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为了降低抗CD3人源化抗体hu12F6对T细胞的激活作用以克服首剂效应 ,构建了恒定区特定位点突变的hu12F6重链表达载体 ,将其与hu12F6的轻链表达载体共转染CHO细胞 .ELISA和RT PCR证实 ,恒定区特定位点突变的人源化抗体hu12F6m在CHO细胞中获得了表达 .竞争抑制实验证实hu12F6m具有与原鼠源抗体及hu12F6相似的特异性和亲和力 .增殖实验表明hu12F6m对T细胞的刺激作用明显弱于hu12F6 .实验结果为深入探讨hu12F6m的生物学特性奠定了基础 .
In order to reduce the activation of T cell by anti-CD3 humanized antibody hu12F6 to overcome the effect of the first dose, a hu12F6 heavy chain expression vector with specific site mutation in the constant region was constructed and cotransfected with hu12F6 light chain expression vector to CHO cells The results of ELISA and RT-PCR showed that hu12F6m, a mutated humanized antibody at a specific site in the constant region, was expressed in CHO cells.Competitive inhibition assay confirmed that hu12F6m has similar specificity and affinity as the original murine antibody and hu12F6. The stimulation of hu12F6m on T cells was significantly weaker than that of hu12F6. The experimental results lay the foundation for further study on the biological characteristics of hu12F6m.