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Updated: Jun 23, 2026
Generation of Monoclonal Antibodies Against Natural Products
Published on: April 6, 2019
Monoclonal antibody production and immunochemical detection of polyether antibiotics
Hyo Jeong Lee1, Seung Sik Cho, Jaya Ram Simkhada
1Department of Complementary and Alternative Therapy, College of Management, Commerce and Social Welfare, Gwangju University, Gwangju, 503-703, Korea.
Abstract:
Polyether antibiotics such as monensin and salinomycin have been primarily used as coccidiostat and growth promoter. Since residues of these antibiotic in food may pose a health risk for sensitive individuals, their use should be carefully monitored. An immunochemical method was developed for the determination of polyether antibiotic using monoclonal antibody (Mab) produced by immunized mice. Conjugates of monensin, salinomycin and laidlomycin were prepared with bovine serum albumin (BSA), keyhole limpet haemocyanine (KLH) and ovalbumin (OVA) by mixed anhydride method and then used as immunogene to produce Mab. Eight hybridoma cell lines were isolated that produced Mabs that competed with polyether antibiotic-protein conjugates in BALB/c-SP2/0 fusion system. Two hybridoma with higher sensitivity, designated as 4G11F and 1C8F1F, were cultured for mass production and then purified from ascites fluid. Antibiotic-protein conjugates were quantitavely analyzed by using the purified Mabs through a competitive enzyme-linked immunosorbent assay (ELISA).
Insights
This study developed a sensitive immunochemical method to detect polyether antibiotics like monensin and salinomycin in food. The new method uses monoclonal antibodies (Mabs) for accurate residue analysis, ensuring food safety.
Area of Science:
- Food Science
- Immunochemistry
- Veterinary Pharmacology
Background:
- Polyether antibiotics (e.g., monensin, salinomycin) are used as coccidiostats and growth promoters in animal agriculture.
- Antibiotic residues in food products pose potential health risks, necessitating sensitive detection methods.
- Current analytical methods may lack the sensitivity or specificity required for comprehensive residue monitoring.
Purpose of the Study:
- To develop a novel immunochemical assay for the sensitive determination of polyether antibiotic residues in food.
- To produce and characterize high-affinity monoclonal antibodies (Mabs) specific to key polyether antibiotics.
- To validate the utility of these Mabs in a competitive enzyme-linked immunosorbent assay (ELISA) for quantitative analysis.
Main Methods:
- Production of monoclonal antibodies (Mabs) against monensin, salinomycin, and laidlomycin using hybridoma technology.
- Preparation of antibiotic-protein conjugates (using BSA, KLH, OVA) as immunogens and for assay development.
- Development and optimization of a competitive enzyme-linked immunosorbent assay (ELISA) utilizing purified Mabs.
Main Results:
- Successfully generated eight hybridoma cell lines producing Mabs capable of competing with antibiotic-protein conjugates.
- Isolated two highly sensitive Mabs (4G11F and 1C8F1F) for mass production and purification.
- Demonstrated the quantitative analysis of antibiotic-protein conjugates using the developed competitive ELISA with purified Mabs.
Conclusions:
- A sensitive and specific immunochemical method for polyether antibiotic detection has been established.
- The developed Mab-based ELISA offers a valuable tool for monitoring antibiotic residues in food.
- This assay contributes to ensuring food safety by enabling accurate quantification of potentially harmful antibiotic contaminants.
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