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  3. Fluorescence-activated cell sorting-based efficient screening of monensin monoclonal antibodies and applications in lateral flow immunoassay

Fluorescence-activated cell sorting-based efficient screening of monensin monoclonal antibodies and applications in lateral flow immunoassay

  • Talanta. 2025 Oct 1:293:128128. doi: 10.1016/j.talanta.2025.128128.
Jingjie Huang 1,  Ao Lin 2,  Yani Gu 2,  Xiaole Pan 3,  XinXin Ma 3,  Yanzhe Qing 2,  Jiancheng Li 4
Affiliations

Affiliations

  • 1 College of Veterinary Medicine, National Key Laboratory of Veterinary Public Health and Safety, Beijing Key Laboratory of Detection Technology for Animal Derived Food Safety, Beijing Laboratory for Food Quality and Safety, China Agricultural University, Beijing, 100193, PR China; Technology Innovation Center for Food Safety Surveillance and Detection, Sanya Institute of China Agricultural University, Hainan, 572000, PR China.
  • 2 College of Veterinary Medicine, National Key Laboratory of Veterinary Public Health and Safety, Beijing Key Laboratory of Detection Technology for Animal Derived Food Safety, Beijing Laboratory for Food Quality and Safety, China Agricultural University, Beijing, 100193, PR China.
  • 3 Technology Innovation Center for Food Safety Surveillance and Detection, Sanya Institute of China Agricultural University, Hainan, 572000, PR China.
  • 4 College of Veterinary Medicine, National Key Laboratory of Veterinary Public Health and Safety, Beijing Key Laboratory of Detection Technology for Animal Derived Food Safety, Beijing Laboratory for Food Quality and Safety, China Agricultural University, Beijing, 100193, PR China; Technology Innovation Center for Food Safety Surveillance and Detection, Sanya Institute of China Agricultural University, Hainan, 572000, PR China. Electronic address: horse20@cau.edu.cn.
PMID: 40222095 DOI: 10.1016/j.talanta.2025.128128
Abstract

Monensin is widely used in livestock and poultry for disease prevention and growth promotion. Still, its improper use can lead to residues in animal-derived foods, posing risks to human health. In immunoassays, monoclonal antibodies (mAbs) remain a preferred choice due to their high sensitivity and specificity. However, traditional hybridoma technology often suffers from a lengthy screening cycle and the risk of losing effective clones. To address these limitations, this study employed fluorescently labeled monensin antigens combined with fluorescence-activated cell sorting (FACS) for rapid screening of hybridoma cells secreting anti-monensin mAbs. Compared to the conventional limiting dilution method, this approach increased the yield of specific hybridoma cells by tenfold and reduced the screening cycle from four weeks to one week. Based on the selected mAb (4E6), a rapid visual lateral flow Immunoassay (LFIA) was developed for on-site detection of monensin, with a total detection time of 10 min. The assay exhibited a half-maximal inhibitory concentration (IC50) of 2.32 μg/kg and a linear detection range of 0.32-10.3 μg/kg. This study provides a promising strategy for efficient hybridoma cell screening and practical monitoring of monensin residues in environmental and food samples.

Keywords

Fluorescence-activated cell sorting; Hapten design; LFIA; Monensin; Monoclonal antibody.

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