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Updated: May 29, 2026
Generation of Monoclonal Antibodies Against Natural Products
Published on: April 6, 2019
[Preparation and identification of OmpW monoclonal antibodies]
Zhen-Ning Ma1, Lei Zheng, Hai-Mei Lin
1Institute of Pharmaceutical Biotechnology and Bioengineering, Fuzhou University, Fuzhou 350002, China. zhenningma2008@yahoo.cn
Aim:
To prepare and characterize the mouse monoclonal antibodies against Vibrio parahaemolyticus OmpW.
Methods:
The OmpW amino acid sequence from three diseased Vibrio was analyzed by Bioinformatics. Mice were immunized with r-OmpW which was highly expressed and purified in E.coli. Five Vibrio(Va, Vp, Vh, Vv, Van) were chosen as antigen for mAb selection.The characters of the anti-OmpW monoclonal antibodies were studied by Western blot, Flow Cytometry, indirect immunofluorescence.
Results:
OmpW was testified a highly conservative membrane protein.Three clones of anti-OmpW mAb was obtained. The Ig subclass of the mAb secreted from fused cell S5C10 was IgG3, which of the titer was 4.6×10(4);. The mAb could specifically recognize Vibrio parahaemolyticus, Vibrio alginolyticus, Vibrio harveyi, Vibrio anguillarum, Vibrio vulnificus, which could not react with Pseudomonas flurosecens, Aeromonas hydrophila, Aeromonas sobria, Aeromonas hydrophila, Escherichia coli.
Conclusion:
The mAb could specially recognize five diseased Vibrio, which is a useful tool for the further study of the diagnosis of Vibrio.
Insights
Researchers developed monoclonal antibodies (mAbs) targeting the OmpW protein of Vibrio species. These highly specific antibodies can detect multiple pathogenic Vibrio strains, aiding in diagnostics.
Area of Science:
- Microbiology
- Immunology
- Biotechnology
Background:
- Vibrio species are significant marine pathogens.
- OmpW is a conserved outer membrane protein in Vibrio.
- Developing specific diagnostic tools for Vibrio infections is crucial.
Purpose of the Study:
- To generate and characterize mouse monoclonal antibodies (mAbs) against the OmpW protein of Vibrio parahaemolyticus.
- To evaluate the specificity and utility of these mAbs for Vibrio detection.
Main Methods:
- Bioinformatic analysis of OmpW sequences from pathogenic Vibrio.
- Immunization of mice with recombinant OmpW (r-OmpW) expressed in E. coli.
- Selection and characterization of anti-OmpW mAbs using Western blot, flow cytometry, and indirect immunofluorescence.
Main Results:
- OmpW was confirmed as a highly conserved membrane protein.
- Three clones of anti-OmpW mAbs were successfully generated.
- The developed mAbs specifically recognized five pathogenic Vibrio species (V. parahaemolyticus, V. alginolyticus, V. harveyi, V. anguillarum, V. vulnificus) but not other bacterial species.
Conclusions:
- Monoclonal antibodies against OmpW demonstrate high specificity for multiple pathogenic Vibrio species.
- These mAbs represent a valuable tool for the further development of diagnostic methods for Vibrio infections.
