A novel fusion protein of the receptor binding domain of VEGF and human IgG1 Fc portion reduces angiogenesis

• Main Authors: Chung, Yo-Shong, 鍾侑松
• Other Authors: Liao, Kuang-Wen
• Format: Others
• Language: en_US
• Published: 2007
• Online Access: http://ndltd.ncl.edu.tw/handle/73057462895264534113

碩士 === 國立交通大學 === 生物科技系所 === 95 === Angiogenesis is a physiological process involving the growth of new blood vessels from pre-existing vessels, rather than through in situ differentiation of undifferentiated precursor cells to endothelial cells. Vascular endothelial growth factor (VEGF) is an angiogenic factor that promotes the growth of solid tumor by inducing angiogenesis. The two VEGF receptors, VEGFR-1 and VEGFR-2, have been shown to be expressed preferentially in the proliferating endothelial cells. Thus, inhibiting these two VEGF receptors may be a promising strategy for treatment of cancer and other angiogenesis-dependent diseases. In this study, we designed a novel recombinant fusion protein composed of a targeting domain and an effector domain. The targeting domain is the receptor binding domain of human VEGF (residues 8-109) and the effector domain is the Fc region of a human IgG1 immunoglobulin that can induce a cytolytic immune response against the targeted cells. The chimeric gene, RBDV-IgG1 Fc, was subcloned into AAV expression vector to produce the fusion protein. We also present an approach for the purification of the fusion protein with high yield and high purity from HEK-293 expression system. The binding of RBDV-IgG1 Fc fusion protein to the VEGF receptors was examined by ELISA and flow cytometry. Furthermore, the proliferation of VEGF-induced human umbilical vein endothelial cells (HUVECs) was inhibited by the RBDV-IgG1 Fc, suggesting an antagonistic role in VEGF/VEGF receptors signal pathway. These results showed that RBDV-IgG1 Fc fusion protein is potential for the suppression of angiogenesis in vivo.