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Semi-automated Biopanning of Bacterial Display Libraries for Peptide Affinity Reagent Discovery and Analysis of Resulting Isolates
Published on: December 6, 2017
Epitope mapping of antibodies using bacterial surface display
Johan Rockberg1, John Löfblom, Barbara Hjelm
1Department of Molecular Biotechnology, School of Biotechnology, Royal Institute of Technology (KTH), AlbaNova University Center, SE-106 91 Stockholm, Sweden.
Nature Methods
|November 26, 2008
Summary
We developed a bacterial display method to map antibody binding sites (epitopes). This technique precisely identifies epitopes for monoclonal and polyclonal antibodies, aiding antibody validation for research and diagnostics.
Area of Science:
- Immunology
- Molecular Biology
- Biotechnology
Background:
- Accurate epitope mapping is crucial for antibody characterization.
- Current methods can be complex and lack whole-proteome applicability.
- Understanding antibody-epitope interactions is vital for therapeutic and diagnostic development.
Purpose of the Study:
- To present a novel method for mapping antibody-recognized epitopes.
- To analyze epitope binding patterns of monoclonal and polyclonal antibodies.
- To demonstrate the utility of bacterial surface display for antibody validation.
Main Methods:
- Bacterial surface display of antigen protein fragments.
- Antibody-based flow-cytometric sorting for epitope identification.
- Analysis of antibodies targeting human epidermal growth factor receptor 2 (HER2), ephrin-B3, and SATB2.
Main Results:
- Monoclonal antibodies recognized single epitopes.
- Polyclonal antibodies identified one to five distinct epitopes per antibody.
- Overlapping binding epitopes were observed between monoclonal and polyclonal antibodies against the same antigen.
- Bacterial display facilitated the generation of epitope-specific antibodies.
Conclusions:
- The bacterial surface display method provides a robust approach for epitope mapping.
- This technique enables systematic validation of antibody specificity and cross-reactivity.
- The method has potential for whole-proteome antibody analysis.
