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Monoclonal antibodies specific for the bundle-forming pilus of enteropathogenic Escherichia coli
1Center for Vaccine Development, University of Maryland School of Medicine, Baltimore 21201, USA.
Abstract:
The bundle-forming pilus (BFP) produced by enteropathogenic Escherichia coli (EPEC) is associated with the presence of a large EPEC adherence factor plasmid and the formation of localized adherence clusters on tissue culture cells. Three mouse monoclonal antibodies (ICA2, ICA3, and ICA4) were produced against BFP purified from EPEC B171 (O111:NM). These monoclonal antibodies reacted in immunoblots with different epitopes of the 19.5-kDa bundlin subunit of BFP of heterologous EPEC. These reagents could serve as diagnostic tools for the identification of EPEC as well as for studying the role of BfpA in the interaction of EPEC with eukaryotic cells.
Insights
Monoclonal antibodies were developed against the bundle-forming pilus (BFP) of enteropathogenic Escherichia coli (EPEC). These antibodies can identify EPEC and aid in understanding BfpA
Area of Science:
- Microbiology
- Immunology
- Bacterial Pathogenesis
Background:
- Enteropathogenic Escherichia coli (EPEC) utilizes the bundle-forming pilus (BFP) for adherence to host cells.
- BFP is linked to the EPEC adherence factor plasmid and localized adherence.
- Understanding BFP structure and function is crucial for EPEC infection research.
Purpose of the Study:
- To generate specific monoclonal antibodies against the BFP subunit.
- To characterize the reactivity of these antibodies with different EPEC strains.
- To evaluate the utility of these antibodies as diagnostic and research tools.
Main Methods:
- Purification of BFP from EPEC strain B171 (O111:NM).
- Production of three mouse monoclonal antibodies (ICA2, ICA3, ICA4) against purified BFP.
- Immunoblot analysis to determine antibody reactivity with the BFP subunit (bundlin).
Main Results:
- Monoclonal antibodies ICA2, ICA3, and ICA4 were successfully generated.
- These antibodies recognized distinct epitopes on the 19.5-kDa bundlin subunit of BFP.
- Antibodies reacted with BFP from heterologous EPEC strains, indicating conserved epitopes.
Conclusions:
- The developed monoclonal antibodies are specific reagents for BFP detection.
- These antibodies can serve as valuable diagnostic tools for identifying EPEC.
- The reagents will facilitate further studies on the role of BfpA in EPEC-host cell interactions.