Preparation and characterization of humanized nanobodies targeting the dimer interface of epidermal growth factor...

Area of Science:

• Oncology
• Biotechnology
• Molecular Biology

Background:

• Epidermal growth factor receptor (EGFR) is a key target in epithelial cancer therapy.
• Current EGFR inhibitors face challenges like low response rates and drug resistance due to mutations and heterodimerization.
• Targeting the conserved EGFR dimer interface presents a potential strategy to overcome these limitations.

Purpose of the Study:

• To investigate the potential of nanobodies targeting the EGFR dimer interface as novel anticancer drugs.
• To screen and characterize nanobodies that specifically bind to the EGFR dimer interface.
• To evaluate the anticancer activity of a selected nanobody against EGFR-overexpressing cancer cells.

Main Methods:

• Screening of nanobodies from a phage antibody library using an EGFR dimer interface peptide (EGFR^237-267).
• Selection and purification of a promising nanobody (EGFR dimer Nb77) expressed in E. coli.
• Assessment of EGFR dimer Nb77 binding to EGFR-overexpressing A431 cells and its tumor cell growth inhibition capabilities.

Main Results:

• A nanobody, EGFR dimer Nb77, was identified and characterized.
• EGFR dimer Nb77 demonstrated specific, dose-dependent, and ligand-dependent binding to EGFR-overexpressing cells.
• The nanobody effectively inhibited tumor cell growth, comparable to a known monoclonal antibody targeting the same interface.

Conclusions:

• Nanobodies targeting the EGFR dimer interface represent a promising new class of anticancer agents.
• EGFR dimer Nb77 shows significant potential for improving anti-EGFR therapies and overcoming drug resistance.
• This study provides the first evidence for the efficacy of nanobodies targeting the EGFR dimer interface in cancer treatment.