scholarly journals Evaluation of the Effects of Human Beta-Interferon Scaffold Attachment Region (IFN-SAR) on Expression of Vascular Endothelial Growth Factor-Fc (VEGF-Fc) Fusion Protein Expression in Chinese Hamster Ovary (CHO) Cells

2020 ◽  
Vol 26 (4) ◽  
pp. 393-398
Author(s):  
Ehsan Naghneh ◽  
Es'hagh Pourmaleki ◽  
Azam Rahimpour
Keyword(s):  
Vascular Endothelial Growth Factor ◽  
Monoclonal Antibodies ◽  
Fusion Protein ◽  
Cho Cells ◽  
Fusion Proteins ◽  
Chinese Hamster ◽  
Vascular Endothelial ◽  
Cell Clones ◽  
Fc Fusion Protein ◽  
Endothelial Growth Factor

Background: Recombinant anti-vascular endothelial growth factor (VEGF) monoclonal antibodies and Fc-fusion proteins have been widely used for the effective treatment of retinal neovascular diseases. In this regard, VEGFR-Fc fusions, which act as strong VEGF inhibitors, have been approved for the treatment of age-related macular degeneration (AMD) and diabetic macular edema (DME). Production of monoclonal antibodies and Fc-fusion proteins relies on mammalian host systems such as Chinese hamster ovary (CHO) cells. Application of genomic regulatory elements including scaffold/matrix attachment regions (SAR/MARs) can profoundly affect recombinant protein expression in CHO cells. Methods: To construct the VEGFR-Fc expression vectors, the enhanced green fluorescent protein (EGFP) gene was replaced by the VEGFR-Fc coding sequence in pEGFP-SAR-puro and pEGFP-puro vectors. Recombinant plasmids were transfected to CHO-K1 cells using TurboFect transfection reagent. VEGFR-Fc expression was evaluated in transiently transfected cells as well as stable cell pools and clones using an enzyme-linked immunosorbent assay (ELISA). Results: IFN-SAR showed no significant effect on transient expression of VEGFR-Fc during 72 h of culture. However, a 2.2-fold enhancement in VEGFR-Fc fusion protein titer was observed in IFN-SAR containing stable cell pools. Further evaluation of the VEGFR-Fc expression level in single-cell clones also indicated that clones with the highest VEGFR-Fc expression belonged to the pools transfected with IFN-SAR construct. Conclusion: Our results indicate that the incorporation of IFN-SAR in expression vector can increase the expression of VEGFR-Fc in stable cell pools as well as single-cell clones. In contrast, transient expression of the fusion protein was not affected by IFN-SAR. More studies are needed to investigate the mechanism underlying this effect, including the analysis of mRNA expression and gene copy number in stable cell pools as well as clonal cells.

Generation of Monoclonal Antibodies to Recombinant Vascular Endothelial Growth Factor

2012 ◽  
Vol 153 (1) ◽  
pp. 139-142 ◽  
Author(s):  
S. A. Shein ◽  
O. I. Gurina ◽  
A. V. Leopol’d ◽  
V. P. Baklaushev ◽  
A. A. Korchagina ◽  
...  
Keyword(s):  
Vascular Endothelial Growth Factor ◽  
Monoclonal Antibodies ◽  
Growth Factor ◽  
Vascular Endothelial ◽  
Endothelial Growth Factor

scholarly journals A Current Review of Targeted Therapeutics for Ovarian Cancer

2010 ◽  
Vol 2010 ◽  
pp. 1-11 ◽  
Author(s):  
Susana M. Campos ◽  
Sue Ghosh
Keyword(s):  
Ovarian Cancer ◽  
Vascular Endothelial Growth Factor ◽  
Monoclonal Antibodies ◽  
Growth Factor ◽  
Kinase Inhibitors ◽  
Recurrent Disease ◽  
Folate Receptor ◽  
Vascular Endothelial ◽  
Targeted Therapeutics ◽  
Endothelial Growth Factor

Difficult to detect, ovarian cancer typically presents at an advanced stage. Significant progress has been achieved in the treatment of ovarian cancer with therapeutics focused on DNA replication or cell division. However, despite sensitivity to induction chemotherapy the majority of patients will develop recurrent disease. Conventional agents for recurrent disease offer little in terms of long-term responses. Various targeted therapeutics have been explored in the management of ovarian cancer. These include monoclonal antibodies to epidermal growth factor receptors, small molecule tyrosine kinase inhibitors, monoclonal antibodies directed at the vascular endothelial growth factor (bevacizumab), and the small tyrosine kinase inhibitors that target the vascular endothelial growth factor receptor. Recently, several other agents have come forth as potential therapeutic agents in the management of ovarian cancer. These include monoclonal antibodies to the folate receptor, triple angiokinase inhibitors, PARP inhibitors, aurora kinase inhibitors, inhibitors of the Hedgehog pathway, folate receptor antagonists, and MTOR inhibitors.

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