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Updated: Aug 19, 2026
A Simple Bioassay for the Evaluation of Vascular Endothelial Growth Factors
Published on: March 15, 2016
Expression and purification of the catalytic domain of human vascular endothelial growth factor receptor 2 for
Li Zhong1, Xiao-Ning Guo, Xiu-Hua Zhang
1Division of Anti-tumor Pharmacology, State Key Laboratory of Drug Research, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, 555 Zu Chong Zhi Road, Zhangjiang, Pu Dong, Shanghai 201203, PR China.
Abstract:
Vascular endothelial growth factor (VEGF), an endothelial cell-specific mitogen, can act in tumor-induced angiogenesis by binding to specific receptors on the surface of endothelial cells. One such receptor, VEGFR-2/KDR, plays a key role in VEGF-induced angiogenesis. Here, we expressed the catalytic domain of VEGFR-2 as a soluble active kinase using Bac-to-Bac expression system, and investigated correlations between VEGFR-2 activity and enzyme concentration, ATP concentration, substrate concentration and divalent cation type. We used these data to establish a convenient, effective and non-radioactive ELISA screening technique for the identification and evaluation of potential inhibitors for VEGFR-2 kinase. We screened 200 RTK target-based compounds and identified one (TKI-31) that potently inhibited VEGFR-2 kinase activity (IC50=0.596 microM). Treatment of NIH3T3/KDR cells with TKI-31 blocked VEGF-induced phosphorylation of KDR in a dose-dependent manner. Moreover, TKI-31 dose-dependently suppressed HUVEC tube formation. Thus, we herein report a novel, efficient method for identifying VEGFR-2 kinase inhibitors and introduce one, TKI-31, that may prove to be a useful new angiogenesis inhibitor.
Insights
Researchers developed a new ELISA method to find inhibitors of Vascular Endothelial Growth Factor Receptor 2 (VEGFR-2) kinase. They identified TKI-31, a potent inhibitor that blocks VEGFR-2 activity and suppresses angiogenesis.
Area of Science:
- Biochemistry
- Molecular Biology
- Pharmacology
Background:
- Vascular Endothelial Growth Factor (VEGF) drives tumor angiogenesis by binding to receptors like VEGFR-2/KDR.
- VEGFR-2 is crucial for VEGF-induced angiogenesis.
Purpose of the Study:
- To develop a non-radioactive ELISA screening method for identifying VEGFR-2 kinase inhibitors.
- To identify and evaluate novel inhibitors of VEGFR-2 kinase activity.
Main Methods:
- Expressed soluble active catalytic domain of VEGFR-2 using Bac-to-Bac system.
- Established an ELISA assay to correlate VEGFR-2 activity with various factors.
- Screened 200 RTK target-based compounds.
Main Results:
- Identified TKI-31 as a potent inhibitor of VEGFR-2 kinase (IC50=0.596 microM).
- TKI-31 blocked VEGF-induced KDR phosphorylation in NIH3T3/KDR cells.
- TKI-31 suppressed HUVEC tube formation in a dose-dependent manner.
Conclusions:
- A novel and efficient ELISA method for VEGFR-2 inhibitor screening was established.
- TKI-31 is a potent VEGFR-2 inhibitor with potential as an anti-angiogenesis agent.
