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Screening and Identification of Small Peptides Targeting Fibroblast Growth Factor Receptor2 using a Phage Display Peptide Library
Published on: September 30, 2019
Development of a Novel Human scFv Against EGFR L2 Domain by Phage Display Technology
Leila Rahbarnia1, Safar Farajnia2, Hossein Babaei3
1Infectious and Tropical Diseases Research Center, Tabriz University of Medical Sciences, Tabriz. Iran.
Abstract:
Epidermal growth factor receptor (EGFR) as a transmembrane tyrosine kinase receptor frequently overexpresses in tumors with epithelial origin. The L2 domain from extracellular part of EGFR is involved in ligand binding and the blockage of this domain prevents activation of related signaling pathways. This study was aimed to develop a novel human scFv against EGFR L2 domain as a promising target for cancer therapy. The L2 recombinant protein was purified and used for panning a human scFv phage library (Tomlinson I). In this study, a novel screening strategy was applied to select clones with high binding and enrichment of rare specific phage clones of the L2 protein. After five biopanning rounds several specific clones were isolated which among them one phage clone with high binding was purified for further analysis. The specific interaction of selected clone against target antigen was confirmed by ELISA and western blotting. Immunofluorescence staining showed that purified scFv binds to A431 cells surface, displaying EGFR surface receptor. In the present study, we isolated for the first time a novel human scFv against EGFR L2 domain. This study can be the groundwork for developing more effective diagnostic and therapeutic agents against EGFR overexpressing cancers using this novel human anti-L2 ScFv.
Insights
Researchers developed a new human antibody fragment (scFv) targeting the L2 domain of the epidermal growth factor receptor (EGFR). This novel anti-EGFR L2 scFv shows promise for developing targeted cancer therapies for epithelial tumors.
Area of Science:
- Biotechnology
- Molecular Biology
- Oncology
Background:
- Epidermal growth factor receptor (EGFR) is a transmembrane tyrosine kinase receptor often overexpressed in epithelial cancers.
- The L2 domain of EGFR is crucial for ligand binding; blocking it can inhibit cancer signaling pathways.
Purpose of the Study:
- To develop a novel human single-chain variable fragment (scFv) targeting the L2 domain of EGFR.
- To establish a potential therapeutic agent for EGFR-overexpressing cancers.
Main Methods:
- Purification of recombinant EGFR L2 protein.
- Panning of a human scFv phage display library (Tomlinson I) using a novel screening strategy.
- Validation of scFv binding using ELISA, Western blotting, and immunofluorescence staining.
Main Results:
- Isolation of a novel human scFv with high binding affinity to the EGFR L2 domain.
- Confirmation of specific interaction with the target antigen.
- Demonstration of scFv binding to EGFR on the surface of A431 cancer cells.
Conclusions:
- Successful isolation of a novel human scFv targeting the EGFR L2 domain.
- This anti-EGFR L2 scFv represents a promising candidate for developing targeted cancer diagnostics and therapeutics.
- Provides a foundation for future development of agents against EGFR-driven malignancies.
