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Updated: Jul 23, 2025
Screening and Identification of Small Peptides Targeting Fibroblast Growth Factor Receptor2 using a Phage Display Peptide Library
Published on: September 30, 2019
Targeting of human fibroblast growth factor receptor 2 by a novel specific nanobody
Mahboubeh Irani1, Mahdi Habibi-Anbouhi2, Mahdi Behdani1
1Venom and Biotherapeutics Molecules Laboratory, Biotechnology Department, Biotechnology Research Center, Pasteur Institute of Iran, Tehran, Iran.
Abstract:
Inhibition of FGFR2 signaling is promising in targeted therapy of FGFR2-related tumors. In this study, anti-FGFR2 nanobodies (Nbs) were isolated through screening of an immune camelid phage display library. Four rounds of biopanning were carried out with commercial human FGFR2 antigen and enrichment was assessed by ELISA and phage titration. The gene of Nb was sub-cloned into the expression vector, and the recombinant vector was transformed into Escherichia coli WK6 cells. The recombinant protein was purified using Ni-NTA affinity chromatography. The anti-FGFR2 Nb (C13) was characterized by SDS-PAGE, western blotting, competitive inhibition ELISA, flow cytometry, MTT, and migration assay. C13 Nb recognized FGFR2 with high specificity and no cross-reactivity was observed with other tested antigens. The affinity of C13 Nb was calculated to be 1.5 × 10-9 M. Results of cytotoxicity showed that C13 Nb (10 µg/ml) inhibited 85% of the proliferation of T-47D cells (p < 0.001). In addition, C13 inhibited the migration of 68% of T-47D toward the source of the growth factor (p < 0.01). The flow cytometry showed that C13 Nb bound to the surface of FGFR2+ cells, T-47D cell line (96%). Results indicate the potential of anti-FGFR2 Nb for targeted therapy of FGFR2-overexpressing tumors after complementary investigations.
Insights
Researchers developed anti-FGFR2 nanobodies (Nbs) for targeted cancer therapy. These nanobodies effectively inhibited FGFR2-positive tumor cell proliferation and migration, showing therapeutic potential.
Area of Science:
- Biotechnology
- Molecular Biology
- Oncology
Background:
- Fibroblast Growth Factor Receptor 2 (FGFR2) signaling is a key target in FGFR2-related tumors.
- Targeted therapies offer promise for treating these malignancies.
Purpose of the Study:
- To isolate and characterize novel anti-FGFR2 nanobodies (Nbs) for potential targeted cancer therapy.
- To evaluate the efficacy of these Nbs in inhibiting tumor cell proliferation and migration.
Main Methods:
- Phage display technology was used to screen for anti-FGFR2 Nbs from an immune camelid library.
- Recombinant Nbs were expressed in *Escherichia coli* and purified.
- Characterization involved SDS-PAGE, western blotting, ELISA, flow cytometry, MTT, and migration assays.
Main Results:
- A specific anti-FGFR2 Nb (C13) was identified with high affinity (1.5 × 10-9 M) and specificity.
- C13 Nb inhibited T-47D cell proliferation by 85% and migration by 68%.
- Flow cytometry confirmed C13 Nb binding to 96% of FGFR2+ T-47D cells.
Conclusions:
- Anti-FGFR2 Nb C13 demonstrates high specificity and potent inhibitory effects on FGFR2-overexpressing tumor cells.
- These findings support the potential of anti-FGFR2 Nbs as a targeted therapy for FGFR2-related cancers.
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