%0 Journal Article %T Production of neutralizing monoclonal antibody against human vascular endothelial growth factor receptor II %A LI Rong %A XIONG Dong-sheng %A SHAO Xiao-feng %A LIU Jia %A XU Yuan-fu %A XU Yuan-sheng %A LIU Han-zhi %A ZHU Zhen-ping %A YANG Chun-zheng %J Acta Pharmacologica Sinica %D 2016 %B 2016 %9 %! Production of neutralizing monoclonal antibody against human vascular endothelial growth factor receptor II %K %X 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. %U http://www.chinaphar.com/article/view/8382 %V 25 %N 10 %P 1292-1298 %@ 1745-7254