TY - JOUR AU - LI Rong AU - XIONG Dong-sheng AU - SHAO Xiao-feng AU - LIU Jia AU - XU Yuan-fu AU - XU Yuan-sheng AU - LIU Han-zhi AU - ZHU Zhen-ping AU - YANG Chun-zheng PY - 2016 TI - Production of neutralizing monoclonal antibody against human vascular endothelial growth factor receptor II JF - Acta Pharmacologica Sinica; Vol 25, No 10 (October 2004): Acta Pharmacologica Sinica Y2 - 2016 KW - N2 - AIM: To prepare neutralizing monoclonal antibody (mAb) against extracellular immunoglobulin (Ig)-like domain III of vascular endothelial growth factor receptor KDR and study its biological activity. METHODS: Soluble KDR Ig domain III (KDR-III) fusion protein was expressed in E Coli and purified from the bacterial periplasmic extracts via an affinity chromatography. Monoclonal antibodies against KDR-III were prepared by hybridoma technique. ELISA and FACS analysis were used to identify its specificity. Immunoprecipitation and [3H]-thymidine incorporation assay were also used to detect the activity of anti-KDR mAb blocking the phosphorylation of KDR tyrosine kinase receptor and the influence on vascular endothelial growth factor-induced mitogenesis of human endothelial cells. RESULTS: A monoclonal antibody, Ycom1D3 (IgG1), was generated from a mouse immunized with the recombinant KDR-III protein. Ycom1D3 bound specifically to both the soluble KDR-III and the cell-surface expressed KDR. Ycom1D3 effectively blocked VEGF/KDR interaction and inhibited VEGF-stimulated KDR activation in human endothelial cells. Furthermore, the antibody efficiently neutralized VEGF-induced mitogenesis of human endothelial cells. CONCLUSION: Our results suggest that the anti-KDR mAb, Ycom1D3, has potential applications in the treatment of cancer and other diseases where pathological angiogenesis is involved. UR - http://www.chinaphar.com/article/view/8382