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San Francisco, CA, United States

Huang L.-C.,Corautus Genetics Inc. | Huang L.-C.,Corautus Genetics Inc. | Chin E.,Corautus Genetics Inc. | Chin E.,Corautus Genetics Inc. | And 2 more authors.
Electronic Journal of Biotechnology | Year: 2010

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. © 2010 by Pontificia Universidad Católica de Valparaíso. Source

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