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Electronic Journal of Biotechnology
Universidad Católica de Valparaíso
ISSN: 0717-3458
Vol. 13, No. 1, 2010
Bioline Code: ej10001
Full paper language: English
Document type: Research Article
Document available free of charge

Electronic Journal of Biotechnology, Vol. 13, No. 1, 2010

 en Development of potency assays for a plasmid containing vascular endothelial growth factor 2
Huang, L.-C.; Chin, E. & Chiang, Y.L.

Abstract

We have developed analytical methods to measure the biological functions of pVGI.1(VEGF2), a naked plasmid DNA product containing vascular endothelial growth factor 2 used in clinical trials for coronary artery diseases (CAD) and peripheral artery diseases (PAD). After being injected into muscles, vascular endothelial growth factor 2 (VEGF-2), presumably expressed in muscle tissues, binds to the endothelial cell receptors VEGFR2 or VEGFR3, triggering the downstream responses including cell proliferation and vascularization. As it is important to make sure clinical material is biological active, we developed a quantitative assay first to measure the receptor binding activity of the pVGI.1(VEGF2) gene product expressed by the transfected host cells, and then a qualitative assay to confirm the cell proliferation promoting activity of the expressed protein. In both assays the signals were plotted directly against input DNA concentrations used to transfect the host cells. We confirmed specificity for both assays. In addition, we demonstrated acceptable levels of spike recovery (86.7-116%), precision (largest relative standard deviation (RSD)=19.3%), linearity and range (60-140% relative potency, 15 - 35 µg/mL) for the quantitative assay. We intend to use the potency assays for routine lot release and stability studies.

Keywords
cell proliferation, plasmid DNA, potency assays, receptor binding, VEGF-2

 
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