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Updated: Jan 18, 2026
An Efficient Method for Adenovirus Production
Published on: June 10, 2021
Expression and Characterization of Human Vascular Endothelial Growth Factor Produced in SiHa Cells Transduced with
N C Parra1, R Mansilla1, G Aedo1
1Recombinant Biopharmaceuticals Laboratory, Department of Pharmacology, School of Biological Sciences, Universidad de Concepción, P.O. Box 160C, Concepción, Chile.
Abstract:
The vascular endothelial growth factor (VEGF) is an essential factor to pathologic angiogenesis. Disruption of VEGF/VEGF receptor interaction in cancer patients inhibits the development of new and pre-existing tumor blood vessels. Consequently, VEGF becomes an important therapeutic target for handling solid tumors. In this work, human VEGF was produced in the culture supernatant of SiHa cells transduced with a replication-defective adenoviral vector (pAdhVEGF121) encoding this molecule. The 35 kDa VEGF121 homodimer was obtained from clarified culture media as a glycosylated protein. VEGF121 expression levels were strictly dependent on the adenoviral viral load used. VEGF121 was produced with purity over 98% after a single step chromatography by immobilized metal affinity chromatography. Additionally, VEGF121 binds Bevacizumab antibody with a KD of 7 nM. Biological characterization by mitogenic assay in HUVEC and ECV-304 cells showed that VEGF121 stimulates cell proliferation in a dose-dependent manner in both cells. Finally, the neovascularization activity of VEGF121 was demonstrated by vascular permeability assays in matrigel plug-bearing mice, showing significantly increased vasculature leakage after treatment with VEGF121. Consequently, transduction of SiHa cells with adenovirus is a suitable alternative for manufacture heterologous proteins of therapeutic interest.
Insights
This study demonstrates a novel method for producing vascular endothelial growth factor (VEGF) using adenoviral vectors. This approach yields high-purity VEGF, a crucial target for anti-cancer therapies.
Area of Science:
- Biotechnology
- Molecular Biology
- Oncology
Background:
- Vascular Endothelial Growth Factor (VEGF) is critical for pathological angiogenesis and tumor growth.
- Targeting VEGF/VEGF receptor interactions is a key strategy in cancer therapy.
- Efficient production of therapeutic VEGF is essential for clinical applications.
Purpose of the Study:
- To develop a method for producing human VEGF using adenoviral transduction of SiHa cells.
- To characterize the purity, binding affinity, and biological activity of the produced VEGF.
- To evaluate the suitability of this method for manufacturing therapeutic proteins.
Main Methods:
- Transduction of SiHa cells with a replication-defective adenoviral vector (pAdhVEGF121) encoding human VEGF.
- Purification of VEGF121 using immobilized metal affinity chromatography.
- Characterization via mitogenic assays, antibody binding assays (with Bevacizumab), and in vivo vascular permeability assays.
Main Results:
- Successfully produced glycosylated VEGF121 homodimer (35 kDa) with >98% purity in a single purification step.
- VEGF121 demonstrated dose-dependent mitogenic activity in HUVEC and ECV-304 cells.
- VEGF121 exhibited significant neovascularization activity in mouse models, confirmed by increased vascular permeability.
Conclusions:
- Adenoviral transduction of SiHa cells is an effective method for producing high-purity, biologically active VEGF.
- This approach offers a viable alternative for the manufacturing of heterologous therapeutic proteins.
- The produced VEGF121 shows therapeutic potential for targeting angiogenesis in solid tumors.
