Nanobody-based magnetic chemiluminescence immunoassay for one-pot detection of ochratoxin A

  • Talanta. 2023 Jun 1:258:124388. doi: 10.1016/j.talanta.2023.124388.
Hu Zuo  1 ,  Xinyang Wang  2 ,  Wentao Liu  2 ,  Zongfen Chen  2 ,  Ruonan Liu  2 ,  Han Yang  2 ,  Chunyan Xia  2 ,  Jinli Xie  2 ,  Tieqiang Sun  3 ,  Baoan Ning  4
Affiliations
  • 1. College of Food Science and Technology, Shanghai Ocean University, Shanghai, China; Tianjin Key Laboratory of Risk Assessment and Control Technology for Environment and Food Safety, Tianjin Institute of Environmental and Operational Medicine, Tianjin, China.
  • 2. Tianjin Key Laboratory of Risk Assessment and Control Technology for Environment and Food Safety, Tianjin Institute of Environmental and Operational Medicine, Tianjin, China.
  • 3. Tianjin Key Laboratory of Risk Assessment and Control Technology for Environment and Food Safety, Tianjin Institute of Environmental and Operational Medicine, Tianjin, China. Electronic address: [email protected].
  • 4. College of Food Science and Technology, Shanghai Ocean University, Shanghai, China; Tianjin Key Laboratory of Risk Assessment and Control Technology for Environment and Food Safety, Tianjin Institute of Environmental and Operational Medicine, Tianjin, China. Electronic address: [email protected].
PMID: 36921368 DOI: 10.1016/j.talanta.2023.124388
Abstract

Ochratoxin A (OTA) contamination seriously threatens food safety and human health and requires sensitive and rapid tools for monitoring. In this study, a convenient enzyme-linked immunosorbent assay based on Avi-labeled nanobody Nb-2G/streptavidin-alkaline Phosphatase and Magnetic Beads (MBS-ELISA) was established for the sensitive detection of OTA, which could be used for one-pot detection without immobilization. After optimization, the 50% inhibitory concentration (IC50) and the lowest limit of detection value of the MBS-ELISA was 1.17 ng/mL and 0.07 ng/mL and the linear range was 248.8 pg/mL-5.28 ng/mL, respectively, which accords with state criteria for food safety. The developed one-step MBS-ELISA was almost 20-times more sensitive than the classic BA-ELISA and could generate results within 15 min, which was significantly less than the classic BA-ELISA at approximately 3 h. The MBS-ELISA indicated good recovery (86.4-114.3%) in spiked sorghum, buckwheat, and mung bean. Thus, MBS-ELISA represents a very promising strategy for the simple, rapid, and accurate detection of OTA and Other toxic and hazardous contaminants.

Keywords
Biotin-streptavidin system; Chemiluminescence immunoassay; Magnetic separation; Nanobody; Ochratoxin.
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