重组人源抗血管内皮生长因子单克隆抗体的制备及鉴定

通过优化密码子获得带有信号肽的人源抗血管内皮生长因子（VEGF）抗体的重链和轻链DNA序列, 分别构建表达载体pcDNA VEGFH和pcDNA VEGFV, 将表达载体共转染HEK 293细胞后, 与C6胶质瘤细胞共培养, 并将所得蛋白进行分离纯化. 实验结果表明: pcDNA VEGFH和pcDNA VEGFV共转染HEK 293F细胞, 可显著抑制C6细胞增殖（P<0.01）; 转染24~144 h后的细胞培养上清液中均有蛋白表达; 所得纯化蛋白和标准品一致; 纯化所得抗体蛋白能明显抑制C6细胞增殖（P<005~001）, 与标准品抑制效果相似.

Preparation and Identification of Recombinant-Humanized Anti VEGF Monoclonal Antibody

DNA sequences of humanized anti VEGF antibody light and heavy chains were optimized to construct pcDNA VEGFH and pcDNA VEGFV expression vectors with signal peptidetide. HEK 293 cells were transfected with pcDNA VEGFH and pcDNA VEGFV,then co cultured with C6 cells, and protein was isolated and purified. The experimental results show that the proliferation of C6cells are significantly inhibited (P<0.01). HEK 293F cells are transfected with pcDNA VEGFH and pcDNA VEGFV, expression of protein is detected in cell culture supernate after 24~144 h.The purified protein is identical with the standard product. The purified antibody protein can significantly inhibit the proliferation of C6 cells (P<0.05—0.01). The results are similar to those of standard products.