扬州大学学报（农业与生命科学版）2011，Vol.32Issue(2)：16-20,5.
人VEGF受体Ⅱ胞外Ⅲ区单克隆抗体研制与生物学活性测定
Development of monoclonal antibodies against extracellular domain Ⅲ of vascular endothelial cell growth factor receptor Ⅱ and its biological activity
摘要
Abstract
A pair of primers was designed according to published gene sequence of human vascular endothelial cell growth factor receptor Ⅱ (KDR). The coding sequence of KDR extracellular domain Ⅲ (KDRD3) was then amplified from human umbilical vein endothelial cells (HUVEC) by RT-PCR and cloned into prokaryotic expression vector Pgex-6P1. The recombinant plasmid was transformed into E. Coli BL21 and induced with IPTG at 37 ℃. As evidenced by SDS-PAGE and western-blotting, recombinant GST-KDRd3 fusion protein was successfully expressed in E. Coli. BALB/c mice were immunized with puried GST-KDRD3 fusion protein, and two KDR specific hybridoma strains were developed using hybridoma technique. Both mAbs could react with natural KDR expressed in HUVEC. The results of HUVEC migration inhibition test and in vitro angiogensis inhibition test showed that interaction between VEGF and KDR could be blocked by any one of KDR mAbs. Therefore, the obtained mAbs have good specificity and neutralization activity to KDR.关键词
血管内皮细胞生长因子受体Ⅱ/单克隆抗体/生物学活性Key words
vascular endothelial cell growth factor receptor Ⅱ/monoclonal antibody/biological activity分类
生物科学引用本文复制引用
孔桂美,张小荣,黄瑶,高巍,廖月霞,卜平..人VEGF受体Ⅱ胞外Ⅲ区单克隆抗体研制与生物学活性测定[J].扬州大学学报（农业与生命科学版）,2011,32(2):16-20,5.基金项目
国家自然科学基金资助项目(30801497) （30801497）
扬州大学研究生创新基金资助项目(2008-17). （2008-17）
