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Updated: Jun 22, 2025
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A Simple Bioassay for the Evaluation of Vascular Endothelial Growth Factors
Published on: March 15, 2016
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A method for rapid and reliable quantification of VEGF-cell binding activity.
Prabuddha Waduge1, Avinash Kaur1, Wei Li1
1Cullen Eye Institute, Department of Ophthalmology, Baylor College of Medicine, Houston, TX, 77030, USA.
Biochemical and Biophysical Research Communications
|July 2, 2024
Summary
We developed a T7 phage display method to quantify human vascular endothelial growth factor (VEGF) isoform binding to cells. The hVEGF206 isoform showed the highest sensitivity for detecting VEGF-cell interactions.
Area of Science:
- Biochemistry
- Molecular Biology
- Cell Biology
Background:
- Vascular endothelial growth factor (VEGF) is crucial for angiogenesis, cell growth, and survival.
- Quantifying VEGF-cell binding is challenging due to reversible ligand-receptor interactions.
Purpose of the Study:
- To quantify and compare the cell binding activity of three human VEGF-A (hVEGF) isoforms using T7 bacteriophage display.
- To evaluate the utility of T7 phage display for rapid ligand-cell binding quantification.
Main Methods:
- Utilized T7 bacteriophage display to generate and present hVEGF111, hVEGF165, and hVEGF206 isoforms.
- Assessed binding of phage-displayed isoforms to immobilized aflibercept, heparan sulfate, and various cell types (HUVECs, HEK293, SK-N-AS).
Main Results:
- All hVEGF isoforms bound equally to aflibercept.
- hVEGF206-Phage exhibited the highest binding to cells and heparan sulfate, while hVEGF111-Phage showed minimal binding.
- hVEGF206-Phage demonstrated the highest sensitivity for VEGF-cell binding detection, despite higher background binding to SK-N-AS cells.
Conclusions:
- The hVEGF206 isoform, presented via T7 phage display, is optimal for quantifying VEGF-cell binding, offering superior sensitivity.
- T7 phage display provides a versatile platform for rapid and convenient quantification of ligand-cell binding activities.
