Related Experiment Videos
Isolation of fibroblast growth factor receptor binding sequences using evolved phage display libraries
S J McConnell1, V J Thon, D G Spinella
1Biological Screening Group, Chugai Biopharmaceuticals, Inc., San Diego, CA 92121, USA. smcconnell@chugaibio.com
Combinatorial Chemistry & High Throughput Screening
|July 27, 1999
Summary
Researchers developed a phage display method to identify fibroblast growth factor receptor (FGFR) inhibitors. Optimized phage clones showed significantly higher binding affinity, leading to a peptide that blocks FGF-FGFR interaction.
Area of Science:
- Biotechnology
- Molecular Biology
- Protein Engineering
Background:
- Fibroblast Growth Factor (FGF) signaling is crucial in cellular processes.
- Dysregulation of FGF signaling is implicated in various diseases.
- Targeting the FGF receptor (FGFR) is a therapeutic strategy.
Purpose of the Study:
- To develop a phage display system for identifying FGFR ligands.
- To isolate and optimize high-affinity FGFR-binding peptides.
- To characterize the inhibitory potential of identified peptides.
Main Methods:
- Construction of a fusion protein: short human fibroblast growth factor receptor (FGFR) and mouse IgG1 heavy chain.
- Screening of four phage display libraries with varying peptide lengths (8, 13, 38, 43 amino acids).
- Oligonucleotide saturation mutagenesis for affinity maturation of selected phage clones.
- In vitro ELISA to assess binding affinity and inhibition of FGF-FGFR interaction.
Main Results:
- Phage clones with specific FGFR binding activity were isolated from 13, 38, and 43 amino acid libraries.
- Affinity maturation increased binding affinity approximately eightfold compared to the parent sequence.
- A consensus sequence (CR/SXLLXGAPFXXXXC) was identified.
- A synthetic peptide based on the consensus sequence inhibited FGF binding to its receptor in vitro.
Conclusions:
- Phage display is effective for isolating high-affinity FGFR-binding peptides.
- Engineered peptides can act as antagonists of FGF-FGFR signaling.
- This approach yields potent inhibitors for potential therapeutic applications.