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Interactome-Seq: A Protocol for Domainome Library Construction, Validation and Selection by Phage Display and Next Generation Sequencing
Published on: October 3, 2018
Phage display selection, analysis, and prediction of B cell epitopes
Natalia Tarnovitski Freund1, David Enshell-Seijffers2, Jonathan M Gershoni1
1Tel Aviv University, Tel Aviv, Israel.
Current Protocols in Immunology
|August 5, 2009
Summary
Phage display libraries identify antibody epitopes using biopanning. This method precisely defines antibody-binding sites and maps epitope profiles for both monoclonal antibodies and polyclonal sera.
Area of Science:
- Immunology
- Molecular Biology
- Biotechnology
Background:
- Antibody epitopes are crucial for understanding immune responses and developing diagnostics/therapeutics.
- Defining antibody epitopes at a molecular level is essential for precise immunological studies.
Purpose of the Study:
- To present a method for discovering antibody epitopes using combinatorial phage display libraries.
- To enable the molecular definition of epitopes for monoclonal antibodies (MAbs).
- To generate epitope profiles for polyclonal serum samples.
Main Methods:
- Utilizing combinatorial phage display libraries of random peptides.
- Employing biopanning for affinity isolation of phage-displayed epitope peptidomimetics.
- Analyzing MAb-specific peptides for computational prediction of B cell epitopes.
Main Results:
- Successful identification and molecular definition of antibody epitopes.
- Generation of MAb-specific peptides for epitope mapping.
- Creation of epitope profiles from polyclonal serum samples.
Conclusions:
- Combinatorial phage display biopanning is an effective strategy for epitope discovery.
- This approach facilitates the molecular characterization of antibody recognition sites.
- The protocols detailed allow for broad applications in immunology and diagnostics.
