关键词: 密码子优化, VEGF单克隆抗体, 纯化, 生物活性

Abstract: 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 C6 cells 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.

Key words: codon optimization, antiVEGF monoclonal antibody, purification, biological activity