Food Chemistry: X (Jun 2024)

Development of broad-spectrum immunoassay with monoclonal antibody to detect five eugenols and study of their molecular recognition mechanism

  • Lin Luo,
  • Rui-Yao Kang,
  • Zhen-Xi He,
  • Bao-Zhu Jia,
  • Zi-Jian Chen,
  • Hao Deng,
  • Zhen-Lin Xu

Journal volume & issue
Vol. 22
p. 101255

Abstract

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In this study, three eugenol fragment-containing haptens were synthesized, and a monoclonal antibody (mAb) selective for five commonly-found eugenol compounds (EUGs, i.e., eugenol, isoeugenol, methyl eugenol, methyl isoeugenol, and acetyl isoeugenol) was obtained. Based on this mAb, a broad-spectrum indirect competitive ELISA for high-throughput detection of five EUGs was developed. The detection limits for eugenol, isoeugenol, methyl eugenol, methyl isoeugenol and acetyl isoeugenol in both tilapia and shrimp samples were 25.3/ 50.6 μg/kg, 0.075/0.15 μg/kg, 0.48/0.96 μg/kg, 0.16/0.32 μg/kg, and 18.16/36.32 μg/kg, respectively. The recoveries for five EUGs ranged from 80.4 to 114.0 % with a coefficient of variation less than 11.5 %. Moreover, homology modelling and molecular docking were conducted to elucidate the interactions mechanism of mAb-EUGs. The work provides a promising tool for high-throughput screening of EUGs in aquatic products, which can serve as a benchmark for designing haptens and developing immunoassays for other small molecules.

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