Fourier transform Infrared (FT-IR) spectroscopy and bicinchoninic acid (BCA) assay. The biosensor has displayed an excellent limit of detection (LOD) of 0.074 ng/mL, good specificity towards OTA in the presence of other mycotoxins (patulin and aflatoxin B1), and interfering molecules in foods such as starch and some tested polyphenols. This sensor has good reproducibility with relative standard deviation (RSD) of 0.735%, is stable for 11 days to efficiently detect OTA and for single use only. Our sensor has displayed a similar detection performance of OTA as the conventional method i.e. ultra high performance liquid chromatography-ultra-violet detection (UPLC-UV) studies in OTA spiked three food samples (corn, chilli, and rice). The swift preparation time, coupled with high sensitivity and selectivity, positions this sensor as a promising tool for enhancing food safety measures and mitigating the risks associated with OTA contamination.

Graphical Abstract