Novel affinity binders for neutralization of vascular endothelial growth factor (VEGF) signaling

Area of Science:

• Biotechnology
• Molecular Biology
• Oncology

Background:

• Angiogenesis, the formation of new blood vessels, is crucial for disease progression in cancer and ophthalmological disorders.
• Inhibiting angiogenesis presents a therapeutic strategy for these diseases.
• Vascular Endothelial Growth Factor Receptor-2 (VEGFR2) is a key mediator of angiogenesis.

Purpose of the Study:

• To develop and characterize novel biparatopic Affibody constructs targeting VEGFR2.
• To evaluate the therapeutic potential of these constructs in blocking VEGFR2 signaling and angiogenesis.
• To assess their suitability for in vivo imaging applications.

Main Methods:

• Generation of biparatopic Affibody constructs with varying linker lengths targeting VEGFR2.
• Assessment of receptor binding affinity and dissociation rates.
• In vitro assays to evaluate blockade of VEGFA binding and inhibition of VEGFR2 signaling.
• Endothelial cell sprout formation assay to measure anti-angiogenic potential.

Main Results:

• Decreased linker length enhanced Affibody dissociation resistance from VEGFR2.
• Biparatopic Affibodies successfully bound VEGFR2-expressing cells and inhibited VEGFA-induced signaling.
• The constructs demonstrated potent inhibition of endothelial cell sprout formation in vitro, comparable to ramucirumab.
• Affibody constructs showed potential for therapeutic and imaging applications.

Conclusions:

• Optimized biparatopic Affibody constructs targeting VEGFR2 exhibit strong anti-angiogenic properties.
• These Affibodies effectively inhibit VEGFR2 signaling pathways.
• The study highlights the potential of biparatopic Affibodies for both therapeutic intervention and diagnostic imaging in angiogenesis-related diseases.